801.
Comparative Effect Of Cidr Based Estrus Synchronization Protocol With Or Without Gnrh In Non-Descript Cows During Low Breeding Season
by Muhammad Bilal (2008-VA-152) | Prof. Dr. Mian Abdul Sattar | Dr. Muhammad Anwar | Dr. Muhammad Rizwan Yousuf | Dr. Muhammad Lateef.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The livestock sector occupies a peculiar position in the national plan of economic development in Pakistan. It subscribed approximately 55.9 percent to the agricultural value added and 11.8 percent to national GDP with a growth rate of 2.7 % in 2013-14 (Anonymous 2014). Dairying has become an important subsidiary source of income for thousands of rural families with an important role in generating earning opportunity. In Pakistan dairy sector is developing and commercializing at a rapid pace to meet increasing requirement of milk and other dairy products (Dongre et al. 2011).
Pakistan owns renowned breeds of dairy buffaloes (Nili-Ravi and Kundi) and cattle (Sahiwal, Red Sindhi, Thari and many others). Cattle in Pakistan belong to genus Bos indicus. According to the latest livestock census (Anonymous 2006), out of 29.6 million cattle, 46% (13.6 million) have been described as non-descript. Non-descript cattle do not fall in any defined breeds of cattle. Milk production of non-descript cows is < 1000 lit per lactation in mountains area of NWEP pakistan (Khan and Usmani 2005). As non-descript breeds make up the largest group of cattle in Pakistan, there is a dire need to work on genetic improvement of these animals. Artificial insemination, the best tool for genetic up-gradation in dairy cattle is applied only in 11.1% cows in Pakistan (Anonymous 2006). The main hindrances are small sized scattered herds and lack of experienced technical manpower in the field. Estrus synchronization of a large number of animals and timed insemination can be used to overcome these hindrances .The technique may also help in
11
reducing a prolonged calving interval and postpartum anestrus and seasonality of breeding in these animals (Zafar et al. 2008).
Estrus synchronization widely practiced in temperate dairy cattle in developed countries (Hansen and Arechiga 1999). Before launching a large scale estrus synchronization program in non-descript cattle, there is a need to assess the efficacy of various synchronization protocols in terms of estrus incidence, intensity and conception rate. Additionaly, distinct differences have also been reported between Bos taurus and Bos indicus in terms of estrus duration and intensity of expression of estrus sign (Mattoni and Ouedraogo 2000). The low estrus intensity and less duration of estrus signs of Bos Indicus are due to smaller diameter of follicle as compared to that of Bos Taurus (Bo et al. 2003).
Developing successful methods for synchronizing estrus and ovulation in cattle has been a major research interest. Ultimately, the goal has been to achieve precise synchronization of ovulation so that cattle can be inseminated without regard to estrus detection. One method to increase conception rates is to use hormonal treatments in zebu breeds for synchronizing ovulation and for timed artificial insemination (TAI). Hormonal programs for synchronizing ovulation to control the follicular and luteal phases and estrus behavior have been used in Bos taurus cows and heifers(Castellanos et al. 1997), and Bos Indicus cows (Pinheiro et al. 1998).
The intensity and duration of estrus behaviors during the estrous cycle is highly variable among individuals. More commonly animals are diagnosed to be in estrus based on the mounting or standing to be mounted, appearance of mucus discharge, and other physical activities (Van Eerdenburg et al. 2002). Scoring system were established on the basis of observed estrus signs and most of them have taken mounting and standing to be
12
mounted behavior as the most reliable signs to predict ovulation time in Bos taurus cows (Roelofs et al. 2005). However, it is now well acknowledged that the expression of estrus behavior change with breed of cows (Naidu and Babu Rao 2006).
Fertility is an important parameter to assess the efficacy of estrus synchronization. Bos indicus cows after treatment with CIDR conception rate in adult cows 40% and in heifers 20% (Singh et al. 2006). CIDR may have ability to overcome the problems in field condition and increasing the reproductive efficiency by minimize the hindrance anestrus postpartum cows. In Pakistan research work have been done to evaluate the efficacy of CIDR for conception rate in indigenous cattle. Until now no study conducted on non- descript cattle. Therefore, it is hypothesized that CIDR+GnRH can provide better in vivo fertility compared to CIDR. Availability: Items available for loan: UVAS Library [Call number: 2248-T] (1).
802.
Occurrence Of Bacterial Contaminants In Poultry Meals And Their Antibiotic Resistance Pattern
by Nayyab Tariq (2009-VA-207) | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Nawaz | Dr. Muhammad Nasir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Poultry is the second largest industry after textile industry in Pakistan. Its consumption
rate is very high as compared to other animal protein sources, as it is cheaper as compared to red
meat. To fulfill increasing demand of poultry, poultry production quality must be improved.
Many factors affect poultry production. One factor is feeding process. Efficiency of poultry
production depends mainly on feeding process which influences both the quality and quantity of
the poultry production (Grepay 2009). The rearing of poultry birds on commercial level requires
use of bulk quantities of poultry feed. Poultry feed costs 60-70% of total cost for production
(Sahraei et al. 2012). The main purpose to increase poultry production is to fulfill nutritional
requirements of human population that largely rely on poultry and poultry by products as a
source of protein(Obi and Ozugbo 2007).
Poultry feeds are food materials designed to contain all necessary feed ingredients for
proper growth, meat and egg production in birds (Obi and Ozugbo 2007). It is a mixture of
various components including plant proteins (cereals and by products, grains etc), animal byproducts,
fats, vitamins and minerals (Ravindran 2013). The major component of poultry feed is
protein which is the key component of eggs and meat. Protein sources in poultry feed are of
plant, marine and animal origin. Plant proteins may lack some of the essential amino acids, thus
are incomplete protein. Proteins of animal origin are better growth promoter than protein of plant
origin, but their safety is a concern. Among plant based proteins, soybean and canola meal are
produced in higher amounts worldwide (Alali et al. 2011). The animal protein sources include
poultry, fish, meat bone and poultry by products meal. Poultry meal is derived from clean tissues
Introduction
2
of slaughtered poultry including bone after the moisture and fat have been extracted in the
rendering process. It may contain whole birds excluding feathers (Anonymus 2014). Among all
protein based meals, poultry meals and poultry by products meal are of superior quality and
provide higher protein content than plant, marine and meat based meals (Samli et al. 2006).
Quality of animal feed has gained importance worldwide. The feeds are found to be
associated with infectious or non-infectious hazards, thus influence human health (Sherazi et al.
2015). Poultry feed can act as carrier of animal and human pathogens (Aliyu et al. 2012). Poultry
feed can get contaminated at any point of harvesting, processing, storage or dispersal of feed.
Primary mode of poultry feed contamination is by dust, soil, water and insects. Poultry meals can
be another source of feed contamination. Poultry meals are added in feed as a source of protein.
Feeds of animal origin like poultry meals are richer in nutrients and water as compared to feed of
plant origin thus are found to have higher microbial load, facilitating the multiplication of
bacteria (Kukier and Kwiatek 2011). Inclusion of contaminated meals in feed increases microbial
load of poultry feed. The contamination of poultry feed not only influences appearance and
nutritional value of feed, but also affects animals and human who consumes it (Maciorowski et
al. 2007). The profitability of poultry production can be greatly affected due to the frequency of
feed contamination and the detrimental effects of the aflatoxins on performance of chickens
(Anjum et al. 2011). Poultry feeds have been implicated in several poultry diseases of viral
(Avian Influenza, Newcastle disease), bacterial (Salmonellosis, Infectious Coryza) and fungal
origin. Many human diseases like Traveler’s Diarrhea and Salmonella Paratyphoid fever have
been associated with consumption of poultry birds that contracted infections from poultry feed
(Obi and Ozugbo 2007).
Introduction
3
The poultry industry relies on ready to use poultry feed prepared by feed mills (Arotupin
et al. 2007). Both bacteria and fungi including mycotoxins usually contaminate feed at different
stages of pre or post processing, depending upon the conditions under which it is handled or
stored (D’Mello 2006). Poultry meals mostly get contaminated post rendering process. The
cooking step in rendering process inactivates bacteria, viruses, protozoa, and parasites(Meeker
and Hamilton 2006) . Still presence of contaminants in meals is attributed to post processing
contamination. Many bacterial pathogens reported in feed are Escherichia coli, Erwinia
herbicola, Salmonella spp., Listeria spp., Enterococcus fecalis, Cl. perferingens and Cl.
botulinum (Aliyu et al. 2012; Lateef and Gueguim-Kana 2014) . The contaminated feed results
in excessive activation of immune system and ultimately decreases poultry production and its
profitability (Kukier et al. 2012). In addition to bacterial contaminants, toxigenic fungi have
threatened quality and safety of feed and have caused severe losses to poultry industry in recent
times. Cereals and grains based poultry feed mostly get contaminated with fungi (Kwiatek and
Kukier 2008). Mycotoxin producing fungal genera that are reported in poultry feed are
Aspergillus, Penicillium and Fusarium (Greco et al. 2014).
As Poultry feed is the first step of the food safety chain in "farm-to-fork" model. Contaminated
feed can also serve as a source of antimicrobial resistant bacteria in poultry meat(da Costa et al.
2007). There are many evidences that pathogens in feed are transmitted to humans through
animals and food of animal origin. It can also become source of some human pathogens in
environment. Feed contamination by fungi is responsible for animal mycotoxicoses and through
consumption of contaminated animal food, results in human intoxications (Kukier et al. 2012).
Birds utilizing toxins containing feed are economical loss for farmers and also affects consumer
Introduction
4
health through its residues (Alam et al. 2012). Poultry feeds containing antibiotic resistant
bacteria results in loss of poultry productivity, making treatment of poultry diseases difficult.
Thus quality of animal food directly depends on usage of nutritionally balanced and safe feed.
Among many feed sources used, poultry meals are gaining importance for their higher nutritional
value, but very less work has been done in world particularly in Pakistan to determine
microbiological safety of poultry meals produced. There is the need to determine various quality
parameters which should be followed to ensure production of safe meal. Availability: Items available for loan: UVAS Library [Call number: 2252-T] (1).
803.
Growth Performance, Body Measurements And Slaughter Characteristics As Affected By Different Patio Strategies And Growth Promoters In Japanese Quail
by Imran Ullah (2013-VA-598) | Dr. Jibran Hussain | Prof. Dr. Muhammad Akram | Dr. Atia Basheer.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Coturnix japonica is the zoological name of Japanese quail, which belongs to order Galliformes, family Phasianidae, genus Coturnix and specie japonica (Beletsky, 2006). It is highly disease resistant with a very short maturity age as well as market life (4-5weeks). Female quail starts laying almost at 7-8 weeks of age, hence getting popular (Ashok and Prabakaran 2012).
Generally the chicks are removed from hatcher only when most of chicks have hatched within the period of hatch window which is almost 36 to 48 hours (Careghi et al. 2005). Many factors affect delay in hatching, the most important being the parent flock age, handling of eggs, time of egg preservation and internal incubation conditions (Decuypere et al. 2001). Little space in the hatcher for hatched chicks, opening of hatcher over many time and late collection of chick can lead to dehydrated and low quality chicks (Bamelis et al. 2005). To reduce above stated risks a concept named “Patio system” was introduced in Netherland, proposed by Kuijpers brothers to improve hatchability and growth performance of chicks. In this system pre incubated eggs (hatching phase) and after hatching chicks (brooding phase) are placed combined in hatchery (http://www.vencomatic.com). Patio system improves body weight, quality of chicks and reduces chick mortality in broilers (Van de Ven et al. 2009). A synergistic effect on growth performance and various economic traits can be attained by using patio system and various feed additives such as probiotics, prebiotics or antibiotics etc. The term probiotic is derived from two Latin words “pro” and “bios”. Probiotics are prepared from different species of microorganisms (Bacillus, Enterococcus, Lactobacillus and Saccharomyces) considered as intestinal beneficial microflora (Dankowiakowska et al. 2013). Probiotics are used to make the products through the use of which, many species of animals improve their intestinal miro-flora (Leeson and Summers 1997). Improved beneficial bacteria
CHAPTER 1
INTRODUCTION
INTRODUCTION
2
of intestine suppress harmful bacteria like E. coli and produce beneficial enzymes and substances that have antibiotic like effects (Montes and Pugh 1993). Probiotics work by two ways. 1. Competitive exclusion 2. Enhancing immune system. In competitive exclusion, bacterias in gastrointestinal tract produce such type of substances that stunt the growth of harmful bacterias and compete for their predilection site (intestinal epithelium) while second mode of action is, to stimulate their immune system as the young one born is sterile, hence, their gastro intestinal tract (GIT) and beneficial bacterias in GIT are not able to produce antibodies against harmful bacterias that aggregate in chick GIT. By using probiotics that attach intestinal mucosa and create barriers for harmful bacteria and enhance immune system (Dankowiakowska et al. 2013). Different probiotics (Bacillus subtili) have different modes of action (competitive inhibition) that utilize oxygen in GIT of birds and convert them in different useful enzymes (subtilin and catalase) by which friendly bacterias (Lactobacilli) colonize in birds GIT and block predilection site of pathogenic bacteria. In addition, Lactobacilli bacteria produce lactic acid that kill many harmful bacteria like, Salmonella and E.coli (Hosoi et al. 2000). Probiotics are also found to enhance activity of immunoglobulins, macrophages, lymphocytes and interferon (Yang et al. 2009). Birds fed on probiotic mixed diet showed higher body weight and increased carcass percentage (Kabir et al. 2004a) but feed intake, proventiculus, gizzard and liver weight were found to be non-significant among different treatments (Dizaji et al. 2012). Similarly in another study, higher body weight and weight gain were observed in ostrich birds fed on probiotic enriched diet (Karimi-Kivi et al. 2015). Using antibiotics as feed additives can also improve health status of birds. The first antibiotic “chlortetracycline” when used in animal feed during early 1940s increased body weight was observed (Dibner and Richards 2005). For more than 50 years, antibiotics have been used in
INTRODUCTION
3
poultry feed as a supplement to improve bird’s growth performance, feed conversion ratio and carcass meat yield (Denli et al. 2003). Antibiotics being used in poultry industry are also used therapeutically in human medicines (Edens, 2003). However, recent progress in poultry industry and consumer concerns have risen the questions on use of antibiotics as growth promoters due to their side effects (Wray and Davies 2000). The European Union (EU) banned some antibiotics as growth promoters because of many bacterial strains showing resistant against them (Edens, 2003) In Pakistan high production cost and market price of quail meat discourage both the producer and consumer. The use of patio system may help reduction in production cost, hence, help in gaining the trust and preference of both quail farmers and consumers. Present study was conducted with the main objectives to study the advantages of patio system in Japanese quail and also provide basic information regarding future use of this technique in quails. It was further tried to understand the utility of different growth promoters in quail production with special emphasis on antibiotics and probiotics. Availability: Items available for loan: UVAS Library [Call number: 2251-T] (1).
804.
Isolation Of Surface Antigen 1 Gene Of Toxoplasma Gondii And Its Cloning In The Expression Plasmid
by Farooq Riaz (2008-VA-231) | Dr. Muhammad Imran Rashid | Prof. Dr. Kamran Ashraf | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Toxoplasma gondii is an obligate intracellular protozoan parasite which comes under the classification of phylum Apicomplexa, subclass Coccidiasina (Cornelissen et al. 1984). Toxoplasmosis is one of the more common parasitic zoonoses world-wide caused by Toxoplasma gondii which is a facultatively heteroxenous, polyxenous protozoon that has developed several potential routes of transmission within and between different host species (Tenter et al. 2000). It is the most important source of toxoplasmosis in humans and animals, with cat as definite host and warm-blooded animals as intermediate host (Frenkel et al. 1970). It was first described by Nicolle, Manceaux and Splendore in 1908 from rodents Ctenodactylus gondii (Black and Boothroyd 2000).
Toxoplasmosis is a worldwide parasitic disease and it is estimated that about one-third total population of the world is seropositive for Toxoplasma gondii (Tenter et al. 2000). Prevalence of infection varies between countries, geographical areas and ethnic groups living within a specific region. In Humans, infection rates range from 50% to 83% in Brazil (Tenter et al. 2000; Dubey et al. 2012). Seropositivity of Toxoplasma gondii in China is about 8% with continuously increase while in USA its 10-15%, 50-70% in France and 20% in UK (Dubey and Jones 2008; Zhou et al. 2008; Jones et al. 2009). Prevalence of toxoplasmosis is higher in males (79%) as compared to females (63.4%) and the age dependent sero-prevalence reaches >92% in age group of 40 to 50 (Coêlho et al. 2003).
Transmission occurs through the ingestion of contaminated vegetable /water with oocysts, as well as the ingestion of contaminated raw/undercooked meat with tissue cysts (Gajadhar et al. 2006). Transmission may also occurs by ingestion of sporulated oocysts, or bradyzoites within cysts present in the tissues of numerous food animals (Esteban-Redondo et al. 1999). In humans, transmission of Toxoplasma gondii happens mainly by eating raw or undercooked contaminated meat, raw cow’s milk and birds eggs, swallowing oocysts dis-charged in feces of infected cats, inoculation of trophozoites through the skin, or by inhalation (Wallace 1971; Wallace 1973; Bannister 1982).
In humans, mostly infections (congenitally or post-natally acquired) are asymptomatic. Congenital infection occurs only when a woman becomes infected during pregnancy. Congenital infections acquired during the first trimester are more severe than those acquired in the second and third trimester (Desmonts and Couvreur 1974). The main clinical signs associated with toxoplasmosis are anorexia, weight loss, lethargy, dyspnea, ocular signs, pyrexia, vomiting and diarrhea, jaundice, myositis, encephalitis and abortion. Humans become infected when they ingest the toxoplasma at infective stages (oocysts and tissue cysts) found in some cat feces and in raw meats.
In addition to being hazardous to livestock animals, the T. gondii infection is also important due to its zoonotic implications (Jittapalapong et al. 2005). Congenital abnormalities in humans, such as microcephaly, hydrocephaly, chorioretinitis, convulsion, cerebral calcification, epilepsy, blindness, deafness, and mental retardation may occur if the mother acquires infection during pregnancy (Jones et al. 2003). In addition to congenital anomalies, T. gondii also causes severe neuropathologic infections in immuno-compromised hosts, such as AIDS and cancer patients receiving chemotherapy (Del Valle and Piña-Oviedo 2005).
Seroprevalence studies of T. gondii among domestic animals in South-Western Pakistan has indicated considerable prevalence (25% in cattle, 2.5% sheep) (Zaki 1995) and suggesting potential transmission to the human community. Small scale study in urban area of Rahim Yar Khan (Punjab), Pakistan has revealed that the overall prevalence of toxoplasmosis in food animals is 19% (Ramzan et al. 2009). Another study has already been published that untreated patients with leprosy in Pakistan have shown significant seroprevalence (29.6%) of antibodies against T. gondii (Hussain et al. 1992).
Vaccine against toxoplasmosis is not available yet with one exception (“Toxovax” for sheep). Vaccine against T. gondii in animals used for human consumption may block the possible transmission to humans (Bhopale 2003). SAG1, among one of the major antigenic components of Toxoplasma gondii is a major surface antigen identified on the surface membrane of this parasite using a monoclonal antibody (Handman et al. 1980). SAG1 is an important surface antigen, expressed by tachyzoite form of T. gondii and is a putative candidate for vaccine and diagnostic against toxoplasmosis (Sharma et al. 1983; Godard et al. 1990). Immunization with SAG1 adjuvanted with saponin Quil A or incorporated in lysosomes provided total protection after challenge (Bülow and Boothroyd 1991; Khan et al. 1991). SAG1 is single copy gene with no introns (Burg et al. 1988), regulates both humoral as well as cellular Th1 immune responses (Liu et al. 2008) and is powerful candidate for vaccine against toxoplasmosis. SAG1 is a potent candidate of diagnostics for detection of serum antibodies against toxoplasmosis in Man and animals (Abu-Zeid 2002).
Availability: Items available for loan: UVAS Library [Call number: 2258-T] (1).
805.
Molecular Diagnosis And Therapeutic Trials Against Bovine Fasciolosis In And Around Okara
by Hafiz Farooq Ahmad (2008-VA-93) | Dr. Muhammad Ijaz | Dr. Waseem Shahzad | Dr. Jawaria Ali Khan | Dr. Shehla Gul Bokhari.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Agriculture sector plays a pivotal role in the economy of Pakistan. At present it
participates 11.4% to GDP. Agriculture provides 45% employment to labour of our country
and contributes in the development of other sectors of the economy. The livestock sector
occupies a distinguishing position in the National Agenda of economic development. It
provides net source of foreign income. In the history livestock has been dominated by small
holders to meet their needs of milk, food safety and cash earnings on daily basis. Besides,
livestock is considered a source of employment generation at rural level. It plays an important
role in poverty mitigation and keeps in elevating the socio-economic situation of our rural
commonalities. Livestock share approximately 55.4 percent to the agricultural value added
and 11.9 percent to national GDP during 2013-14. Anonymous (2013-14).
Parasitism is one of the major problems lowering livestock productivity round the
earth Vercruysse and Claerebout et al. (2001). The significance of helminthes infections is
increased manifold in developing countries like Pakistan where 65.2% population is rural
(Population Census Organization (2007) depending upon livestock for their incomes and
where parasitism acts as a serious trouble for livestock economy Chaudhry et al. (1984).
Among helminthes infections, fasciolosis commonly called as liver fluke disease is of vital
importance due to its wider spectrum of ultimate hosts Rondelaud et al. (2001) causing acute
and chronic infections Sampaio-Silva et al. (1996). The disease is primarily caused by
Fasciola hepatica and Fasciola gigantica Soulsby et al. (1987). Fasciolosis is a parasitic liver
infection of wild and domestic ruminants caused by genus Fasciola, which has worldwide
distribution Soulsby et al. (1986). Fasciolosis causes economic losses as a result of
mortalities, abortions, retard growth, reduced milk and meat production, condemnation of
infected liver and emaciated carcasses and cost of animal treatment Gracey and Collins et al.
(1992). Fasciolosis caused by F. hepatica is a cause of important economic losses in
ruminants. The disease is widely spread throughout in Africa, Asia and the Pacific. Although
mostly a problem of young stock, the disease is also found in older animals where it
adversely affects health, growth rate and development. Apart from its great veterinary
importance throughout the world, fasciolosis caused by both F. hepatica and F. gigantica has
recently been shown to be a re-emerging and wide spread zoonosis affecting a number of
human populations Mas-Coma and Bargues et al. (1997); Esteban et al. (2003).
Prevalence of fasciolosis infestation in buffaloes is 49.01%. The infestation of
fasciolosis was optimal from January to September, while least during October to December.
Fecal samples indicated F. hepatica prevalence 65%. Whereas prevalence of F. gigantica
was observed higher in adult cattle as compared to cattle calves by fecal egg counts Khalil et
al. (2009). 4% Buffalo calves infested with trematodes. Availability of intermediate host and
the grazing habits of the final host determines the epidemiology and seasonal pattern of
infection with trematodes Pfukenyi et al. (2005); Bhutto et al. (2002). The life cycle of F.
hepatica includes many stages which develop in the environment or the intermediate host.
Metacercariae which is the infective stage encysted and ingested by grazing herbivores.
Temperature and rainfall are the important factors affecting the life cycle and the prevalence
of F. hepatica infection McCann et al. (2010). Many studies revealed that F. hepatica is
endemic in buffaloes, cattle, goats, sheep and humans in Pakistan Ijaz et al. (2009); Iqbal et
al. (2007); Khan et al. (2010) and Qureshi et al. (2005).
Fasciolosis is the disease by which about 250 million sheep and 300 million cattle are
potentially affected worldwide Boray et al. (2005), and more significantly infects all
ruminants Iqbal et al. (2007). F. hepatica and F. gigantica are the two liver flukes commonly
reported to cause fasciolosis in ruminants Walker et al. (2008). Infected cattle can exhibit
Introduction
3
poor weight gain and dairy cattle have lower milk yield, and possibly metabolic diseases
Mason et al. (2004). Losses due to fasciolosis in the United Kingdom and Ireland alone are
more than ￡18 million a year Mulcahy and Dalton et al. (2001); €52 million a year or €299
per infected animal in Sweden Schweitzer et al. (2005); 0.26 million USD annual due to
fasciolosis associated liver condemnations in cattle slaughtered in Kenya Kithuka et al.
(2002). In Pakistan, 35697 USD (3141360 PKR) are reported in only one tehsil of Punjab
province, the Sargodha T.U. Rehman et al. (2013). While Fasciolosis is prevalent in whole
Punjab province of Pakistan but the marshy areas such as regions of Gujranwala, Lahore,
Multan, Faisalabad, Jhang, Muzaffargarh, Sheikhupura, and Sargodha are of major concern.
Fascioliasis in ruminants causes substantial economic losses, estimated at US$ 2
billion per annum worldwide, to rural agricultural communities and commercial animal
producers due to death of infected animals, condemnation of affected livers and production
losses associated with reduced feed conversion efficiency Urquhart et al. (1996); Spithill and
Dalton et al. (1998). In tropical countries, fascioliasis is prevalent up to 90% and is
considered the most important helminthes infection of cattle Spithill and Dalton et al. (1998).
Most important, human can also become infected with Fasciola spp. and the disease is
referred to as human fasciolosis. It is estimated that 2.4 million people in more than 60
countries are infected and the number of people at risk is more than 180 million throughout
the world Haseeb et al. (2002); Mas-Coma et al. (1999); Ishii et al. (2002). Khalil et al.
(2009) recorded 49.01% prevalence of fasciolosis infestation in buffaloes. He also observed
that optimum infestation was seen from January to September, while minimum during
October to December. Whereas Bhutto et al. (2002) recorded 4% buffalo calves infested with
trematodes.
Introduction
4
Morphological identification of Fasciola species requires significant parasitological
skills and is not an accurate method of characterization, especially for the ‘intermediate’ form
Kendall et al. (1965); Lin et al. (2007); Le et al. (2008). Hence, different molecular tools have
been developed during the last decade for the accurate identification of Fasciola spp.
Marcilla et al. (2002); Velusamy et al. (2004); Cucher et al. (2006); Magalhaes et al. (2008);
Ai et al. (2010). Conventional techniques like fecal examination used for its diagnosis are not
comprehensive because of Species similarity in distribution in many countries of East and
North Africa, and Southeast and Central Asia, and are similar in egg morphology.
Coprological methods continue to be the most widely used approach for the detection and
quantification of Fasciola and other helminth eggs in laboratory animals, domestic and
sylvatic animals and humans Cringoli et al. (2004, 2010); Bergquist et al. (2009), with a more
recently developed method that is currently undergoing broad-scale validation the FLOTAC
techniques Cringoli et al. (2010) for the detection and quantification of F. hepatica eggs in
feces obtained from experimentally-infected rats before and after drug administration.
FLOTAC has been used successfully in the diagnosis of F. hepatica infections in naturallyinfected
sheep, which underwent treatment with standard Cringoli et al. (2006). However
PCR based diagnosis using specie-specific primers has been proved more accurate and
successful in diagnosis and distinction of Fasciola species Le TH et al. (2012). The molecular
technique such as PCR is used for the recognition of parasite. These molecular techniques are
largely accepted all over the world. These are more exact methods than traditional in
epidemiological studies Heckeroth and Tenter et al. (1999).
Herbal drugs have been used since ancient times to cure diseases and medicinal plants
have been used to treat fasciolosis among these Nigella sativa and Allium sativum have been
used to treat the worm infestation. No report of resistance development to plant anthelmintic
although many have been used in veterinary medicine for many years Ashaal et al. (2010);
Introduction
5
Waller et al. (1995); Koko et al.(2000); Thienpont et al. (1979); Keiser et al. (2011). Herbal
(indigenous) drugs have been used since ancient times to cure diseases, and several medicinal
plants have been used to treat fasciolosis Satyavati et al. (1987). Among these, Nigella sativa
has been used to treat worm infestation Nadkrani et al. (1954; Said et al. (1969); Akhtar et al.
(1988). N. sativa can be used as a therapeutic agent against helminthes Khan et al. (2013).
Neem leaf powder can be used as therapeutic agent against gastrointestinal parasites of
camel, equine, cattle, buffalo and small ruminants Mehmood et al. (2013). Garlic powder can
also be used as herbal product against the gastrointestinal helminthes of livestock Ijaz et al.
(2008). Availability: Items available for loan: UVAS Library [Call number: 2256-T] (1).
806.
Effect Of Nigella Sativa On Embryonated Eggs Experimentally Infected With Newcastle Disease Virus
by Arif Ullah Khan (2013-VA-561) | Dr. Muhammad Yasin Tipu | Dr. Raheela Akhtar | Dr. Ali Ahmad Shiekh.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: In Agriculture Industry of Pakistan poultry sector plays a vigorous, animated and a well
known role. There are more than 1.5 million peoples whose financial gain and employments are
related directly or indirectly to this sector which provides livelihood to about 60 % rural
inhabitants. Meat and eggs are the products of poultry industry. In Gross Domestic Products its
constant cost factor is 1.2%, in livestock is 10.4% and in agriculture it contributes 5.76%.In the
total meat production quantity of the country, poultry meat contribution is 26.8 % (Anonymous
2013).
The member of family Paramyxoviridae, Newcastle disease virus (NDV) contains six genes,
single stranded RNA genome without segment and have different strains arranged as virulent
which is velogenic, intermediate virulent mesogenic and less virulent lentogenic. The Newcastle
disease or pseudo-fowl pest is complete destructive having high death ratio in poultry. The NDV
attacks on respiratory system causes distress, produce nervous disorders, disturb gastrointestinal
tract and causes hemorrhagic lesions in intestine, downward change in level of egg production
and high mortality (Alexander 2000).
In embryonated eggs the cells attached with allantoic cavity help to grow and develop the
NDV virus which is inoculated for the purpose of propagation. In approximately 24 hours post
inoculation the virus reach higher concentration. First of all the virus destroys the live cells in
which it is growing and then it is widely spread by the allantoic fluid. If the inoculated strains of
virus are virulent i:e velogenic, embryos die within two days post inoculation mostly. The
embryos which are dead or alive show histopathological and gross changes remarkably caused
by the inoculated NDV (Al-Garib et al. 2003).
Introduction
2
To evaluate the value or determine the pathogenicity of the virus or to propagate Newcastle
disease virus the embryos of chicken have been usually used. The NDV strains which are
virulent means velogenic, kill the embryo within 48 hours, and lentogenic or less
virulent(mesogenic) take 5 to 7 days to kill either the embryos or may not kill at all (Lam et al.
1995).
It always had an inherited pattern of thoughts to advance the progress in the process of
healing by using herbal plants in all the civilizations. All over the world, to provide treatment for
diseases in daily life medicinal herbal plants and seeds have been used since hundreds of year
ago and in ahead of times. Plants play important role and serves as a base in the development of
drugs (Ates and Turgay 2003).
For the promotion and well being of health of both animals and humans, since thousands of
year ago black cumin seeds which is commonly known as black seed are used. It is stated by
Prophet Muhammad (P.B.U.H) that black seeds are treatment for every disease except the final
state the death. Its scientific name is Nigella sativa and belongs to the family Ranunculaceae.
The black seeds contain the main active and wider in scope ingredient named as crystalline
nigellone. The composition of other ingredients which are present in these seeds are proteins,
vitamins B1, B2, B3, oleic acid, myristic acid, linolic acid, folic acid, thymoquinone, palmitic
acid, beta sitosterol and stearic acid. In black seed the main elements are phosphorous, iron, zinc,
calcium, and copper. Anti-bacterial, anti-parasitic, anti-tumor, anti-hypertensive, immunestimulator,
anti-inflammatory and anti-histamine are the main effects of black cumin seeds
(AitMbarek et al. 2007).
Black cumin seeds are a hypothetical remedy for all ills, diseases and disorders, but not to
stop growing old and death from happening. In Bible it is identified as black cumin and tending
Introduction
3
to restore health (Worthen et al. 1997).
The antiviral drugs are not usually used in poultry industry due to economic problems
and may cause toxicity, but some drugs shows effect against virus which effect poultry. The
antiviral drug Ribavirin was used to compare its effect with a herbal plant to see which one is
effective. Higher doses produce toxicity and also were the cause of death. The 20μl/ml dose
showed the significant effect and the embryoes remain live after experimentally infected with
Newcastle disease virus (Omer et al. 2014). Availability: Items available for loan: UVAS Library [Call number: 2255-T] (1).
807.
Comparative Effect Of Alpha Lipoic Acid And Butylated Hydroxytoulene On Post Thaw Quality Of Buck Semen
by Muhammad Khurram Shahzad | Prof. Dr. Mian Abdul Sattar | Dr. Mushtaq Ahmad | Dr. Muhammad Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Amongst different livestock species, goats and sheep are the major source of livelihood for over a million livestock farmers in Pakistan. Total goat population in Pakistan is estimated to be 66.6 million. These animals are mostly kept by small holders for whom these are only source of their livelihood. Milk production from goats is 0.822 million tonnes while mutton production from both sheep and goats is 0.657 million tonnes (Anonymous 2014). Pakistani people mostly prefer the goat meat over sheep.
All irrigated areas of Punjab including district Faisalabad, Sahiwal, Sargodha, Jhang, Jhelum, Lahore and Multan are the habitat of Makhi Cheeni Beetal goats. The color of its body coat is red spotted or golden brown with white patches. Its body is very well developed and compact. Males have long spiraled horns while females have shorter. It has roman nose with pendulous broad and long ears. It has long teats and well developed udder. Female weighs 37kg and males 46kg. Twins or triplets births are more than 50%. In 130 days of lactation period, there is 290 liters milk yield (Shah et al. 2001).
Some breeds of goats especially dairy goats have more demand than the others and these bucks are not available everywhere. To cope with this situation artificial insemination techniques is necessary. Artificial insemination plays a great role in increasing the economics by spreading the superior genetics within a short period of time.
Semen is processed by different methods but cryopreservation is considered to be the best method. Cryopreservation has been reported to compromise the quality of processed semen resulting in the loss of sperm motility, viability, in-vivo fertilizing ability, deterioration of plasma membrane and acrosomal integrity, apoptosis and damage of deoxyribonucleic acid (DNA) (Medeiros et al. 2002; Purdy 2006a). Sperm damage may occur due to various factors like osmotic stress, oxidative stress, low-temperature exposure and combination of different factors (Sarıözkan et al. 2009). Thawing of semen may also cause osmotic changes and the sperm quality is further decreased. It is generally accepted that sperm viability is reduced by as much as 50% during the process of semen cryopreservation (Watson, 2000).
Extension of buck semen with egg yolk containing extender may be more injurious to sperms. This is due to the presence of coagulating enzymes of bulbourethral origin named as egg yolk coagulating enzymes (EYCE). EYCE decreases the tenacity of chilled or frozen semen (Roy, 1957). EYCE also catalyze the conversion of egg yolk lecithin into lsolecithin and fatty acid, thus sperm membrane become more fusogenic due to hydrolysis. So there is increase in chromatin decondensation and acrosomal reaction that is harmful for sperm (Leboeuf et al. 2000). Due to excess of poly unsaturated fatty acids (PUFA) in sperms, they are more susceptible to lipid peroxidation (Cassani et al. 2005). Lipid peroxidation of PUFA lead to production of reactive oxygen species (ROS) (Alvarez et al. 1995). Small amount of ROS are normally involved in capacitation, acrosmal reaction and ultimately fertilizing ability of sperms. But when the ROS are produced in excess
Introduction
3
amount, these may compromise the enzymatic function and sperm fertility (Baumber et al. 2000). At 4-5 ºC the motility and plasma membrane integrity is decrease with the passage of time which ultimately leads to decrease in fertility. One of the cause of this decrease is production of ROS by the lipid peroxidation of spermatozoa’s membrane (Storey et al. 1998). Major decrease in sperm motility and fertility occur during phase transition from liquid crystalline to gel phase (Chakrabarty et al. 2007). Lipid peroxidation leads to irreversible loss in motility and damage to DNA of sperm (Maxwell et al. 1996). Motility of sperm is adversely effected with ROS, when the ROS harm the plasma membrane and acrosomal integrity which ultimately leads to fragmentation of DNA. Sperms have their own antioxidants system which include the glutathione (GSH) , GSH peroxides, superoxide dismutase, catalase and chelators of transferrin, lactoferrin and ceruplasmin (Agarwal et al. 2002). Normally the ROS production and scavenging are in equilibrium but during the semen preservation the excessive production of ROS (superoxide, hydroxyl, hydrogen peroxide, nitric oxide, peroxynitrile) with low level of scavenging system and antioxidants leads to oxidative stress. During the process of freezing and thawing the natural antioxidants systems are unable to stop lipid peroxidation. Therefore a powerful antioxidant system should be used to avoid the cryo-injuries and lipid peroxidation (Irvine 1996).
Different antioxidants are being used i.e. fetuin (F), amino acid (AS), cysteine (CY) taurine, glutathione (GSH) glutathione peroxidase (GSH-PX), catalase (CAT), superoxide dismutase (SOD) glutamine, hyaluronan, trehalose, Alpha lipoic acid (ALA) and Butylated Hydroxytoulene (BHT) (Atessahin et al. 2008; Bucak et al. 2009; Taşdemir et al. 2014). Addition of antioxidants to semen extenders are considered to improve the quality of semen (Rao et al. 2013). ALA is a short chain fatty acid which act as an antioxidant in both aqueous and lipid environments, its therapeutic effects in other tissues like brain (Piotrowski et al. 2001), heart, kidneys and testicles has already been
Introduction
4
discussed. It is called as universal antioxidant because of its effect in different parts of body. It is not only involve in scavenging the ROS but also activate the body antioxidants systems against ROS. ALA reduced to dithiol form called dihydrolipoic acid (DHLA) which is an excellent antioxidant (Handelman et al. 1994). ALA also regenerates vitamin C from reduced vitamin C in the presence of glutathione (GSH) which also enhance the antioxidant activity (Ibrahim et al. 2008). BHT, a phenolic lipophilic antioxidant that has antiviral activity, have the ability to relieve the cold shock in spermatozoa from several animal species. It stops the auto oxidation by converting the peroxide radical to hydroperoxide as it is also called as synthetic analogues of Vit E (Memon et al. 2011). BHT acts as a membrane lipid protectant which reduces the changes in permeability of sperm plasma membrane in cold shock (Graham et al. 1992). BHT minimizes the effect of cold shock on semen (Shoae et al. 2008), boar (Roca et al. 2004) and goat (Khalifa et al. 2008). Availability: Items available for loan: UVAS Library [Call number: 2254-T] (1).
808.
Expression And Mutational Analysis Of Breast Cancer Susceptibility Gene 1 (Brca1) And Cyclooxygenase-2 (Cox-2) Gene In Feline And Canine Tumours
by Haleema Sadia (2007-VA-567) | Dr. Muhammad Wasim | Prof. Dr. TahirYaqub | Dr. Abu Saeed Hashmi.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Cancer is the first cause of death in cats and dogs while in human it is the second most cause of death (Jemal et al. 2008). According to an estimation, cancer related deaths in the world are 13% and 70% of these deaths are in poor countries (World Health Organization 2012). Such natural cases of cancers in cats and dogs especially, in dogs offer an opportunity to use the dogs for comparative cancer studies and as an animal model for anticancer drug development (Pawaiya 2008). Inu a series of more than 2000 autopsies, it was found that almost forty five percent dogs that lived for ten or more years expired because of cancer (Bronson 1982). Dogs are affected by skin cancer 35 times more often than humans. They are also affected 4 times more often by mammary gland cancer, 8 times more often by bone cancer, and twice more often by leukemia, than humans (Cullen et al. 2002). The regulation of cell proliferation, genome stability and programmed cell death are important for systemic homeostasis.
1.1Historical perspective on cancer causation
Hippocratic and Galenic medicine attributed the spread of black bile (one of the four humours) in the tissue as the cause of the cancer (Diamandopolus 1996) is an idea survived intact through the Middle Ages and Renaissance. With the discovery of the lymphatic system by Gasparro Aselli in 1662, the black bile theory was superseded by the idea that cancer was an inflammatory reaction to extravasated lymph; a theory modified 150 years later by John Hunter who introduced the notion that contaminated coagulating lymph was the origin of the cancer (Kenneth 2003). A German pathologist Johannes Muller first time demonstrate that cancer is made up of cells (1838) but he also gave an idea that cancer cells were originated from a bud called Blastema instead of normal cells (Kardinal and Yarbro 1979). Following Schleiden and Schwann's cell theory of tissues,it was Rudolf Virchow (Muller’s student) who in 1855 demonstrated that every cell was derived from another cell (omnis cellula e cellula), including cancer cells (Mazzarello 1999; Porter 1999). In 1867 Wilhelm Waldeyer supported the theory of the normal cell for the origin of cancer and he believed that metastasis resulted from transportation of cancer cells by blood or lymph (Porter 1999).
Around the turn of the twentieth century the beginning of tumour transplantation experiments led to the new view of the cancer cell as an autonomous cell. The first successful tumour transplants were described in 1876 by the Russian veterinarian Mstislav Aleksandrovich Novinski (Novinski 1876). He reported in his thesis entitled “On the Question of the Inoculation of Malignant Neoplasms” the first successful serial passage of tumours through transplantation in dogs. Novinski's transplantation experiments were based on the inoculation of canine transmissible venereal tumour (CTVT) in puppies. Novinski stated that successful tumour transplantation depends on the inoculation of a living element of the tumour and that the transplantation of the element of a cancerous tumour to healthy tissue acts as an infecting agent. In 1888 Wehr repeated Novinski's transplantation experiments in dogs with similar results (Shimkin 1955). It is interesting to note that the dogs used for transplantation of CTVT did not come from a single breed and were therefore not highly inbred. The allo-transplantation of tumours seemed less surprising in the late 19th Century than it does today with our modern knowledge of histo-incompatibility.
The successful results obtained with CTVT served as model for tumour transmission in other animals. Hanau in 1898 inoculated two rats with vulvar epidermoid carcinoma and observed growth of the tumour in the recipients (Shimkin 1955). In 1901 Leo Loeb supported the transplant ability of tumours in rats (Witkowski 1983; Brent 1997). In 1903 a Danish veterinarian Carl O. Jensen determined the successful growth of transplanted tumours in mice by heredity (Brent 1997). The discovery that the tumour could be successfully transplanted into (Witkowski 1983; Brent 1997) other mice, led the scientists to use rodent system to supply tumours for experiments. The observation that a single tumour could be expanded through many generations exceeding the life span of the laboratory mouse led Leo Loeb to the "cancer immortality" concept (Witkowski 1983).
The earliest observations reported by John Hill in 1759 and by Percival Pott in 1775 on the association of a specific tumour to a specific profession or work, led to the idea that some chemicals can cause cancer (Greaves 2000). In 1918 Yamagiwa and Ichikawa induced cancer by applying coal tar to rabbit skin(Greaves 2000; Luch 2005). After the discovery of the X rays by Wilhelm Conrad Roentgen in 1895, Frieben published data in 1902 indicating that cancer rates were increased among persons working with X-rays (Cassileth 1983; Greaves 2000)
1.2 Tumour Progression
The first detailed characterization of the dynamic nature of cancer was described by Leslie Foulds (Foulds 1949). Foulds showed that tumours progress (evolve) through different stages, characterized by the acquisition of different phenotypic traits such as increased growth rate, hormone dependence, invasiveness, formation of metastasis (Foulds 1949; Fould 1954; Foulds 1957). With the progress of molecular biology the phenotypic view had been replaced with the somatic mutation theory, where cancer evolved through the accumulation of different mutations in several genes (Greaves 2000). The accumulation of mutations in somatic cells implicated the presence of different cells bearing different mutations and also the presence of natural selection, which selected the cells with advantageous mutations. One of the questions arising from the somatic mutation theory was whether a tumour had a single or a multiple origin.
This observation was supported by a karyotype analysis in chronic myelogenous leukaemia (CML) by Peter Nowell and David Hungerford in 1960 (Nowell 2002). They described the presence of an unusually short chromosome 22 in all CML tumour cells analyzed, and the absence in the normal cells from the same patients. This observation suggested that this mutation was a somatic mutation that occurred in one cell in the bone marrow, which gave it a selective advantage to expand as a clone. Nowell postulated that a tumour develops by a Darwinian evolutionary process, where cells with mutations conferring a growth advantage are selected and expanded (Nowell 1976; Greaves 2002). In 1954 Peter Armitage and Richard Doll analyzed human cancer incidence over the age, and showed that chances of cancer increased in older people (Armitage and Doll 1954).
The concept that cancer might be contagious also recurs throughout the past 300 years.In the 17th and 18th centuries, physicians Daniel Sennert and Zacutus Lusitanus supported the hypothesis that cancer was contagious. In fact in 1779 a hospital in Paris was directed to move the cancer patients from the city (Cassileth 1983; Kenneth 2003).
1.2.1 Exogenous and endogenous factors
In 1844 the Italian physician Domenico Antonio Rigoni-Stern noted that cancer of the cervix was frequent among married ladies, rare among unmarried ladies and absent in Italians nuns. In contrast, breast cancer was more frequent among nuns (Greaves 2000). These observations led to the hypothesis that cervical cancer was sexually transmitted, and we now know that the cause is a papilloma virus (Hausen 2002).In 1908 Wilhelm E and Olaf B, transferred the leukemia in chicken by tissue filterates (Wyke 2003). In 1911, Peyton Rous demonstrated that viruses were the cause of solid tumours (Sarcoma) in chickens but it took many decades before his data were accepted (Dulbecco 1976). The notion that viruses can cause cancer was a discovery that brought back the fear that cancer was a contagious disease.
There are many exogenous and endogenous risk factors that affect the tumor suppressor genes and oncogenes (Todorova 2006). Tumour viruses (Bishop 1980), chemical carcinogens (Loeb et al. 2000), natural chemicals, (Ames et al. 1990), herbicides (Glickman et al. 2004), physical carcinogens like radiation (Upton 1978) are exogenous factors while inherited genetic defects, immune system (Rosenthal 1998) and hormonal factors (Rodney 2001) are among endogenous risk factors.
Although tumour cells are generally described as independent evolving units, recent results suggest that tumour cells are able to stimulate stromal cells to produce growth factors that increase tumour proliferation (heterotypic stimulation) (Kinzler and Vogestein 1998; Skibe and Fuseing 1998; Iyengar et al. 2003). It has been demonstrated that cells involved in the immune response to tumours may produce factors such as inflammatory chemokines that may also promote the tumour proliferation (Pollard 2004; Wyckoff et al. 2004)
1.2.2 Two hit hypothesis
Retinoblastoma is a tumour that becomes manifested early in life. Retinoblastoma can be inherited or sporadic. According to the two hit hypothesis in the inherited form a single mutation in the Retinoblastoma (Rb) gene is present in the germ line which gives the genetic predisposition to develop cancer, but a second mutation in the normal Rb allele which occurs in the retinoblast must be acquired to develop cancer (Knudson 2001). In the sporadic form the two mutations in the Rb alleles occur in the somatic cells. Although the epidemiological and molecular observations have consolidated the multistage theory of cancer, the number of mutations and in which sequential order they have to be acquired to develop cancer is still an open question (Hanahan and Weinberg 2001; Hahn and Weinberg 2002b).
1.2.3 Oncogenes
Early experiments involving transforming retroviruses and the transfer of genes from tumour cells into established rodent cells allowed the identification of several cancer causing genes called oncogenes. The result of these experiments suggested that cancer could be induced by the mutation of one proto-oncogene. However, the rodent cells used as recipient in the gene transfer experiments were not normal, but were immortalized, thus acquiring the ability to proliferate indefinitely. When the normal rodent cells were used, the transfer of a single oncogene failed to induce transformation, while the transfer of two oncogenes resulted in transformation. Human cells require more mutations than rodent cells and that there are differences also between cell types within the same species (Rangarajan et al. 2004).
1.3 Cancer Hallmarks
Despite the enormous variety of tumours affecting different types of tissues in animals and humans, research over the past 50 years has revealed that all malignant cancers share the same essential alterations (Hanahan D and Weinberg RA 2000).
These hallmarks include:
Immortalization
Evasion from programmed cell death (apoptosis)
Independence from growth stimulation
Resistance to growth inhibition
Angiogenesis
Invasion and metastasis
Genetic instability.
These hallmarks are briefly described below.
1.3.1 Immortalization
Telomeres contain DNA sequence repeats and protein. The repeat sequence consists of hexameric motifs such as GGGTTA in humans, extended for 10 –20 kilobases. The 3’ end has a 100-400 nucleotide over-hang (Mathon and LIoyd 2001). Telomeric DNA is generated by an enzyme called Telomerase Reverse Transcriptase (TERT) which has two subunits, RNA and catalytic protein subunit. This RNA binds the telomeres DNA ends thus acting as template for telomere elongation. The chromosome ends are protected by several proteins: TRF-1, TRF-2, and POT–1 (Mathon and LIoyd 2001; Hahn and Weinberg 2002a). Several experiments have shown that senescence is activated when the telomeres are shortened down to 5 kb and that senescence is triggered by the shortest telomere present in the cell (Hemann et al. 2001).
Many reports have suggested that the replicative senescence is not activated by the erosion of the double strand repetitive sequence, but by the degradation of the 3’end single strand overhang, resulting in loss of protective capping (Stewart et al. 2003). Telomere length is maintained by the activation of telomerase or by an alternative mechanism called alternative lengthening of telomeres (ALT), where the telomeres are regenerated through recombination-based inter chromosomal exchange of sequence information (Bryan et al.1997; Dunham et al. 2000). In the normal cell telomerase is transiently expressed, since it can be detected only in S phase, but in neoplastic cells its expression is increased and is detectable throughout the cell cycle (Mathon and Lloyd 2001). In tumour cells the senescence and crisis barriers are avoided by the activation of telomerase regenerating the telomeres and by the inactivation of tumour suppressor and pro-apoptotic genes (Hanahan and Weinberg 2000; Hahn and Weinberg 2000b).
1.3.2 Apoptosis.
The sensors detect the intra- and extra-cellular signals. The intracellular signals include DNA damage, hypoxia and oncogene overexpression (Evan and Littlewood 1998). The extracellular signals monitor the cell-cell and cell-matrix homeostasis (Aoshiba et al. 1997; Prince et al. 2002; Alberts et al. 2002a). The signals detected by the sensor are mainly conveyed to the mitochondria, where a series of cytoplasmatic proteins of the Bcl2 family control the release of cytochrome C from the mitochondria (Alberts et al. 2002a). The release of cytochrome C activates an array of intracellular proteases called caspases causing protein and DNA degradation (Hanahan and Weinberg 2000). The caspases can be directly activated by extracellular proteins such as FAS ligand, which binds to the death receptor FAS (Houston and O’ Connell 2004).
Once the caspase cascade is triggered it cannot be inactivated (Alberts et al. 2002a). It has been reported that the tumour suppressor p53 can trigger the caspase cascade by the overexpression of the Bax protein, a member of the Bcl2 family, which in turn increases cytochrome C release thus inducing apoptosis (Hanahan and Weinberg 2000). In CTVT it is likely that expression of c-myc is up-regulated, due to insertion of a LINE-1 element as discussed later. Ectopic c-mycexpression can promote tumour growth and survival, as seen, for instance, in immunoglobulin gene c-myc chromosome rearrangements in Burkitt's lymphoma (Hemann et al. 2005).
1.3.3. Independence from growth stimulation
1.3.3.1. Growth factors
Thus the proliferation of a cell is dictated by the needs of the cells around it (Hanahan and Weinberg 2000). In contrast, a tumour cell escapes from the external dependence to become an autonomous evolving unit, by producing its own growth signals.
1.3.3.2 Growth factor receptors
Another mechanism selected by tumour cells is the overexpressions of growth factor receptors, which induce the tumour cells to become sensitive to concentrations of growth factor that normally, do not trigger proliferation (Hanahan and Weinberg 2000). Proliferation can also be induced by a mechanism independent of the growth factor, for example the alteration of the cytoplasmic tail of growth factor receptor causes self-activation of the receptor, which therefore becomes independent from the external microenvironment (Alberts et al 2002b).
1.3.4 Resistance of growth inhibition
Like growth signals, the anti-proliferative signals derive from soluble factors or surface proteins that are produced by neighbouring cells, or are induced by components of the extracellular matrix (Hanahan and Weinberg 2000; Alberts et al. 2002d). These external inhibitory signals activate different intracellular pathways that regulate the cell cycle (Alberts et al. 2002c).
The Rb protein and its related proteins, p107 and p130 play a key role in controlling this transition (Weinberg 1995). The association of Rb with the transcription factor E2F inhibits the transcription of genes involved in the G1-S progression (Alberts et al. 2002c). The hyper-phosphorylation of the Rb protein induces the dissociation with E2F, therefore allowing progression to S phase (Alberts et al. 2002c). Normally complexes of cyclin and cyclin dependent kinase (CDK) induce the phosphorylation of the Rb protein (Alberts et al. 2002c). Many tumours can avoid the antigrowth signals by altering Rb activity or the proteins involved in Rb phosphorylation (Mittnacht 2005).
1.3.5 Angiogenesis
Although the majority of the new vessels in adult tissues are derived by sprouting from existing vessels, many evidences indicate that progenitor endothelial cells are derived from the bone morrow contributing to the vessel growth (Zhang et al. 2000; Contreras et al. 2003; Nishimura and Asahara 2005; Religa et al. 2005). Although endothelial cells are highly proliferative in response to several angiogenic factors, they have long half-lives up to several years (Carmeliet 2003). In order to adapt the vascular system to the tissue's requirements, several mechanisms regulate the process of angiogenesis (Carmeliet 2003). A key molecule involved in the angiogenesis process is the vascular endothelial growth factor (VEGF) (Carmeliet 2003). In addition it has been demonstrated that tumours can activate or inactivate pro- and anti-angiogenic factors respectively present in the extracellular matrix by producing several proteases (Gately et al. 1997; Harlozinska 2005).
1.3.6 Metastasis
In cancer during tumour progression, some tumour cells acquire the ability to migrate and form new colonies at secondary sites and these cells then make new tumour cells (Hanahan and Weinberg 2000). It has been estimated that 90 % of mortality associated with cancer is due to metastasis (Sporn 1996). Results show that few cells in the primary tumour acquire the ability to grow in the secondary sites and that the tendency to metastasise is acquired in the early steps of tumour progression (Van’t Veer and Weigelt 2003).
Progressive alteration of normal tissue homeostasis by tumour and stromal cells, allow tumour cells to move throughout degraded matrix, and to invade surrounding tissues (Hanahan and Weinberg 2000). Tumour cells are also aided to migrate by soluble factors (chemotaxis) and bound adhesion molecules (haptotaxis) (Nguyen 2004). In order to invade new organs, circulating tumour cells need to stop and exit the systemic circulation. In an unspecific manner, the extravasation may be due to the fact that large arteries progressively narrow in to arterioles and then capillaries and tumour cells can be trapped in this small vessel, thus allowing the migration in the new organ (Nguyen 2004). Although the exact mechanism behind the tumour homing is not completely understood, recent results suggest that the selective homing of cancer cells may be due to three mechanisms: 1) presence in the target tissue of specific growth factors or appropriate extra-cellular matrix that favour the selective tumour growth, 2) presence in the target organ vessel endothelium of specific adhesive proteins that interact with the tumour cells, favouring the tumour invasion, 3) production of a chemotaxis soluble factor by the target tissue that attract the tumour cells ( Fidler 2003).
1.3.7 Genetic instability
Over the past 25 years numerous genetic alterations have been described in human and animal tumours. These genetic alterations can affect the DNA sequence and the chromosomes (Lengauer et al. 1998). The mutations of DNA include: substitution, deletion, translocation and insertion and they can affect one or more nucleotides. The necessity to transmit genetic information faithfully between generations demands genetic stability (Eisen and Hanawalt 1999)
In normal conditions the genome is affected by spontaneous mutations caused by physiological DNA instability and by imprecision of the DNA polymerase proofreading activity during the DNA replication (Alberts et al. 2002e). In eukaryotic cells, several enzymes have been described with DNA polymerization activity, and five are the most important DNA polymerases involved in DNA replication and repair, alpha, beta, gamma delta and epsilon. To date the only polymerase involved in mitochondrial DNA replication is polymerase gamma. In vitro studies on the fidelity of DNA duplication has shown that the nucleotide mis incorporation rate varies among polymerases, with one in 5000 bases for beta and one in 10 000 000 for delta and epsilon polymerases (Umar and Kunkel 1996; Loeb and Loab 2000). To avoid non-complementary nucleotide incorporation, polymerase delta, gamma and epsilon contain a proofreading activity (Kunkel and Alexander 1986). Normally DNA replication is carried out by delta polymerase, but recent reports show that in some tumours this priority is shifted in favour of less accurate polymerases, thus increasing the mutation rate (Loeb and Loeb 2000). Environmental agents such as ultraviolet light, ionizing radiations and toxic substances in the dietary uptake can induce mutations (Loeb and Loeb 2000).
1.3.7a Single Base Excision Repair
When a mutation effects on a single nucleotide then base excision repair take place. BER employs enzymes called DNA glycosylases, which are specific in removing a specific mutated base (Krokan et al 2000).
1.3.7b Nucleotide excision repair
The nucleotide excision repair (NER) system is able to repair DNA damage induced by UV. In contrast to BER, the NER system recognizes altered nucleotides by scanning the DNA for a conformational alteration (bulky lesion) (Wood 1996).
1.3.7c Mismatch repair
The mismatch repair (MMR) pathway includes a series of proteins that are involved in correcting errors that escape the DNA polymerase proofreading activity during DNA replication. They are also involved in suppressing recombination between non-identical sequences both in mitosis and meiosis (Kolodner and Marsischky 1999). Unlike BER and NER, MMR does not act on damaged or mutated sequences, but it targets only the newly synthesized DNA strand.
Inactivation of the MMR system produces microsatellite instability (MSI) (Atkin 2001).
1.3.7d. Homologous recombination
Homologous recombination repairs double strand breaks by using an intact and homologous DNA molecule as a template. In eukaryotes several proteins are involved in the homologous recombination process (Kanaar et al. 1998; Haber 2000).
1.3.7e. Non-Homologous End Joining
Non-Homologous End Joining (NHEJ) is the more important repairing mechanism when there is break in DNA double strand and it is very important mechanism in mammals (Khanna and Jackson 2001). During the NHEJ process small deletions are generated. Given that majority of the mammalians genome is composed of non-coding regions, the probability that in normal situations the NHEJ process induces mutation in genes is low (Alberts et al 2002e). However, if there are multiple break points NHEJ increases the occurrence of illegitimate recombination
(Rothkamm et al 2001).
1.3.7f Chromosome Instability (CIN)
The cell reproduces by a series of events that allow DNA replication and cell division in a process known as the cell cycle. In order to check the correct order of events that take place in the cell cycle, a complex cell-cycle control system has evolved (Alberts et al 2002c). This system checks normal cell cycle progression by a series of stage-specific sensors known as checkpoints that are able to induce the arrest of the uncompleted stage until it is completed.
The two fundamental processes in the cell cycle are the duplication and the division of the chromosomes, which take place during the Synthesis (S) and Mitosis (M) phase respectively. To prevent the possibility that two daughter cells have non-identical genomes, there are two checkpoints known as DNA replication and DNA damage checkpoints before mitosis, and one known as spindle-attachment checkpoint during mitosis (Alberts et al 2002c).
Chromosome instability (CIN) is also associated with structural alteration of chromosomes, which include reciprocal and non-reciprocal translocations, amplifications, deletions and insertions (Cairns 2005). Structural chromosome instability, resulting from DNA breaks and rearrangements, is due to alteration of cell cycle checkpoints, DNA damage response and telomere integrity (Gollin 2005). Structural alterations may results in altered gene expression or produce fusion or chimeric proteins with dysregulated or new properties (Greaves and Wiemels 2003). Studies have shown that a large proportion of human tumours with chromosome instability have a high rate of loss of heterozygosity (Rajagopalan and Lengauer 2004). Therefore it has been argued that chromosome instability could accelerate the rate of inactivation or activation of tumour suppressor genes or oncogenes respectively (Rajagopalan and engauer 2004).
CIN-associated genes can be classified on the basis of the mutations (Michor et al. 2005). Class I genes of CIN, e.g Mitotic Arrest Deficient gene (MAD-2 ) boost up CIN in case one allele is mutated or deleted. Class II genes of CIN e.g. Human Budding Uninhibited by Benzimidazoles (hBUB-1) gene boost up CIN if mutation is in one allele in a dominant negative fashion. Both Class I and Class II genes are required at the spindle assembly checkpoint (Amon 1999; Hoyt 2001). Class III genes of CIN e.g. Breast cancer gene BRCA1 and another Breast cancer gene BRCA2 boost up CIN if both alleles are mutated. BRCA genes have very important role at checkpoint and it is involved in DNA repairing and recombination (Yarden et al. 2002).
1.4 Evolutionary Dynamics of Tumour Development
According to clonal evolution theory, cancer is the result of somatic mutations selected during tumour evolution (Nowell 1976). It has been argued that tumour cells cannot acquire the mutations needed for tumour progression at a physiological mutation rate, but that the tumour cell must acquire an increased mutation rate (Cairns 1998; Loeb and Loeb 2000).
In order to induce cancer the mutations must affect a variety of genes that restrain somatic conflict (Frank and Nowak 2004). These genes are known as cancer related genes and can be subdivided in three categories: Gatekeeper, Caretaker, and Landscaper (Michor et al 2004). Gatekeeper mutations increase the cellular proliferation rate by the alteration of oncogenes, tumour suppressor genes and apoptotic genes (Michor et al 2004). Caretaker mutations increase genome instability by inactivating genes involved in maintaining genome integrity (Lengauer et al 1998). Landscaper mutations increase tumour proliferation by affecting genes involved in regulating the external cellular microenvironment (Bissel and Radisky 2001).
While mutations affecting oncogenes behave in a dominant way, because only one mutated allele can induce a tumour phenotype, mutations affecting tumour suppressor genes can be neutral if the normal allele compensates the mutant allele, disadvantageous if the mutant allele triggers apoptosis, and advantageous if the mutated allele is inactivated and the second allele is insufficient to balance the wild type allele (Michor et al. 2004). In small compartments the inactivation of the two alleles of a tumour suppressor gene, is unlikely, unless the mutation rate is increased by genetic instability (Nowak et al. 2005). Loss of heterozygosity increases with chromosome instability (Michor et al. 2004).
1.5 Tumours of Feline and Canine included in this study
1.5.1Mammary Tumours
Mammary gland tumours are most frequent in dogs (Moulton 1990) while in cats it is third in prevalence, after haemopoietic and skin tumours (Misdorp et al. 1999). The average age of peak prevalence of tumours in cats is approximately 9.3 years (Roccabianca et al. 2006). Mammary tumours can also affect male cats and dogs, with the average age for them being 12.8 years (Rutterman et al. 2000). Siamese has twice the risk in comparison to other breeds of cat (Weijer et al. 1972). Same predisposition was observed with our data, that all five cases collected in this study were belonging to Siamese breed. Mammary tumours are more prevalent in Pakistan and all the cats and dogs were between 5-11 years old. This suggests that there are more chances of mammary tumours in older cats and dogs. Mammary tumours included in this study were 23% all 22 tumours studied.
1.5.2 Canine Transmissible Venereal Tumours (CTVT)
Canine transmissible venereal tumours first reported by Blaine in 1810 (Blaine, 1810) is a transmissible cancer in dogs. Studies found that CTVT was transmitted by transplantation of living cells (Novinski 1876), confirming it as a transmissible cancer. CTVT is of clonal origin, originating from a founder dog 11,000 years ago (Katzir et al. 1985; Murgia et al. 2006; Rebbeck et al. 2009; Murchison et al. 2014). It is one of only two transmissible cancers known (Murchison 2008) and is spread by allogeneic transfer of cells between dogs, usually during coitus. It manifests as a tumour, associated with the external genitalia of both male and female dogs, although tumours can also arise in the mouth, nose or skin. It is purported to be of histiocytic origin (Mozos et al. 1996; Mukaratirwa and Gruys 2003), and usually remains rather localised, except for rare cases of metastatic spread.
Recorded cases of metastasis include involvement of the lymph nodes (Higgins 1966), skin (Dass 1986) and eye (Barron et al. 1963), among others. Experimental transplants of CTVT tumours into subcutaneous sites in experimental dogs are characterized by progressive and regressive phases. This is seen as a rapid volume increase, followed by tumour shrinkage, and eventually complete regression accompanied by serum-transferable immunity to reinfection (DeMonbreun 1934). In this project we collected 6 samples for BRCA1 and COX-2 studies in different tumours while 16 more samples for Department of Veterinary Medicine, University of Cambridge, UK. Although the prevalent rate of CTVT is in second number, many attentions were paid to collect CTVTs. For BRCA1 and COX-2 studies the 28% were CTVT out of 22 different tumours.
1.5.3 Perianal adenomas/Adenocarcinomas
There are many glands present around the anus of dogs. These are sebaceous and non-secretary glands, while anal sac glands are positioned at 4 and 8 o clock to the anus and secrete their secretions into the lumen of theanal track (Yang Hai-Jie et al. 2008). Perianal adenomas are more frequent than adenocarcinomas (malignant form). In this study 3 tumours were collected which were 14% of total canine tumours collected. These tumours are mostly common in medium to older age dogs.
1.5.4 Granuloma
Granuloma is also called as lick granuloma in dogs it is a type of skin cancer It typically results from the dog’s urge to lick the lower portion of one of her or his legs. This study reported 9% of total tumours included in this study.
1.5.5 Oral Tumours (Squamous cell Carcinoma)
Oral tumours are 4th common cancers in canines. Male dogs have 2.4 times greater risk of developing oral tumours than female dogs (Dorn et al. 1968). This study reported 9% oral tumours in a period of 2 years.
1.5.6 Lymphoma
Lymphoma is the second most prevalent intra –ocular tumours of dogs. Basic cause of lymphoma in dogs is unknown but genetic (chromosomal segregation), environmental and infectious factors such as retroviruses play vital role in developments of this cancer (Fighera et al. 2002). This study reported 9% Lymphomas of total collected tumours.
1.6 Rationale behind selection of genes
1.6.1 BRCA1 gene
BRCA1 gene is tumour suppressor gene, it is involved in repairing the DNA double strands breaks and in case of failure it leads the cells towards apoptosis (Starita. 2003). BRCA1 forms BRCA1 Genome Surveillance Complex (BASC) when it combines with different types of tumour suppressor genes, DNA damage sensors and signal transducers (Wang et al. 2000). It is involved in Ubiqutination, transcription regulation (Friedenson 2007; Friedenson 2008). In humans BRCA1 was first identified at chromosome 17 (Hall et al. 1990) and it was isolated in 1994 (Miki et al. 1994). It is present at 17q21 with a length of 100 Kb. In canine it is located on chromosome 9. BRCA1 has 22 exons in canines and felines; it encodes a protein of 1882 amino acids in canine and 1871 amino acids in feline. Many scientists from different research showed that women who have famililal mutations in the BRCA1 or BRCA2 (BRCA1/2) genes have increased risk of breast cancer (Struewing et al. 1997).
Fig 1: BRCA1 mechanism in DNA repairing. http://www.publichealthunited.org/leading-by-example-angelina-jolie-and-the-brca1-gene-mutation/
1.6.2 Cyclooxygenase-2 Enzyme (Prostaglandins, COX-2).
Cyclooxygenase-2 enzyme (Cox-2) is also called as Prostaglandins Endoperoxide synthase (PTGS). It is involved in the synthesis of prostaglandins which act as biological mediators in many body functions. It was first isolated from prostate gland that’s why it is called as Prostanglandin. Cyclooxygenase enzymes have two types, cyclooxygenas-1 and cyclooxygenase-2. Cyclooxygenase-1 is constitutively produced in the cell while cyclooxygenas-2 is inducible and it is constitutively produced only in kidneys, seminal vesicles and central nervous system. Its high expression has been recorded in many different types of tumours, it has been involved in anti-apoptosis, cell proliferation, tumour angiogenesis, cell invasion and immune suppression activities. In canine COX-2 is present on chromosome 7 having 604 amino acids and 10 axons. This correlation of cyclooxygenase-2 in cancer development suggests using new therapeutics against it. Studies have shown cycoloxygenase-2 high expression in number of different tumours (León-A 2008), such as intestinal, pancreatic, ovarian, prostatic, nasal cavity, oral cavity and mammary tumours of dogs (McEntee et al. 2002; Mohammed et al. 2004; Borzacchiello et al. 2007; Eplattenier et al. 2007; Mullins et al. 2004; Pireset al. 2010; Dore et al. 2003).
Fig 2:COX-2 mechanism of actionhttps://www.google.com.pk/search?q=cox+2+mechanism+of+action.
1.6.3 DLADQA1 (MHCII gene) (Additional work performed at Department of Veterinary Medicine, University of Cambridge, UK).
The Major Histocompatibility Complex (MHC) is a cell-surface protein mediating immune recognition through its interactions with T cells (Fig 3). There are three classes of MHC molecules in mammals - the classical MHC-I and II, and non-classical MHC-III Table 1). MHC-I interacts with CD8+ cytotoxic T cells, whilst MHC-II binds to CD4+ helper T cells. MHC molecules mediate antigen presentation to T cells. MHC-I typically presents self- peptides, whilst MHC-II presents foreign peptides. MHC molecules are extremely variable and polymorphic across the population, with a huge number of alleles at each MHC locus.
This allows MHC molecules themselves to behave as antigens in transplant rejection, with the graft MHC peptide recognized as non-self by the recipient, and thus rejected. It would be expected that CTVT, an allergenic graft, should be rejected for two reasons: host MHC will present tumour antigens as foreign non-self to the host immune system, and tumour MHC will present a mismatch to the host immune system as a foreign antigen itself (Fig 3). This project focuses on the DLADQA1 locus (Wagner et al. 2002), a classical MHC-II gene on dog chromosome 12. There is high level of MHC allelic variability in any population (Niskanen et al. 2013).
Fig 3:MHC is involved in graft rejection.
This rejection (represented by the red arrow) occurs according to two principles. Firstly, host T cells may recognize the host MHC presenting a foreign peptide that should activate an immune response. Secondly, host T cells would also be able to recognize the tumour MHC presenting any peptide as foreign, since it is not self-MHC. It is thus surprising that CTVT is able to persist as an allogeneic graft. MHC expression was previously characterised molecularly by Murgia et al through RT-PCR of a MHC-I (DLA88) and MHC-II (DLADRB1) gene (Murgia et al. 2006). They found that there was downregulation of expression of both these MHC genes. DLA88 showed low levels of tumour-specific expression, whilst there was no detectable tumour expression of DLADRB1. Murgia et al. also performed MHC genotyping for a number of CTVT samples and confirmed that all CTVTs shared the same haplotype (Murgia et al. 2006). They identified two clusters at the DLADQA1 locus, with some CTVTs appearing to be haploid the locus, whilst others remained diploid. This is in contrast to evidence that suggests the DLADQA1 locus had undergone a copy-neutral loss of heterozygosity (LOH) (Murchison et al. 2014).
1.6.4 Technologies used in this research work.
Different technologies are being used in cancer research such as PCR, flow cytometry, immunohistochemistry (IHC), in- situ hybridization (FISH, CSH) and microarray for diagnosis (Pawanet al. 2010). Here, I used Real time PCR for gene expressional analysis of BRCA1 and COX-2 and DLADQA1 (MHCII). Histopathology (Hematoxyline and Eosin staining) was performed for the diagnosis of tumours. CTVT diagnostics qPCR was also performed to measure the allele’s quantity of LINE-myc gene and CDKN2A gene. Conventional PCR measures at End-Point, while Real-Time PCR collects data during the PCR shows the data and quality of data during exponential growth phase also it has increase dynamic range of detection, it is very sensitive and no need for post PCR processing. Immunohistochemistry was performed to find out the expression of MHCII antigens in CTVTs. The serum protein electrophoresis and serum biochemistry was also measured. Western blotting was performed to detect antibodies in CTVTs (protein expression). It is a very good technique to measure the gene expression at protein level in fluidic material of cells. We performed capillary electrophoresis to find the mutations/SNPs in our genes of interest (BRCA1, COX-2 and DLADQA1). Genetic analyzer was used to find the sequence variations in our genes of interests. Other methods used for sequence variation studies, like SSCP, DGCG and HPLC miss the mutations (Rassi 2009). So the sequencing by capillary electrophoresis was the best option for this study.
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809.
Development And Evaluation Of Mozzarella Cheese Influenced By Type Of Starter Culture And Fat Content Of Milk
by Imran Taj Khan (2013-VA-866) | Mr. Muhammad Junaid | Dr. Muhammad Ayaz | Mr. Muhammad Saadullah.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is the 4th largest milk producing country in the world having 50.99 million tons/year in which buffalo and cow add 61% and 35% share, respectively to total milk production in Pakistan, while the rest 5% of the total milk is contributed by other species like small ruminants and camel. Out of the total produced milk only 7-8% of the milk is processed and passes via milk supply chain while the remaining is distributed as raw milk (GOP, 2013-14). Processing of milk into cheese is a small segment in Pakistan, moreover the eating habits are not developed in Pakistani population along with the increase in price of local as well as imported cheese.
The manufacturing of cheese is one of the best examples of preserving milk, dating back from 6000-7000 BC till now. Cheese like other value added dairy products are among the highly nutritious diet of the world. Cheese comes in great variety of tastes and flavors, in a wide array of shapes, and are highly healthful and nutrimental (Walstra et al. 2006). The utilization of cheese in the form of pizza garnishes, cheese mixes, salads, sandwiches, stuffing has expanded because of dietary resemblance, ease and extensibility being used and cheese quality (Fox et al. 2000). Cheese manufacturing is increasing worldwide at the rate of 4.9% every year (Valhovic et al. 2014).
Cheese is the value added dairy product acquired by the seepage (of fluid) after coagulation of milk proteins through utilization of rennet and starter. Cheese is a mainstream nourishment because of its differing qualities in application, wholesome quality, accommodation and appealing taste. The assorted qualities are because of an expanding information of the innovation of cheese making and the natural chemistry and microbiology of cheese maturing (Sulieman et al. 2013). There are 500–800 assortments of cheese accessible in the universal business sector (Razzaq, 2003).
Mozzarella cheese is placed in the group of “Pasta filata” or stretched cheeses. Stretching is a treatment that gives the curd elasticity which is a dominating functional characteristic of Mozzarella cheese. Melting and stretching are the most important characteristics of Mozzarella cheese which is highly acknowledged in the making of Pizza as it is a chief ingredient (Owni et al. 2009).
Mozzarella cheese is a standout amongst the most prominent cheese varieties on the planet, in light of its essential use in the pizza topping (Kindstedt et al., 2004). The funtional attributes of Mozzarella cheese like shape, composition, meltability, stretchbility and color are changed by the components like milk composition, culture source or type and maturing environment dominating in the midst of the cheese availability (Luecy et al. 2003).
Mozzarella cheese is produced utilizing a paired lactic acid bacteria starter cultures of Streptococcus salvarius ssp. thermophilus and Lactobacillus delbrueckii ssp.bulgaricus (Ahmed et al. 2011).
At a certain point, Mozzarella was made just from water buffalo milk. Presently, it is normally produced using milk of bovine, goat and sheep. There are two types of Mozzarella cheese; i.e. regular Mozzarella which is accessible in low-fat and nonfat structures and has a semi-delicate in texture, versatile composition and is drier than fresh Mozzarella while, Fresh Mozzarella is produced using entire drain and has a gentler texture and sweet pleasing flavor and is commonly pressed in water or whey (Ibrahim, 2003).
Mozzarella cheese also has numerous therapeutical advantages; it is a decent wellspring of protein, vitamins and minerals. Utilization of Mozzarella cheese may secure against gout, an agonizing condition that outcomes in the development of uric acid stones in the joints. The calcium found in Mozzarella cheese additionally has its commitment in body weight reduction and gives insurance against breast tumor and metabolic disorder, which is a gathering of conditions that build the danger of creating cardiovascular disease or stroke (Ibrahim, 2003).
Low fat Mozzarella cheese is a nice wellspring of protein and calcium, furthermore it is non calorie-dense and less in saturated fat, which can prevent from cardiovascular illness, type 2 diabetes, joint inflammation, and memory loss, also block weight reduction endeavors upon consumption. Eating low fat ("part-skim") or fat-free Mozzarella cheese is an extraordinary approach to gain by its alimental advantages while minimizing unfortunate additional items. The calcium in Mozzarella cheese aides keep up healthy pulse rate and blood pressure (Bauer, 2014).
Mozzarella cheese production in the country in demand based being its most use in the pizza sector while its limited use as fresh product. Moreover the industry is using commercial imported culture for production of this cheese. The high price of the culture is one of the factor in the high price of the final product. Therefore the present study was designed with an objective to utilize the local culture for production of Mozzarella cheese. The idea was to maintain the quality of product at its higher regime. Furthermore its quality attributes were compared with the standard cheese produced from commercial culture. In addition the effect of change in fat level of milk on the quality and overall acceptability of the produced Mozzarella cheese was assessed.
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810.
Antimicrobial Potential Of Bovine Lactoferrin Against Foodborne Pathogens
by Ammarah Khatoon (2012-VA-631) | Prof. Dr. Muhammad Ayaz | Mr. Ishtiaque Ahmed | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Health is recourse of everyday life, but not the object to live. It is positive to give special importance to personal and social resources. However, in Pakistan and other developing countries conditions are different, most people have low income and they live in un-sanitized environment. They eat un-hygienic food and also lack safe drinking water. People do not adopt any preventive measure to minimize the risk of contamination. Food storage is also un-hygienic. These conditions lead towards contamination and result in foodborne infections and gastro-enteritis. Foodborne illnesses are always a serious health issue in the Pakistan and throughout the world. Individual’s record for foodborne illnesses is impossible but it is reported that 7 out of 10 people suffer from foodborne illness caused by different microbes each year worldwide (WHO survey 2012).
Foodborne illness is caused by eating contaminated food with pathogenic bacteria. Some common pathogens are Escherichia coli, Campylobacter jejuni, Clostridium botulinum, Clostridium perfringens, Listeria monocytogens, Bacillus cereus, Staphylococcus aureus and Salmonellaspecies. Incubation period for onset of symptoms of food poisoning ranges from hours to days. Nausea, vomiting, abdominal cramps, fever and diarrhea are symptoms which appear commonly in most of food poisoning. However, foodborne illnesses if left untreated can lead severe dehydration, imbalance of intestinal micro flora, digestive disorders and even death in some cases. It was recorded that 2.2 million people killed from foodborne illness globally every year and the burden arising from foodborne diseases is larger (Kuchenmuller et al. 2009).
Antibioticsare massively used to overcome food poisoning; however, from health point of view they badly affect thenormal micro flora of gut but also microbes become antibiotic
resistance. The problem needs to be dealt with some other way like adding bio preservatives or antimicrobial agents in food.
To control microbes in foods, numerous methods have been adopted including the use of synthetic and natural antimicrobial agents. Scope of natural antimicrobial agents are increasing day by day and different natural sources are being utilized to get these agents. Among these natural sources milk is best and widely utilized source from long times. Milk contains many biologically active compounds among which lactoferrin is one of them.
Lactoferrin is a multifunctional globular glycoprotein from transferrin family, an iron-binding protein. It is part of innate immune system and has antibacterial activity known as far back as 1930. It was first isolated in 1939 from cow milk (Charrondiere et al. 2011). Lactoferrin belongs to the transferrin family having ability to bind iron two times higher than other transferrin proteins. Its molecular weight is 80 kDa and has about 700 amino acids depending upon species e.g. cow, buffalo, goat and sheep (Adlerova et al. 2008). Lactoferrin molecule consists of simple polypeptide chain folded into two symmetrical and highly homologous lobes (N and C) connected by a hinge region. Both lobes bind two metal ions in synergy with carbonate (CO32-). Not only Fe2+ and Fe3+ ions but Cu2+, Zn2+ and Mn2+ ions can also bind. Lactoferrin can bind Fe3+ reversibly so it can exist as free of Fe3+ (Apo-Lf) or in association with Fe3+ (Holo-Lf) and exhibits different three dimensional structure depending upon binding to Fe3+. Apo-Lf has an open structure and holo-Lf has closed which provide resistance to proteolysis. At iron-binding site Aspirin, two Tyrosine, and Histidine amino acids are directly involved in each lobe and Arginine is bound to CO32- ions. Number and position of Cystine-residues allows intermolecular disulfide bridges and Asparagine-residues in both lobes provide several sites for N-glycosylation (Farnaud and Evans 2003).
Lactoferrin is produced by mucosal epithelial mammary cells of human, cows, buffaloes, goat, horses, many other mammals and fish. It is widely distributed in body tissues and present in mucosal surfaces, specific granules of leukocytes and in biological fluids like tears, saliva, digestive fluids, seminal fluids and most abundant in milk comprising the second highest protein in human milk after casein. Concentration of lactoferrin in different species is for cow milk (80-500 mg/L), buffalo milk (50-320 mg/L), camel milk (200-728 mg/L), goat milk (98-150 mg/L) and sheep milk (20-140 mg/L) (Krol et al. 2011).
Many physiological functions of lactoferrin have been attributed. It plays an important role in iron regulation, non-specific immune response, regulation of cells growth and differentiation, protection from cancer, anti-inflammatory, anti-oxidant and strong antimicrobial activity against bacteria, fungi, yeast, viruses and parasites (Conneely et al. 2005). Another dominant role of lactoferrin is during involution of mammary gland. Concentration of lactoferrin increased dramatically from 0.1-0.3mg/ml in normal milk to 20-30mg/ml by 30 days in dry period. It is particularly important for bacteriostatic properties and non-specific defense against invading bacteria. Lactoferrin also affects phagocyte function and limit oxidative degeneration of cell components during inflammation and involution (Welty et al. 1976).
Lactoferrin exhibits strong antimicrobial activity against different bacteria, virus, protozoa, fungi and yeast (Hancock and Janssen 2009). The antibacterial activity of lactoferrin is due to two mechanisms; by binding the iron at infection sites, making it unavailable to bacteria and direct interaction of N-terminal of lactoferrin with micro-organism (Cruz et al. 2009, Orsi 2004). Lactoferrin acts differently with Gram-positive and Gram-negative bacteria (Sharma et al. 2013). It damages Gram-positive cell wall through interaction with negatively charged lipoteichoic acid causing reduction in negative charge on cell wall and favor contact between lysozyme and inner peptidoglycan (Fayad 2012). Gram-negative bacteria are destroyed by interaction of lactoferrin with external lipopolysaccharides by preventing contact with Ca2+ and Mg2+ ions which cause release of lipopolysaccharides, increase permeability and ensures damage (Ochoa and Cleary 2009, Ekins et al. 2004).
Milk and milk products are one of main diet in Pakistan and all over the world. During manufacturing different milk products, a number of by-products are obtained. Among them, cheese whey is produced in high volumes. It is commonly dumped off into sewerage which cause serious environmental problem as it contain high organic matter as well as loss of valuable nutrients it contain. Whey has Biological Oxygen Demand (BOD) ranges from 40,000 to 60,000 ppm (Sayadi et al. 2006) while permitted limit for BOD of domestic sewerage is 200 to 300 ppm. In order to overcome this problem there is need of effective and permanent way for treatment of whey. However, conversion of whey into non-food items like biogas is unreasonable as it is rich in unique nutrients.
Now-a-days there is an interest growing on to find new ways of whey utilization throughout the world. One option is to use the whey in processes in which saleable food or pharmaceutical products can be obtained. Whey could be subjected to different techniques to isolate different components like lactose, lysozyme and immunoglobulin. Likewise, lactoferrin can be isolated from cheese whey by cation exchange chromatography without loss of its biological properties in single step method and about 90% purity (Wu et al. 2011, Moradian et al. 2014).
In this study, we anticipated to use lactoferrin from bovine milk as natural antimicrobial agent. It has been shown that lactoferrin hasstrong antimicrobial activity against different bacteria, fungi, yeast, viruses and parasites (Conneely et al. 2005). In our country, very little work has been carried out onlactoferrin as natural anti-microbial agent. In fact, all over the world, the research scenario is now changing and concentrating toward the extraction of natural agents for product safety and health improvement. The lactoferrin has a potent anti-microbial activity against common foodborne pathogens. Due to the negative health effects of synthetic anti-microbial agents, the uses of natural sources are being encouraged all over the world. Our main focus of this study is to check the anti-microbial activity of lactoferrin against three pathogenic bacteria Escherichia coli, Staphylococcus aureus and Salmonella enteritidis isolatedduring our previous study.
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811.
Effect of Curcuma Longa on Embryonated Eggs Experimentally Infected With Avian Influenza Virus
by Syed Iftikhar Ali Shah (2013-VA-436) | Dr. Muhammad Yasin Tipu | Prof. Dr. Asim Aslam | Prof.Dr.Muhammad Sarwar Khan.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: CD Error Availability: Items available for loan: UVAS Library [Call number: 2264-T] (1).
812.
Effects Of Short Term Anti-Stressor Feeding On Bone Characteristics And Tissues Minerals In Heat Stressedbroilers
by Vishal Sharoon (2013-VA-567) | Dr. Muhammad Shahbaz Yousaf | Dr. Muhammad Quaid Zaman | Dr. Saif-Ur-Rehman Kashif.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Broiler chicken is the fastest growing meat type nowadays. Increased broiler’s growth rate increases their body heat production. When the body becomes overheated, a condition of heat stress exist which results in decreased growth rate primarily due to reduced feed intake (Hurwitz et al. 1980). Heat stress in birds cause suffering that leads to death and to reduce this problem of heat stress, heat production needs to be reduced, which can be done by evaporative heat loss through respiratory tract. Many other techniques have been developed to alleviate heat stress like upgrading the house padding (Nilipour 2000; Yahavet al. 2004), setting upcooling systemssuch as evaporative cooling (Donald 2000), reducing bird density, providing cool drinking water, and feeding birds during the cooler times of the day like in the morning hours or when the sun sets (Lott 1991; Yahav and Hurwitz 1996).
Minerals are essential for the naturalfunctioning of the body. Manganese, zinc, copper, calcium, and phosphorus are a few minerals that are important for the growth and development of bone (Watkins and Seifert 1997). Heat stress increases mineral excretion because digestibility is affected intestinal motility is increased (Smith 1987). El-Husseiny and Creger (1981) reported that heat stress decreases rates of calcium, iron, potassium, magnesium, sodium and zinc preservations in broilers. Similarly, Sahin et al. (2006) found that heat stress decreases tibial ash, calcium, phosphorus, and magnesium in quails. It also decreases length and widths of tibia, and promotes mineralization which in turn reduces bone strength of broilers (Vakili et al. 2010).
Heat stress also has negative effects on concentrations of potassium, sodium, phosphorus, copper, and zinc in serum. Their reduction is reported under heat stress conditions in broilers (Belay and Teeter 1996).Heat stress has also shown decreased iron, and chromium concentrations in serum. Heat stress also effects liver minerals and has shown reduction in the liver concentrations of iron, zinc, and chromium (Sahin et al. 2001).
As it is quite expensive to keep animal buildings cool, different methods of dietary manipulation are mostly considered. During heat stress, synthesis of vitamin C, E, zinc and chromium is decreased. So poultry diet is supplemented with these minerals due to their anti-stress effects (Sahin and Kucuk 2001). Melatonin supplementation also restores the decreased concentrations of vitamin C, vitamin E, vitamin A, Fe, Zn, Cu, Cr, in serum induced by heat stress (Sahin et al. 2004). Genistein supplementation is also helpful in increasing apparent nutrientdigestibility and bone mineralization. The reduced amount of minerals is excreted out of the body whereas the mineral concentration in tibia ash increases (Sahin et al. 2006). A supplemented dietincluding chromium at 1200 ppb can lessen the detrimental effects of heat stressed broiler. Feed intake, body weight and improved feed effectivenessincreases as a result of increased chromium supplement (Sahin et al. 2002). Another dietary practice to reduce heat stress of broiler is to supplement poultry feed or drinking water with various electrolytes which will increase their water intake. Addition of Dietary fibre in feed also has beneficial effects on broilers (Lee 1994). ASI supplementation (ASI; Arg =49.47%, silicone = 8.2%, inositol = 25%) considerably enhanced bone mineralization in quails and had no effect on weight gained, feed effectiveness and feed consumed (Sahin et al. 2006).During heat stress, KCl at level of 1.5% and NaHCO3 at level of 0.5%, improvedweight gain, and significantly increased serum potassium and bicarbonate level during heat stress. The results showed that combination of KCl and NaHCO3 supplementation decreased the negative effects of heat stress (Naseem et al. 2005).
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813.
Application Of Microsatellite Markers For Genetic Diversity Analysis Of Endangered Punjab Urial (Ovis Orientalis Punjabiensis) In Pakistan
by Anam Aftab (2012-VA-534) | Dr.Tanveer Hussian | Dr. Wasim Shehzad | Dr. Muhammad Tayyab .
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Biological diversity is now recognized as common concern of mankind and genetic diversity is the major driver of variation within and across breeds, which helps populations to adapt to environmental changes. There is very little importance is given to conserve wild sheep in previous years and its genetic diversity is decreasing day by day. Every now and then breeds are being haunted for crossing to some other imported breed without attempting to see if such efforts will be sustainable. For any breed development efforts thus, available genetic resources need to be characterized both at phenotypic and genetic levels (Khan et al. 2007).
Among the three levels of Biodiversity, one is Genetic variation which is suggested bythe International Union forNatureconservation (IUCN) for preservation (Mc Neely et al. 1990). The reason for it is that firstly; genetic diversity favors the changes as the environment changes and secondly; it prevents inbreeding depression (Reed and Frankham 2003). In this way genetic diversity increase the survival status and increase fitness of individuals.
Among many other wild animals present in Pakistan, there are 6 to 9 species of wild sheep (Ovis orientalis) are present which have different color and size of their winter neck ruff of males, saddle patches and horns color. Urial is a picture of Marco Polo in texture and hue. In Pakistan, ladakh urial, Blanford urialand Punjab urial are found in Gilget, Baluchistan and Punjab respectively. The discrepancy lies in the color of ruff among these 3 sub species (Roberts, 1977). Urial is among those precious wild animals that were hunted severely for trophy and other purposes in the past, that’s why included in red list of IUCN in vulnerable category (IUCN 2000).
Punjab Urial (A type of wild sheep – Ovis vignei punjabiensis) belongs to family bovidae which is the large family consisting of 140 species (Glazko et al. 2011) is facing serious threat of extinction in Pakistan is a medium-sized wild sheep which is included in IUCN red list of endangered animals (IUCN 2002). Urial is inhabitant in Western Central Asianregion stretching from northeast side of Iran and west side of Kazakhstan to Balochistan (Pakistan) and Ladakh regions of North India. The local name of Urial is Shapo, Arkar and Gad. Reddish-brown outstretched pelt that achromatizes during the winter is one of the distinct traits of urial (Aleem 1977; Schaller 1977). Urial is gregarious and sexually dimorphic as males are called rams and females as eves (Awan 2001). Male have weightof 40 kgand have large spiralled horns having height of 80 to 100 cm and females have comparatively less weight and height of 25 kg and 12 cm respectively and have uncurled horns.Females give birth to 1 or 2lambs in recent days of April (Awan, 2001). Males have a black ruff expanded from the neck to the trunk and notably longer horns. Table 1.1:Some physical features of the Punjab Urial (Awan et al. 2001)
Features Urial
Body weight 40 kg male; 25 kg female
Shoulder size 31-35 inches or 80-90 cm
Horn size 39 inches or 80-100 cm long male; 12 cm long female
Urial are found in moderate to very arid habitats, especially grasslands including agricultural fields and woodland areas (Valdez, 1982). Urial is herbivorous and eats grasses, shrubs and grains. The patch of salt range of Pakistan which fall in the area of Pind Dadan Khan, Choa Saidan Shah and Kallar Kahar is considered like a paradise on earth for the wild fauna. The fascinating hills of these areas are covered with thick trees and different wild plants are also the sanctuary of urial.
Table 1.2: The details of these sub species in IUCN list of endangered mammals (IUCN 2002).
Subspecies Citation in IUCN list
Ovis vegnei blanfordi VUC 1 Appendix ll
Ovis vegnei punjabienses ENA1cde,c1+2a Appendix ll
Ovis vegnei vegnei VUC 1 Appendix ll
In Pakistan, Punjab Urial dispersed throughout the Kala Chitta and Salt Range in a very little number(Hess et al, 1997). The Afghan Urial inhabits Baluchistan, Khyber Pakhtunkhwa, and Sindh Provinces. In Chitral District, little segregated populations of Ladakh urial are stillextensively distributed near the west bank of the Kunar river from Chitral southwards to Drosh. Ladakh Urial existence at the east bank of Kunar river and north of Chitral are not proved (Malik 1987). Total population estimate of urial is recorded by conducting several surveys. Reasonably 2,500 to 3,000 urial existsin Baluchistan (Hess et al. 1997). In 1993 the overall population assessment of Northern Areas was four hundred to five hundred Urial (Rasool 1999). In Pakistan, there were seemingly less than 600 Ladakh urial (Hess et al. 1997;NWFP 1992; Schaller 1971, 1977) but the number of urial decreased to 200 to 300 urial in all over northern areas (Rasool 1999). Based on the facts mentioned above there is dire need to conserve the Urial population in the country. The urial is one the precious fauna of Pakistan and provides us with wool and meat (fat, flesh or any eatable part). It is also important in economic way and in maintaining of ecosystem balance. In the beginning, sheep were reared for meat, milk and skin (Ensminger and parker, 1986). After 3500 B.C. men learnt to spin wool and so used wool in textile industry (Smith et al. 1997). So, because of increasing world population, there is great demand of these products and increasing day by day. That’s why we have to conserve is natural resource of Pakistan.Habitat fragmentationleads to the risk of exaggerated genetic drift and inbreeding in isolated population. So, there is need to save urial from these threats by enforcement of law and conserving it for future.
In all over the euchromatic genome Microsatellite markers are present. These markers are highly polymorphic (Ellegren 2000; Schlotterer 2000).A lots of polymorphic microsatellites have been analyzed in ruminants like domesticsheep, cattle etc. (de Gortari et al. 1997,1998 ;Hayes et al.1996; Jenkings et al. 1997) aiding the use of these in parentage testing.
Microsatellite markers are among the most reliable molecular markers for genetic characterization studies in animal species (Sunnucks, 2001) and are simple sequence repeats (SSRs) of 1-6 base pairs, repeated tandemly in coding as well as noncoding portion of DNA in prokaryotes and eukaryotes (Weber and May, 1989; Toth et al., 2000). Microsatellite markers have often used for genetic diversity studies because they areabundant, unbiased, widely distribution all over the DNA, highly polymorphic, easy in assessment like genotyping of these markers (Canon etal. 2001).Microsatellite markers aid in genetic differentiation and conservation studies (Peter et al. 2007;Rendo et al. 2004; Arranz et al. 2001).These are considered very useful markers for assessment of genetic diversity, parentage confirmation, genome mapping, disease research population genetic studies and conservation genetics. These are also reported to be efficient enough to identify within and among breed differentiation and population sub structuring in cattle (Glowatzki-Mullis et al. 1995; Ciampolini et al. 1995; Garcia-Moreno et al. 1996; Jarne and Lagoda, 1996; MacHugh et al. 1998).Therefore the conservation activities are very important to save Punjab Urial from extinction and the study is designed to explore its genetic diversity.
Availability: Items available for loan: UVAS Library [Call number: 2261-T] (1).
814.
Seroprevalence And Associated Risk Factors Of Leptospirosis In Sheep And Goat In And Around Lahore
by Muhammad Awais Akram (2008-VA-230) | Dr.Muhammad Hassan Saleem | Dr. Muhammad Avais | Dr. Hassan Mushtaq.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Sheep and goats, although representing an important source of animal protein in third world countries such as Pakistan, seem to have benefited little from veterinary care and production improvement. Sheep and goats are often the main source of daily meat and are used in ceremonial festivities throughout the country. Small ruminants (sheep and goats) are ubiquitous, with important contributions to the subsistence, economic, and social livelihoods of many humans, particularly in developing countries. According to FAO, (2010), approximately 95.7% of all goats and 63.3%of all ewes worldwide are located in developing countries and represent more than 70% of total animal production. Among the various factors that may lead to low productivity in tropical countries, infectious diseases may be very prevalent, impairing milk and meat production.
Leptospirosis is an outstanding neglected disease, and since it is usually silent, its effects on livestock are often underestimated. As an example that may be considered for other tropical areas of the world, it was recently described as the most frequent and potentially the major infection impairing productivity in small ruminants. Unfortunately, a definitive diagnosis of leptospirosis is difficult to make. Most of diagnostic laboratories do not attempt to isolate leptospires because of their fragile nature, cost and complexity of the isolation media, and prolonged incubation period. Therefore, recognition of leptospiral infection has been based generally on serological evidence. A wide variety of serological tests, which show varying degrees of serogroups and serovar specificity, have been described. Two tests have a role in veterinary diagnosis: the microscopic agglutination test (MAT) and the enzyme-linked immunosorbent assay.
A total of 180 serum samples were examined in this study. The animals were included in this study from various sources representing the diverse livestock production system / management conditions i.e. rural subsistence, peri-urban and semi commercia, sheep and goat farms in and around the Lahore. The blood samples were collected from randomly selected animals as well as on the basis of Leptospirosis- like symptoms or any other indication of the disease. The blood samples were collected in clean sterile vacutainers having no anticoagulant. From each animal 5-10 ml blood sample were collected by phlebotomy. For this purpose the area on jugular vein was sterilized with 70% alcohol and blood was collected in the vacutainer. The blood samples were put in slanted position in the refrigerator for two hours. Upon appearance of serum in the tubes usually after three hours of refrigeration, samples were centrifuged at 4000 revolution per minute (RPM) for five minutes. The sera were separated using a sterile pipette for each sample and clean sterilized vials were stored at -20°C in the freezer till used. The standard hygienic measures were adopted during collection and processing of blood samples. The ELISA is performed by the procedure that is described by the ELISA kit manufacturer.
The sheep were divided into three categories that were healthy, pregnant and aborted, which account for 0, 3.34 and 26.6%, respectively, positive samples. Whereas, The goat were also divided into three categories that were healthy, pregnant and aborted, which account for 0, 6.67 and 30%, respectively, positive samples. The highest percentage were observed in aborted animals that indicated that the leptospirosis had contribution in the abortion of the goat and sheep. The sheep were divided into four categories that were urban, peri urban, semi-commercial and commercial, which account for 13.3, 6.67, 6.67 and 10%, respectively, positive samples. Whereas, the goat were also divided into four categories that were urban, peri urban, semi-commercial and commercial, which account for 16.7, 13.4, 6.67 and 10%, respectively, positive samples. The highest percentage were observed in urban areas where the sheep and goats were raised together that indicated that the leptospirosis can be spread from animal to animal.
Conclusion:
From the finding of the current study suggested that leptospirosis can be difficult to diagnosis properly. The proper diagnosis can helpful for the controlling the leptospirosis. The urban area, and physiological conditions, of sheep and goat, are the major risk factors.
Suggestion and Recommendations:
Proper diagnosis and good management can reduce the risk of leptospirosis in sheep and goat. The infected animal must be isolate and treat with proper medications. The further studies can helpful for more proper disease diagnosis and control. Availability: Items available for loan: UVAS Library [Call number: 2274-T] (1).
815.
Comparison Between Aspiration And Slicing Methods For Retrieval Of Oocytes In Bovine
by Muhammad Husnain (2008-VA-281) | Prof. Dr. Mian Abdul Sattar | Dr. Qaiser Shahzad | Dr. Amjad Riaz | Dr. Muhammad Avais.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Livestock contribution to agriculture stood at 55.9 percent while it contributes 11.8 percent to the national GDP during 2013-14. Buffalo, cattle, sheep and goat population in Pakistan is 34.6, 39.7, 29.1 and 66.6 million numbers during 2013-14. Total milk production from buffalo and cattle as major milk producing animals is 31,252 and 18,027 (000 tons) (Economic Survey of Pakistan 2013-14).
Advanced biotechnologies coming from different areas of biological sciences exhibit great promise to enhance the efficiency of livestock production. From these technologies one such biotechnology is the use of in vitro maturation of follicular oocytes and in vitro fertilization for production of livestock embryos in laboratory. Proper oocytes recovery and their selection in the laboratory are of great importance for successful in vitro embryo production. Total one hundred and forty four ovaries (n=144) from cattle (72 ovaries) and buffalo (72 ovaries) were collected and 223 oocytes were retrieved from these ovaries. Average oocytes per ovary were 1.66 + 0.43 oocytes per ovary were obtained via aspiration and 1.89 + 0.00 average oocytes per ovary through slicing method from cattle ovaries. Average 1.55 ± 0.55 oocytes per ovary via aspiration and 1.53 ± 0.20 oocytes per ovary through slicing from buffalo ovaries.
Overall grade-A oocytes were 28 (40) percent with aspiration in cattle and 25(36.76) through slicing method. In buffalo overall grade-A oocytes retrieval was obtained in percentage as 20 (44.44) and 26 (52) through aspiration and slicing methods respectively. Grade-B oocytes recovery obtained was in percentage as 23 (33.82) with slicing and 19 (31.67) through aspiration technique from cattle ovaries.
Summary
26
Commonly used methods of recovery of oocytes from slaughterhouse animals are aspiration and slicing. Recovery rate of oocytes is different from slaughterhouse ovaries. Aspiration is the best method for retrieval of good quality oocytes from slaughterhouse bovine ovaries because it gave more good quality oocytes in less time than slicing method. In this study, it is found that weight of ovary and no. of follicles/ovary in cattle have strong correlation of 71% existed between weight of ovary and no. of follicles /ovary in buffalo was observed. Correlation between average number of follicles on ovary and weight /ovary was stronger in cattle. The more the number of follicles present on the ovaries and more weight of the ovary, the more will be the recovery of oocytes.
In cattle average number of follicles was 10.09 ± 0.30 and when it was checked in buffalo, differed significantly and it was found as 7.16 ± 0.19 on an average per ovary. Likewise weight of buffalo in this study was differed significantly from cattle 4.04 ± 0.10 and 7.62 ± 0.15 respectively. It is suggested that oocytes retrieval should be done in buffalo using aspiration method to retrieve better quality oocytes.
It is concluded that aspiration is the suitable method for retrieval of good quality oocytes from slaughterhouse buffalo ovaries because it gave more good quality oocytes in less time than slicing method. But both methods have minor difference between recovery rates but aspiration is more convenient than slicing and it yields more quality oocytes. It is also found that there is very strong correlation existed between average weight of ovary and number of follicles per ovary and the both parameters play a great help for more quality and quantity oocytes. Availability: Items available for loan: UVAS Library [Call number: 2279-T] (1).
(1 votes)
816.
Effect Of Taurine Supplementation On Post-Thaw Quality Of Sahiwal Bull Semen
by Muhammad Irfan (2013-VA-599) | Dr. Aijaz Ali Cheema | Dr. Muhammad Younas | Dr. Amjad Riaz | Dr. Muhammad Haroon Akbar.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: It is well recognized that cryopreservation of bovine semen results in decreased spermatozoal viability due to cryodamages. Oxidative stress is produced due to Reactive Oxygen Species (ROS) during freez-thaw process.Extreme production of ROS during cryopreservation has been associated with decreased post thaw %age motility, viability, and membrane integrity and sperm fertility capability. Basically antioxidants works to reduce or, taking up the formation of ROS. In recent years, taurine has been used as anti-oxidant in semen extenders and has been used in the cryopreservation of sperm from many species to improve post-thaw quality of spermatozoa by inhibiting lipid peroxidation and protecting cells against accumulation of ROS. Therefore, supplementation of taurine in semen extendercontaining 0,15,30,45 and 60mM concentration was used to decrease the harmful effects of cryopreservation on Sahiwal bull spermatozoa. Three Sahiwal bulls (4-8years) were used in the present study. The bulls were maintained at Semen production unit, Qadirabad and were offered good quality seasonal fodder (at the rate of 10% body weight), supplemented with concentrates (2-4kg/day). The semen was collected from each bull twice a week. Initially the semen was assessed for volume, motility, and concentration. Then the semen was pooled from all bulls and divided into 5 aliquots (150 ul each). Each aliquots diluted with 5groups extender which contain 0, 15, 30, 45 and 60mM taurine concentration. The semen was filled in 0.5ml straws cooled to 5c for 4 h, then frozen and stored in liquid nitrogen (-196c). Post-thaw motility, viability, plasma membrane integrity, acrosome and DNA integrity was evaluated and statistically analyzed by one way ANOVA. The group supplemented with 60mM concentration of taurine decreased all parameters of post thaw quality ofSahiwal bull spermatozoa. However the group supplemented with 15mM concentration of taurine show higher significantly (p<0.05) results than control and other groups. The post-thaw motility of the group supplemented with 15mM taurine concentration (54.50±2.1) higher significantly (p<0.05) than control (45.50±0.90) and other groups. The treatment group of 15mM taurine concentration show signifantly (p<0.05) higher viability (58.60±1.58 vs50.80±0.70) , plasma membrane integrity (57.10±1.43 vs 49.00±0.65) acrosome integrity (56.80±0.59 vs48.10±1.66) and DNA integrity (98.80±0.23 vs 97.54±0.39) as compared to the control and other groups. It is concluded that the maximum beneficial effect of addition 15mM taurine in tris-based egg yolk extender gives better post-thaw parameters. Availability: Items available for loan: UVAS Library [Call number: 2269-T] (1).
817.
Effect Of Alpha Lipoic Acid On Post Thaw Quality And In Vitro Incubation Of Nili Ravi Buffalo Bull Semen
by Muhammad Hammad Fayyaz (2008-VA-171) | Dr. Mushtaq Ahmad | Dr.Muhammad Younas | Prof. Dr. Nasim Ahmad | Prof. Dr. Muhammad Sarwar Khan.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Cryopreservation is the freezing of cells or tissues to subzero temperatures, typically -196 º C. Many benefits have resulted from the process of cryopreservation. Damage induced by cryopreservation has been results cold shock, oxidative stress, osmotic changes, and formation of ice crystal and lipid–protein reorganizations within the cell membrane. Oxidative damage caused by reactive oxygen species (ROS) leads to impaired cell functions. Free radicals, includes ROS and RNS, are normal pro - oxidant molecules in aerobic metabolism. Alpha lipoic acid is a non-vitamin coenzyme that helps in significant metabolic and antioxidant functions in the body. Alpha lipoic acid has been reported to have extra functions by which they are able to synthesize vitamin C from its reduced form in the presence of glutathione. It is matchless among biological antioxidants, because it is equally lipid and water soluble. This allows it to nullify free radicals almost everywhere in the body, inside as well as outside the cells. Therefore, the objective of present study is to determine the effect alpha lipoic acid on post thaw quality and in vitro incubation of buffalo bull semen. Alpha lipoic acid scavenge on reactive oxygen species formed in semen during the process of cryopreservation, so it maintained good semen quality during post thaw and in vitro incubation. Three mature Nili-Ravi buffalo (Bubalis bubalis) bulls (4-8 year age) kept at SPU, Qadirabad Sahiwal Pakistan were used in the study. These bulls are being used as regular donors at SPU. There semen was collected with artificial vagina of temperature 42c; three ejaculates (one from each) was pooled and diluted (30 million sperms/ml) with extender of different inclusion levels (0.0, 0.5, 1, 1.5, 2, 2.5 mmol/ml) of alpha lipoic acid. Straws were filled and extended then semen was cooled for 2 hours and equilibrated for two hours. Semen was placed in Liquid nitrogen vapors for 10 minutes. Finally semen straws was put in liquid nitrogen, Total five replicates were performed. Now post thaw quality was checked in
Summary
39
which various tests were performed, like %age motility, Acridine orange assay for DNA integrity, HOST for plasma membrane integrity, Fitc-PNA/PI for viability and acrosomal integrity. Longevity test was performed by in vitro incubation of frozen thawed semen sample in SOF and evaluating it at 1.5, 3 and 4.5 hour interval in Carbon dioxide incubator. It was expected that Alpha lipoic acid shown positive effect on post thaw quality and in vitro incubation of buffalo bull semen, in the meaning of increased percentage motility, Less DNA damage during cryopreservation and incubation, Increased acrosomal and plasma membrane integrity. So alpha lipoic acid shown positive effect by counter acting on ROS during cryopreservation and in vitro incubation. Results acquired from this study shown that an increase in sperm motility, plasma membrane integrity, DNA integrity, Acrosomal integrity, viability and survival was caused by ALA competences in energy production and anti-oxidant properties, when used at the concentration of 0.5mM and 1mM. In summary, based on the results of our study, it can be concluded that an optimal concentration (0.5mM and 1mM) of ALA improved PMI, sperm motility and viability, minimize DNA damage and improved sperm survival. Availability: Items available for loan: UVAS Library [Call number: 2268-T] (1).
818.
Comparison of The Effect of Ovsynch and G6G Synchronization Protocols on Ovulation and Pregnancy Rate in Nili-Ravi Buffaloes
by Muhammad Waqas (2008-VA-289) | Dr. Muhammad Usman Mehmood | Dr. Qaiser Shahzad | Prof. Dr. Mian Abul Sattar | Dr. Muhammad Afzal Rashid.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Thesis submitted with blank CD Availability: Items available for loan: UVAS Library [Call number: 2267-T] (1).
819.
Detoxification Of Aflatoxins Using Different Organic Acids
by Sana Ejaz (2013-VA-14) | Dr. Mateen Abbas | Dr. Muhammad Tayyab | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: From global prospective of food safety and food security, mycotoxin contamination of foods has gained much attention as potential health hazards for humans and animals. Cereals and other crops are exposed to fungal attack in the field or during storage and this attack may result in mycotoxin contamination of crops. Animal feed is basic necessity for all the live stock, poultry and other animals. AF is the most important for human and animal health perspective and in developing countries such as Pakistan where climate conditions favor the formation of these toxic metabolites. Governments and private organizations of international level have established maximum residue levels (MRIs) which usually guide to control AF in feed. Therefore, the current study was planned to detoxify AF by using different organic acid treatments in animal feed collected from different dairy farms of Punjab.
The samples of cotton seed cake, maize oil cake and animal feed were collected and checked the presence of AFB1 qualitatively by TLC and quantitatively by HPLC. The samples which gave positive results were treated with different acidic treatments applied on it. Firstly checked the results of citric acid, acetic acid and lactic acid on feed sample qualitatively by TLC. TLC plates were checked under UV box and the samples which showed the detoxification of AF were quantitatively analyzed by HPLC in Toxicology Laboratory, QOL, UVAS, Lahore, Pakistan.
The average concentration of AFB1 found in the cotton seed cake, maize oil cake and mixed feed were 279.8 ppb, 34.2 ppb and 25.5 ppb, respectively much greater than permissible levels proposed by European Union. Treatments of varying concentration of citric acid, acetic acid and lactic acid were applied on positive samples (≥20 ppb) and checked their effect on rate of detoxification.
All the above mention treatments applied on the feed samples in order to obtained in vitro detoxification of AFB1. Sprayed different concentration of acetic acid, citric acid and lactic on positive samples by varying volumes and placed them over night then extracted and analyzed.
It has been observed that 1N concentration of citric acid, acetic acid and lactic acid showed complete detoxification. However, when these samples were treated with 0.5N solution of organic acids then variation was seen in rate of detoxification.
Statistically these results were analyzed by ANOVA which showed that effect of these treatments on rate of detoxification was highly significant (P<0.05). In vitro detoxification of AF by these organic acids was proved beneficial in order to reduce the animal and human health risks. However, in vivo detoxification of aflatoxin by using these organic acids should be studied in future.
Availability: Items available for loan: UVAS Library [Call number: 2283-T] (1).
820.
Isolation And Characterization Of Antibiotic Resistant Lactic Acid Bacteria From Poultry Gastrointestinal Tract
by Nabeea Saleem (2008-VA-234) | Dr. Muhammad Nawaz | Dr. Aamir Ghafoor | Dr. Aqeel Javeed.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Lactic acid bacteria (LAB) are heterogeneous group of bacteria which are fastidious in
nature. LAB has acquired status as Generally-Recognized-as-Safe (GRAS) status. Over-use and
misuse of antibiotics in veterinary and human clinical setups give rise to antibiotic resistant lactic
acid bacteria in gastrointestinal tracts which act as antibiotic resistance reservoir. Acquired and
transferable antibiotic resistance in lactic acid bacteria is a newly identified safety concern in
poultry. So, it is dire need of time to determine the situation of antibiotic resistance in lactic acid
bacteria of poultry gastrointestinal tract (GIT).
The present study was conducted to isolate lactobacilli from indigenous and broiler
gastrointestinal tract of chicken. For this purpose, chicken feces, cloaca and caecum samples
(n=20 each) were collected from Lahore. Lactic acid bacteria were isolated on MRS medium.
Isolates were identified by phenotypic characteristics including Colonial morphology, Gram
staining and Catalase test. While molecular identification of lactobacillus spp. was done by PCR
at an annealing temperature of 55°C using the primers XB-5 and LbLMA-1 with an expected
product size of 250bp. Minimum inhibitory concentrations of different antibiotics such as
ampicillin, erythromycin, tetracycline, ciprofloxacin, cephradine, cefuroxime, ofloxacin,
levofloxacin were determined by the broth micro dilution method following the EFSA
guidelines. Antibiotic resistance genes, including erythromycin arm (B) and tetracycline Tet (M)
were amplified by polymerase chain reaction.
The comparison of % antibiotic resistance pattern between broiler and indigenous
lactobacillus spp. against different antibiotics was analysed for chi-square test using SPSS
……………………………………………………………………………………………Summary
67
version 16.0. The study provided data on antibiotic resistance pattern of transferable resistance
genes in lactic acid bacteria of poultry gut.
Conclusion:
From the present study it is concluded that a high level of resistance was shown by lactobacillus
spp. against tested antibiotics. Lactobacillus spp. were screened by PCR for known resistance
genes and thus were able to determine the presence of erm(B) and tet (M) genes in all
lactobacillus spp. using erm(B) and tet (M)-specific primers. All lactobacillus spp. were also
phenotypically resistant to erythromycin and tetracycline. Thus, the present study indicates that
such erm (B) and Tet (M) genes occur among different LAB genera and species therefore it is
the need of time to study other resistance determinants to ensure the safety of poultry meat and
spread of resistance determinants. Availability: Items available for loan: UVAS Library [Call number: 2282-T] (1).
821.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Opuntia Dillenii (Ker-Gawl) Haw. Leaves Against Common Poultry Pathogens
by Sadaf Raana | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: This project was designed to evaluate the antibacterial efficacy of hexane, chloroform, ethanol and aqueous extracts of Opuntia dillenii Haw. stems against common poultry pathogens. Pathogens used were Staphylococcus aureus, Escherichia coli, Salmonella, Clostridium perfringens type A and Haemophilus species.
This study was conducted to assess antibacterial and cytotoxic activity of O. dillenii Hexane, chloroform, ethanol and water extracts were prepared and antibacterial activity was evaluated by agar well diffusion method in which zones of inhibition were measured. Minimum inhibitory concentration (MIC) of plant extracts was evaluated by micro broth dilution method. The extracts which showed the antimicrobial activity were evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell culture media was prepared and cell lines were propagated, monolayer was formed. Monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated.
O. dillenii stems extracts inhibited the growth of both Gram negative and Gram positive bacteria. Chloroform and ethanol extracts of O. dillenii showed significant antibacterial activity against all the pathogens studied as compared to hexane and aqueous extracts. Hexane extract showed maximum zone of inhibition against Haemophilus species (13mm), for chloroform extract maximum zone of inhibition was obtained for C. perfringens (25.6mm), for ethanol extract maximum zone of inhibition was obtained for C. perfringens (23.0mm) and for aqueous extract maximum zone of inhibition was obtained for C. perfringens (23.0mm). Minimum inhibitory concentration for chloroform extract was lowest for all the tested strains. For S. aureus, C. perfringens type A and S. enterica MIC was 1250μg/mL. For E. coli and
CHAPTER 6
SUMMARY
Summary
88
Haemophilus species MIC was 2083.3 and 2916μg/mL, respectively. The extracts were further investigated to test cytotoxic effect on Vero cell line using MTT assay. Only ethanol extract was observed to be cytotoxic.
Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA). The results of antibacterial activity and MTT assay were evaluated for significance of difference using analysis of variance (ANOV). The homogeneity of groups was verified by Duncan’s test at an alpha level equal to 5%.
Chloroform extract of O. dillenii stems possess antibacterial activity and can be used to design traditional medicines for the development of therapeutic agent which will be more safe, effective and economical. Availability: Items available for loan: UVAS Library [Call number: 2281-T] (1).
822.
Antinematodal Efficacy Of Ivermectin (Oral) And Extracts Of Coriandrum Sativum In Sheep
by Memrez Khushal Gigyani (2013-VA-564) | Dr. Muhammad Ovais Omer | Dr. Muhammad Mushtaq | Mr. Qamar Niaz | Dr. Nisar Ahmad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: This project was designed to evaluate the anthelmintic efficacy of Coriandrum sativum plant extracts against sheep nematodes. Anthelmentic resistance (AR) is one of the major problems all over the world. This project was designed to evaluate the anthelmintic efficacy of Chloroformic and methanolic extracts of Coriandrum sativum against sheep nematodes.
For this purpose sixty sheep positive for nematodal infection in BLPRI Kherimurat (Punjab) were selected after fecal examination. Experimental animals were divided into 6 groups (Group A, Group B, Group C, Group D, Group E and Group F) having 10 animals in each group. Group A was Un-treated control. Group B was given Ivermectin (orally), Group C and D was treated with Chloroformic extract of Coriandrum sativum at 50 and 100 mg/kg body weight respectively by oral route. Group E and F was treated with methalonic extract of Coriandrum sativum at 50 and 100 mg/kg body weight respectively by oral route. Percent efficacy of Group A on day 7, 14 and day 28 post treatment was 0%. The percent efficacy of the Group B was calculated on day 7 was 81.4 %, on day 14 was 87.17 % and on day 28 was 92.60 %. The efficacy of Group C on day 7, 14 and 28 was 0. Similarly the efficacy of Group D on day 7, day 14 and day 28 was also 0. Percent efficacy of Group E i.e 50 mg/kg body weight Methanolic extract of Coriandrum sativum was 20.81 % on day 7, 27.14 % on day 14, and 33.48 % on day 28. Percent efficacy of Group F i.e 100 mg/kg body weight Methanolic extract of Coriandrum sativum was 49.76 % on day 7, 56.27 % on day 14 and 60.69 % on day 28.
CONCLUSION
It is concluded that the methanolic extract of the Coriandrum sativum has good anthelmintic effect against nematodes in sheep. Availability: Items available for loan: UVAS Library [Call number: 2280-T] (1).
823.
Effect Of Oiling And Packaging On Shelf Life Of Eggs Stored At Two Different Temperatures
by Marium Munir (2008-VA-388) | Dr. Muhammad Nasir | Dr. Sanaullah Iqbal | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: In Pakistan, poultry industry is playing a vital role in the economy of the country. As eggs are perishable so it must be handled with safety. It is imperative to handle and store the eggs at appropriate conditions. But improper storage of eggs is a problem in our country which affects its quality and there is chance of microbial contamination in eggs. Oiling and packaging has variable impact on shelf life of eggs at different storage temperatures (Matt et al. 2009).Raw eggs have many benefits, they contain essential nutrients for the brain, nerves, glands and hormones, they are nutritionally balanced. raw eggs also contain an abundance of other vital substances including protein, essential fatty acids along with niacin, riboflavin, biotin, choline, vitamins A, D and E, magnesium, potassium, phosphorous, manganese, iron, iodine, copper, zinc and Sulphur. Egg yolks are one of the few foods that contain vitamin D(Watkins, 2002). As eggs are perishable food stuff,so the purposes of present research work are to analyze the effect of oiling and packaging on shelf life of eggs at two different temperatures. For this a total of 864 eggs were collected. The four different treatments were applied along with two different temperatures. Each category was further divided into four treatment strategies (108 eggs in each strategy) i.e. eggs without any treatment, oil coated eggs, eggs packed (air-tight) in white polythene bags, oil coated eggs packed (air-tight) in white polythene bags.Eggs undergone each treatment strategy were analyzed for six parameter i.e. sensory evaluation, microbial load, Physical parameters (weight, pH, egg shell percentage and haugh unit) using 18 eggs for each further divided into three replicates (6 eggs for each replicate). All the eggs were stored for 1, 7, 14, 21, 28 and 35 days.
Summary
76
Data was analyzed statistically by the 2- way ANOVA (Analysis of Variance) with 5% probability. Means was compared by DMR test.At the end of this study we were able to assess the shelf life of eggs with respect to their oiling, packaging and storage conditions. Availability: Items available for loan: UVAS Library [Call number: 2289-T] (1).
824.
Incidence Of Dog Bite Injuries Reported In Tertiary Care Hospitals
by Ambreen Shahzadi (2012-VA-440) | Dr. Mamoona Chaudhry | Dr. Shakera Sadiq Gill | Dr. Muhammad Ijaz.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Animal bites people should be considered an important public health problem. Moreover, the number of pets, specifically the dog, is increasing. (Palacio J et al., 2003).Animal bites, particularly dog bites, are a major public health problem throughout the world (Morgan and Palmer, 2007). Animal bites and scratches, even when they are minor, can become infected and spread bacteria to other parts of the body.
Dog attacks, by street or domestic dogs, with injuries from very minor to significant and severe to fatal, are not uncommon. According to WHO report, ten million people are bitten by animals (especially Dogs) around the world, considered for prophylaxis and treatment against rabies and almost (55,000) people die from this disease annually (Mohdjunaid et al., Oct 2012).
6.2 Hypothesis:
The incidence of dog bite injuries is high in densely populated towns of Lahore when compared to less populated towns of Lahore.
6.3 Methodology:
General information about patients and possible causes was collected on a structured questionnaire. All the dog bite victims that were come to emergency ward during three months period were included in the study. Questionnaire include closed questions about the demographics of the victims, circumstances of bite incidents, body parts injured and the degree of injury, type of dog, history of previous bites, the level of knowledge about rabies, post bite home treatment (washing of bite wound etc.) prior to visited the hospital for medical treatment and post exposure treatment at hospital. The densely populated area (ravi town, data ganj bakhsh, shalimar town, samanabad etc) and less populated area (gulberg, azizbhati town, wagah, Allama iqbal town, & nishtar) was be compared.
6.4 Statistical Design
Cumulative incidence was calculated as described by Leon Gordis (2008). Chi square test was applied on the data by using SPSS (version 19.0).
6.5 Results
In this study, cumulative incidence is 4.653 per 1000 popluation, most of the dog bite victims were male 79.9 % as compare to female 20%.5.7% cases visited Mayo Hospital’s emergency department during the study period, 4.5% cases were reported in Services Hospital and 89.9% cases from IPH Lahore. In this the higher percentage was the patients who were illiterate (47%) and very few patients were those who had intermediate and above education (6%). The occupation of the patients who were attended Hospitals, were farmers (31.56%). The 24% patients were those who had bitten once before in previous years. In those 24% patients 19.2 % those who were bitten by dog and 5% were bitten by others (cats, horse, donkeys, etc). Most of the victims were bitten in rural area (65%) and most of the injuries were from stray dogs (65.9%). Most of the victims had provoked bite (74%) and the high frequency were recorded that the cases had type II wound category (deep scratch but no bleeding) 82%. The cases received treatment in different hospitals were RIG (18%), Tetanus toxoid (32%), Antibiotic (78%) and Suturing (7.9%).
Most of the patients were from Ravi town Lahore that is one of the highly densely populated areas of Lahore (38%) and very few cases reported from Nishtar town Lahore (0.5%). The demographic location and the hospitals had no association as the results were insignificant (p-value>0.05). Most of the cases who were bitten by dog and they already bitten by dog in previous years, significant in relation as p-value <0.05. The association between dog status and biting animal was significant, as mostly biting dogs were stray dogs and most of the bite was provoked, there was significant association between biting animal and biting type (p-value<0.05).
Availability: Items available for loan: UVAS Library [Call number: 2285-T] (1).
825.
Comparative Efficacy Of Anthelmintics Against Gastrointestinal Nematodes In African Lion (Panthera Leo)
by Muhammad Usman Munir (2008-VA-141) | Dr. Aneela Zameer Durrani | Dr. Muhammad ijaz.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: This study was conducted to compare the efficacy of Zental ( Albandazole) and Vermox (Mebandazole) against gastrointestinal nematodes like toxocara canis, toxocara mystax and toxascaris leonina in captive african lion (Panthera leo) kept in Lahore zoo, Lahor zoo safari, Bahawalpur zoo and Zain wildlife breeding farm Lahore because captive African lions are prone to different parasitic infestations.
A survey was carried out to estimate the prevalence of gastrointestinal nematodes in captive African lion and it was found that toxocara canis, toxocara mystax and toxascara leonina are the most commonly prevailing gastrointestinal nematodes in captive African lions in above mentioned places. The overall incidence rate of toxocara canis was 82.5 %, of toxocara mystax was 60% and of toxocara leonina was 50% respectively.
The Anthelmintics trials were conducted on 14 animals which were divided into 2 groups A and B each having 7 animals.
Group A was treated with Zental (Albandazole), group B was treated with Vormox (mebendazole), The percentage efficacy of each drug was determined on the basis of reduction on number of eggs per gram of discharged in faeces at day 7, 10, 13, 16 and 22 after deworming.
In group A, decrease in EPG of toxocara canis in group A was 21.1% at day 7, 37.2% at day 10, 54.7% at day 13, 74.6% at day 16 and 88.8% at day 22. Decrease in EPG of toxocara mystax was 15.9% at day 7, 38.1% at day 10, 56.6% at day 13, 70.6% at day 16 and 82.7% at day 22.similarly decrease in EPG on toxocaris leonina was 15.6% at day 7, 29.2% at day 10, 45.6% at day 13, 61.1% at day 16 and 83.1% at day 22 respectively
In group B, decrease in EPG of toxocara canis in group A was 7.24% at day 7, 14.57% at day 10, 23.26% at day 13, 31.27% at day 16 and 38.08% at day 22. Decrease in
Summary
25
EPG of toxocara mystax was 16.38% at day 7, 22.4% at day 10, 29.6% at day 13, 34.3% at day 16 and 41.2% at day 22.similarly decrease in EPG on toxocaris leonina was 13.6% at day 7, 21.29% at day 10, 30.8% at day 13, 39.1% at day 16 and 47% at day 22 respectively.
Results showed that efficacy of Zental (Albendazole) against gastrointestinal nematodes in African lion was higher as compared to that of Vermox (Mebendazole). Also it was more effective than Vermox (mebendazole) against different parasitic stages of nematodes as evident from EPG count at various days after medication. Availability: Items available for loan: UVAS Library [Call number: 2270-T] (1).
826.
Development Of Cheddar Cheese By Using Crude Flowers Extract Of Citrus Aurantium (Sour Orange) As A Milk Coagulant
by Usman Mir Khan (2009-VA-510) | Mr. Ishtiaque Ahmad | Prof. Dr. Muhammad Ayaz | Mr. Hifz-ul-Rahman.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Presently in Pakistan a few dairy companies are producing Cheddar cheese. Several
alternative proteases obtained from different vegetable and fruit sources such as fruits,
roots, latex and flowers have been recommended as rennet coagulant replacer in cheeses
processing. As there is now decrease in slaughtering of young calves, natural rennet
shortage occurs, so to fill it, sometimes alternates of rennet used in cheese production
technology.
This research project was designed to implement and introduce the use of Citrus
aurantium crude flower extracts (CFE) as an economical, easily available and rennet
substitute in Cheddar cheese production. CFE was evaluated for its first time usage as
coagulant in Cheddar cheese production and compared with rennet for its physicochemical
characteristics and sensory evaluation.
Cheddar cheese was made from buffalo milk. Standardized buffalo milk was pasteurized
and then cooled to inoculation temperature. It was inoculated with addition of 2% of
starter culture and given stay of 20-50 minutes. Then control sample was coagulated with
0.002% rennet and other samples were coagulated by Citrus aurantium crude flower
extracts (CFE) at different levels of 1%, 2 %, 3% and 4% at coagulation time. After
curdling, curd was cut, stirred and whey was drained. Then milling and cheddaring of
cheese blocks was done. Salting was done at the rate of 2.5%. After pressing, cheese was
stored in hygienic packaging and left for ripening at 10°C for 2 to 3 months.
Acceptability of newly developed Cheddar cheese using crude flower extracts (CFE) as a
coagulant was evaluated by sensory evaluation using criteria of color, Cheddar cheese
Summary
49
samples were analyzed for physico-chemical characteristics and sensory evaluation at 63
days of storage. The statistical analysis was carried out by using one way Analysis of
Variance (ANOVA) techniques under (CRD) Completely Randomized Design (Steel et
al. 1987). Duncan’s Multiple Range (DMR) Test was used for significant difference
comparisons (SAS 9.1 Statistical Software).
The cheese prepared from using 1% and 4% CFE cheese showed higher fat contents from
0 to 63 days while CFE cheese prepared with 1% and 3% have significant fat contents
and less than control sample prepared with rennet. Cheddar cheese with %, 2% and 3%
showed similar 25% protein content respectively but less than 4% and rennet coagulated
standard Cheddar cheese. Cheddar cheese made from 2% and 1% CFE showed slightly
higher moisture content while 3% and 4% showed moisture content of 33% almost
similar to the rennet coagulated Cheddar cheese which showed 32% of moisture content.
The cheese prepared from using 4% CFE cheese showed highest pH of 5.57 at 0 days
while after storage at 63 days pH decreased to 5.52. CFE Cheddar cheese prepared with
1%, 2% and 3% showed pH of 5.55 at 0 days and decreased to 5.52 which is standard pH
of the Cheddar cheese. Salting of all cheese samples was done at the rate of 2.5%. The
cheese prepared from using 2% and 3% CFE showed decrease in salt but it was higher
than 1% and controlled Cheddar cheese. While 4% showed lower salt contents than all
cheeses. Results showed that cheeses were made with 1% and 2% of CFE had a longer
and slightly softer texture. While cheeses contained 3% and 4% CFE had semi-hard
textural properties of curd similar to rennet added cheese which is similar to the standard
Cheddar cheese made with rennet. Cheddar cheese with 3% and 1% were preferred by
consumers instead of 2% and 4% for their better taste, texture/appearance and overall
Summary
50
acceptability but it was less preferred over standard Cheddar cheese (controlled) due to
the presence of bitter and intense aroma compounds of Citrus aurantium flowers.
It will give a benefit to the cheese industry by introducing a new economical, nutritional
and easily available rennet substitute source of milk coagulation. Pakistan is producing
largest amount of Citrus aurantium, so it will improve its export in other countries for
usage as an alternate of rennet and to provide economic benefits to our country.
Furthermore, it will open new ways for researchers to find out the characterization of
extract and enzyme of CFE. Availability: Items available for loan: UVAS Library [Call number: 2302-T] (1).
827.
Chemical Characteristics Of Trans Free Margarine Enriched With Omega Fatty Acids Through Chia (Salvia Hispanica L.) Oil
by Muhammad Ajmal (2009-VA-495) | Prof. Dr. Muhammad Ayaz | Dr. Muhammad Nadeem | Dr. Muhammad Hayat Jaspal.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Omega fatty acids has been related with low cholesterol level in blood, reduce the blood pressure, decreases the risk of heart attacks. Omega-3 PUFA is anti-carcinogenic, anti-atherogenic, anti-lipogenic, prevent the hypertension. Margarine was prepared by blending milk fat 70%, palm oil different concentration 30%, 27.5%, 25%, 22.5% and 20% T0 to T4 respectively .Chia seed oil in various proportions 0%, 2.5%, 5%, 7.5% and 10% T0 to T4 respectively. Milk fat, Palm oil and chia Seed oil was characterized for free fatty acids, moisture content, saponification value, iodine value, refractive index. Solid fat index was determined at 0, 10, 20, 30 and 40˚C by the dilatometric method, melting point was determined by open capillary tube technique.Color, peroxide value, anisidine value, conjugated dienes and trienes was determined. Fatty acids composition of margarines were determined by the transformation into fatty acid methyl esters. Margarine was stored at -6±1oC, for 60-days, iodine value, peroxide anisidine values, free fatty acids, conjugated dienes and trienes was determined. Induction period of margarine was determined by oxidizing the samples under a steady stream of oxygen (20L/hour) at 120oC, on a Rancimat. Sensory evaluation was performed by a panel of ten trained panel of judges, samples was evaluated for color, smell, taste and overall acceptability on a 9-point Hedonic scale.All treatments were replicated three times, every sample was analyzed three times and data were expressed as Mean ± SD. The data was analyze by one way and two way analysis of variance technique, the difference among the treatments was made by Duncan Multiple Range Test
Free fatty acids increased in all the treatments during the entire storage period from 0 day to 60 days, the content of fatty acid during storage period depend upon the degree of unsaturation. . Peroxide value increased in trans free margarine enriched with omega fatty acids through chia oil the during the storage of margarine. The rise in peroxide value at all the
determination frequencies was in the order of T4> T3> T2> T1> control. Iodine value increases in trans free margarine enriched with omega fatty acids through chia oil during the storage time. The decline in iodine value of all the treatments and control was in the order of T4> T3 > T2 > T1 > control. The treatment having more unsaturated fatty acids, underwent more oxidation and yielded the higher extents of oxidation products. Anisidine value of all the treatments and control increased throughout the storage period, all the determination frequencies revealed an increasing trend, however, the rise in magnitude of oxidation products was different in all the treatments and control.Values of conjugated dienes and trienes numerically increased during the storage period, all the measurement intervals revealed an increasing trend, however, the rise in extent of oxidation products was mainly dependent upon the fatty acid composition and treatments having higher concentration of unsaturated fatty acids suffered more oxidation.Addition of chia oil in margarine did not have any impact on moisture content of margarine, moisture content of all the treatments and control was not different from each other (P>0.05). Non-significant changes melting point during storage time in margarine. Color of all the experimental margarines and control were not different from each other (P>0.05). Saponification value non-significant in treatments during storage time from 0 day to 60 days. Analysis of variance indicated that treatments had significant effect on SFI, whereas, the effect of storage and the interaction between treatments and storage was non-significant. The content of C18:3 (omega fatty acid) in margarine is significantly increased from T1 to T4 due to chia oil because it contain 68 % alpha linolenic acid.
Availability: Items available for loan: UVAS Library [Call number: 2301-T] (1).
828.
Prevalence, Associated Risk Factors And Treatment Of Cryptosporidium Parvum In Foals
by Choudhry Usman Rasheed Butter (2008-VA-253) | Dr. Muhammad Avais | Prof. Dr. Aneela Zameer Durrani | Dr. Shakera Sadiq Gill.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Cryptosporidium parvum is an emerging waterborne zoonotic disease prevalent
throughout the world. There are different strains of cryptosporidium but most important is
C.parvum.It is intestinal protozoon belongs to coccidian family that causes damage to intestinal
epithelium that leads to villous atrophy so decrease absorption sites from intestine. It is not host
specific it can equally infect humans, calves and foals. It is cross transmissible among mammals.
In horses, cryptosporidiosis is most commonly seen in foals (most frequently 1–4 weeks of age)
and is associated with diarrhea and weight loss. Immuno-compromised foals (including foals
with severe combined immunodeficiency syndrome) are particularly at risk. The present study
was conducted in different studs farms in and around Lahore keeping in view the importance of
C.parvum in foals because it is gaining attention as a most important cause of diarrhea in foals so
that effective therapeutic measures should be adopted to control infection. There are 323 samples
are collected among different stud farms and veterinary hospitals and analyzed by using fecal
floatation method and modified Ziehl-neelsen technique. Infection rate of C. parvum in foals in
this study recorded was (12.30%) from different stud farms and veterinary hospitals. Infection
rate (27%) is higher in foals of age group range between 1day to 3 months while the least
infection rate (2.46%) was observed in 9-12 months age group. Infection rate is reciprocal to age
as the age increase infection rate decrease. Infection rate is independent of sex. There is no
relationship between sex and infection rate. Diarrhea is an important risk factor related to
infection rate. In diarrheic foals infection rate is (22.5%) while in case of non-diarrheic foal’s
infection rate is (6.5%) which showed that infection rate in diarrheic foals is three time more as
Summary
36
Compared to non-diarrheic foals. A significantly higher infection rate was observed in those
foals having contact with other animals like rodents, ruminants and dogs. Infection rate (15%)
was observed in those foals having contact with other animals and infection rate (6.5%) observed
in foals having no contact with other animals. Purpose seems to very important in this study.
Game horses have more infection rate which was (15.45%) as compared to draught horses
(4.4%). Game horses have three times more infection as compared to draught horses because
game horses remain in close confinement and foals have more prone to infection. Management is
an important factor infection rate was more in those stud farms having poor management as
compared to those have good management. Infection rate is poorly manage farms was (20.97%)
and (5.5%) in properly manage farms. The results of the comparative efficacy of nitazoxanide,
furazolidone and garlic showed that the Nitazoxanide was the most effective of the three in
treating cryptosporidium infection under field conditions. Efficacy of Nitazoxanide in treating
C.parvum in foals was (88%) during the study followed by furazolidone which have comparative
efficacy of (77%) than garlic with the efficacy of (70%).
In conclusion from above discussion reveals that C. parvum is prevalent in stud
farms in and around Lahore. Associated risk for C. parvum in foals are age less than 6 months,
immune-deficiency, contact with other animals like ruminants and poor management. Infection
rate is more in game horses as compared to draught horses because game horses are manage in
close confinement so the chances of infection are more. Treatment of C.parvum has long course
and effective drug for the treatment of cryptosporidiosis in foals is Nitaoxanide than
furazolidone. Availability: Items available for loan: UVAS Library [Call number: 2300-T] (1).
829.
Effect Of Different Treatment Trials On The Carrier Status Of Streptococcus Equi In Horses Recently Recovered From Strangles
by Muhammad Afzal (2007-VA-108) | Dr. Muhammad Hassan Saleem | Prof. Dr. Aneela Zameer Durrani | Dr. Hassaan Bin Aslam.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Strangles is characterized by upper respiratory tract infection, dysponea, anorexia, regional suppurative lymphadenitis causing high morbidity and low mortality in horses as well as in mules and it is an infectious problem of equine. Considering the significance and utilization of equines in our country and the substantial losses rendered by Strangles, the present project was designed to study epidemiology, diagnosis and chemotherapy of strangles in Lahore, Okara and Sargodha districts of the Punjab province in Pakistan. Streptococcus equi subspecies equi is highly prevalent in animals recently recovered from strangles and antibiotics along with immune stimulants and non steroidal anti-inflammatory drug became helpful in the elimination of carrier status of animals for S. equi. Samples were collected from animals recently recovered from Strangles. Samples were collected with the help of sterile cotton swabs dipped in normal saline from nasopharynx & oropharynx followed by culturing of samples on blood agar plates which were incubated anaerobically for a time period of 24-48 hours respectively and S. equi were isolated on the basis of colonies characteristics and growth pattern. Streptococcus equi was confirmed with the help of Gram staining and biochemical tests Catalase reaction, Methylene blue reduction test and Sugar fermentation test. In vitro antibiotic sensitivity test were also performed to select three antibiotics showing best efficacy against S. equi. Carrier animals were subjected to treatment with the help of antibiotic along with combination of immune stimulants and non steroidal anti-inflammatory drug. Carrier status was considered eliminated with disappearance of S. equi from nasopharynx. Blood samples were collected from carrier as well as from healthy animals to check out hematological parameters such as TLC (total leukocytes count), TEC (total erythrocytes count) and MHC (mean hemoglobin concentration) etc. Out of
Summary
47
hundred samples collected from Sargodha, Lahore and Okara districts of Punjab the prevalence of S. equi in horses and mules was 15%. Highest prevalence in equines was found at Remount Depot Mona, was recorded which is 18.3% followed by Okara 13.33%. No case was found to be positive for Strangles out of ten samples collected at UVAS, Lahore.
In-vitro antibiotic sensitivity test was performed on above isolates and it was found that ceftiofur Na was found to be most effective drug followed by norfloxacine and ampicillin. In vivo treatment trials showed that Group C animals treated with ceftiofur Na, norfloxacine, and ampicillin along with Vit E & Selenium supplemented by Phenyl butazone were found to be negative for post treatment carrier status. This study had aided in diagnosis as well as in treatment of strangles and was also provided us with the understanding of hematological parameters.
Statistical analysis:
Data on prevalence of S. equi in carrier animals were analyzed by Chi square test.
While comparison of different treatments trials was done by Z test.
Hematological parameters were analyzed by mean ± SED using SPSS software 16.0. Availability: Items available for loan: UVAS Library [Call number: 2299-T] (1).
830.
Prevalence And Chemotherapy Of Gastro- Intestinal Helminths In Camels Of Cholistan Area Of Bahawalpur
by Fariha Mariam (2008-VA-97) | Dr. Muhammad Ijaz | Dr. Khalid Mehmood | Prof. Dr.Muhammad Sarwar Khan | Prof. Dr. Azhar Maqbool .
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Gastrointestinal helminths are responsible for wide range of health problems, economic losses in camels and are characterized by impaired milk, meat, infertility, low calving rates, decreased working efficiency and even death of the camel.
To study the gastrointestinal helminths, 384 camels of different age, sex and breed was examined coprologically. For this, five gram of fresh fecal sample was carefully collected into a sealed container from each camel and was transferred to Medicine lab, UVAS, Lahore in containers with ice packs. Fecal samples were scored 1-3 based on the consistency. The individual samples was triturated in saturated salt solution, sieved and examined for helminths eggs by using different techniques i.e. Direct Smear Method, Sedimentation technique and Floatation technique while the eggs count was performed by McMaster technique. Among the camel population, the current study indicates that in Marrecha breed the prevalence of Nematodes was Trichostrongylus 1.77%, Haemonchus 8.44%, Nematodirus 3.11% and Trichuris 7.11% respectively. Mixed infestation of all the nematodes was found 26.66% in infested camels, while the prevalence of Camelostrongylus, Trichostrongylus, Strongyloides, Haemonchus, Nematodirus and Trichuris in Barella breed was 4.40%, 1.88%, 2.51%, 6.91%, 2.51% and 7.54% respectively. Mixed infestation of all the nematodes was found 25.78% in infested camels. The prevalence of Fasciola hepatica was found higher in Marrecha breed as compared to Berrela breed. It was 18.66% in Marrecha breed and 14.46% in Barella breed while the overall prevalence of Fasciola hepatica in the camels irrespective of the breeds was 16.93%. While the prevalence of Fasciola gigantica was also found higher in Marrecha breed than Barella breed. It was 16% in Marrecha breed and 10.69% in Barella breed while the overall prevalence of Fasciola gigantic in the camels irrespective of the breeds was 13.80%. The overall prevalence of fascioliosis in camel is 30.31. The prevalence of Moniezia expansa was found higher in Marrecha breed as compared to Barella breed. It was 10.22% in Marrecha breed and 8.80% in Barella breed while the overall prevalence of Moniezia expansa in the camels irrespective of the breeds was 9.63%. The sex wise prevalence of gastrointestinal Nematodes in males was 39.28% which includes Camelostrongylus 8.03%, Trichostrongylus 3.57%, Strongyloides 6.25%, Haemonchus 11.60%, Nematodirus 4.46% and Trichuris 5.35% respectively while in the female it was 25.37% which includes Camelostrongylus 4.41%, Trichostrongylus 2.20%, Strongyloides 2.94%, Haemonchus 6.25%, Nematodirus 2.57% and Trichuris 6.98%. The overall prevalence of Nematodes in Male was found higher as compared to Female which was 39.28% and 25.37% respectively. Sex wise prevalence of GI Trematodes in Camels was determined through collected samples. The prevalence of Fasciola hepatica was found higher in Female as compared to Male. It was 16.91% in Female and 15.18 % in Male while the overall prevalence of Fasciola hepatica in the camels irrespective of the sex was 19.53%. The prevalence of Fasciola gigantica was also found higher in Female than Male. It was 12.13% in Female and 11.61% in Male while the overall prevalence of Fasciola gigantica in the camels irrespective of the sex was 11.98%. Sex wise prevalence of GI Cestodes in Camels was determined. The prevalence of Moniezia expansa was found higher in Female as compared to Male. It was 14.34% in Female and 13.39% in Male while the overall prevalence of Moniezia expansa in the camels irrespective of the sex was 14.06%. The overall prevalence of Nematodes in age group >10 yrs. was higher 30.25% as compared to age group 5-10 yrs. 20% which includes Camelostrongylus, Trichostrongylus, Strongyloides, Haemonchus, Nematodirus and Trichuris 3.36%, 2.52%, 6.72%, 5.88% 4.20% and 7.56 in age group >10 yrs and 1.88%, 1.51%, 1.88%, 4.90%, 2.64% and 7.16 respectively, in age group 5-10 yrs.
The prevalence of Fasciola hepatica in age group 5-10 yrs was found higher as compared to age group >10 yrs. It was 30% in age group 5-10 yrs and 16.99% in age group >10 yrs while the overall prevalence of Fasciola hepatica in the camels irrespective of the age groups was 18.75%. The prevalence of Fasciola gigantica in age group 5-10 yrs was found higher as compared to age group >10 yrs. It was 9.33% in age group 5-10 yrs and 7.55% in age group >10 yrs while the overall prevalence of Fasciola gigantic in the camels irrespective of the age groups was 8.59%.
The prevalence of Moniezia expansa in age group >10 yrs was found higher as compared to age group 5-10 yrs. It was 10.92% in age group >10 yrs and 10.18% in age group 5-10 yrs while the overall prevalence of in Moniezia expansa the camels irrespective of the age groups was 7.81%.
To study the effect of gastrointestinal helminths on various blood parameters of camels, blood samples were collected from 5 infected and 5 healthy camels. Samples were analyzed using Hematology Analyzer and results were compared.
For therapeutic trials, a total of 30 camels positive for helminths was taken and divided into three groups A-C each group comprising of 10 animals. A fourth group D was selected comprising of 10 uninfected camels. The camels of group A were treated with dry Neem (Azadirachta indica) leaf powder at a daily oral dose of 100 gm/camel for five days, group B was treated with Albenzole granules® (Albendazole) at a dose rate of 15mg/kg bd. wt. PO once. Group C (untreated) was designated as positive control, and the camels in Group D as negative control. Efficacy was determined on the reduction of eggs in feces at day 3, 7 and 14 (post-treatment). In the current study Neem leaves and Albendazole gave following results in the camels affected with Helminths. Neem leaves cured 20% of the animals on day 3rd after the drug administration while 40% and 60% of the animals cured on the day 7th and 14th respectively. Albendazole also gave good results against helminths affected camels. On the day 3rd of the treatment 30% animals cured while 60% animals were cured on the 7th day and on 14th day 80% animals recovered.
Data regarding prevalence was measured by non-parametric, chi-square (χ2), while therapeutic trails were analyzed by repeated measures one way ANOVA, using SPSS (Statistical package for social sciences), P < 0.05 was considered significant.
Availability: Items available for loan: UVAS Library [Call number: 2298-T] (1).
831.
Effect Of Citrullus Colocynthis On Serobiological Parameters In Alloxan Induced Diabetic Rats
by Farah Javed (2012-VA-398) | Dr. Muhammad Quaid Zaman | Dr. Imtiaz Rabbani | Prof. Dr. Asim Aslam .
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Diabetes mellitus is one of the most common endocrine disorders affecting almost 25% of the world's population.The pretentious worldwide increase in the incidence of diabetes mellitus is posturing a huge health problem in both developed and developing countries. Diabetes mellitus is a metabolic disorder which is manifested by polyuria, polyphagia, polydipsia, hyperglycemia and dyslipidemia and is still one of the most leading causes of disability and death. Oral hypoglycemic agent and insulin are common treatment of diabetes but these treatments have prominent side effects. In the recent years the use ofherbal medicines has increased for the treatment of diabetes and fascinated the consideration ofmany researchers all over the world. Citrullus colocynthis is used commonly in different parts of the world for the treatment of a number of diseases including diabetes, jaundice, leprosy, cancer, asthma, bronchitis, joint pain and mastitis. In the present study I evaluate the anti-diabetic effects of roots of Citrullus colocynthis and also the dose dependent anti-diabetic effects of medicine in alloxan induced diabetic rats.
Twenty-five adult male rats were divided into five groups; Negative control, Positive control, and 3 groups for different treatment dose of roots of Citrulllus colocynthis (200 mg/kg body weight, BW), Citrulllus colocynthis (300 mg/kg BW) and Citrulllus colocynthis (500 mg/kg BW). Diabetes induction had done in four groups, other than negative control (normal saline injected), by subcutaneous administration of alloxan (120 mg/kg BW). Blood glucose level of rats reached above 250mg/dl considered as hyperglycemic. Treatment was given to all groups excluding control negativefor 21 days. Body weight of rats of all groups was recorded weekly. After completing 21 days of treatment with different doses of roots of Citrullus colocynthis blood samples were collected in fasting condition from rats of each group by cardiac punctureunder general anesthesia. Serum was collected from blood to measure serum glucose level, serum lipid profile, liver function test and renal function test.
Result data was analyzed by using SPSS software. Data was analyzed by using one-way ANOVA. The group differences were compared by the Duncan’s Multiple Range Test. Differences was considered significant at P < 0.05.
The obtained results showed that roots of Citrullus colocynthis has efficiency to control the diabetes mellitus by reducing serum glucose levels as well as the increasing dose decreased the serum glucose levels. Only 500mg/kg body weight dose is efficient to reduce the muscle wastage due to diabetes in alloxan induce diabetic rats. This dose also works tomaintain the serum ALT, AST, urea, creatinine, total cholesterol and triglycerides, HDL-C LDL-C levels.
The data obtained from this study also show the dose dependent anti-diabetic activity of medicine as the dose of 500mg/kg body weight is more effective to control the diabetes as compared to other two doses; 200mg/kg body weight and 300mg/kg body weight.
Availability: Items available for loan: UVAS Library [Call number: 2297-T] (1).
832.
Study On Pathogenesis Of Mixed Infection Of Infectious Bronchitis Virus (Ibv) And Avian Influenza Virus (H9n2) In Experimentally Infected Broiler Chicken
by Arsala Khan (2013-VA-605) | Prof. Dr. Asim Aslam | Dr. Muhammad Yasin Tipu | Prof. Dr. Khushi Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Avian influenza and infectious bronchitis are the most devastating contagious and viral diseases, affected poultry in Pakistan and many other Asian countries. Regarding the virulence, Avian influenza may be low, or high pathogenic. Mainly due to its pandemic potential and contagious nature, the low pathogenic avian influenza virus (H9N2) is of major concern in poultry industry especially in Pakistan. Co-infection of poultry with more than one bacterial and /or viral agent is common in field especially in our country due to inadequate biosecurity measures which causes high mortality and confusing the diagnosis of these viruses. Hence the present project was designed to study the pathogenesis of mix infection of IBV and H9N2 through gross, histopathological lesions and antibody titer in experimentally infected broiler chicken. For this purpose a total of 80 specific pathogen free chicks were procured, randomly divided into four groups A, B, C and D; and raised under uniform managmental condition in experimental sheds of UVAS Lahore. Group A and B were intarnasally challenged with 0.2 ml infectious bronchitis virus (EID50 = 104.5 per 0.1 ml) at 23rd day of age, likewise group B and C were inoculated with 0.2 ml of H9N2 (EID50 = 106 per ml) at 26th day of age while, group D served as a negative control group. After infection the clinical signs, feed consumed and body weight gain were regularly monitored, the serum samples taken on days 23rd, 26th, 29th, 35th, and 40th day of age were tested for HI antibody titer simultaneously, the necropsy of birds (n=3) sacrificed were conducted to observe the gross lesions, tissue samples including lungs, liver, kidneys and trachea were collected for histopathological evaluation. In IBV infected group, respiratory distress i.e., tracheal rales, nasal discharge, coughing, sneezing, gasping and reduced feed intake were observed during early stages, later up to 10 days post infection watery diarrhea with ruffled feathers were observed. In mix infected group clinical signs manifested rapidly and were persistent with
Summary
59
high severity. Gross lesions in mixed infection were more profound, including; severely congested and hyperemic lungs, tracheitis with catarrhal exudates in lumen; pale, swollen and enlarged kidneys with urates deposition in tubules. Six birds died in mix infected group, revealed caseous exudate in trachea extended up to lower bronchi while, in IBV infected group lesions were mild and confined to trachea, lungs and kidneys. Mortality was high in mix infected group (30%) followed by IBV infected group in which two birds died. While, in H9N2 infected and control group none of the bird died. Histopathological lesions in mix infected group were aggravated markedly tracheal epithelium degeneration and sloughing; congestion, interstitial nephritis, leukocytes infiltration, tubular degeneration and necrosis were observed while, in lungs pneumonia of peribronchiolar area and interstitium with lymphocyte and macrophages infiltration, additionally degeneration and vacuolization of hepatocytes with focal necrotic areas in liver were also noted. In IBV and H9N2 infected group microscopic lesions were of mild degree. GMTs against both IBV and H9N2 in mix infected group were significantly different at P>0.05 than individually infected birds. Among the groups, statistically significant increase in FCR of birds in mix infected group was observed. On the basis of these findings it might be conclude that mix infection of IBV and H9N2 causes severe disease as compared to single infection. Availability: Items available for loan: UVAS Library [Call number: 2296-T] (1).
833.
Prevalence Of Tick Borne Hemoparasites And Identification Of Ticks With Their Treatment In Dogs
by Muhammad Shehriyar Khan (2013-VA-438) | Dr. Muhammad Ijaz | Dr. Muhammad Avais | Prof. Dr. Muhammad Arif Khan.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Dogs are one of the most commonly owned companion pets worldwide. They are kept for various factors such as hunting, guarding, and amusement or for animal assisted therapy. Tick infestation is a common problem in dogs and its transmitted infections usually tick borne hemoparasitic diseases are emerging as a worldwide problem not just for animals but also for humans. Due to this phenomenon, ticks are now recognized as a major cause of various diseases in of urban environments. The current research has been conducted to study the prevalence of tick infestation, associated risk factors and chemotherapeutic trials of topical acaricides were used in order to study comparative efficacy, were conducted in dogs. Furthermore, prevalence of tick borne hemoparasitic diseases was also observed by performing microscopic blood smear examination in dogs of Lahore district.
A total of 300 dogs (n= 200 positive for tick infestation; n= 100 negative for tick infestation) having tick infestation were included in the present study. Dogs of different sex, age and breed were selected for tick collection. The ticks were collected with the help of forceps from the target sites of each dog, while adopting all necessary measures to avoid damage to the mouth parts of the ticks and skin of the host. The collected ticks were preserved in plastic containers containing 70% ethanol, which was separate for each specimen. Ticks were identified under stereoscope by observing morphological characteristics of each tick, according to the key. Blood sample for tick borne hemoparasitic diseases was drawn from cephalic vein of each dog using disposable syringe and was poured into EDTA vacutainer. Thin blood smear slide was then prepared from the blood of the animal, dried in air and was stained using Geimsa’s staining method. The hemoparasites observed in the blood of the dogs were then characterized and
Summary
51
identified by their morphological characteristics. For chemotherapy, a total of 30 tick infested dogs of different breeds viz. Rottweiler, German shepherd and Labrador was used in therapeutic trials. Each breed of dog was divided into 2 subgroups which were then treated with topical ivermectin and fipronil 1500μg/kg b.w and 15mg/kg b.w respectively. Efficacy of the drugs was then measured on the basis of disappearance of clinical signs and reversal of tick infestation at day 2, 4 and 6 of post-medication. Data regarding the prevalence of tick borne hemoparasitic diseases and tick infestation was analyzed by using Non-parametric, Chi-square test whereas data on therapeutic trials was analyzed by Z-test, using statistical package for social science (SPSS). P < 0.05 was considered significant.
The results obtained showed us that the study area is at a risk of tick infestation and tick borne hemoparasitic diseases. In dogs, Rhipicephalus sanguineus (brown dog tick) species was observed and identified as a major tick infesting dogs of the study area during the course of study. Various determinants including tick density and its genus, host age, sex, predilection site host breed and season significantly affected the tick infestation in the study area. The prevalence for tick infestation was found highest in Pastoral dogs as compared to other breed types found in dogs. Among breeds German shepherd showed highest prevalence for tick infestation. Adolescence age group among dogs was more prone to tick infestation. The number of males affected by ticks was highest as compared to females in dog population. Among the dog population, majority of the animals infested by ticks were categorized under mild infestation. The most favorable predilection site for tick infestation in dogs was ear pinnae, followed by head and then neck. Monsoon was the most favorable season for tick infestation in dogs. September was found to be highest favorable month for tick infestation in dogs.
Summary
52
In tick borne hemoparasitic diseases, babesiosis was the highest prevalent disease in dogs (63.33%), followed by trypanosomiasis (0.66%) and mixed infection (0.33%). Age group among dogs showed non-significant factor affecting prevalence for TBHD’s. The number of affected dogs for babesiosis was greater in adolescence age group as compared to other age groups. On the other hand, trypanosomiasis and mixed infection adult age group was found highly susceptible. Males of canine group were slightly more susceptible than females in case of babesiosis. Similarly for trypanosoniasis and mixed infection males were more prone than females. For babesiosis, monsoon was the most favorable season in dogs. Whereas for trypanosomiasis and mixed infection, post monsoon was found to be favorable season in dogs. In breed type of dogs, prevalence for babesiosis was found highest in Pastoral dogs. In dogs the prevalence for babesiosis was found highest in German shepherd breed, whereas for trypanosomiasis and mixed infection Cross breed was the most susceptible breed. For trypanosomiasis and mixed infection Cross breed was the most susceptible breed type. The prevalence for babesiosis was found highest in the month of September. For trypanosomiasis and mixed infection, the month of October was found to be highest susceptible month for dogs.
Chemotherapeutic trial of topical acaricides against tick infestation in dogs revealed that Fipronil was more efficacious drug as compared to Ivermectin.
Such type of epidemiological data of tick infestation, TBHDs and its chemotherapeutic trial will help to control tick infestation and tick borne hemoparasitic diseases to save the life of companion animal. Availability: Items available for loan: UVAS Library [Call number: 2303-T] (1).
834.
Effect Of Zinc Oxide Nanoparticles On Serum Mineral Profile And Bone Health Of Broilers
by Sidrah Shafique (2013-VA-845) | Dr. Muhammad Quaid Zaman | Dr. Muhammad Shahbaz Yousaf | Dr. Saif ur Rehman Kashif.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Background: Zn is an essential mineral of biological importance and in the past processed and unprocessed forms of Zn was used with uncertain composition. Zn is very important from nutritional point of view and is often supplemented in poultry diet. Zn promotes the growth, skeletal development, mineral retention, Zn concentration in tissues, production of neutropphils and antibodies and it improves the functions of macrophages. Zn deficiency causes reduction in weight gain, skeletal malformations, poor bone mineralization and immunological dysfunctions. ZnO nanoparticles (ZONP) are mostly used due to its advantages over conventional forms. Current study is aimed to evaluate the effects of ZONP on serum minerals profile, tibial bone minerals profile and bone health in broilers.
Hypothesis: ZONP may enhance bone health and serum minerals in broiler.
Methodology: The study included 120 day-old broiler chicks divided into four groups, each with 6 replicates. Group A served as control group and group B, C and D served as treatment groups supplemented with 80mg/kg ZnO, 40mg/kg ZONP and 80mg/kg ZONP respectively. At the end of 35 days, two birds from each replicate were randomly selected and slaughtered to collect tibial bone samples and blood samples. The parameters reported were bone health i.e. bone weight, bone length, robusticity index and tibiotarsal index and minerals in serum and bones were analysed by atomic absorption spectrophotometry and flame photometry.
Statistical analysis: The data was analyzed using one way Analysis of variance (ANOVA). The group differences were compared by the Duncan’s Multiple Range Test. Differences were considered significant at P < 0.05.
Results: This study helped us to determine the bone health and minerals level of bones and serum in response to ZONP in broiler. The results showed no change in serum and bones level of Zn, Fe, K and Na in Zn supplemented groups compared to control group but serum and bones Ca level showed significant increase in Zn supplemented groups compared to control group. The results showed no significant difference in bone dry weight, wet weight, length, bone diameter, medullary canal diameter, robusticity index and tibiotarsal index of broilers supplemented with Zn compared to control group.
Conclusion: It is concluded that Zn supplementation, either ZnO or ZONP, has no effect on bones health, bone minerals and serum minerals. However, Ca level of bones and serum was increased by Zn (either ZnO or ZONP)
Availability: Items available for loan: UVAS Library [Call number: 2304-T] (1).
835.
Genotoxic And Mutogenic Study Of Formaldehyde, Sodium Hypochlorite And Cresol
by Ann Fatima (2012-VA-995) | Prof.Dr. Muhammad Ashraf | Dr. Muhammad Adil Rasheed | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Disinfectants are used to control, prevent or destroy harmful microorganisms on inanimate surfaces. These chemicals are being used to dispose the contagious hospital wastes like disposable plastics and microbiological waste. Various factors like temperature, contact period concentration of disinfectant, organic soil and nature of water used for dilution affect of disinfection process. So, disinfectants must be tested prior to any specific applications for its proper effectiveness. This study has been designed to study the genotoxicity and mutagenicity of three commonly used disinfectants, formaldehyde, sodium hypochlorite and cresol alone and in combination.
Different dilutions of formaldehyde (1, 0.3, 0.1, 0.006, and 0.003%) sodium hypochlorite (8, 4, 2, 1, and 0.5%) and cresol (7.6, 3.8, 1.9, 0.95, and 0.475%) alone and in combination were investigated for mutagenicity as well as genotoxicity in vitro. Mutagenicity was investigated by Ames Salmonella/Microsome assay with and without metabolic activation system; S-9 with the help of two strains of Salmonella typhimurium, TA 100 and TA 98 and genotoxicity was checked by Comet assay using peripheral blood lymphocytes. The results were analyzed by statistical package of Social Sciences; results were presented as mean ± S.D and the data analysis was done by using one-way analysis of variance. Differences were considered significant at P < 0.05.
Summary
83
Formaldehyde, sodium hypochlorite & cresol showed significantly mutagenic potential against TA 100 & TA 98 strains of Salmonella with and without metabolic activation system and genotoxic effects. A higher concentration showed more significant results.
Formaldehyde, sodium hypochlorite & cresol has both mutagenic and genotoxic potential. But this mutagenicity and genotoxicity has been observed more with higher concentrations as compared to low concentration. Availability: Items available for loan: UVAS Library [Call number: 2305-T] (1).
836.
Genetic Characterization Of Pakistani Wild Quails Using Mitochondrial Coi Gene
by Wajiha Shakil (2012-VA-817) | Dr. Ali Raza Awan | Dr. Muhammad Yasir Zahoor | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: A latest taxonomic tool termed as DNA barcoding is being used to genetically characterize animals. DNA barcoding is helpful in determining evolutionary relationship among species. Being a small sized genome and agile enough to show rapid mutation, mtDNA has been used as a pertinent marker of molecular biodiversity. DNA barcode works as an efficient tool in food manufacturing industry, diet investigation, forensics analysis, preventing unlawful trade and felonious poaching. The aim of this study was to develop DNA barcode for genetic characterization of Pakistani wild quail. Pakistani wild quail is important due to its demand for eggs, meat production, experimental purposes and gaming as well. Japanese quail was also included in this study because this quail is excessively produced in Pakistani farms. Japanese quail is present throughout the year and is comparatively bigger in size than wild quail. It has longer lifespan; farmers can easily breed this species in farms. It is suitable in poultry due to better meat yield.
COI gene (500bp) was used as a molecular marker for identification at species level. DNA was extracted from blood samples of ten wild quails (Coturnix coturnix and fifteen japanese quails (Coturnix japonica). Reported bird universal primers were used to amplify COI region from the extracted mtDNA samples using PCR. Amplicon were then sequenced by Sanger sequencing method (Sanger et al. 1977). Forward and reverse DNA Sequences were aligned with the reference sequence using nucleotide BLAST on NCBI to observe the dissimilarity among the sequences. Consensus sequences generated were used to construct their phylogenetic tree to see their evolutionary relationship with other bird species. Japanese quail which is thought to be domesticated from Japan, its Pakistani population showed close relation with sequences
Summary
90
generated in Japan for this particular species. Pakistani wild quail species showed its closest linkage with C. coturnix.
In conclusion, COI barcode proved as an authentic tool for species identification and phylogenetic inference of Pakistani wild and farm grown quails. Wild quail species has been characterized using partial COI gene sequences. This study has provided a specific genetic marker which can differentiate Japanese quail from wild quail at molecular level as most of the time both species are confused with each other. It can be helpful to the farmers and bird fanciers because they can select the birds of their choice correctly. This is the first study reporting DNA barcode of this Pakistani quail species. It would help researchers to study about phylogenetic and taxonomic status and also assist quail fanciers and quail farmers to unaffectedly identify their species of interest in farming. Identification of quail species is also important for conservation of biodiversity as it helps in preservation and identification of endangered species by generating their barcodes from even minimal evidence available. Availability: Items available for loan: UVAS Library [Call number: 2311-T] (1).
837.
Prevalence And Risk Analysis Of Coxiella Burnetii In Soil Of Faisalabad And Gujranwala Districts
by Zia Ul Hasnain (2007-VA-290) | Dr. Muhammad Zubair Shabbir | Dr. Arfan Ahmed | Dr. Muhammad Yasir Zahoor.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Coxiella burnetii is a causative agent of Q-fever, a widespread zoonosis. The effective adaptation of C. burnetii to intracellular existence is in contrast with its ability to survive in the environment outside the host cells and its resistance to chemical and physical agents. Besides nutrients and minerals, soil is aggregate of number of pathogens. Many of those organisms are of zoontoic importance and have significant threat to public health. One of these is Coxiella burnetii that has been reported from other countries including the neighboring to Pakistan. Its occurrence in soil, clinical significance and importance to human and animal health has been reported; nevertheless nothing is known of C. burnetii in Pakistan particularly in rural setup where human and animals are in close proximity to each other as well as the fact that how different risk factors can be implicated in its spread and survival in the soil. PCR helps to identify the organism on the basis of its genome and it is highly preferable over other conventional detection assays. Soil borne C. burnetii has not shown any association with different risk factors. The factors include presence or absence of pathogens with or without animal interaction, distance from animal market, main road, canal, animal and human density in a village under study. PCR technique was used to identify C. burnetii in the soils of Faisalabad and Gujranwala district. Soil samples (n= 730) were collected from each village of the both districts and processed for genome extraction using commercial soil DNA extraction kit. The extracted DNA from the soil samples was run further for PCR analysis of transposase IS1111a followed by standard gel electrophoresis technique. Only 6 (0.82%) samples were positive out of 730. Furthermore pathogens prevalence was geographically mapped in relation to roads, canals,
-----------------------------------------------------------------------------------------------------------------------Summary
45
rivers, drains, animal and human population to determine the risk areas according to intensity of identified pathogens for both districts. Odd Ratio was calculated to access the association in terms of absence or presence of pathogens with particular risk factors, which did not show any kind of association between pathogen and risk factors. The phylogenetic analysis of C. burnetii shows different convergence percentage with the isolates of worldwide i.e., Namibia (99.53%), Brazil (100%), Taiwan (99.53%), India (99.07%).
Study was contributed to understand about the previously unrevealed prevalence of C. burnetii in soil of district Gujranwala and Faisalabad together with risk factor analysis implicating possible health significance as well as survival in the soil. Availability: Items available for loan: UVAS Library [Call number: 2312-T] (1).
838.
Isolation And Characterization Of Avian Isolates Of Lactobacilli Species And Their Antisalmonella Activity
by Anum Shaukat (2009-VA-220) | Dr. Muhammad Nawaz | Dr. Jawad Nazir | Prof. Dr. Mansur-Ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Poultry industry is second largest industry of Pakistan. Poultry industry is cheaper source
of protein and provides jobs for more than 1.5 million people. It is facing several problems due
to microbial diseases. Salmonella is one of the leading causes of diseases in poultry. These are
being treated with antibiotics but misuse and overuse of antibiotics result in antibiotic resistant
strains of microorganisms. We need some alternatives for treatments. Lactobacilli are one of the
alternatives to antibiotics used as probiotics.
It has been found that the Lactobacilli of poultry origin have antimicrobial activity
against Salmonella.
Lactobacilli was isolated from the droppings, cloaca and caecum of rural poultry birds
using deMan Rogosa Sharpe (MRS) medium. The isolates were screened for anti-Salmonella
activity against S. enterica along with their properties to resist low pH and bile acids, antibiotic
sensitivity, auto-aggregation and co-aggregation. The isolates showed anti-salmonella activity
were identified using microscopic characters and biochemical profile. The isolates were
confirmed by PCR using species specific primers and sequencing 16S rRNA gene.
The data was analysed using one-way ANOVA at significance level P value <0.05 by
using the statistical software Graph Pad Prism version 5.3.
The study was conducted on a total of 60 samples including caecal swabs (n=20), cloacal
swabs (n=20) and dropping (n=20) of indigenous poultry. From these samples, five isolates were
Summary
57
selected based upon the tests performed. Isolates namely CLB-41, CLB-45, PDL-13, PDL-26
and PDL-33 showed best results. Further characterization was done by PCR and sequencing.
Availability: Items available for loan: UVAS Library [Call number: 2313-T] (1).
839.
Combine Effect Of Ionomycin And Strontium Chloride To Induce The Parthenogenetic Activation Of Mouse Oocytes
by Muhammad Ashraf (2013-VA-13) | Dr. Amjad Riaz | Dr. Mushtaq Ahmad | Dr. Muhammad Imran Rashid.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Parthenogenesis is a phenomenon in which the development of oocyte oocur without fusion of male gamete. During fertilization spermatozoa trigger intracellular Ca+2 oscllation in M-II stage oocyte which initiates the embryonic development. The rises of intracellular calcium (Ca2+) ions is the basic step for the parthenogenesis. During parthenogenetic activation calcium channel open from endoplasmic reticulnum or depletion of calcium store and facilitate the calcium (Ca2+) from extracellular environment. Parthenogenetic technique is applied in cloning and production of embryonic stem cell lines for used to treat different diseases. Many scientists used different chemicals agents for artificial activation such as strontium, Ionomycin and Ethanol. Strontium chloride has been used widely for parthenogenetic activation of mouse oocyte, but its result to blastocyst development is poor. The objective of present study is to improve parthenogenetic activation and embryo development by combination of Ionomycin with strontium. Hypothesis of my study was Addition of Ionomycin in Strontium based activation protocol improves embryonic development.
The present study was conducted in embryology lab of department theriogenology, university of veterinary and animal sciences, Lahore.Six to eigth week old female mice (n=100) were super ovulated with intra-peritoneal injections of eCG (5iu) followed by hCG injection (5iu) at 48 hrs interval. 14 hrs post hCG, the cumulus oocyte complexes were collected from oviduct of the mice. In experiment 1, the oocytes were activated by using Ionomycin with concentration of 5, 10 and 15 µmol/l for 5 and 10 followed by this activation with strontium chloride (10mmol/l). In experiment: 2, The oocytes were activated by activation medium having strontium (10 mM/l) and Ionomycin (5, 10 or 15 µmol/l) in combination. CZB medium were used for oocyte cultured in CO2 incubator of 5% CO2 at 37°C. Number of activated oocytes were analyzed by cleavage rate to blastocyst stage. In-vitro developmental potential of the activated oocytes were assessed by blastocyst. In experiment: 3, Zygotes were collected 18 h post-hCG and treated with the optimum concentration to check the toxicity effects on embryo development.
In experiment 1, There were insignificant results observed on the bases of cleavage rate in each groups and time of activation as compared to control group. The tendency of morula and blastocysts formation rate was higher (p<0.05) in the 15µM for 10 min activation time as compared to other treatment groups and control group. In experiment 2, The tendency of cleavage rate was significantly higher in the 10 µM and 15µM groups as compared to other treatment group. The blastocyst formation rate was no statistically difference in all treatment and control group. While the toxicity experiment, there was no toxic effect of Ionomycin with Strontium Chloride.
In conclusion, there was higher cleavage rate, 4 cells, morula and blastocyst formation rate in 15µM concentration of Ionomycin for 10 min with Strontium Chloride, there was no toxic effect of Ionomycin with Strontium Chloride on embryos and Ionomycin improved the activation rate and embryo development in combination with strontium chloride.
Availability: Items available for loan: UVAS Library [Call number: 2319-T] (1).
840.
Impact Analysis Of Quality Control Practices In Medical Diagnostic Labs Operational In District Lahore
by Tahira Naz Saif (2009-VA-204) | Dr. Ali Ahmed Sheikh | Dr. Ali Ahmed Sheikh | Dr. Muhammad Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Medical laboratory services are essential to patient care as laboratory results influence 70% of diagnosis. When tests are performed, there is always some level of inaccuracy. The challenge is to reduce the level of inaccuracy as much as possible. In orders to reduce these errors throughout the lab testing cycle, it is very important to maintain the quality of diagnostic lab. For this purpose standards should be set in the laboratory. These standards have the general requirement for effective management and competent testing.
In this study 5 public and 4 private sector medical diagnostic labs in district Lahore were selected. Permission from the competent authorities was taken and information was gathered through questionnaire including parameters such as lab building design, human resource of the lab, equipment and consumables, quality control measures adopted in the labs, environmental monitoring, waste management and customer care covering factors like recruitment policy of staff, Job descriptions of the staff, trainings conducted in the lab, SOPs, quality control and quality assurance program, feedback system, corrective and preventive action implemented in lab, result reporting etc.
The data was analyzed statistically through frequency distribution by using Statistical Package for Social Sciences (SPSS) version 18.0 for development of Graphs and Tables.
This study was conducted to help to evaluate the quality system of medical diagnostic labs and to identify the critical points which affect the quality of the testing and diagnosis and are being ignored. Information regarding different parameters was summarized in tabulated form by calculating percentage values. Two types of comparison were done. Firstly percentage value for each lab is calculated to check which labs fulfill maximum requirements of the parameters described. Second comparison was done to find out the percentage of the labs out of total selected labs, which fulfill different factors of the parameters described. After statistical analysis using chi square results showed that there is no significant difference between private sector labs and public sector labs.
In the first parameter, lab building design lab 1 and lab 9 got 100% quality points and lab 6 and lab 8 got minimum scores of 20%. In second parameter, human resource of lab 1, 2 and 3 got 100% quality points. Lab 6 and 8 got minimum quality points of 33%. When third parameter equipment and consumables was investigated it was found that lab 7 and lab 9 100% fulfill this parameter and lab 6 and lab 8 fulfill this parameter only 16.6%. In fourth parameter, quality control practices implemented in labs; lab 2 got 86% scores and lab 1 got 82% while lab 8 got minimum quality points of 22%. In fifth parameter, waste management of the lab; lab 4 got maximum quality points of 83% and all other labs got the same score of 66.6% in this parameter. When sixth parameter, environmental monitoring was investigated. It was found that lab 2 and lab 9 monitor the environment of the lab only 66.6% and in lab 8 environmental monitoring is not done. In seventh parameter, customer care lab 9 got 100% scores and labs 3, 4, 5 and 6 got 50% scores.
Conclusion:
Results show that there is no significant difference between government and private sector labs. Labs do not pay attention to the quality control measures and use same approaches of quality control in their system.
Availability: Items available for loan: UVAS Library [Call number: 2317-T] (1).
841.
Prevalence And Risk Analysis Of Coxiella Burnetii In Soil Of Sheikhupura And Attock Districts Of Punjab
by Sidra Akram (2009-VA-246) | Dr. Muhammad Zubair Shabbir | Prof. Dr. Masood Rabbani | Dr. Waseem Shahzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Background:
Besides nutrients and minerals, soil is aggregate of number of pathogens. Many of them are of
zoonotic importance and have significant threat to public health. Of these is Coxiella burnetii
that has been reported from other countries including the neighboring to Pakistan. Its occurrence
in soil, clinical significance and importance to human and animal health has been reported;
nevertheless nothing is known of C. burnetii in Pakistan particularly in rural setup where human
and animals are in close proximity to each other as well as the fact that how different risk factors
can be implicated in its spread and survival in the soil. PCR helps to identify the organism on the
basis of its genome and it is highly preferable over other conventional detection assays.
Methodology:
PCR technique was used to identify C. burnetii in the soils of Sheikhupura and Attock
districts.Soil samples were collected from each village of the both districts and processed for
genome extraction using commercial soil DNA extraction kit. Following genome extraction, the
samples were run further for PCR analysis followed by standard gel electrophoresis technique.
Later the pathogens prevalence has mapped in relation to roads, canals, rivers and drains for both
districts.
Summary
47
Outcome:
Contribute to the understanding about previously unrevealed prevalence of Coxiella burnetii in
soil of district Sheikhupura and Attock together with risk factor analysis implicating possible
health significance as well as survival in the soil. The distribution among two districts showed a
close association of gene IS1111 positivity and the land use. Positive samples were mostly found
along the roads and water bodies (canals, drains, river etc.). Availability: Items available for loan: UVAS Library [Call number: 2314-T] (1).
842.
Evaluation Of Current Food Safety Practices At Various Food Establishments In Lahore
by Muhammad Shahbaz (2013-VA-600) | Dr. Muhammad Nasir | Dr. Zubair Farooq.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Food Safety is a scientific discipline describing handling, preparation and storage of foods in ways that prevent food borne illnesses. In present era there is much advancement in the process of product control and practices which are aimed to produce wholesome and safe food. Governments have imposed very strict quality and food safety assessment parameters for food establishments. All food establishments have not adopted practices to ensure food safety.In the present study the survey of 500 food establishments of Lahore were conducted to evaluate current food safety practices adopted by various food establishments. The evaluation was done by using structured questionnaire/checklist. The questionnaire/checklist detailed of food establishment, management and personnel, personal hygiene, establishment design and physical facilities, maintenance and sanitation of establishments, control of operations and consumer awareness with product information. SPSS version 16 was used for all data analyses. Descriptive statistics including frequencies, means, and standard deviations were calculated for all variables as appropriate.The project highlighted the situation of current food safety practices and gaps in implementation adopted by various food establishments in Lahore. Evidence from the literature suggests that majority of Pakistani foods are drastically implicated with contaminants. Food hygiene training as a means of improving food safety, strong consumer protection groups, awareness of food safety, and the right to acquire safe food, strict adherence to food laws, new legislation with changing food safety perspectives, and the recognition of this challenging problem by all stakeholders are some of the practical, systematic, and aggressive steps that can be taken to curtail the risk of microbiological and chemical foodborne diseases and to minimize the health consequences associated with the consumption of unsafe foods in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 2331-T] (1).
843.
Assessment Of Genotoxicity Of Propofol, Thiopental And Ketamine In Patients Under Balanced Anesthesia With Isoflurane
by Maidah Mehtab (2013-VA-597) | Dr. Muhammad Adil Rasheed | Dr. Tanveer Akhter Butt | Dr. Muhammad Ovais Omer | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Exposure of anesthetic agents to the patients and operating room staff may involve a genotoxic risk so the knowledge of their effects on genetic material can give valuable support to anesthesia care providers to make better treatment performance and improve patient safety.
Comet assay was used to study the genotoxic actions of three IV anesthetic agents (propofol, thiopental and ketamine) that were used for induction during balanced anesthesia with inhalational anesthetic isoflurane. Three groups consisted of total18 patients who were undergone elective abdominal procedure lasted about 2 hours. Intravenous samples of blood were obtained before anesthesia induction (T0 —baseline), immediately after anesthesia induction (T1), 10 min (T2), 60 min (T3) 120 min (T4), 6 hours (T5) and 12 hours (T6) after anesthesia induction. Lymphocytes were isolated and single-cell gel electrophoresis/comet assay was used in which the cell suspension on agarosed slides was lysed in high salts and detergents containing lysing solution, exposed to alkaline buffer solution for DNA unwinding and then following electrophoresis at 24 volts and 300 mA and stained with ethidium bromide. These preapared slides were analyzed under fluorescent microscope. The anesthetics induced damage to DNA on 50 cells per sample per patient was measured as total comet length (i.e. damage index) categorized as undamaged to highly damaged (class0- class3) cells.
The data collected was analyzed by analysis of variance (ANOVA) Post Hoc Test LSD using Statistical Package of Social Sciences (SPSS).
By comparing the genotoxicity of propofol, thiopental and ketamine, it can be concluded that propofol causes the least or no genotoxicity during balanced anesthesia with isoflurane and could be the best choice for induction when isoflurane is used for maintenance.
Availability: Items available for loan: UVAS Library [Call number: 2323-T] (1).
844.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Zingiber Officinale Rhizome Against Common Poultry Pathogens
by Shumaila Nawaz (2013-VA-442) | Dr. Muhammad Adil Rasheed | Dr. Aqeel Javeed | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Plants produce a diverse range of bioactive molecules, making them rich source of
different types of medicines. Calotropis procera, a giant milk weed, is known for its
pharmacological importance for centuries. This shrub has been known to possess analgesic,
antitumor,
antihelmintic,
antioxidant,
hepatoprotective,
anti-diarrhoeal,
anticonvulsant,
antimicrobial, oestrogenic, anti-nociceptive and anti-malarial activity. A very little information is
available regarding the antibacterial and cytotoxic activity of Calotropis procera so the present
study is designed to evaluate the antibacterial and cytotoxic activity of this plant. This study was
conducted to access antibacterial and cytotoxic activity of Calotropis procera.
Hexane, chloroform and ethanol, aqueous extracts were prepared by sequential extraction
method and antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli,
Salmonella enterica, Clostridium perfringens type A and Haemophilus paragallinarum by agar
well diffusion method in which inhibitory zones were measured. The extracts which showed the
antimicrobial activity were evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell
culture media was prepared and cell lines were propagated, monolayer formed. Monolayer was
exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival
percentage was calculated.
Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for
windows version 16, SPSS inc, Chicago, IL, USA). Results will be compared using one way
ANOVA analysis.
102
SUMMARY
Chloroform and ethanol extracts of Calotropis procera leaves have antibacterial activity.
It may help to design traditional medicines for the development of therapeutic agent which will
be more safe, effective and economical.
Availability: Items available for loan: UVAS Library [Call number: 2330-T] (1).
845.
Efficiency Of Nitrogen Utilization In Response To Level Of Crude Protein Supplies In Multiparous Lactating Nili-Ravi Buffaloes
by Rizwan Munawar (2008-VA-114) | Dr. Muhammad Naveed Ul Haque | Mr. Murtaza Ali Tipu | Prof. Dr. Anjum Khalique | Dr. Qamar Shahid.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Livestock has a vital role in pakistanʾs economy. Livestock play a key role in alleviating poverty from our rural areas. Nutritional management is of keen importance in livestock business. Optimum supply of nutrients (protein and energy) is very important for precise farming. Establishing protein and energy requirement for animals has a vital role in livestock farming for guaranteeing the optimum supply of these nutrients. Under feeding and over feeding of these nutrients may negatively affect the performance of animal and also may have some adverse effect on environment by production of different types of hazardous gases. Protein requirement of buffalo depends on milk production and milk protein contents together with efficiency of conversion of dietary N to milk N. buffalo is reported to be less efficient converter of dietary N to milk N as compared to cows, however average dietary N conversion to milk still needs further investigation.
This study was conducted at Buffalo Research Institute, Pattoki. Three levels of crude protein were supplied to check their effect on production performance of buffaloes. Design for the experiment was 3×3 latin square including two animals in each treatment. Experiment was completed in 63 days (21 days in each period). During the study different parameters were determined by using different laboratory techniques. Milk parameters included milk yield, milk fat, milk protein and milk lactose. Blood parameters included urea, total protein, triglycerides (TG) and glucose. Fecal sampling was done to check N digestibility and feed analysis were also performed by using standard procedures. Data were analyzed using GLM procedures of SAS.
DMI was similar at all the treatments. DMI at low CP diet, medium CP diet and high CP diet was 14.3, 14.4 and 14.1 kg/d respectively. Milk yield was also identical at all the protein supplies and was 9.90, 10.0 and 9.82 kg/d for low protein supply, medium and high protein
Summary
41
supply respectively. Milk protein contents also showed no response and were 3.99, 3.94 and 3.99 % at low CP, medium CP and high CP respectively. Milk fat contents were also same at all the treatments and these were 6.63, 6.58 and 6.67 % at low, medium and high CP level of diet. Milk lactose concentration was 5.30, 5.26 and 5.31 % at all the three diets. Similarly protein yield, fat yield and lactose yield showed no response of this dietary variation of protein. Body weights of the animals were increased by increasing CP in diet. Body weights of the buffaloes were 478.3, 498.5 and 499.1 kg at low CP, medium CP and high level of CP in diet, respectively. Nitrogen intake was significantly higher at higher CP supplies and was 248.4, 297.3 and 314.3 g/d at low protein, medium protein and high protein levels. Similarly, CP intake was higher at higher protein supplies. CP intake increased from 1.553 to 1.964 kg/d by varying protein from low to high level. Nitrogen efficiency was decreased by increasing level of CP in diet and was decreased from 24.94 to 19.64% by increasing CP from low to high level in diet. BUN was increased linearly by increasing CP in diet, whereas blood glucose and triglycerides showed no response of this dietary manipulation.
In current feeding situation, no effect of increasing protein supply was observed on production status of these buffaloes. Blood urea nitrogen was increased whereas efficiency of nitrogen utilization was decreased by increasing CP in diet. So the recommended level of CP in ration for milk yield of 10 kg with 4% protein and 6.6% fat is 10.9%. But this still needs further investigations. Current experiment served as an opportunity to explore the effect of varying levels of CP on nitrogen utilization efficiency in buffaloes. Further research is required in high forage based diets. Availability: Items available for loan: UVAS Library [Call number: 2327-T] (1).
846.
Genetic Association Study Of Apolipoprotein A-V (Apoa5) And Sortilin (Sort1) Genes With Risk Of Coronary Artery Disease
by Irfan Basharat (2012-VA-802) | Dr. Akhtar Ali | Dr. Wasim Shehzad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: In developed countries cardiovascular disorders are prominent cause of death. One third deaths in the world are due to cardiovascular disorders. Among cardiovascular disorders coronary artery disease responsible for one in five deaths in USA. Its main reason is the lipids values particularly cholesterol and triglycerides in the blood. An estimation made by WHO indicated that 9 million people die per year due to hypercholesterolemia.
100 blood samples were collected from patients of coronary artery disease and from normal patients with no myocardial history. Allele specific primers for SORT1 gene and APOA5 genes were designed using Primer 3 software web facility. Genomic DNA will be amplified by PCR then genotyping will be carried out and DNA will also be sequenced.
Hardy-Weinberg principle and Fisher Exact test were used to assess the allele frequency and significant variations from results
When patient of MI and normal group were genotyped and sequenced we find out that there are 34 AA homozygous, 1 GG homozygous and 12 heterozygous persons in case of APOA5. The SORT1 person shows 24 GG homozygous and 3 AA homozygous and 13 heterozygous persons.
Our study shows a definite association between APOA5 and SORT1 with respect to MI disease persons. This study shows a significant association of single nucleotide polymorphism in APOA5 and SORT1 genes with coronary artery disease. Availability: Items available for loan: UVAS Library [Call number: 2326-T] (1).
847.
Studies On Growth Performance, Morphology, Reproductive Traits And Behavioral Aspects Of Ring Necked Pheasants In Captivity
by Sana Ashraf | Dr. Arshad Javid | Dr. Khalid Mehmood Anjum | Prof. Dr. Muhammad Ashraf | Prof. Dr. Muhammad Akram.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Besides ecological importance, pheasants also have aesthetic values which mainly contribute to their decline in population due to poaching in their native habitat. Among 49 species of pheasants in the world, 5 are endemic to Pakistan with distribution in the Himalaya and remote northern parts of the country. Due to increase in human population, intrusion, poaching, and habitat disturbance pheasants are threatened. However, much of the work on captivity is based on production rather morphological, ornamentation or reproductive traits which could have important implications for the management of wild and captive populations as a whole. Some studies still remain to be explored.
Present study was conducted to find out the relationship of egg weight with egg quality parameters and growth traits of ring-necked pheasant Phasianuscolchicus. Total of 450 eggs were collected and were divided into three egg weight categories viz. light (20.0-26.0g), medium (27.0-32.0g) and heavy (33.0-40.0g) egg weight eggs. Fifty eggs for each of the egg category were reserved for the evaluation of internal egg quality parameters. External egg quality parameters i.e. egg length, breadth, egg volume and surface area varied significantly (P<0.05) between all the three egg weight categories. Similarly, significantly higher albumen and yolk weight were recorded in heavy weight egg category while non-significant relationship of egg weight was observed for shell and membrane thickness, yolk percentage, yolk index, yolk pH and albumen pH of the egg. The hatching percentage for the remaining 300 egg kept in incubator was 47.33%. Forty chicks from each of the egg weight category were selected and chick weight, wing length and wingspan were taken at the time of hatching and thereafter increase in these parameters were noted on weekly basis. The effect of egg weight on chick weight, live weight gain, wing length and wingspan was significant (P<0.05) from 1st to 12th month of age. Our studies revealed that egg weight has strong influence on external and internal characteristics of the eggs and the growth parameters in P. colchicus chicks.
The present study was planned to evaluate the time budgets of ring necked pheasants Phasianuscolchicus in captivity. The birds were kept in cages of 5 ft × 5 ft × 3 ft (length × width × height) and were housed in a 20 ft × 20 ft (length × width) well ventilated room at Department of Wildlife and Ecology, Ravi Campus, University of Veterinary and Animal Sciences, Lahore. Captive birds were divided into three categories viz. adult male, adult female and chicks and were placed into separate cages. Behavioral parameters viz. jumping, aggression, preening, feather pecking, walking, standing, sitting, litter pecking, drinking, feeding, body shaking, voice call and feather flapping were assessed for 30 birds from each of the three categories through scan sampling. Statistically significant variations were recorded in behavioral aspects among all the three categories. Male birds spent significantly higher times in aggression (155.26±3.10 sec), preening (74.04±3.05 sec), walking (1370.93±54.45 sec), drinking (74.00±3.18 sec), body shaking (24.92 ±3.11 sec), voice call (20.08±3.17 sec) and feather flapping (15.42±2.73 sec) while female P. colchicus spent significantly higher times in sitting (364.57±3.74 sec). Similarly, the chicks spent significantly higher times in jumping (36.17±2.75 sec), feather pecking (265.19±3.17 sec), standing (1230.13±23.86 sec), litter pecking (234.89±2.97 sec) and feeding (115.44±3.11 sec) as compared to the adult female and male birds.
Fecal and blood samples of ring necked pheasants, Phasianuscolchicus were analyzed to record the parasitic prevalence in these pheasants. A total of 1000 samples, 500 blood and 500 fecal samples were collected from Captive Breeding Facilities for Birds, Department of Wildlife and Ecology, Ravi Campus, University of Veterinary and Animal Sciences, Lahore. Parasitic genera identified from blood samples of P. colchicus include Leukocytozoon, Plasmodium and Haemoproteus. Prevalence of Leukocytozoon was 16 % while the prevalence of Haemoproteus was 14.3%. Parasitic genera identified from fecal samples of P. colchicus include Eimeria, Isospora, Trichomonas and Giardia. Eggs of five species of nematodes viz. Capillaria, Syngamus trachea andAscaridia,Heterakisisoloncheand Heterakisgallinarum were also identified from the fecal samples. The ectoparasites include one species of burrowing mite Knemidocoptesmutansand two species of chewing lice i.e. Amyrsideaperdicis and Lipeurusmaculosus.
Variations in hematological parameters during different life history stages were recorded in ring-necked pheasants (Phasianuscolchicus) for a period of 1 year. Thirty birds were selected for analysis of selected hematological parameters viz. red blood cells count, white blood cells count, hemoglobin, mean corpuscular hemoglobin concentrations, total serum protein and leucocyte count. These birds were kept in cages, each cage having separate drinking and feeding facilities. Five birds per cage were confined and these cages were housed in a well-ventilated 20 × 20 feet (length × width) room. Blood samples were taken from ulnar vein and variations in blood parameters were recorded on monthly basis. Significantly, lower RBC’s count was observed during 2nd month of age while during same month significantly higher WBC count was noticed. Significantly lower values of hemoglobin were observed during 1st and 2nd month of age. Significantly higher mean corpuscular hemoglobin concentration percentage (MCHC%) was recorded during 6th and 7th month of age. Lower packed cell volume (PCV) values were observed during 2nd and 6th month of age while the total serum protein concentrations were recorded maximum during 10th month of age. Significantly, higher heterophils count was recorded during 2nd and 10th month of age while maximum lymphocyte count was observed during 2nd and 7th months of age. Significantly, higher concentrations of monocytes were recorded during 11th, 12thand 10th month of age. The eiosinophils count varied from minimum (110 ± 13.50) during 3rd month to maximum (902 ± 93.22) during 11th month of age. Similarly, significantly higher values of basophils were recorded during 1st month of age. It can be concluded from the present study that the blood profile of the pheasants changes with age.
Variations in hematological parameters for adult male and female ring-necked pheasants (Phasianuscolchicus) were recorded. Forty adult ring-necked pheasants (20 ♂, 20 ♀) were kept in separate cages, each cage having separate drinking and feeding facilities. Five birds per cage were confined and these cages were housed in a well-ventilated 20 × 20 feet (length × width) room. Blood samples were taken from ulnar vein. Different blood and serum chemistry parameters such as red blood cells (RBCs), white blood cells (WBCs), hemoglobin (Hb ) concentrations, mean corpuscular hemoglobin concentration (MCHC), packed cell volume (PCV), heterophils, lymphocytes, monocytes, eosinophils, basophils, ALP, uric acid, cholesterol, total serum protein, albumin and creatinine were determined among adult male and female pheasants. Non-significant differences in RBCs, WBCs, heterophils, eosinophils, MCHC and Hb values were observed among male and female pheasants. Significantly, higher values of lymphocytes, monocytes and PCV were observed in males while higher basophil count was observed in female as compared to male birds. Significantly higher values for ALP, cholesterol, total serum protein and creatinine were observed in males while higher uric acid values were observed in females as compared to male P. colchicus. However, non-significant differences in albumin were recorded among male and female birds.
Availability: Items available for loan: UVAS Library [Call number: 2350-T] (1).
848.
Analysis Of Genetic Polymorphism In Exon 6 & 11 Of Glucosidase Beta Acid (Gba) Gene In Gaucher Diseased Patients From Punjab, Pakistan
by Aysha Arshad (2009-VA-571) | Dr. Muhammad Yasir Zahoor | Dr. Muhammad Imran | Dr. Imran Altaf.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Gaucher disease (GD) is amajor predominant heterogenic, inherited and metabolic lysosomal storage disorder. It is prompted by an alteration in glucosidase acid beta (GBA) gene. GBA gene encodes a 497 amino acid glucocerebrosidase enzyme. It is a lysosomal hydrolase, present in all mammalian cells membrane that carries the catalysis of complex ubiquitous sphingolipids called glucocerebrosides (GlcCer) into smaller and simpler molecules of sugar and ceramide. The human glucocereborside (GBA) gene is present in highly gene dense area on q arm of 21 chromosome and its fragment length is 7.8kb comprising of 11 exons. A pseudogene is also present in vicinity of GBA gene which shares 96% homology of sequence with functional gene. Genetic recombination and gene conversion among these two GBA genes are responsible for 10-20% GD mutations. >300 mutations of GBA have been described till 2014. GD has three different clinical forms depend on its heterogeneity. These are characterized by the age of onset and with or without the participation of CNS.
In this study, 10 blood samples were collected of GD patients from repository at Molecular and Genomic Laboratory located at IBBT department, UVAS Lahore and from Children Hospital Lahore. DNA extraction was done by using organic method from blood samples. Amplification of GBA gene exons 1, 6 and 11 was performed using PCR. PCR products were sequenced using Sanger di-deoxy sequencing method. Different bioinformatics tools were applied for the sequence analysis of exon 1, 6 and 11. We found two variants of GBA gene. A deletion of CT nucleotide repeat in intron 1 was found. We also found a substitutional change of nucleotide T>A in intron 8. Availability: Items available for loan: UVAS Library [Call number: 2334-T] (1).
849.
Variation Analysis Of Hepatitis C Virus Gene Encoding E2 Glycoprotein
by Saimoon Theeen (2009-VA-565) | Dr. Muhammad Imran | Dr. Sehrish Firyal | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Hepatitis C virus (HCV) major cause of liver infections was discovered in 1989. It is positive stranded RNA virus and belongs to Flaviviridae family. Its genome shows high rate of variations due to which, its rate of infection is high. As in Pakistan 3% to 6% population and 170 million people worldwide are affected by it. Due to variations in its genome it is classified into 6 major genotypes and these genotypes are further classified into many subtypes. Size of HCV is about 9500 bps which only encodes single polyprotein. This 3000 to 3300 amino acids polyprotein is processed by cellular and viral proteases to generate 10 polypeptides consisting of 4 structural (Core, E1, E2 and P7) and 6 non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, NS5B). Whereas E2 is considered the most immunogenic gene from all the genes. It involves in the interaction with the host cell and easily escape from the immune system of host due to the presence of hypervariable regions in E2 gene.
To isolate the E2 gene RNA extraction was done using the kit method and then it was converted to the cDNA which is then followed by the PCR amplification. The amplification products were then purify and sent for the sequencing to CAMB. Then the bioinformatics tools were applied on the results. In which the protein structural analysis and epitope mapping was done. Then the conserved epitopes were predicted using the IEDB conservancy analysis tool. The conserved B-cell epitopes (TElAILPCSFTPMPAL and RGERCDIEDRSEQH) and T-cell epitope (TPMPALSTG) are now considered valuable to produce the antibodies against E2 protein.
For diagnosing HCV genotype 3a, these conserved epitopes may be highly useful and may also help in developing a successful vaccine that can target 3a genotypes. Availability: Items available for loan: UVAS Library [Call number: 2333-T] (1).
850.
Comparison Of Antifungal Activity Of Human Salivary Histatin Between Diabetic And Nondiabetic Individuals
by Farid-Ul-Haq (2013-VA-555) | Prof. Dr. Tahir Yaqub | Dr. Ali Raza Awan | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Histatins are antimicrobial proteins found in human saliva. These proteins have also been
observed to have the ability to aid in wound healing in various organisms. The genes HTN1 and
HTN3 have been studied to govern these proteins. Histatin proteins have a vast array of
antimicrobial properties. While a fungus, Candida albicans or C. albicans is a part of the human
normal gut flora, it is a threat to people who have a compromised immune system. An
overgrowth of the fungi belonging to the Candida family leads to candidiasis in humans, and oral
candidiasis has been reported to a large extent namely in diabetic patients. The antifungal
activity of histatin proteins laid the basis of the current research work.
In this study, the antifungal activity of saliva from a total of 64 healthy and diabetic
human samples against Candida albicans has been evaluated. The samples of both healthy and
diabetic human samples belong from different age ranges: 15-25, 25-35, 35-45 and 45-55 years
in order to change in antifungal activity with respect to age of an individual. Antifungal activity
was observed through both agar well and agar disk diffusion methods, with agar disk diffusion
methods showing positive results. According to the outcomes of this study at least 120μL of
healthy saliva sample is required to create a zone of inhibition. Saliva from diabetic individuals
showed no antifungal results.
This occurrence led to the next part of this study involving amplification of HTN3 gene.
The nucleotide sequences of both healthy and diabetic individuals were compared together and
showed that the absence of antifungal activity in diabetic individuals might have reasons other
than a genetic one, according to this study. The results observed from the present study indicate
that healthy human saliva possesses antifungal activity against Candida albicans. In accordance
Summary
39
to these results, the naturally occurring antimicrobial activity of histatin proteins present in
human saliva can have immense use in the field of medicine. Availability: Items available for loan: UVAS Library [Call number: 2341-T] (1).
