1.
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).
2.
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).
3.
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).
4.
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).
5.
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).
6.
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).
7.
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).
8.
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).
9.
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).
10.
Effects Of Physico-Chemical Properties Of Diluents On The Infectivity Titers Of Freez Dried Ppr Virus Vaccine
by Fariya Yaqub Baig (2009-VA-240) | Dr. Jawad Nazir | Prof Dr. Aftab Ahmad Anjum | Dr. Waseem Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: 6.1. Introduction.
Peste des Petits Ruminants (PPR) is a febrile viral disease of sheep and goats. The disease is
responsible for low productivity in small ruminants and great economic losses to the farmers in
Africa and Asia including Pakistan. There is no specific treatment for PPR and prevention is only
possible through the use of live attenuated vaccines. Being an enveloped virus, PPRV is heat
sensitive. Poor immunological responses have been observed after the vaccination of PPR.
Reasons may be disturbance in maintenance of cold chain, improper handling and route of
vaccination as well as reconstitution in inappropriate diluents. Physiochemical properties
(temperature, pH, osmotic pressure, salinity and UV light) of diluents effects the infectivity of
PPR freeze dried vaccine as live virus vaccines work properly after reconstitution within the
recommended time interval.
6.2. Experimental Design.
Effect of three diluents (normal saline, phosphate buffer saline and distilled water)
adjusted to various pH conditions (5.00, 6.00. 7.00, 8.00, and 9.00) on the infectivity of PPRV
was evaluated. Contents of the freeze dried PPR vaccine vials were diluted with one ml of the
respective diluent adjusted to above mentioned pH conditions. Virus infectivity from the vials
was measured immediately following reconstitution. One set of vials was kept at room
temperature (25 ºC ±2) and virus infectivity was measured afterwards at 30, 60, and 120 minutes
as described in the section 3.3.2. While other set of the vials was kept at refrigerated temperature
(4 ºC ±2) and virus infectivity was measured at 30, 60, 120, and 180 minutes as described in the
Summary
46
section 3.3.2. The change in pH of each vial following reconstitution was also measured
accordingly. Each set of experiment was repeated three times independently.
6.3. Results.
PBS and NS gave equivalent results and proved better than distilled water to restore the
infectivity of PPRV. For a better comparison of virus stability in the PPR freeze dried vaccine
following reconstitution in various diluents adjusted to various pH conditions the infectivity titer
of the virus in the vaccines were measured at various time points. The serial data thus obtained
was analyzed by linear regression model to calculate T-90 values (Time required for 90 %
reduction in virus infectivity). The higher T-90 values indicate better stability of the virus at a
designated condition. At both of the temperatures the T-90 values were equivalent and relatively
higher for all of the diluents adjusted to pH 7.00 and 8.00 while these values are lower at extreme
pH conditions. Minimum T-90 values (126± 56) were observed at pH 9.00 in normal saline kept
at ambient temperature while maximum T-90 (448 ± 49.7) values were observed at pH 7.00 in
phosphate buffer saline kept at refrigerated temperature.
6.4. Conclusion.
Results of the present study suggest that virus infectivity in the live attenuated PPRV vaccine can
be better stabilized following reconstitution in PBS adjusted to neutral or slightly alkaline pH.
Chilled vaccine diluent is preferable to the one kept at room temperature and vaccine should be
administered within 30 minutes of reconstitution. Availability: Items available for loan: UVAS Library [Call number: 2645-T] (1).
11.
Affect Of Temperature, Cell Density And Multiplicity Of Infection On Biological Titer Of FMDV Type “O”
by Qazi Ithram Ul Haq (2009-VA-104) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Imran.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: FMD is a transmissible viral disease of animals. It is causing very highly economical loses in Pakistan and all over the world. Through vaccination FMD is being controlled in Pakistan. Inactivated virus is used in vaccines. FMD virus grows on BHK-21 cell lines. FMDV show good adoptability on these cell lines. For good and high titer FMDV needs few physical factors to grow on BHK-21 cell lines. These factors include Temperature, Cell density and Multiplicity of infection (MOI)was considered in this research. The FMDV strain “O” was grown on BHK-21 cell line. The cells monolayer was propagated for conduction of effect of these factors on the virus. The mentioned factors were studied to get optimum level of virus titer in in vitro cell lines. The effect of 35°C, 37°C and 39°C was evaluated on the virus growth. Maximum virus propagation was noted at optimum temperature 37°C. The viral concentration at 37°C was significantly (P<0.05) higher than at 35°C and 39°C. The effect of cell density was studied on the virus concentration. Flask of three different densities 25cm2, 175cm2 and 275cm2were utilized in the current study. The virus concentration in all three different densities was not significantly different (P>0.05) from each other. Another factor Multiplicity of infection (MOI)was investigated in the study. Five different volumes 10ul, 20ul, 30ul, 40ul and 50ul of the FMDV strain “O” were used to investigate the effect of factor on the virus concentration. The results revealed highest viral harvest concretion at 50ul volume with MOI of 7.1, %age of cells infected with single virus and 6.3 × 1079. The MOI at 50ul was significantly higher (P<0.05) than the other four concentration of the virus. It was concluded from the study that the optimum temperature for the maximum FMDV concentration harvest is 37°C. The density of cell has no significant effect on the growth of virus that is flask of any density may be used to grow the FMDV. Multiplicity of infection (MOI) of 14.2 give maximum
SUMMARY
34
TCID50 Optimizing the conditions for the cell culture and virus cultivation helps in maximum virus harvest achievement. From the present study it may be suggested that the physical factors may be optimized for the remaining strains of FMD and other vaccine viruses to attain maximum virus grow. Availability: Items available for loan: UVAS Library [Call number: 2681-T] (1).
12.
In Vitro Activity Of Selected Biocides Against Fungal Isolates From Production Area Of Pharmaceutical Industry
by Sana Ilyas (2009-VA-238) | Dr. Muhammad Nawaz | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Ovais Omer.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pakistan pharmaceutical industries have grown to grab their position amongst top ten pharmaceutical industries of Asia Pacific region. These are serving with 80% of pharmaceutical needs. The industry on the other hand faces some challenges in terms of sterile pharmaceutical product manufacturing. The fungal contamination causes spoilage to pharmaceutical products, cosmetics, and food products. The fungal contamination to pharmaceutical products has resulted in direct losses to human health and to economy.
A total of 50 air samples were collected from clean area of a pharmaceutical production unit by exposing sabouraud dextrose agar (SDA) plates by settle plate method (4 hours exposure). Fungal colonies were purified by sub-culturing and later identified macroscopically and microscopically. Selected biocides included isopropyl alcohol (70%), chloroxylenol (20%), chlorhexidine gluconate (20%), and benzalkonium chloride (20%) were used in this study. A 100 μl of spore suspension of each fungal contaminant (1.0 × 106 to 5.0 × 106 spores/mL) was exposed to 9.9 mL of biocide preparation for 15 and 30 minutes while exposure was stopped by adding 1 mL of mixture (spores exposed to biocide) into 9 mL of respective neutralizing agents The enumeration of colonies was started immediately after the growth was visible and expressed as Mean±S.D. and converted to log10. Antifungal activity of biocides was expressed as log10 reduction and different biocides‟ activity was compared using ANOVA technique by graphed prism 5.0 statistical software.
Total 204 colony forming units (CFU) were identified from filling area (36), solution room (47), and buffers (121). The antifungal activity in terms of log reduction was lowest by isopropyl alcohol at 15 minutes and highest was shown by chlorohexidine gluconate at 30 minutes against
Summary
64
Aspergillus flavus. In case of Aspergillus fumigatus all the biocides presented significant difference of antifungal activity at 15 minutes. The response of Aspergillus niger against different biocides at 15 minutes and 30 minutes was same as was in case of Aspergillus flavus while each biocide‟s antifungal activity was found significantly increased with increase in time of exposure. The similar response of antifungal activity of different biocides at both exposure times was noted against Saccharomyces cerevisiae. The antifungal activity of all biocides against penicillium was found significant different at 15 minutes and 30 minutes exposure time. Similarly, each biocide‟s antifungal activity increased with increase in time of exposure. On overall basis, isopropyl alcohol was found less effective while benzalkonium chloride and chlorohexidine gluconate presented comparatively higher efficacy against fungal isolates. Availability: Items available for loan: UVAS Library [Call number: 2705-T] (1).
13.
Isolation And Characterization Of Phytase Producing Fungi For Poultry Feed
by Ali Ahmad (2002-VA-121) | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Masood Rabbani | Prof. Dr. Kamran Ashraf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Isolation And Characterization Of Phytase Producing Fungi For Poultry Feed Availability: Items available for loan: UVAS Library [Call number: 2770-T] (1).
14.
Evaluation Of Antimicrobial Activity Of Essential Oil And Extracts Of Nigella Sativa Against Antibiotic Resistant Salmonella Enterica Isolates Of Human And Poultry Origin
by Sadia ashraf(2011-VA-402) | Prof. Dr. Aftab Ahmad Anjum | Dr. Ali Ahmad Sheikh | Dr.Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The research was designed to evaluate the efficacy of methanolic and aqueous extracts of Nigella sativa, Black seed oil and thymoquinone against antibiotic resistant molecular characterized Salmonella enterica isolates of human and poultry origin (n=5 each). The compounds that have shown the antibacterial activity was also checked for their cytotoxicity by MTT assay.
Salmonella is causative agent of invasive diseases in poultry and humans, results in high mortality. Salmonellosis is a disease caused by Salmonella enterica with serious health issues related to food borne illness and most of world’s population is suffering from it. Mostly infections are treated by antibiotics but now a day’s resistance developed by Salmonella enterica. So it is need of time to develop some alternate ways to combat the problem caused by resistant bacterial pathogens. Use of essential oils and extracts of seeds are good weapons against resistant bacteria.
Salmonella enterica isolates of human and poultry origin (n=5 each) were taken from Department of microbiology UVAS Lahore and identified by colony morphology, microscopic characters, biochemical testing (Indole production test, Methyl red test, Voges Proskaeur test, Citrate utilization test and Urea utilization test) and polymerase chain reaction (PCR). For PCR product 1.5% agarose gel was run by gel electrophoresis. The biochemically identified and molecular characterized S. enterica isolates were screened for antibiotic susceptibility by Kirby Bauer disc diffusion method against amoxicillin, ampicillin, cefixime, ceftriaxone, ciprofloxacin, gentamicin, nalidixic acid, co-trimoxazole, ofloxacin and tetracycline and resistant pattern was 100% against ampicillin and Nalidixic acid and isolates shown 60% resistant against co-trimoxazole, amoxicillin and tetracycline, 80% and 40% resistant found against ofloxacin and ciprofloxacin while all isolates sensitive to cefixime and ceftriaxone. Aqueous and methanol were
CHAPTER 6
SUMMARY
used as solvents for extraction from Nigella Sativa. Seeds were dried, mixed, centrifuged, filtered and filtrate evaporated to obtained extracts. Percentage yield of methanolic extract was more than aqueous extract. Commercially available black seed oil, thymoquinone, water and methanol extracts of black seed would be evaluated for antibacterial activity by well diffusion method. Zones were measured in millimeters. All compounds gave the zones of inhibition except aqueous extract against Salmonella enterica isolates. The minimum inhibitory concentration (MIC) was determined by micro broth dilution method and then methanolic extract, black seed oil and thymoquinone used in MTT assay to evaluate their cytotoxicity. Cell survival percentage was calculated by a formula.
Data were analyzed by one way analysis of variance (ANOVA) followed by Duncan’s multiple range tests (DMRT) using statistical package for social sciences (SPSS version 20.0). Antimicrobial activity of essential oils, thymoquinone, water and methanol extract would be compared by graph pad prism 5.0 statistical software. Availability: Items available for loan: UVAS Library [Call number: 2827-T] (1).
15.
Spatial Ecology And Distribution Of Soil Borne Burkholderia Mallei In Punjab, Pakistan
by Muhammad Asad Ali (2002-VA-73) | Prof. Dr. Khushi Muhammad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Mansur-Ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Burkholderia mallei is a causative agent of glanders, the disease of equines. The disease is characterized by pulmonary, nasal and cutaneous forms. B. mallei is excreted through nasal discharge, lacerated skin/wounds and expiration. Diseased animals shed bacteria through the discharges contaminating soil, water, fodder and other susceptible animals in its vicinity. The present study was designed to map and investigate the association of different physical factors and soil chemistry analytes with persistence of B. mallei genome in soil of 10% percent villages (n=456) from eight selected districts of Punjab province, Pakistan.
Eleven (0.48%) out of 2, 280 soil samples were positive for B. mallei genome in varied locations of Punjab. Higher prevalence (2.37%) for genome was detected in Sheikhupura district followed by Chakwal district (2.10%). None of the samples from Gujranwala, Sahiwal, DG Khan, Attock, Faisalabad and Sargodha districts were found positive for B. mallei genome. The genome of B. mallei was distributed in 25% study districts of Punjab, Pakistan. In Chakwal district, the genome of B. mallei was strongly associated with moisture (p=0.008) in all positive samples ranging from 0.80 to 39.20%, Phosphorous (p=0.050) ranging from 1.74 to 21.75 mg/Kg. While, this association in Sheikhupura district soil samples was with Sodium (p=0.018) and moisture (0.026) ranging from 1.90 to 133.59 mg/Kg and 0.80 to 39.20%, respectively. The odds of detecting DNA of B. mallei were recorded higher (1.4, 6.8, 5.0, 2.8 and 10.6 ) when soil sample sites were < 500 meters away from vehicular traffic roads, < one kilometer from animal markets, < 100 meters from canal, animal density < 1,000 animals and human population < 300 houses/village. While the odds of detecting DNA of B. mallei were 0.1, 0.3, 0.4, 0.2 and 0.5 when soil sample sites were > 500 meters from vehicular traffic roads, > one kilometer from animal markets, > 100 meters from canal, animal density > 1000 animals and human population > 300 houses/village, respectively. Soil-borne B. mallei DNA is more likely to be detected in areas closer to roads with vehicular traffic along the interstate routes in Punjab and soil containing low level of moisture.
It was concluded that soil of two districts out of eight selected was positive for B. mallei genome in Punjab province. Odds of less distance from main road to animal farm and high animal density at farm were positively associated with B. mallei DNA persistence in soil. Moisture, sodium and phosphorus were positively associated with persistence of B. mallei DNA in soil.
Availability: Items available for loan: UVAS Library [Call number: 2900-T] (1).
