1.
Role Of Non-Antimicrobial Agents In Reversal Of Antibiotic Resistance In Escherichia Coli
by Kalim Ullah | Prof. Dr. Muhammad Ashraf | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1839,T] (1).
2.
Isolation And Molecular Characteracterization Of Staphylococcus Aureus From Raw Milk
by Ibrar hussain | Prof. Dr. Muhammad ayaz | Dr. Imran javed | Prof. Dr. Aftab ahmad anjum.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1853,T] (1).
3.
Genetic And Evolutionary Characterization Of Pakistani Pigeons And Parrots Through Mitochondrial D-
by Sehrish firyal | Dr. Ali raza awan | Prof, Dr. Aftab | Prof, Dr. Tahir yaqub.
Material type: ; Format:
print
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1873,T] (1).
4.
Isolation Characterization And Growth Optimization Of Starch Hydrolyzing Fungi From Soil Of Livestock Farms
by Saba Sana | Prof. Dr. Aftab ahmad anjum | Dr. Muhammad Nawaz | Prof. Dr.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1898,T] (1).
5.
Plasmid Mediated Analyses And Plasmid Curing Of Previously Isolatedmulti-Drug Resistant Eschetichia Coli From Retail
by Mawra gohar | Dr. Ali ahmad sheikh | Dr.Tanveer | Prof, Dr. Aftab ahmad anjum.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1923,T] (1).
6.
Genotoxic Mutagenic And Cytotoxic Potential Of Metformin And Celecoxib Alone And In Combination
by Asad ullah | Prof/ Dr. Muhammad Ashraf | Dr. Aqeel javeed | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1989,T] (1).
7.
Isolation Characterization And Optimization Of Potential Probiotic Bacteria From Poultry Droopings
by Muhammad Hashim khan | Prof. Dr. Aftab ahmad anjum | Dr. Jawad nazir | Prof. Dr. Mansur-ud-din.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1991,T] (1).
8.
Isolation And Characterization Of Antibiotic Resistant Lactobacilli From Fermented Food Products
by Shahgull | Dr. Muhammad Nawaz | Prof. Dr | Prof. Dr. Aftab anjum.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2002,T] (1).
9.
Cytotoxicity Mutagenicity And Genotoxicity Potential Of Carvedilol And Celecoxib Alone And In Combination
by Ali attiq | Prof.Dr. Muhammad Ashraf | DR. Aqeel Javeed | Prof. DR. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2004,T] (1).
10.
Comparative Efficiency Of Coprological Identificatiocn With Sensitive Detection Of Cryptosporidium By PCR In Domestic and Commercial Chickens
by Hafiz fahad nazir | Dr. Nisar ahmad | Prof DR. Azhar maqbool | Prof. DR. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2010,T] (1).
11.
Comparison Of Ameliorative Potential Of Sacccharomyces Cerevisiae And Bentonite Clay On Pathological Effects Induced By Aflatoxin in Broilers
by Muhammad Saqlain | Dr. Ishtiaq ahmad | Dr. Gulbeena saleem | Prof. dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2014,T] (1).
12.
In Vitro Antibacterial Activity Of Star Anise (Illicium Verum) Oil Against Common Food Borne Pathogens And Its Utilization in Cookies Preparation
by Shamim khalid | Dr. Naureen naeem | Dr. Sanaullah iqbal | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2033,T] (1).
13.
Phylogenetic Analysis Of Plasmodium Species In Sparrows And Domestic Chicken
by Ghanwa ahmad | Dr. Haroon Akbar | Dr. Muhammad lateef | Prof Dr. Aftab.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2075,T] (1).
14.
Detection Of Bacterial Load In Quail Meat Available In Lahore Market
by Muhammad Rameez akram | Dr. Naureen naeem | Ms. farasat rizwan | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2078,T] (1).
15.
Assessment Of Microbial Load In Vegetables Grown In Sewage Polluted Water In Lahore Surroundings
by Asifa | Dr. Sanaullah Iqbal | Dr. Naureen Naeem | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2119,T] (1).
16.
Synergistic Effect Of Antimicrobial Agents In Combination With Colistin Against Bacterial Isolates From Patients
by Sahar Safdar | Prof. Dr. Muhammad Ashraf | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2123,T] (1).
17.
Determination Of Microbial Contaminants Of Canned Fruit Products Available In Local Markets Of Lahore Pakistan
by Muhammad Waseem Akram | Dr. Muhammad Nasir | Dr. Zubair Farooq | Prof. Dr. Aftab.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2143,T] (1).
18.
Prevelance Of Brucellosis In Aborted Women Visiting Tertiary Care Hospitals Of Lahore City
by Saba Yasmin (2009-VA-211) | Prof. Dr. Aftab Ahmad Anjum | Dr. Tayyaba Ijaz (Co Supervisor) | Prof. Dr. Khushi Muhammad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is an agriculture based country whose rural population depends upon livestock for livelihood. Contribution of livestock to agriculture sector is 55.9 percent while 11.8 percent to the national GDP during 2013-14 (GOP 2013-2014). A number of infectious diseases hamper the growth of livestock sector. Some of the livestock diseases are zoonotic in nature and threat to human health. Brucellosis is considered among major zoonotic diseases throughout the world. The Mediterranean Basin, south and Central America, Eastern Europe, Asia, Africa, the Caribbean and the Middle East are considered as high-risk countries (Memish 2001).
Brucellosis in human beings is a major concern of community health. It causes acute and chronic illness, physical incapacity and loss of health. Bacterial species involved include Brucella abortus, Brucella melitensis or Brucella suis. Brucellosis is acquired by human beings from infected animals by close contact with vaginal secretions, urine, feces, blood, aborted fetus, or consumption of unpasteurized milk or other raw milk products. Shepherds, milkmen, butchers, knackers, veterinary assistants and abattoir workers are at high risk (Agasthya et al. 2007). Prevalence of brucellosis recorded by Mukhtar and Kokab (2008) in abattoir workers of Lahore Pakistan was 21.7 percent. Higher prevalence of brucellosis was observed in females (37.06%) than males (24.2%) in patients admitted at Peshawar, Pakistan (Shahid et al. 2014).
Symptoms of disease vary among human patients, ranging from non–specific, flu-like symptoms (acute form) to undulant fever (chronic form). Some of the serious complications of skeletal system, cardiovascular and central nervous systems may develop. Other important signs observed include arthritis, orchitis, epididymitis, abortion, retained placenta and stillbirth (Baba et al. 2001; Grilló et al. 2006). In animals, brucellosis in most of the cases results in abortion, birth of weak calves, death of young stock, infertility in males and reduced milk yield in females (Maadi et al. 2011; Abubakar et al. 2012).
There is actual need for teamwork between public health officials and veterinary officers to reduce communication of brucellosis between animals and human in endemic areas (Jelastopulu et al. 2008; Makis et al. 2008). Clinical picture of brucellosis is nonspecific and may vary from patient to patient. Therefore, laboratory diagnosis by isolation and culture or recognition of specific anti–Brucella antibodies is essential for confirmation of brucellosis (Al-Attas et al. 2000).
Diagnosis of brucellosis by culture and phenotypic description is time-consuming. Furthermore, risk of infection to worker is always there. Serological tests are commonly preferred for brucellosis in cattle and small ruminants, especially at farm level screening. Chance of cross-reactions with other gram negative bacteria is a major problem. Rose Bengal Plate Agglutination Test (RBPT) and Slow Agglutination Test (SAT) are extensively used for detection of anti-Brucella antibodies (Halling et al. 2005). Enzyme Linked Immunosorbent Assays (ELISA) have been developed to resolve suspected samples by RBPT. ELISA is more sensitive, so it can detect Brucella carriers which are negative by RBT, SAT and CFT (Aert et al. 1984). Molecular techniques are more reliable and specific than serological tests. Final confirmation of brucellosis is carried out using polymerase chain reaction (PCR), a molecular technique. Real-time PCR offers enhanced sensitivity, specificity and rapidity of performance when compared to conventional PCR (Gwida et al. 2012). Availability: Items available for loan: UVAS Library [Call number: 2225-T] (1).
19.
Status Of Awareness Among Zoo Workers About Zoonotic Diseases
by Tahir Khan (2012-VA-806) | Prof. Dr. Mansur Ud Din Ahmed | Shelly Saima Yaqub | Dr. Shakera Sadiq Gill | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: A zoo is a place where wild animals are kept for exhibition purposes to the public.It includes: aquaria, sanctuaries, bird gardens and safari/wildlife parks. These are centers for wild animal’sconservation and for public recreation and education (Cuaron2005). Epidemiologists, wildlife biologists, veterinarians and conservationists used these for research purpose.According to an estimate Pakistan is maintainingapproximately 27 zoos, deer parks, etc.(Walker 2014).
Zoonotic diseases are those which are naturally transmitted from animals to human beings and vice versa. The word Zoonosis is derived from the Greek word zoon (animal) and nosos (disease). The diseases which are transferred from human beings to animals are known as Zooanthroponotic (Greek “Zoon” = animal, “anthrópos” = man, “nosos” = disease) diseases e.g. tuberculosis, measles, giardiasis and amoebiasis. On the other hand the diseases which are transmitted from animals to human beings are known as anthropozoonotic diseases e.g. anthrax, AIDS, psittacosis and rabies (Epstein and Price 2009).
Zoonosis can be classified according to their circulation in the ecosystem. These are either classified as synanthropic zoonosis, with an urban (domestic) cycle in which the source of infection are domestic and synanthropic animals (e.g. cat scratch disease, urban rabies and zoonotic ringworm) or exoanthropic zoonosis, with a sylvatic (feral and wild) cycle in natural foci outside human habitats (e.g. wildlife rabies, arbovirus, lyme disease and tularemia). Some zoonotic diseases can circulate in both urban and natural cycles (e.g. chagas disease and yellow fever).
A review study identified that 1415 species of infectious organism are pathogenic to human beings. This includes 217 viruses and prions, 538 bacteria and rickettsia, 307 fungi, 66 protozoa and 287 helminthes. Out of these, 868 (61%) are zoonotic in nature (Taylor et al. 2001). More than 60% of the emerging human infectious diseases are zoonotic in nature and 70% of their reservoirs are wild animals (Cutler et al. 2010).
The reservoirs of several zoonotic diseases are wild animals whose causative agents are viral, rickettsial, chlamydial, bacterial, parasitic and mycotic(Bengis et al. 2004). Zoonotic diseases like tuberculosis, plague and rabies have badly affected the mankind since ancient times and the reservoirs of all of these are wild animals (Stone et al. 2009). Some zoonotic diseases in human beings are self-limiting whose signs range from few days to a long term illness e.g. gastroenteritis caused byGiardia, Cryptosporidium, and Salmonella species.Some zoonotic diseases may cause abortions (Toxoplasmosis) and fatal encephalitis (Japanese encephalitis). Whereas some zoonotic diseases may causes high mortality e.g. Marburg hemorrhagic fever(MacNeil and Rollin 2012). Zoonotic diseases cause death not only in their natural hosts but also in endangered wild animal species near to extinctione.g. Ebola virus cause high mortality in monkeys (Nunn et al. 2008). It is clear from various studies in different zoos that both anthropozoonotic and zooanthroponotic transmission can occur (Adejinmi and Ayinmode 2008).
Zoonotic agents have potential to be used for bioterrorism. The bioterrorism attack is aimed to cause fear, destabilization, stress, illness and death in people, animals and plants. (Lin 2014). Air, water and food may be the warfare biological vehicles for its spread. During World War 1, anthrax was used as a biological warfare in animal populations. Glanders and typhoid were also used for bioterrorism attack in 1910 and 1970, respectively. Several cases of bioterrorism also occurred in the United States due to anthrax in September and October 2001 (Spencer 2007).
A Zoo worker should haveknowledge of the transmission of the disease to avoid its transmission. The common ways of the transmission are direct mode (ingestion, animal bites, inhalation, needle prick injuries and skin contact) and indirect mode (vector borne, fomite, long distanceand airborne transmission). In zoo management, the role of veterinarians is extremely useful. Their job exposes them to several health-related threats during routine operations. e.g. animal bites, needle prick injuries, back injuries, exposure to anesthetic gases and even mortality in certain cases (Hill et al. 1998; Kabuusu et al. 2010).
The personal protective equipment’s are not used during restraining, treatment, necropsy and cleaning the animal enclosures. It may increases the chances of zoonotic diseases to zoo workers and veterinarians. The disposal of wild animal carcasses, organs, unused food, feces and urine by unscientific methodsenhances the process of pathogens transmission(McLaughlin 2002). Laboratory personnel can also be infected with zoonotic diseases due to lack of good laboratory practices in wildlife disease diagnostic laboratories(Rietschel 1998).
Therefore, prevention and control of zoonosis must be an important part of zoo occupational health and safety measures. Preventive measures can be either general or specifically designed for a particular disease. It is possible to prevent many of the zoonotic diseases by following basic hygiene and sanitation procedures.The present study was conducted to determine knowledge, attitude, practice and experience levels about zoonosis among zoo workers of district Lahore(Lahore Zoo, Jallo Wildlife Park and Lahore Safari Zoo).
Availability: Items available for loan: UVAS Library [Call number: 2229-T] (1).
20.
Modulation Of Antibiotics Resistance Pattern In Escherichia Coli By Different Plant
by Bushra Chaudary (2009-VA-232) | Dr.Muhammad Nawaz | Prof. Dr. Aftab Ahmed | Dr. Naureen Naeem.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Escherichia coli (E. coli) is Gram negative microorganism belonging to family
Enterobacteriaceae. It is part of normal micro flora of gastrointestinal tract of human and all
warm blooded animals (Kaper et al. 2004). Escherichia coli is source of many infectious diseases
in human as well as in animals. Common E. coli infections are enteritis, urinary tract infection,
septicemia and neonatal meningitis. In pets and farm animals, E. coli is associated with diarrhea
(Allocati et al. 2013). Poultry industry is facing huge annual losses due to infection of avian
Pathogenic E. coli (APEC) in broilers (Oosterik et al. 2014). E. coli causes a variety of
syndromes in poultry including yolk sac infection, respiratory tract infection, swollen head
syndrome, septicemia and cellulitis (Buys et al. 1989)
Antibiotics are chemical agents which inhibit the microbial growth and used to eradicate
infections. Mechanisms of action of antibiotics provide a base to categorize antimicrobial agents.
Most important classes of antibiotics act as inhibitors of cell wall synthesis, protein synthesis
(tetracyclines and macrolides), nucleic acid synthesis (fluoroquinolones), metabolic pathway
(trimethoprim-sulfamethoxazole) and cell membrane (polymyxins). Bacteria may have intrinsic
or acquired resistance to antimicrobials (Tenover 2006).
Urinary tract infections are mostly caused by E.coli. Antibiotics generally used for the treatment
of E. coli infections include ampicillin, nitrofurntion, cephalosporin, sulphonamides
(trimethoprim-sulfamethoxazole) and quonolones (neladixic acid, ofloxacine, ciprofloxacin and
levofloxacine) (Lin and Lin 2010).
Extended use and misuse of antibiotics lead to the development of resistant bacteria. Resistant
E. coli strains are common source of hospital born and community acquired infections. Ease of
Introduction
2
international travelling is one of the major spreading factor for antibiotic resistance. Resistant
bacteria got opportunity to move from one geographical area to another (van der Bij and Pitout
2012). New strains of E. coli resistant to carbapenems (New Delhi metallo-β-lactamase 1 (NDM-
1) are major global health issue (Kumarasamy et al. 2010). Antibiotic resistance has become a
serious public health problem. Currently, world is facing great difficulty in treatment of many
infectious disease of human and animals. One of the reasons of treatment failure is emergence of
resistant bacteria (Levy 2002).
To develop new strategies for treatment of infectious diseases, it is necessary to understand the
mechanisms of resistance. Efflux pump inhibitors, enzymatic degradations and alteration of
target sites are major strategies by which bacteria acquire or develop resistance to antibiotics
(Sibanda and Okoh 2007).
Scientists are looking for alternatives of antibiotics such as bacteriopheges, naturally
antimicrobial compounds and some non antimicrobial agents (Worthington and Melander 2013).
Probiotics (Lactobacillus and bifidobacterium) can be a prophylactic measures against E. coli
and may be used to treat intestinal tract infections of E. coli and other bacteria (de Vrese and
Schrezenmeir 2008).
Phytochemicals, secondary metabolites of plants, have antibacterial activity against many
pathogenic organisms. These phytochemicals in combination with antibiotics may show
synergistic effect. Phytochemicals and plant extracts can be a source of antibiotic resistancemodifying
agents (RMAs) (Abreu et al. 2012). Plant extracts shown antibacterial activity
because of phytochemicals like alkaloids, tannins, flavonoids, phenolic compounds and steroids
(Gobalakrishnan et al. 2013). Plant extracts are used as traditional medicine for the treatment of
many diseases. Plant extracts like Zingiber officinalis (Ginger) Gymnema sylvestre (Gurmar buti), Astragalus (goat’s thorn), Calotropis procera (apple of Sodom) and oputia dillenii (cactus)
have antimicrobial activity (indu et al. 2006 and Kumaar et al. 2013). Plant extracts also have
antibiotic resistance modulation potential (Mako et al. 2012).
Availability: Items available for loan: UVAS Library [Call number: 2247-T] (1).
21.
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).
22.
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.
Availability: Items available for loan: UVAS Library [Call number: 2265-T] (1).
23.
In Process Quality Control Factors Affecting The Quality Of Locally Prepared Salmonella Gallinarum Antigen
by Zahra Malik (2009-VA-245) | Dr. Arfan Ahmad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Asim Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Fowl typhoid is a septicaemic disease caused by S. gallinarum biovar gallinarum has major economic significance in many parts of the world. It is an acute or chronic septicaemic disease that usually affects the birds (mostly adult birds). Eradication of disease is normally done by identifying the infected flocks and eliminating the reactor birds by using serological tests, but diagnosis of the disease is much expensive because antigen used for this purpose is imported. The study, therefore, has been proposed to prepare and evaluate the stained antigen of S. gallinarum using local isolates.
A total of 15 isolates were procured from Poultry Research Institute (PRI) Rawalpindi, University Diagnostic Lab (UDL) and Department of Microbiology, UVAS Lahore, which were identified by Biochemical testing and further confirmed by Polymerase Chain Reaction. Among all 15 isolates two isolates were confirmed as S. gallinarum and proceeded to prepare local antigen of S. gallinarum. Locally prepared antigen was checked with known positive and negative sera, Effect of different preservatives (Sodium azide and Thiomersal sodium) and different storage temperatures (4°C, 25°C and -20°C) was also studied after every fifteen days post storage upto 6 months to observe the stability and shelf life of local antigen. On the end of study both preservatives i.e. Sodium azide and Thiomersal sodium was found equally effective for antigen activity, whereas 4°C proved best storage temperature to be used for the antigen preservation.
Activity of locally prepared antigens was also compared with the imported antigen (Charles, River, USA) stored at different temperatures regularly throughout the six months, which showed that local antigens was almost as good as the imported antigen.
Summary
51
CONCLUSION
Locally prepared S. gallinarum antigen was found as effective as imported antigen. Both the test preservatives (Sodium azide and Thiomersal Sodium) had the same effect on antigen preservation. Among all three test temperatures, 4°C was accepted as best storage temperature for the long term preservation of local antigen with either of the preservative. Availability: Items available for loan: UVAS Library [Call number: 2278-T] (1).
24.
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).
25.
Genetic Diversity Among Different Isolates Of Pasteurella Multocida From Poultry
by Arslan Sardar (2013-VA-282) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Fowl cholera is an acute bacterial disease of broiler breeders and layer breeders caused by Pasteurella multocida. In the present study, 10 isolates from different areas of Punjab were purified. These samples were confirmed by API Kit. Different molecular techniques like PCR and RFLP were used to investigate variation at the molecular level among 10 isolates collected from different areas of Punjab. Different mutations were observed among 10 field isolates at different mutation sites by sequencing. Phylogentic tree was also made using MEGA6 software that supported the sequencing results. ‘Msp1’ endonuclease cleaved bacterial whole genome at different cutting sites, all 10 isolates collected from different districts of Punjab cleaved into 3 to 5 fragments ranging from 600 to 10000 base pairs which showed the genetic variation among 10 isolates of P.mulocida. Availability: Items available for loan: UVAS Library [Call number: 2315-T] (1).
26.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Zingiber Officinale Rhizome Against Common Poultry Pathogens
by Ghalia Qayyum (2013-VA-779) | Dr. Aqeel Javeed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Antimicrobial compounds having plant origin inhibit bacteria through different mechanisms and can be used for the treatment of infections against resistant microbes. Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the present study is designed for antibacterial and cytotoxic evaluation of different extracts of Zingiber officinale rhizome against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale were prepared by soxhlet extraction. Antibacterial activity of these extracts was determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of inhibitions were determined by well diffusion method. MICs of plant extracts were determined by micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero cell lines
The zone of inhibitions showed by hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale against Staphylococcus. aureus were 12.33mm, 13.67mm, 16.33mm and 14mm; against Clostridium perfringens type A were 12mm, 16.33mm, 14mm and 8.33mm; against Escherichia coli were 14.33mm, 13.33mm, 14.33mm and 12mm; against Salmonella enterica were 17mm, 17.33mm and 12mm; against Haemophillus paragallinarum were 12.67mm, 13mm and 14mm respectively. Hexane extract showed no zone of inhibition against Salmonella enterica and aqueous extract was ineffective against Haemophillus paragallinarum.
MICs values of hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale rhizome against Staphylococcus. aureus were 2500µg/ml, 625µg/ml, 2500µg/ml and 2083.33µg/ml; against Clostridium perfringens type A were 2500µg/ml, 312.5µg/ml, 1250µg/ml and 5000µg/ml; against Escherichia coli were 5000µg/ml, 1250µg/ml, 5000µg/ml and 5000µg/ml; against Salmonella enterica were 312.5µg/ml, 5000µg/ml, 5000µg/ml; against Haemophillus paragallinarum were 2500µg/ml, 1458.33µg/ml and 2500µg/ml respectively. MIC was not performed against hexane extract of Salmonella enterica and aqueous extract of Haemophillus paragallinarum as no zone of inhibition observed against them. Hexane extract of Zingiber officinale rhizome was cytotoxic at concentration ≥ 750µg/ml, chloroform extract at concentration ≥ 1500µg/ml and aqueous extract at concentration ≥5000µg/ml. Ethanol extract at concentration ranging from 1500µg/ml to 2.92µg/ml was not cytotoxic to cell.
The indigenous plant Zingiber officinale have antibacterial activity against common poultry pathogens and helpful to develop new drug from plant origin.
Availability: Items available for loan: UVAS Library [Call number: 2324-T] (2).
27.
Production Of Single Cell Protein By Using Banana Peels As Substrate And Its Biological Evaluation In Broiler Chicks
by Muhammad Sheraz Yasin (2012-VA-603) | Miss Shagufta Saeed | Dr. Muhammad Tayyab | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The term single cell protein (SCP) refers to dead, dry microbial cells or total proteins extracted from pure microbial cell culture and is produced using a number of different microorganisms including bacterium, fungus and algae. It can also be called biomass, bioprotein or microbial protein.
Besides high protein content (about 60-82% of dry cell weight), SCP also contains fats, carbohydrates, nucleic acids, vitamins and minerals.
Fermentation media containing grinded banana peel as substrate was used to check the production of single cell protein for the selected Arachniotus sp. Different parameters were optimized for higher production of SCP e.g: Incubation period, pH, volume of inoculum, carbohydrate source, concentration of corn steep liquor and ionic salts concentration.
The biomass yield was estimated for total protein content by Lowrymethod. Biomass produced from fermentation was used for biological evaluation in feed trials of broiler chicks.
It is found that Arachniotus sp gave maximum single cell protein 7.49 g/L using 10 g banana peels at 72 hours incubation period. And protein concentration increased 7.58 g/L by optimizing volume of inoculum 2ml. It is observed in present study carbohydrate source also increases the protein concentration 8.41 g/L when carbohydrate source was optimized (glucose 3%).
Later on it was found that nitrogen source also enhance the protein production upto 12.61 g/L by using 2% corn steep liquor. Results also revealed that ionic salt concentration also play important role in the production of biomass protein, addition of 0.075% CaCl2.H2O produced 14.45 g/L single cell protein using above mentioned optimized conditions. 0.050 %
K2HPO4 produced 15.06 g/L. Addition of 0.050% MgSO4.7H2O produced maximum protein 15.86 g/L.
Biological evaluation in broiler chicks of this biomass protein shown there is no deleterious effects on weight gain, feed conversion ratio, protein efficiency ratio and net protein utilization. Maximum weight gain observed 215.6 grams in the group (C) in which 50% sunflower meal was replaced with biomass protein.
Feed conversion ratio in group (C) was 2.64 in which 50% sunflower meal was replaced by biomass protein and in group (B) was 2.51 in which 25% sunflower meal was replaced. And in control group (A) feed conversion ratio was 2.41.
Protein efficiency ratio was observed with non-significant value. And same results were shown by Chaves et al (1988) who reported non-significant differences among the standard and test diet when Chaetominumcellulolyticum biomass was fed to chicks. Net protein utilization observed in present study gave significant P value among the groups.
So it is concluded that single cell protein produced by this method is cheap and can be used in the food industry as food supplements and can also be included in poultry feed. The study findings suggested that microbial biomass produced by Arachniotus sp using banana peels as substrate can be replaced upto 50% of the protein supply by sunflower meal without any deleterious effects on growing broiler chicks. Moreover, it will also help in the reduction of pollution by using waste i.e. banana peel for useful purpose.
Availability: Items available for loan: UVAS Library [Call number: 2347-T] (1).
28.
Status Of Brucellosis And Its Effect On Hemogram And Serum Biochemistry In Indigenous, Cross-Bred And Exotic Dairy Cattle Herds
by Muhammad Hareem Afzal (2008-VA-250) | Dr. Muhammad Avais | Dr. Jawaria Ali Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Brucellosis mainly infects food animals such as cattle, buffalo, goats and sheep. Brucella abortus is the principal cause of brucellosis in cattle and is shed from the infected animal at or around the time of calving or abortion. The present study was conducted on 450 animals on three different strains/breeds of cattle i.e. Exotic (150), Cross-bred (150) and local cattle (150) from 10 different privately owned livestock farms of varying holdings of district Lahore. An epidemiological questionnaire focusing on herd traits as well as husbandry and sanitary practices that could be associated with the risk of Brucellosis infection was completed. Serum samples were collected and analyzed using Rose Bengal Plate Test (RBPT). The serum samples positive for Brucellosis through RBPT further subjected to Serum Agglutination Test (SAT). To check the effect of Brucellosis on hemogram, blood samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis were collected and processed for TLC, DLC, RBC, Hb, MCV, MCHC MCH and platelets using automated haematology analysed at UDL, UVAS, Lahore. Similarly, to see the effect of Brucellosis on Serum biochemistry, serum samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis collected and analysed for glucose, total protein, albumin, Creatinine, Alanine Aminotransferase (ALT), Aspartate Aminotranferase (AST) and Sorbitol Dehydrogenase (SD) using commercially available kits.
Summary
62
RBPT revealed overall prevalence 17.7% higher than SAT 10.6%. Prevalence of brucellosis is higher in Cross-Bred (22.7%) followed by local cattle (18.9%) and exotic (12%).
Hemato-boichemical results showed that increase in TLC, MCV While slight changes in Hb, MCHC, RBC and values of MCV stays within normal range. On the other hand serum biochemistry increase in AST while decrease in ALT and SD found. Availability: Items available for loan: UVAS Library [Call number: 2348-T] (1).
29.
Proteomic And Genomic Analysis Of Methicillin-Resistant Staphylococcus Aureus And Efficacy Of Indigenous Medicinal Plants Essential Oils
by Sarwat Ali Raja | Prof. Dr. Muhammad Ashraf | Dr. Tayyaba Ijaz | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: A Cohort study (prospective and observational) was performed to study the prevalence of Methicillin resistant Staphylococcus aureus from the healthy individuals of community, hospitalized patients and associated health-care workers and indigenous plants essential oils were screened as new, improved & potent antibacterial/s against resistant strains of MRSA.
The method involved isolation and identification of MRSA from surgical wounds of hospitalized patients & associated health care workers in a tertiary care hospital in Lahore and healthy volunteers from the community. Plant essentials oils & extracts were evaluated for their antibacterial activity against selected MRSA isolates. Oils were recovered by steam distillation using an all-glass distillation assembly. Then in vitro sensitivity and MICs of plant essential oils were determined using vancomycin and linezolid as commercial standards. The essential oils were screened further for the active constituents by column chromatography using various solvents and identification of compounds were performed by GC/MS analysis and the fractions which showed prompt results were evaluated for antimicrobial activity against the MRSA isolates in quest to find new therapeutic options. Finally effective essential oils and their active fractions were studied for their toxicity using in vitro Genotoxic assays such as Ames and Comet assays. To further ensure their beneficial effects antimutagenic effect of the essential oils were also studied.
Prevalence of S. aureus among patients was 52.9%, in HCWs 86.5% and in community 74% with an overall percentage of 72.6%. Among S. aureus those declared as MRSA were 91.8% from patients, 50.6% from HCWs and 59.5% from community with an overall percentage of 62.2% MRSA. Among the isolated MRSA overall 90.6% were Coagulase positive and 75.2% were biofilm positive.
SUMMARY
211
The pattern of MRSA resistance against current antibiotics have shown an overall increase in the resistance with maximum shown for lincomycin followed by tetracycline, ampicillin, fusidic acid, amoxicillin and piperacillin with tazobactam. The most effective options among current regime were tigecyclin, amikacin and meropenem showing an overall least resistance. Resistance against linezolid was observed with an overall percentage of 25.6 % and vancomycin 33.3% by disc diffusion method.
The MRSA isolates resistant to one or more groups of antibiotics were declared as MDRs. Among patients and health-care workers all were declared as MDRs where as in community 93.1% isolates were MDRs.
Upon Protein profiling using whole cell proteins 44 bands of the polypeptides were produced with molecular size 10-200kDa from the three sampling groups and were categorized into 5 clusters showing an overall significance correlation with each other explaining an interesting fact that all these strains were interlinked establishing the fact of flow of hospital acquired MRSA in the community and vice versa. This analysis also gave an insight in explaining the fact of horizontal transmission of infection within the hospital.
Keeping in view the raise in resistance among current available antibiotics indigenous medicinal plants essential oils were screened for active constituents exhibiting anti-bacterial effects against MRSA isolates.
Maximum yield was obtained from Carum copticum followed by Cuminum cyminum and minimum yield was obtained in case of Zingiber officinale. Upon qualitative analysis of all five essential oils Carum copticum essential oil showed zones of inhibition greater than the standards vancomycin and linezolid followed Cuminum cyminum and Zingiber officinale in all three
SUMMARY
212
sampling groups. Anethum sowa and Myristica fragrans essential oils showed no activity against MRSA.
Minimum inhibitory concentration of the three essential oils determined by micro broth dilution method indicated that Carum copticum showed least value in all three types of MRSA isolates followed by Zingiber officinale and Cuminum cyminum.
Effective essential oils were further fractioned using silica gel gravity columns. All the fractions obtained were screened for the anti-bacterial activity against all three types of MRSA isolates. Only fraction F1 of Carum copticum showed activity greater than pure essential oil and the two commercial standards of vancomycin and linezolid.
For the identification of active constituents GC/MS analysis was performed on all three essential oils and their respective fractions. In case of fraction F1 the most dominant constituents were Carvacrol, p-Cymene, Ʈ-Terpinene and Apiol. In other two plants none of the fractions were effective. Therefore it was concluded to use pure essential oils in case of Zingiber officinale and Cuminum cyminum rather than their individual fractions and incase of Carum copticum Fraction F1 has shown superior activity.
Finally these essential oils were tested for possible mutagenic effect using bacterial reversion mutation assay and Comet assay. No mutagenic effects were observed at MIC and above doses. These effective essential oils were also evaluated for possible antimutagenic effect. Both Carum copticum and Zingiber officinale essential oils showed strong antimutagenic effects and weak antimutagenic effect by Cuminum cyminum. Upon analysis of nuclear damage none of the plants essential oils and fraction F1 of Carum copticum showed genotoxic effects and indicated to be safe. Thus from the study it was concluded that Carum copticum essential oil and its fraction F1 were the most effective to be further investigated as an alternative treatment for MRSA infections. Availability: Items available for loan: UVAS Library [Call number: 2410-T] (1).
30.
Sero-Screening Of Camels For Different Infectious Diseases
by Mazia Khalid (2008-VA-358) | Dr. Aamir Ghafoor | Prof. Dr. Aftab Ahmed Anjum | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Camel is the precious and important animal in Pakistan. Camel is the most well adapted livestock species, survives and produces in climatic extremes and is well appreciated for its significance in the pastoral economy of the province. The camel being an important livestock species uniquely adapted to hot and arid environments and therefore contributes significantly to the food security of the nomadic pastoral households. Although camel being hardiest animal is less susceptible to diseases as compared to other livestock animals but literature shows that some diseases are still prevalent in camels. In view of the significance of camel as livestock animal as well as the symbol of cultural heritage of the nomadic pastoralists, there is a need to combat different diseases to which camels are susceptible and then appropriate control strategies should be applied.
Present study was designed to check the percentage positivity of different major diseases in camels that may pose serious issue relating to camel health and its importance as an important livestock animal. The diseases included in this study are Q fever, Brucellosis, FMD, CBPP and Neosporosis. ELISA is used to detect antibody prevalence by using specific kit based protocol for each disease whereas in case of Brucellosis RBPT is also used as basic screening test.
And it was found that Q fever has highest percentage seropositivity in both districts as compared to other diseases whose presence in camels was found to be almost seronegative.
So it was concluded that camel is still resistant to many diseases though some diseases are still prevalent in camels and these diseases should be controlled through public awareness and routine screening. Availability: Items available for loan: UVAS Library [Call number: 2401-T] (1).
31.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Eucalyptus Globulus Leaves Against Common Poultry Pathogens
by Asma Iqbal (2013-VA-563) | Dr. Muhammad Adil Rasheed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Phytomedicines mark the major component of health care as natural medicines have always provided the strong foothold for the discovery and manufacturing of synthetic drugs. So plants are a rich source of bioactive compounds having many therapeutic activities and majority of them are still untapped. Eucalyptus globulus is a medicinal plant known for its value to cure asthma, respiratory infections, cough and allergic reactions. The antimicrobial activity, insecticidal and hypoglycemic activity have also been credited to the plant. Most of the studies have been conducted on the essential oils of Eucalyptus globulus and little work has been reported on extracts. Whereas, sequential extracts has not been employed yet.
Hexane, chloroform and ethanol, aqueous extracts were prepared by the sequential extraction on Soxhlet apparatus and antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli, Salmonella enterica, Clostridium perfringens type A and Haemophilus paragallinarum by agar well diffusion and micro broth dilution method. The zones of inhibition and minimum inhibitory concentration were determined. The extracts showing antimicrobial activity were further evaluated for cytotoxicity by using MTT assay on Vero cell line. The cell culture media was prepared and cell lines were propagated to form monolayer then monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated.
The statistical analysis was conducted with help of Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA) and results were compared using one way ANOVA.
Summary
89
The zones of inhibitions showed by hexane, chloroform, ethanol and aqueous leaf extracts of Eucalyptus globulus against Staphylococcus aureus were 0.0, 19.3, 20.3 and 23.3mm; against Clostridium perfringens type A were 14, 22.3, 14.0 and 15.3mm; against Escherichia coli were 0.0, 12.6, 13.3 and 15.6mm; against Salmonella enterica were 10, 12.3, 18.6 and 21mm; against Haemophilus paragallinarum were 0.0, 8.6, 14 and 18mm respectively. Hexane extract showed no zone of inhibition against Staphylococcus aureus, Escherichia coli and Haemophilus paragallinarum.
The MICs values of hexane, chloroform, ethanol and aqueous leaf extracts of Eucalyptus globulus against Staphylococcus aureus were 0.00, 104.1, 32.55 and 312.5 μg/ml; against Clostridium perfringens type A were 52.08, 39.06, 16.27 and 312.5 μg/ml; against Escherichia coli were 0.00, 78.12, 260.4 and 625.0 μg/ml; against Salmonella enterica were 13.02, 104.1, 130.2 and 416.6 μg/ml; against Haemophillus paragallinarum were 0.00, 104.1, 260.4 and 416.6 μg/ml respectively. MIC was not performed against Staphylococcus aureus, Escherichia coli and Haemophilus paragallinarum for hexane extract as no zone of inhibition was observed against them.
Hexane extract of Eucalyptus globulus was cytotoxic at concentration ≥ 312.5μg/ml, chloroform extract at concentration ≥ 375μg/ml, ethanol extract at concentration ≥ 625μg/ml and aqueous extract was cytotoxic at concentration ≥312.5 μg/ml.
The indigenous plant Eucalyptus globulus has antibacterial activity against common poultry pathogens and can be helpful for development of new drugs of plant origin. Availability: Items available for loan: UVAS Library [Call number: 2429-T] (1).
32.
Effect Of Bio-Stimulation On Estrus Expression And Pregnancy Rate In Cidr Based Synchronization Protocol In Nili-Ravi Buffalo
by Abdul Waheed (2009-VA-133) | Dr. Aijaz ali Channa | Dr. Syed Murtaza Hassan Andrabi | Prof. Dr. Mian Abdul Sattar | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Our water buffalo (Bubalus bubalis) has much potential for production of milk. But this
animal has some problems regarding reproduction including delayed puberty, poor estrus behavior,
silent heat, long postpartum period and low conception rate by artificial insemination. This leads
to poor reproduction and hence great economic loss. Therefore, the requirement is to address these
problems efficiently and formulate more effective techniques for improvements. Researchers have
devised many estrus synchronization protocols (PGF2α, P4, GnRH, eCG, hCG etc.) that help
bringing many animals in heat and hence improve the reproductive performance when fixed time
artificial insemination is combined with them. But these protocols give inconsistent results when
they are applied on buffaloes making it necessary to improve the techniques.
This study was planned on the hypothesis that presence of bull (bio-stimulation), at the
time of synchronization, may play an important role in enhancement of estrus intensity and fertility
rate in Nili-Ravi buffaloes. Seventy one adult buffaloes were randomly selected from different
areas of field conditions and LRS (NARC) and subjected to CIDR based heat synchronization in
combination of either bio-stimulation or non-stimulation. The animals were observed for
behavioral estrus signs twice a day starting after 12 hours of CIDR removal till 96 hours.
Pregnancy diagnosis was done by rectal palpation 60 days post CIDR removal. Estrus response
and pregnancy rate were analyzed by Chi-square test using MINITAB version 15. Estrus signs and
total estrus intensity were compared by Mann Whitney U test. Difference was considered
significant at probability level of (P < 0.05).
In peri-urban areas, more animals from bio-stimulated group showed better behavioral
estrus signs, more total intensity score and significantly higher pregnancy rate as compared to nonSUMMARY
63
stimulated group of animals. At LRS (NARC), more animals from non-stimulated group were
found in behavioral estrus but intensity of heat signs was high in bio-stimulated animals.
Pregnancy rate was also higher in non-stimulated animals but the difference was not significant.
Overall, in this study, we got higher pregnancy rate in bio-stimulated animals than non-stimulated
group which indicates a positive response of bull stimulation on reproductive performance of Nili-
Ravi buffaloes who were synchronized with CIDR based estrus synchronization protocol. Availability: Items available for loan: UVAS Library [Call number: 2469-T] (1).
33.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Astragalus Membranaceus Roots
by Sadia Alvi (2013-VA-595) | Dr. Aqeel Javeed | Dr. Muhammad Ovais Omer | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The present study was designed to evaluate antibacterial and cytotoxic evaluation of different extracts of Astragalus membranaceus root against common poultry pathogens. Sequential extraction with hexane, ethanol, chloroform and aqueous solvents was prepared and antibacterial activity was evaluated by using agar well diffusion. Minimum inhibitory concentration (MIC) of plant extracts was evaluated by micro broth dilution test. The extracts exhibiting antimicrobial activity were further evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell culture media was prepared and cell lines were propagated, monolayer was formed. This 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 of antibacterial activity and MTT assay were compared using DMR posthoc test. Growth of Clostridium perfringens, Escherichia coli, Haemophilus species, Salmonella enterica and Staphylococcus aureus inhibited by all extracts of Astragalus except aqueous extract which shows no zones of inhibition against C. perfringes. MIC values were higher for aqueous extract against all selected bacteria and lowest for chloroform against E. coli, S. enterica and Staph. aureus (208.3ug/ml, 156.25ug/ml, 78.125ug/ml respectively) for hexane against Haemophilus species (833.3ug/ml) and for all three extracts against C.perfringes (1250ug/ml). Hexane, chloroform and ethanol extracts were appeared to be safe at all concentrations except ≥ 2000μg/ml, ≥1000μg/ml and ≥3000μg/ml respectively while aqueous extracts showed cytotoxicity at concentrations ≥625μg/ml. Astragalus membranaceus
SUMMARY
104
showed antibacterial activity against all selected pathogens. Chloroform and hexane extracts showed greater antibacterial activity than ethanol and aqueous. Cytotoxicity values for chloroform extract are safer than rest of three extracts. Astragalus membranaceus may 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: 2444-T] (1).
34.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Ocimum Basilicum Leaves Against Common Poultry Pathogens
by Shomaila Naz (2013-VA-1001) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Antimicrobial compounds having plant origin inhibit bacteria through different mechanisms and can be used for the treatment of infections against resistant microbes. Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the present study is designed for antibacterial and cytotoxic evaluation of different extracts of Ocimum basilicum seeds against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Ocimum basilicum leaves and seeds were prepared by soxhlet extraction. Antibacterial activity of these extracts was determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of inhibitions were determined by well diffusion method. MICs of plant extracts were determined by micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero cell lines. All the results were statistically analyzed by one way ANOVA and compared means by Duncan’s multiple range of posthoc test at significance level of P≤0.05.
The results of zone of inhibitions showed by Ocimum basilicum leaves and seeds extracts ranging from 11.33-20.0 mm values of MIC results ranging from 4.889 μg/ml-2500 μg/ml of hexane, chloroform and ethanol. The aqueous extract of Ocimum basilicum have no activity against any bacterial pathogen. Ethanol extract of Ocimum basilicum leaves was cytotoxic at 500 μg/ml. Hexane extract of Ocimum basilicum seeds was cytotoxic at concentration ≥625 μg/ml, chloroform at concentration ≥19.53 μg/ml and ethanol extract at concentration ≥750 μg/ml.
The indigenous plant Ocimum basilicum have antibacterial activity against common poultry pathogens and helpful to develop new drug from plant origin. Availability: Items available for loan: UVAS Library [Call number: 2443-T] (1).
35.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Glycyrrhiza Glabra (Liquorice) Roots Against Common Poultry Pathogens
by Javaria Arooj (2013-VA-596) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the
present study is designed for antibacterial and cytotoxic evaluation of different extracts of
Glycyrrhiza glabra Linn. roots against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Glycyrrhiza
glabra Linn. roots were prepared by soxhlet extraction. Antibacterial activity of these extracts was
determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens
type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of
inhibitions were determined by well diffusion method. MICs of plant extracts were determined by
micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero
cell lines.
The zone of inhibitions showed by hexane, chloroform and ethanolic extracts of
Glycyrrhiza glabra Linn. roots against Staphylococcus. aureus were10.3mm, 13.0mm, 11.6mm;
against Clostridium perfringens type A were20.0mm, 17.3mm, 17.3mm; against Escherichia coli
were11.6mm, 19.3mm, 16.0mm; against Salmonella enterica were13.6mm, 14.0mm,14.0mm;
against Haemophillus paragallinarum were13.0mm, 15.0mm, 17.0mm respectively. Aqueous
extract showed no zone of inhibition against any test bacteria.
MICs values of hexane, chloroform and ethanolic extracts of Glycyrrhiza glabra Linn.
roots against Staphylococcu aureus were 13.0μg/ml, 312.5μg/ml and 104.1μg/ml; against
Clostridium perfringens type A were 9.766μg/ml, 71.61μg/ml and 520.8μg/ml; against
Escherichia coli were 65.1μg/ml, 52.8μg/ml and 156.25μg/ml; against Salmonella enterica were
Summary
86
19.5μg/ml, 130.2μg/ml and 78.12μg/ml; against Haemophillus paragallinarum were 91.1μg/ml,
29.2μg/ml and 130.2μg/ml respectively. Aqueous extract showed no MIC value as no zone of
inhibitions wereobserved against them. Hexane extract of Glycyrrhiza glabra Linn. roots was
cytotoxic at concentration ≥ 650μg/ml, chloroform extract at concentration ≥ 2500μg/ml and
ethanolic extract was not cytotoxic to cell.
The indigenous plant Glycyrrhiza glabra Linn. roots have antibacterial activity against
common poultry pathogens and helpful to develop new drug from plant origin. Availability: Items available for loan: UVAS Library [Call number: 2442-T] (1).
36.
Evaluation Of Microbiological Quality Ofshawarma Sold By The Street Vendors Of Lahore
by Sana Adrees (2013-VA-896) | Dr. Zabair Farooq | Dr. Muhammad Nasir | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Food safety is a scientific discipline describes handling, preparation and storage of food that prevents food borne diseases.It includes a number of ways that must be followed to avoid potentially severe health hazards.The tracks within this line of thought are safety between the market and consumer. With the fast pace of life the consumption of fast food is going to increase day by day in Lahore. It is imperative to prepare food at good hygienic conditions because of its perishability. The usual thougt is that food should be safe in the market and the concern is safe delivery and preparation of food for the consumer.
Microorganisms affecting food comes from natural micro flora or are introduced by manufacturing steps ranging from processing storage and distribution. In some cases these micro flora have no effect on the food and can be consumed without consequence, but those that are introduced during course of processing depending on type and level of contamination can spoil the food and cause food borne illnesses.
Food can transmit diseases from one person to another as well as serve as growth medium for bacteria that can cause food poisonoing. In the developed countries there are standards for food, whereas in less developed countries the main issue is the availability of safe water supply which is one of the critical item.
The present research work was undertaken to investigate the microbial count in shawarma which is a vended food. Samples collected from Anarkali,Wahdat road and Islampura showed high microbial count which shows contamination due to poor handling and hygienic conditions and improper storage conditionswhile the samples taken from Fortress, Gulberg having less microbial count as compared to these areas.
High microbial load in street foods occur due to improper food handling, unhygienic food preparation and processing, cooking and storage at inappropriate temperature. Consumer, food vendors and all type people should be aware of food hygiene, public health, implications of consuming contaminated foods, causative diseases. Food safety rules and implementation of food regulatory laws in food preparation, serving and preservation should be strongly maintained to avoid contamination problems and food-borne diseases.
Following points should be strongly observed and maintained for future work and further investigative study to improve the quality of shawarama.
1. Ensuring regular inspection and periodical check on these food preparation procedures.
2.Continuous lab test and analytical lab analysis to check the unwanted presence of any new
harmful agents in these foods to ensure food safety for consumers.
Availability: Items available for loan: UVAS Library [Call number: 2455-T] (1).
37.
Evaluation Of Microbial Quality Of Burger Sold By Roadside Fast Food Centers In Various Regions Of Lahore
by Nimra Khalid (2013-VA-900) | Dr. Zubair Farooq | Dr. Muhammad Nasir | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Food safety actions need to cover the entire food chain, from production to consumption. The implementation of food safety include safety between industry and the market and then between the market and the consumer. In considering industry to market practices, food safety considerations include the origins of food including the practices relating to food labeling, food hygiene, food additives and pesticides residues, as well as policies for certification systems for foods.
Food industry regarding safety issue is one of the most unseen areas of policy in developing countries, especially in Pakistan. As a result, food systems in these countries are not always as well controlled and comprehensive as in the mechanized world. This situation is exacerbated by an ever increasing population, rapid urbanization and, most importantly, a lack of the economic and practical resources needed to hold a sound food safety system. As a result, people in developing countries are continuously facing a wide range of potential food safety risks.
Food borne illness is a disease caused by consuming contaminated food or drink.There are more than 250 known food borne diseases. Food can transmit disease from individual to individual as well as serve as a growth medium for bacteria that can cause food poisoning.
Today, people try to save as much time as possible due to the requisite to cope with many tasks within restricted time. In many cases people who have busy schedule don’t have enough time to cook food therefore prefer to eat fast food like burger, shawarma, fries etc. from fast food centers.
The present research was undertaken to investigate the microbial load of the chicken burger samples (n=96) collected aseptically from different regions of Lahore. Samples were collected in sterile plastic bags and analyzed for microbial load immediately after collection in Microbiology lab of University of Veterinary and Animal Sciences.
Nutrient agar was used to perform Total Plate Count, Mackonkey Agar was used for total coliform count, Manitol salt agar was used to count Staphylococcus Aureus and SalmonellaShigella agar was used to detect Salmonella.
The findings of the current study showed that samples taken multinational restaurant have less contamination than from local fast food centers. While among local fast food centers Liberty, Fortress, Anarkali and Akbarimandi have lesser microbial count than samples collected from Ichra, moon market, Mori gate. The difference in microbial load is due to improper food handling, processing, cooking and storage temperature, time and manufacturing practices of the food handlers.
After conducting my research I concluded that following points should be strongly observed and maintained to improve the quality of chicken burger in local fast food centers
• Ensuring regular inspection and periodical check on these food centers
• Continuous lab test and analytical lab analysis to check the unwanted presence of any new harmful agents in these foods to ensure food safety for consumer.
Availability: Items available for loan: UVAS Library [Call number: 2456-T] (1).
38.
Characterization And Thermostability Of Phytase Produced By Indigenous Aspergillus Niger Isolates
by Madeeha Tariq (2010-VA-293) | Prof. Dr. Aftab Ahmad Anjum | Dr. Jawad Nazir | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Phytase enzyme now becomes more important commercially. Presence of phytate in food
and feed make them less nutritive due to phytate complexes mainly with mineral ions and
proteins. Phytase in monogastric animals and human stomach either produced in small amount or
not. This leads to phosphorous Pi deficiency. Supplementation of food and feed with phytase
enzyme full fill this deficiency through degradation of phytate complexes and release of Pi.
Degradation of phytate complexes makes phosphorous other mineral ions and amino acids
available for growth and development. It was proved that feed conversion rate in poultry
increased due to supplementation of phytase in poultry feed. Feed of monogastric animals mostly
at industrial level pelleted to give it a shape or to kill microorganisms (sterility). At industrial
level enzyme production and processing cost about 2 billion. So this demands a thermostable
phytase to use at industrial level or its cost effective production.
Aspergillus niger have been used industrially for production of beneficial enzymes. A.
niger isolates procured from department of microbiology were confirmed through macro and
microscopic characteristics as A. niger. These isolate were screened for phytase production on
phytase screening medium PSM agar. Positive isolates identified through noval staining using
2% cobalt chloride, 6.25% ammonium molybdate and 0.42% ammonium vanadate for contrast.
Positive isolates next proceeded for phytase enzyme production in broth media (pH 5.6) using
0.5% sodium phytate as substrate. Incubation was done at 30oC for 5-7 days in shaking incubator
150rpm. After production quantification of enzyme was carried out through enzyme activity
assay. There maximum (274.99±10.14 FTU/ml) and minimum (68.88±2.55 FTU/mL) activity of
phytases from isolate PASN01 and PASN08 was observed. Phytases characterized through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) to know protein
molecular weights. Highest molecular weight 107.82kDa was PASN06 and lowest was 35.21kDa
of PASN 01.
Aspergillus niger spores subjected to steam heat treatment at 30oC, 45oC, 60oC, 75oC and
90oC for 15, 30, 45, 60minutes to identify thermostability. At 30oC and 45oC temperature, spores
of A. niger isolates found to be thermostable. But at 60oC, 75oC, or 90oC treatment spores
become inactivated or there 6.0 logarithmic reduction in spore count was observed.
Thermostability of phytases was found at 60oC, 75oC, 90oC for 15, 30, 45, and 60
minutes treatments. Enzyme from A. niger PASN01 and PASN08 observed as thermostable at
60, 75 and 90oC. Phytases from PASN01 and PASN08 showed 160.55±42.96 and 00±.00
FTU/mL decreased in activity after 45 minutes of treatment at 60oC temperature, respectively.
PASN01 phytase displayed 163.88±23.35, 172.77±7.52 and 171.66±7.26 FTU/mL decreased in
activity after 60minutes treatment at 60, 75 and 90oC. In case of PASN08 phytase at 60, 75 and
90oC temperature after 60minutes treatment, 13.33±10.41, 16.66±6.00 and 23.88±41.37 FTU/mL
decreased in activities were observed, respectively. PASN08 phytase observed more
thermostable than other phytases of A. niger isolates. Enzyme can bear pelleting and pre
pelleting temperatures. Enzyme from PASN08 also observed stable during storage at room
temperature.
Conclusion:
A. niger PASN08 spores inactivated or killed and phytase observed stable at 60oC
temperature, after 60mins treatment. Temperature 60oC may be used industrially for cost
effective thermostable phytase production from indigenous A. niger isolate PASN08. Availability: Items available for loan: UVAS Library [Call number: 2475-T] (1).
39.
Isolation And Antibiotic Resistance Profiling Of Enterococcus Faecium Recovered From Retail Fish In Lahore City
by Maria Butt (2010-Va-281) | Dr. Ali Ahmad Sheikh | Prof. Dr. Aftab Ahmad Anjum | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Enterococcus faecium is an enteric, gram positive and lactic acid bacteria which belongs to genus enterococcus and inhabit the intestinal tract of human, fish and other warm blooded animals. Due to irrational use of antibiotics in human and veterinary sector, antibiotic resistance has been developed in commensal bacteria including Enterococcal species. These resistant bacteria are released in environment through human and animal waste and transfer resistant genes to susceptible bacteria present in wetlands making them antibiotic resistant. E. faecium is considered to be involved in transmission of resistance genes, present on mobile genetic elements through conjugation to other bacteria. The resistant bacteria can be transferred to human through food chain. The present study was designed to evaluate the prevalence of E. faecium recovered from retail fish samples collected from various areas of Lahore city. Antibiotic resistance profiling of the isolates against commonly used antibiotics was also determined.
In current study 65 fish samples (intestinal swabs) were processed for isolation of E. faecium through standard culturing and biochemical reactions. Out of 65 swab samples, 30 samples (47.69%) were found positive for Enterococcus faecium. Antibiotics resistance profiling showed that the isolates were resistant to antibiotics mentioned as below: Ampicillin (100%) > erythromycin (56.6%) > rifampicin (53.3%) > Chloramphenicol (30%), ciprofloxacin (30%) > tetracycline (20%), vancomycin (20%) > Teicoplanin (13.3%) > Doxycyclin (6.6%) > Fosfomycin (0%). E. faecium isolates showed resistant to at least 2 or 3 antibiotics of different group. In conclusion it is observed that retail fish is the carrier of antibiotic resistant Enterococcus faecium and
Summary
51
could transfer resistant genes to wetlands and other aquaculture from where it could be transferred to human body. Efforts should be made to use antibiotics wisely and hygienic practices should be followed during slaughtering and processing of fish meat to avoid bacterial spread from animal source to human beings. Availability: Items available for loan: UVAS Library [Call number: 2493-T] (1).
40.
Antibiotic Resistance Pattern Of Staphlococcus Aureus And Its Resistance Modulation Using Medicinal Plant Extracts
by Iqra Asif (2010-VA-279) | Prof. Dr. Aftab Ahmed Anjum | Prof. Dr. Khushi Muhammad | Ms. Tehreem Hussain.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This project was designed to evaluate the antimicrobial efficacy of Chloroform and
ethanol extracts of Calotropis procera (C .procera) and Eucalyptus globulus (E. globulus)
against Multiple Drug Resistant (MDR) Staphylococcus aureus isolated from human origin. This
study was conducted to evaluate the antimicrobial potential of C. procera and E. globulus alone
and in combination with antibiotics to check synergism between medicinal plants and resistant
antibiotics.
S.aureus is a major pathogen which causes various infections. Infectious diseases affect
millions of people around the world and in the history these diseases are major cause of mortality
and morbidity across the globe. In past few decades rate of mortalities are continuously
increasing because of acquired resistance of S. aureus against multiple drugs, thus it is utter need
of time to discover some alternatives to antibiotics so that we can resolve this dilemma of
antibiotic resistance. Plant extracts are hope for this purpose as they have many compounds
which have potential to lower down the number of micro-organisms. Plants have benefits over
other as they are non toxic, non-reactive and have least side effects.
Total 20 samples of human origins were procured from Department of Microbiology,
UVAS Lahore and were subjected to check their antibiotic resistance profile against
Erythromycin, Amoxicillin and Ciprofloxacin by Kirby Bauer disc diffusion assay. Out of 20,
nine resistant isolates were separated. Among them three were resistant to Erythromycin, three to
Amoxicillin and three to Ciprofloxacin.
Summary
72
Leaves of C. procera and E. globulus were processed in Chloroform and ethanol
Solvents. Antimicrobial activity was evaluated by agar gel well diffusion assay in which zone of
inhibitions were measured. Minimum inhibitory concentration (MIC) of plant extracts was
evaluated by micro broth dilution method. Best antimicrobial activity was observed by ethanolic
extract of E. globulus.
Then combine effect of sub-inhibitory concentrations (SICs) of plant extracts and
minimum inhibitory concentration of antibiotics were determined by Well Diffusion assay. Four
different sub inhibitory concentrations of plant extracts i.e.10μg/ml, 20μg/ml, 40μg/ml and
80μg/ml were used in combination with fixed concentration of antibiotics i.e.100μg/ml to check
combinational effect of both. At selected sub-inhibitory concentration plant extract alone did not
show any antibacterial activity. Two of the isolates had shown modulation when amoxicillin and
plant extracts combination was used against them. The isolate labeled as S.aureus 4 showed
modulation with the use of Ethanolic extract of Calotropis procera and S.aureus 5 had shown
modulation with the use of chloroform extract of Calotropis procera. For further confirmation
two more concentrations of 160μg/ml and 320μg/ml were used along with100μg/ml Amoxicillin
against same isolates S.aureus 4 and 5. Zone of inhibition was observed with increased diameter
indicating modulation of two isolates. While erythromycin and ciprofloxacin resistant isolates
didn’t show any modulation.
Data of antibiotic resistance and resistance modulation using plant extracts was analyzed
by one way analysis of variance (ANOVA) followed by Duncan’s multiple range(DMR)
posthoc test using Statistical package for social sciences (SPSS) 17.0 Statistical software at α <
0.05. Availability: Items available for loan: UVAS Library [Call number: 2501-T] (1).
41.
Evaluation Of Antibacterial Effect Of Gymnema Sylvestre Species Cultivated In Pakistan
by Muhammad Tahir (2011-VA-339) | Dr. Muhammad Adil Rasheed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This study was conducted to determine the use of medicinal plants as an antibacterial agent and its potential to as an alternative medicine against bacterial infections. For this purpose Sequential extracts (i.e. Hexane, Chloroform, Ethanol and Aqueous) of Gymnema sylvestre R.Br. were tested against S. aureus, E. coli, S. enterica, C. perfringens type-A, H. paragallinarum. Of each bacterium 3 isolates were tested by using well diffusion method. The results were obtained by determining the ZOI by well diffusion method and MIC by using 96 well ELISA plate.
The mean ZOI and mean MIC values of G. sylvestre leaves extracts showed that chloroform and ethanolic extracts have more antibacterial activity against all five microorganisms. Only chloroform and ethanolic extracts showed antibacterial activity against all 5 microorganisms while hexane extract showed antibacterial activity against S. enterica, S. aureus, H. paragallinarum, C. perfringens type- A but no activity was observed against E. coli. On the other hand aqueous extract have showed antibacterial activity only against C. perfringens type-A but no antibacterial activity against remaining four bacteria under study. While analyzing results based upon MIC, the chloroform extract has more antibacterial effect when compared with hexane. Hexane extract was more potent than aqueous extract whereas ethanolic extract was the least potent.
When overall antibacterial effect of all the extracts was evaluated against all bacterial strains, it was observed that C. perfringens type-A was the bacterium most vulnerable to antibacterial activity of sequential extracts of dried leaves of G. sylvestre as it responded
Summary
94
to all four sequential extracts and gave maximum zone of inhibition (10-22mm range) while no other bacteria showed such bigger zone of inhibition.
On the basis of MIC, it can be assumed that chloroform extracts have more antibacterial components as compared to hexane extract. Hexane extracts have more antibacterial components as compared to ethanolic extracts. The activity of aqueous extracts is negligible as it showed response against only one bacterium.
MTT assay was performed on supersaturated solutions of sequential extracts of dried leaves of G. sylvestre. Results revealed that small concentrations of these extracts are not toxic. Cell survival percentage (CSP) values below 50% were given at concentrations of 5800μg/ml (38.76%), 7225μg/ml (43.71%), 8150μg/ml (44.90%) and 3125μg/ml (41.84%) by hexane, chloroform, ethanolic and aqueous extracts respectively. Finally, on the basis of MIC and CSP for all of four sequential extracts, it is concluded that chloroformic extract is the most active and safe extract against all of 5 experimental bacteria, while hexane extract is safe against only C. perfringens type-A and ethanolic and aqueous extracts are cytotoxic on their MIC values for all the experimental bacteria. Statistical analysis showed that ZOI and MIC values were significantly different between the groups while within the same group they were non-significant.
Finally it can be concluded that the leaves of plant Gymnema sylvestre R.Br. cultivated in Pakistan has considerable antibacterial activity and considerable safety profile so it must be further studied, characterized, purified and chemically isolated so that may be converted to proper dosage form and this miracle plant may be used therapeutically to cure various ailments including bacterial infections especially poultry infections. Availability: Items available for loan: UVAS Library [Call number: 2503-T] (1).
42.
Amelioration Of Pathological Effects Of New Castle Disease By Aloe Vera
by Sayyed Raza Ali Shahid (2014-VA-515) | Prof. Dr. Zafar Iqbal Chaudhry | Dr. Gulbeena Saleem | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Poultry industry has many threats from the infectious diseases. Newcastle disease is the most important disease of the poultry as it is distributed all over the world and it can cause huge economic losses in poultry industry. It is caused by the Newcastle disease virus (NDV) that can infect about 240 species of birds. Newcastle disease cause immune suppression in birds. It is reported that supplementations of Aloe vera enhances the immune status and reduce inflammation. So this research project was design to observe the effect of Aloe vera on lymphoid organs, growth performance and antibody response in Newcastle disease challenge birds. For this research a total of 120 broiler chicks were divided into four groups A, B, C, and D. Group A was control group while B and C were treated with 2 percent aqueous extract of Aloe vera. Group C was also vaccinated against New Castle disease. Aloe vera was given to group B and C from day one to end of study trial. Both of the groups were challenged with ND virus at day 21. Group D was vaccinated against ND and was challenged with ND virus at day 21 without supplementation of Aloe vera. Blood samples were collected at day 1, 7, 20, 24, 26 and 28 to determine the antibody titer against ND. Highest antibody titers were observed in group C as compared to all other groups which was vaccinated against ND along with supplemented with Aloe vera. For gross pathology and histopathology, lymphoid organs were collected at day 24, 26 and 28..The average feed intake of group A and D was significantly higher than group B and C before challenge of virus but the body weight gain of 2% Aloe vera supplemented broiler was significantly (p<.05) higher than without treatment of Aloe vera. The FCR of birds supplemented with Aloe vera treatment was significantly different from the birds without Aloe vera treatment. The FCR value of group C and B was higher than A and D.
lxxv
A significant difference was observed in the weight of lymphoid organs of birds treated with Aloe vera as weight of organs was less in group C followed by group B, group A and group D. This was due to anti-inflammatory effects of Aloe vera. Microscopic examination revealed congestion, depletion of lymphocytes, dysplasia of thymic lobules, thinning of cortex, focal necrosis, disappearance of lymph follicles and inter-follicular edema like lesions within lymphoid organ of the groups challenged with Newcastle disease virus. However, cellular hypertrophy and decreased lymphocytes population were prominent changes in lymphoid organs of broiler treated with 2% Aloe vera. To check the virucidal effects of Aloe vera, a separate experiment was conducted in which 9 day old embryonated eggs were inoculated with ND virus along with 2 percent Aloe vera gel extract after incubation at 37Cº for an hour in group A while only ND virus was inoculated in group B. Candling was performed to see the survival of embryos in both groups which revealed a significant difference i.e. 16 percent embryos were found dead in group A while 80 percent was found dead in group B. Later on the amnioallantoic fluid of the eggs was used for spot Haemagglutination test. Group A showed less agglutination activity then group B. From this study it was concluded that Newcastle disease caused immune suppression and damage of vital organs in broiler while Aloe vera have immunomodulatory, anti-inflammatory and antiviral effects as it raised antibody titer against Newcastle disease virus and lowered the inflammatory processes along with inactivation of ND virus. It also promotes growth performance of broilers and helps the birds to survive against lethal ND disease. Availability: Items available for loan: UVAS Library [Call number: 2525-T] (1).
43.
Pathogenesis Of Field Isolates Of Mannheimia Hemolytica In Experimentally Infected Rabbits
by Syeda Fakhra Waheed (2014-VA-10) | Prof. Dr. Zafar Iqbal Chaudhry | Prof. Dr. Asim Aslam | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Shipping fever is one of the most economically important infectious diseases of ruminants with a wide prevalence throughout the continents. The disease is characterized by an acute febrile course with severe fibrinous bronchopneumonia. Infected animals may die within a few days of the onset of clinical signs, but those which survive the acute attack may become chronically infected. Both Mannheimia and Pasteurella species are commensally resident in the respiratory tract of healthy ruminants and are capable of causing infection in animals with compromised pulmonary defense system. Bovine respiratory disease (BRD) is the most common and costly problem encountered in stocker or feedlot calves. BRD also called “shipping fever”, accounts for major economic losses to the producer by reducing average daily gain, feed efficiency, and overall performance of beef calves. The aim of present study was isolation of M.haemolytica from cattle. The identification of organism was performed through biochemical tests and confirmation by polymerase chain reaction. The nature of disease was evaluated through gross and microscopic lesions.
A total of 50 tissue samples (25 lungs and 25 pharynx) were collected from Punjab Agriculture and Meat company Lahore and brought to the Department of Pathology UVAS, Lahore and were analyzed for biochemical and molecular detection of M .haemolytica. For studying the pathogenesis of the disease, experimental infection was given to rabbits in Department of Pathology, UVAS Lahore. Rabbits were randomly divided into Group A, Group B and Group C with nine rabbits (n=9) in each group. Experimental infection of field isolated M. hemolytica was given intratrachealy to the rabbits. Rabbits of group A and B were infected with 0.5 mL bacterial inoculum having 103 and 106 CFU/mL respectively. The rabbits of Group C served as control group. Rectal temperature of each rabbit was recorded daily. On postmortem,
CHAPTER 6
SUMMARY
Summary
67
gross and microscopic lesions were recorded.
The results showed that rabbits of control group not showed any gross or microscopic change. There was significant increase in rectal temperature of infected rabbits as compared to uninfected rabbits. The gross lesions were specific for the organism which was prominently observed in lungs of rabbits. The microscopic lesions revealed that there was severe consolidation, congestion and fibrin exudation in lungs of rabbits of group A which were given less number of organism and they developed clear signs of disease. The rabbits of Group B showed less prominent signs compared to group A due to early death of rabbits. There were multiple hemorrhages, of varying sizes and hyalinization of myocardial cells in infected rabbits. The severity of changes was significantly more different in Group A, as compared to Group B.
It can be deduced by this study that the rabbit can be used as a model for further studies exploring the pathogenesis of the disease as the lesions resemble to shipping fever caused by M. hemolytica in ruminants. The lesions, which developed, could be descending infection resulting in typical lesions of bronchopneumonia or lobular pneumonia. Availability: Items available for loan: UVAS Library [Call number: 2517-T] (1).
44.
Antibacterial Activity Of Plant Extracts Against Antibiotic Resistant Pseudomonas Aeruginosa And Their Cytotoxicity Profile
by Hafiza Farah Asghar (2010-VA-276) | Prof. Dr. Aftab Ahmed Anjum | Dr. Ali Ahmad Sheikh | Muhammad Nasir.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2016Dissertation note: Pseudomonas aeruginosa is a common opportunistic pathogen of respiratory tract
and cause both hospital and community acquired infections. For the treatment of
infections antibiotics are used but due to random selection of commonly used antibiotics,
resistance in P. aeruginosa has developed. This problem may leads to the high morbidity
and mortality rate. Different medicinal plants have antibacterial activity in their
secondary metabolite. Secondary metabolites are terpens, flavonoids, alkaloids and
alcohols etc. So the plant extract could be the alternative therapy for the treatment to
reduce the antibiotic resistance problem.
Isolates of P. aeruginosa was procured from the main clinical laboratory of Mayo
Hospital, Lahore and identified biochemically according to bergey’s manual of
determinative bacteriology. Antibiotic resistance pattern of identified P. aeruginosa was
evaluated by Kirby Bauer disc diffusion assay against selected antibiotics includes
ciprofloxacin, levofloxacin, meropenem and imipenem. Measure the zone of inhibition
and isolates marked as resistant, intermediate and sensitive. Resistant strains were
alienated for further evaluation.
Leaves of Eucalyptus globulus (Tasmanian blue gum) and Calotropis procera
(apple of Sodom) proceed for extraction and the plant extracts was obtained by using
solvent chloroform and ethanol. Percentage yield of both plant extract was calculated.
High percentage yield was obtained from Eucalyptus globulus and less percentage yield
was gained from Calotropis procera in comparison The obtained extract was dried and
the resultant material was used in well diffusion assay to evaluate the antibiotic
CHAPTER 6
SUMMARY
Summary
66
sensitivity of resistant P. aeruginosa against selected plants. Stock of plant extracts was
prepared by dissolving 1g of plant extract in 1ml of DMSO. Well diffusion assay was
performed and zones were measured in millimeter and categorized as resistant, sensitive
and intermediate. Isolates that are susceptible to plant extracts were separated and
Minimum inhibitory concentration of susceptible isolates was determined by broth micro
dilution assay and cytotoxicity profiling was done by 3-(4, 5-dimethyl-2-thiazolyl)-2,5-
diphenyl-2H-tetrazolium bromide (MTT) assay and cell survival percentage was
calculated.
Data recorded during the study was analyzed by one-way analysis of variance
(ANOVA) followed by Duncan’s test using the SPSS statistical software program.
Differences were considered significant at P < 0.05. Availability: Items available for loan: UVAS Library [Call number: 2545-T] (1).
45.
Evaluation Of The Microbiological Quality Of Ice Cream Sold At Local Shops In Lahore
by Muhammad Bilal (2009-VA-492 | Dr. Naureen Naeem | Dr. Sana Ullah Iqbal | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Ice cream is a delicious, wholesome, nutritious frozen dairy food. It is noticed that generally manufacturing of ice cream on small scale production units like local shops doesn't totally follow the standard procedures of ice cream production. Ice cream can be contaminated with microorganisms if some ingredients have been added after pasteurization or by means of improper sanitation of the equipment and environment. Microbiological quality of ice cream reflects hygienic practice in production and is an indication of food safety. There was variation in quality of locally produced ice cream in different areas of Lahore.
Food safety is a scientific discipline describes handling, preparation and storage of food that prevents food borne diseases. It includes a number of ways that must be followed to avoid potentially severe health hazards. The tracks within this line of thought are safety between the market and consumer with the fast pace of life the consumption of fast food is going to increase day by day in Lahore. It is imperative to prepare food at good hygienic conditions because of its perishability. The usual thought is that food should be safe in the market and the concern is safe delivery and preparation of food for the consumer.
Microorganisms affecting food comes from natural microflora or are introduced by manufacturing steps ranging from processing storage and distribution. In some cases these micro flora have no effect on the food and can be consumed without consequence, but those that are introduced during course of processing depending on type and level of contamination can spoil the food and cause food borne illnesses.
Summary
49
Food can transmit diseases from one person to another as well as serve as growth medium for bacteria that can cause food poisoning. In the developed countries there are standards for food, whereas in less developed countries the main issue is the availability of safe water supply which is one of the critical item. Foodborne illness is a problem resulting from the consumption of contaminated food, bacteria, viruses, or parasites, natural toxins, chemicals that contaminate food.
For this study total 108 locally produced samples of ice cream were carried from different areas of Lahore. Mughalpura, Sadar, Model town, Gulberg, Town ship and Johar town. 6 shops from each area and 3 samples from each shop were purchased and put in the sampling box with ice packs. Samples were analyzes for microbial load in microbiology Lab of University of Veterinary and Animal Sciences.
Each sample was analyzed for microbiological analysis (Enumeration of Total Viable Count (TVC), Enumeration of Total Coliform Count (TCC), Enumeration of Total Staphylococcal Count (TSC) and detection of salmonella spp.
Nutrient agar was used to perform Total plate count, Mackonky agar was used for Total coliform count, Mannitol salt agar was used to count S.aureus , Salmonella Shigella agar for Salmonella detection . Each analysis was performed 3 times.
The samples collected from Mughalpura, Sadar and Model town were highly contaminated having more microbial count which shows contamination due to poor handling and hygienic conditions while Gulberg, Johar town and Town ship having less contaminated. Availability: Items available for loan: UVAS Library [Call number: 2543-T] (1).
46.
Comparative Quality Evaluation Of Raw And Pasteurized Milk
by Hafiza Saima Ghaffar (2009-VA-230) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmed Anjum | Dr. Sana Ullah Iqbal.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This particular project was designed to evaluate the overall quality of raw and pasteurized milk available at different areas of Lahore. The parameter which was checked includes microbiological analysis, adulterants, physicochemical properties and the effect of temperature on vitamin C in milk samples. Raw samples were collected from ten different towns of Lahore, whereas pasteurized milk samples belong to ten different brands. Ten samples were collected under control conditions from animals in sterilized containers.
For microbiological analysis four parameters were selected including total plate count (TPC), total coliform count (TCC), total psychrotrophic count (TPSC) and total yeast and mold count (TYMC) whereas, different adulterants like adulteration test was done such as urea, starch, hydrogen peroxide, detergent or soap, sorbitol, quaternary ammonium compound, boric acid, cane sugar, sodium chloride, formalin and hypochlorite were checked by using the milk adulteration kit in QOL. Milk contains casein and whey proteins. Whey protein being added in the milk to increase its density which is considers being an adulterant. In this project whey protein was estimated in all milk samples by titration method. Physicochemical characteristics of milk are an important parameter to judge the quality of milk. These physicochemical properties include fat%, SNF%, density kg/m3, lactose%, solid/ash, protein% and pH. Physicochemical properties were evaluated mechanically by Milkoscan. Heat treatment is an important method to reduce the microbiological contamination of milk. These treatments may include pasteurization and UHT etc. During the heat treatment some of the micronutrients may deteriorate thus compromising the quality of milk. Vitamin C is among those heat labile micronutrients. Vitamin C was checked quantitatively in market and self-collected samples by using titration method.
It was concluded that total plate count TPC, TCC, TPSC and TYMC of raw milk samples were above the standard value indicating the poor quality of the milk. As far as the pasteurized milk samples were concerned ninety percent of the samples showing higher values for TCC, TPSC and TYMC. Total plate counts of all self-collected raw milk from a healthy animal were found within the standard value. Counts were in range of 3.8x 103 – 8.9x103 CFU/mL of all milk samples. TPC of all self-collected raw milk from a healthy animal were found within the standard TCC were found within permissible value (102 CFU/mL .TPSC were negative for all milk samples. TYMC were in range of 2.6x101 -7.2x101 CFU/mL. Among milk samples (n=10), three samples (30%) were positive for TYMC were while remaining samples (70%) were negative and showed no growth.
Physicochemical factor show that 50 percent of raw milk have low nutritional value as compared to the standards which are buffalo and cow milk contains 7.6, 4.5% fat, 3.8, 3.8 % protein, 5.1, 4.9% lactose, 0.78, 0.72% ash and 17.0, 13.9% total solid respectively. In raw milk mean of fat (%), solid not fat (%), lactose (%), Solid/ash (0%), protein(%) and pH were 4.50±0.03, 7.915±0.06, 23.05±0.055, 3.893±0.06, 3.85±0.05, and6.9±0.0.02 respectively. In pasteurized milk mean value for fat, SNF, lactose, ash, protein and pH were 3.48 ±0.13, 7.24±0.10, 3.60±0.05,0.5 ±0.06, 2.82±0.05, 7.2±0.20 respectively. Pasteurized milk is good for consumption.
Different adulterant such as urea, starch, hydrogen peroxide, Sorbitol, QAC, Boric acid, Cane sugar, NaCl, Carbonate, Formalin, hypochlorite, whey protein, Added water and soap /detergents were evaluated in all milk samples. Among these adulterant water (66%) was found in majority of milk samples, followed by whey protein (15%), starch (13%), (10%) NaCl and
(8%) cane sugar were detected in raw milk samples. n Pasteurized milk samples only added water (49%) and whey protein (31%) was detected.
Among the raw milk samples the maximum and minimum concentration of vitamin C was observed 0.33±0.02 and 3.33 ±0.02 mg/100ml and for pasteurized milk maximum and minimum concentration of vitamin C was observed 2.54mg/100ml and 0.32 ±0.02 mg/100ml respectively. In self- collected samples the minimum and maximum concentration of vitamin C was observed 5.25±0.02 and 8.34 ±0.04 mg/100ml respectively and after pasteurization in laboratory minimum and maximum concentration of vitamin C was observed 3.48±0.04 and 5.83 ±0.02 mg/100ml respectively. These observations had showed that pasteurization treatment decreased Vitamin C quantity. Availability: Items available for loan: UVAS Library [Call number: 2536-T] (1).
47.
Assessment Of Afflatoxins Contamination In Peanuts
by Zanib Hashmi (2009-VA-512) | Dr. Naureen Naeem | Dr. Sanaullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Peanut is the most important agricultural crop of Pakistan. Peanut is a dicotyledonous, herbaceous, pubescent, rigid or low growing plant and the only species cultivated is (Arachishypogaea L.). Peanut is rich in protein, fat and carbohydrates, some percentage of Ca, K, P, Mg and vitamin E is also present. Peanut is an excellent source of edible oil as it contains about 50 to 53 percent good quality oil used in ghee, margarine and salad. There is high risk of contamination of peanuts with aflatoxins(AFB1, AFB2, AFG1 and AFG2) because of fungal attack during the drying of peanut pods. Out of all these aflatoxins AFB1 is most important. Aflatoxins are toxic, carcinogenic secondary metabolites of Aspergillusflavus, Aspergillusparaciticus and Aspergillusnomius. Aflatoxins can cause illness to human results in Aflatoxicosis. Aflatoxins are carcinogenic compounds that are causative agents in human hepatic and extra hepatic carcinogenesis. The chief attacking organ for aflatoxins B1 toxicity and carcinogenicity is liver. From the safety point of view aflatoxin management is important for the production of safe and excellent quality peanuts.
For this purpose present study was conducted to determine the level of aflatoxins in peanuts (roasted, un-roasted). Samples will be collected/purchased by simple random collection technique from local markets and vendors from different areas ( Sabzazar, Wahdat road , Shad bagh, Data darbar, Akbarimandi, Beaden road, Lohari gate, Ek-moria pull, Liberty, Firdous market, Siddiqiacoloney, Mughal pura, Faizbagh, Rehmanpura, Gulberg, Model town, Islam pura, Shahdara, Rang mahal, Muslim town, Township, Iqbal town, Awan town, Niazbegh, Mozang, Outfall road, Sanatnagar, Cantt, Secretriate and Shad man) of Lahore. The samples were analyzed by thin layer chromatography (TLC) to check the presence of aflatoxins (B1, B2,
G1 and G2). TLC analyses were further confirmed by high performance liquid chromatography (HPLC) to verify the accuracy of TLC. These analyses were performed in the Department of Food Science and Human Nutrition and WTO labs, University of Veterinary and Animal Sciences, Lahore. As out of 120 total samples of peanuts 60 samples were taken from vendors with 2 categories of roasted and unroasted while 60 samples were collected from shops with the same categories. Out of 120 samples, 55 (45.8%) were contaminated. In these 55 samples 48 (87.2%) samples were contaminated with aflatoxin B1.Aflatoxin G1 is also present in 3 samples (5.45%), aflatoxin B2 in 3 (5.45%) samples and Aflatoxin G2 is present only in one samples collected from vendors, and we can say that 1.8% samples were contaminated with aflatoxin G2.
Present study will be supportive for the investigation of aflatoxins in peanuts. Peanuts are widely consumed all over the world and occurrence of aflatoxins in this commodity is a major concern to human health. The present situation is too much worse about the levels of aflatoxins which are higher than the prescribed limit by the regulatory authorities. It was observed that TLC technique is good for the determination of aflatoxins in developing countries where the facilities of sensitive instruments are not accessible. Furthermore to quantify levels of aflatoxins by using sensitive instruments like HPLC, GC-MS and LC-MS is required for accurate detection of Aflatoxins in peanuts in markets to protect the consumers from exposure of aflatoxins high level which are carcinogenic and hepatotoxic.
Availability: Items available for loan: UVAS Library [Call number: 2614-T] (1).
48.
Effect Of Garlic And Ginger Extract On The Shelf Life Of Fish
by Dure-e-Shahwar (2009-VA-439) | Dr. Naureen Naeem | Dr.Sanaullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The fish is highly perishable food which contains high protein and omega3 fatty acids. It contain enzyme which cause autocatalysis of muscles after harvesting. Due to lack of Knowledge and poor storage and handling practices cause fish spoilage and deterioration of fish.
Ginger and garlic are spices, also contain a variety of bioactive substances which are of considerable use from the standpoint of food science and technology. Ginger and garlic shows excellent inhibition against food pathogens such as Staphylococcus aureus; Bacillus spp., Escherichia coli and Salmonella spp.
Antimicrobial properties of garlic and ginger may control the microbial growth of fish and is able to minimize fish spoilage. Fish was taken from fish farm then washed and cleaned, cut the fish and left at room temperature for water dropping then weighed it. Each sample was containing 20gm weight.
Then dipped samples in extract of ginger and garlic that have doses 15%, 20 %, 25%for ninety minute, then was wrapped in polythene bag and put in refrigerator for 5 months.
Aerobic plate count was performed after fortnightly by the method of standard plate count and assessed sensory condition of fish by sensory evaluation after one month.
In control group, the Bacillus cereus significantly increased with time (during storage) While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life and Bacillus cereus significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. Garlic was more effective then ginger in separately treatment.
In control group, staphylococcus significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life of staphylococcus significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. In comparison to garlic, ginger was observed most efficient in controlling staphylococcus growth in fish samples.
In control group, Salmonella significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life and Salmonella significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. Seprately20 % garlic and ginger show same result.
In control group, Streptococcus significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life and Streptococcus significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. But garlic showed better results as compared to the ginger in respective concentrations.
In control group, Shigella significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life Shigella significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group.
The sensory evaluation results showed that with increasing concentration of ginger and garlic separate and in combination of both have profound effects on sensory parameters. It is evident
Summary
63
from the results after five months of trial that garlic and ginger can be used to control microbial growth in fish samples and their acceptability on sensory scale is better than the control samples. Treated samples were more liked and observed acceptable according to grading scale. By comparing the whole results of sensory evaluation it has become very easy to access the positive outcomes of the applications of ginger and garlic in different concentrations and in combination. Ginger and garlic in combination were more liked and maintained their color, juiciness, flavor, tenderness and oiliness level.
Data was statistically analyzed by applying 2 Way ANOVA. There was mean score difference (p<0.05) among garlic treatment, ginger treatment and combination of garlic and ginger treatment with bacterial count. But ginger has least effect as compare to garlic but in combination they became more effective against bacterial count. There was mean score significant difference (p<0.05) among treatment and time with sensory evaluation.
This study shows that combination of both spices 25% ginger & garlic is more effective then separately ginger & garlic. Garlic shows better result against control of bacterial count Streptococcus and Bacillus cercus. Ginger shows better result against control of bacterial count in Staphylococcus and Shigella. Both spices show almost same control of bacterial count against Salmonella. Availability: Items available for loan: UVAS Library [Call number: 2611-T] (1).
49.
Isolation, Molecular Identification And Antibiotic Resistance Pattern Of Salmonella Enterica From Fancy Birds
by Aqeela Kousar (2010-VA-303) | Mr. Muhammad Asad Ali | Prof. Dr. Aftab Ahmed Anjum | Prof. Dr. Mansur-ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Salmonellosis is a disease with serious health issues related to food borne illness and
most of world’s population is suffering from it. Early diagnosis in case is very important for
treatment of disease. Salmonellosis may hidden as a carrier state, acts as zoonotic components
for transmission of disease. Therefore the test with more diagnostic value needs to be developed
like Polymerase chain reaction after culturing and microbiological examination.Salmonella
enterica infections continue to pose a significant risk for poultry industry and fancy birds.
Salmonella infections have been controlled by antibiotics but in recent times antibiotic resistance
in microorganisms especially in Salmonella is a global health issue. Antibiotic resistant
Salmonella has further compounded the problem. Poultry isolate of Salmonella enterica (n=150)
were procured from Jallo park, Safari park and household pets which are taken to Pet Centre
University of Veterinary and Animal Sciences Lahore then brought to Department of
microbiology UVAS Lahore and identified by biochemical testing, morphology, staining
characters and genus specific PCR. Antibiotic Susceptibility was checked by disc diffusion
method against amoxicillin (30μg), ampicillin (10μg), cefixime (5μg), , ceftazidime (30μg),
ceftriaxone (30μg), ciprofloxacin (5μg), gentamicin (10μg), nalidixic acid and tetracycline
(30μg) and resistant pattern was 100 % in ampicillin and tetracycline and 41.18% and 58.82% %
in gentamicin and ciprofloxacin respectively while antibiotic show 0% resistance. Fancy birds
are carriers of drug resistant Salmonellae.
A total of 150 samples collected from Zoo Lahore, safari park and household pet fancy
birds each of n=50. Samples will enriched by non-selective and selective media, After isolation
on selective media macroscopic, biochemical analysis and microscopic examination done. DNA
Summary
53
extracted from culture isolated from cloacal swabs and polymerase chain reaction performed
using primers. Amplication will be observed using Agarose gel electrophoresis.
Research highlighted the prevalence of Salmonella in fancy birds and its possibility of
transmission to human beings. Research also provided data on antibiotic resistance in
Salmonellae from fancy birds and its possible role in ever increasing problem of antibiotic
resistance. Availability: Items available for loan: UVAS Library [Call number: 2615-T] (1).
50.
Chemical, Microbiological And Toxicological Evaluation Of Textile Dyeing Industry Wastewater
by Muhammad Furqan Akhtar (2011-VA-265) | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Exposure to complex mixtures like textile effluent poses risks to animal and human health such as mutations, genotoxicity, pathological lesions and oxidative damage. The aim of the present study was to quantify metals and identify organic pollutants in untreated textile dyeing industry wastewater, to determine the bacterial load of wastewater, isolate and identify heavy metals tolerant bacteria and to determine its mutagenic, genotoxic and cytotoxic potential, influence on normal physiology and effects on oxidative stress biomarkers in effluent exposed rats.
Metal analysis through AAS revealed presence of high amounts of zinc, copper, chromium, iron, arsenic and mercury in industrial effluent. Various organic pollutants such as chlorpyrifos, cucurbitacin-b and phthalates were identified by screening through GC-MS.
Microbiological evaluation of textile dyeing industry wastewater revealed a high bacterial load. Different bacteria isolated from wastewater such as Staphylococcus aureus, Pseudomonas aeruginosa, Corynebacterium xerosis, Bacillus megaterium, Staphyoloccus epidermidis and Micrococcus varians exhibited resistance to Cr and Cu salts and antibiotics to varying degree.
Ames test with/without enzyme activation and MTT assay showed strong association of industrial effluent with mutagenicity and cytotoxicity respectively. Bacterial reverse mutation assay revealed that the mutagenicity of textile dyeing industry wastewater decreased with increase in dilution of wastewater.
In-vitro comet assay revealed the evidence of high oxidative DNA damage induced by textile wastewater. Wastewater exhibited concentration dependent genotoxicity in sheep
SUMMARY
147
peripheral lymphocytes. When Wistar rats were exposed to industrial effluent in different dilutions for 60 days, then activities of total superoxide dismutase and catalase and hydrogen peroxide concentration were found to be significantly lower in kidney, liver and blood/ plasma of effluent exposed rats than control. Vitamin C at a dose of 50mg/Kg/day significantly reduced oxidative effects of effluent in rats. Industrial effluents may decrease activities of T-SOD and CAT and concentration of H2O2 in liver, kidney and blood/plasma of Wistar rats. Vitamin C may have a possible ameliorating effect on industrial effluent induced oxidative stress in Wistar rats.
Wastewater exposed rats exhibited necrosis of epithelial cells of nephron, pulmonary emphysema, and inflammation of the lungs, degradation and infiltration of cardiac myocytes, fibrosis of the liver, damage to the intestinal mucosa and sloughing off epithelial cells from the intestinal lumen.
This study concludes that untreated textile dyeing wastewater being a complex mixture of inorganic and organic pollutants may be highly eco-toxic and may contaminate of the environment via continuous release of various organic and inorganic pollutants. Availability: Items available for loan: UVAS Library [Call number: 2580-T] (1).
