1.
Comparitive Immunogenic Evaluation Of Purified And Whole Culture Vaccine Of Pasteurella Multocida B2 Of Bovine Origin
by Anika Khalid (2006-VA-357) | Dr. Imran Altaf | Dr. Jawad Nazir | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pasteurella multocida is responsible for causing Heamorrhagic Septiceimia which is a fatal and acute disease mainly associated with bovines.
Pasteurella multocida consists of two important antigenic structures i-e Outer membrane protein(OMP) and Lipopolysaccharide(LPS) that forms capsule.These two structures played vital role in pathogenicity as well as in the immunogenicity.
In our present project we study the role of LPS,OMP alone and in combination as an immunogens.We compared the immunogenic behaviour of whole culture vaccine(consists of LPS and OMP) purified vaccine(consists of OMP),LPS vaccine (consists of LPS only) by IHA and CFT (using OMP and LPS both as an Ag separately) and the overall results showed that whole culture HS Alum precipitated vaccine produced better antiboby titer against both outer membrane proteins (OMP) and lipopolysacharrides (LPS) when checked against respective antigens as compared to the vaccines containing OMP and LPS alone.
So it was concluded that LPS no doubt act as an immunogen but its immunogenicity increases many times when combine with protein (OMP) as in whole culture vaccine. Availability: Items available for loan: UVAS Library [Call number: 2316-T] (1).
2.
Antibiotic Resistance Pattern Of Clostridium Perfringens Type A From Poultry And Its Resistance Modulation Using Medicinal Plant Extracts
by Arifa Jabeen (2008-VA-398) | Dr. Ali Ahmad Sheikh | Prof. Dr.Aftab Ahmad Anjum | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Avian acute necrotic enteritis (NE) is the burning issue in poultry sectors worldwide resulting in huge economic losses. Cl. perfringens; causative of NE; is an anaerobic, rod shaped Gram positive, spore former, catalase negative bacteria which produce characteristic double zone of hemolysis on 5 % sheep blood agar.
A total of 100 (n=100) intestinal samples were collected from ten different poultry farms in and around Lahore. Intestinal contents were enriched in fluid thioglycollate medium (FTM), isolated and purified on the tryptose sulfite cycloserine (TSC) agar which is highly selective and recommended medium for Cl. perfringens; identified by microscopic examination and typical biochemical tests (catalase and blood hemolysis).
Results showed that fifty six out of 100 samples were positive for the presence of Cl. perfringens with the overall positivity ratio of 56 % that is the highest percent prevalence in broilers and type was confirmed on the detection of cpa alpha toxin producing gene resulting in the product band size of 128 bp through PCR.
Five PCR confirmed isolates were subjected to antibiotic resistance studies. According to the results and CLSI standards, ampicillin and amoxycillin were susceptible to Cl. Perfringens while tetraacycline, ciprofloxacin and streptomycin were found resistant to all of the five isolates.
By agar well diffusion method plant extracts (Astragalus, Zingiber officinale, Calotropis procera and Gymnema sylvestre) were tested for their anti-clostridial activity. All produced zones of inhibitions of varying diameters except the aqueous extracts of 1st three plants that
Summary
75
produced no any zone of inhibition on agar plates against any of the five isolates tested while those of Gymnema sylvestre produced good zones. MIC of plant extracts were determined against isolates. The extracts of Gymnema showed the lowest MIC values among all and in between sequential extracts of plants, the chloroform extracts showed relatively low MIC values in comparison to others statistically using One way ANOVA. MIC values of plant extracts were determined in combination with the specific MIC breakpoint concentrations of antibiotics too for their resistance modulatory effects against the isolates. Statistical result findings were very effective in combination with the resistant antibiotics as MIC values significantly reduced in comparison to performed solely with plant extracts. Especially the modulatory results for ethanol and chloroform extracts of nearly all four plants were noticeable as MIC values were rapidly declined below 100 in comparison to plant extracts alone where MIC values were higher to 500. All of the results data obtained through whole experiment were analyzed using SPSS versions 20.0.
Hence on the basis of above findings with the current study, it is concluded that medicinal plant extracts may prove effective herbal therapeutic agents against Cl. perfringens type A; the potent cause of necrotic enteritis in poultry. The findings of the study might be helpful in renewing and rescheduling the antibiotic administration plan with the use of medicinal plant extracts to modulate the action of antibiotics. Availability: Items available for loan: UVAS Library [Call number: 2276-T] (1).
3.
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).
4.
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).
5.
Antibiotic Resistance Profiling Of Enterococcus Faecium Recovered From Retail Chicken Meat From Lahore City
by Amina Habib (2010-VA-313) | Dr. Ali Ahmad Shiekh | Dr. Fareeha Akhtar | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Enterococcus faecium is gram positive bacteria which are normal intestinal flora of warm blooded animals and humans beings. It is responsible for various types of diseases such as neonatal meningitis, endocarditis, urinary tract infections and intra-abdominal or pelvic wounds. Retail chicken could be a source of Enterococcus faecium. Irrational use of antibiotics in chicken rearing can lead to emergence of antibiotic resistant Enterococcus faecium. The chicken meat gets contaminated at the time of slaughtering and resistant bacteria may transfer to human beings through food chain.
In present study prevalence of E. faecium recovered from retail chicken meat samples collected from various areas of Lahore city was estimated. A total 43 chicken sample (leg or wing) were processed for isolation of E. faecium. Identification of E. faecium was made using standard culturing and biochemical reactions. Out of 43 samples, 30 samples (69%) were found positive for Enterococcus faecium. Antibiotics resistance profiling showed that the isolates were resistant to following antibiotics mentioned as below: Ampicillin (100%) >Tetracycline (73%) > Erythromycin (53%) > Ciprofloxacin (46%) > Chloramphenicol (40%) > Rifampicin and Vancomycin (36%) > Teicoplanin (33%) > Doxycycline (20%) > Fosfomycin (0%). From the study it is concluded that retail chicken is the carrier of antibiotic resistant Enterococcus faecium and could transfer resistance to humans. Efforts should be made to use antibiotics wisely and hygienic practices should be followed during slaughtering and processing of chicken meat to avoid bacterial spread from animal source to human beings. Availability: Items available for loan: UVAS Library [Call number: 2489-T] (1).
6.
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).
7.
Studies On Biological Control Of Salmonellosis In Poultry
by Kiran Imtiaz (2010-VA-296) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Contaminated poultry food products are the main cause of Salmonellosis all over the
world. Salmonella may enter in different poultry farms through vertical or horizontal
transmission. Previously, Salmonella infection and number were controlled in poultry and
their products by using different methods such as usage of hazard analysis critical control
point (HACCP), antibiotics, symbiotics, etc. But not any method gives the better result that
shows the reduction of Salmonella species in poultry that cause infection. Therefore, the
attention was diverted to find the alternative therapies such as the usage of bacteriophage,
Bacteriophage specific to Salmonella used in reduction of Salmonella number as a biocontrol
agent in poultry origin. It proves effective for Salmonella reduction.
In the current study samples for isolation of Salmonella and bacteriophages were
collected. Salmonella was isolated from liver, caeca and lungs of infected chickens collected
from different infected poultry farms. After processing of sample according to the literature
method, confirmation of salmonella from samples were done by inoculating them separately
on S.S agar by streaking method. Further confirmation of Salmonella was done on the basis
of biochemical testing.
Bacteriophage was isolated from sewage water near to the poultry farms. For their
isolation, sewage sample was centrifuged and then supernatant was collected in separate tube.
Filtration was done of Supernatant, and then Filtrate was used as a bacteriophage source.
Bacteriophages in filtrate were confirmed by spot test method. After confirmation of both, in
vivo study was performed to evaluate the effect of bacteriophage on Salmonella when
inoculated in combination or separately in chickens groups.
51
Summary
Statistical analysis:
The data will be transferred on spreadsheet using MS Excel 2010. The data will be
analyzed through one way ANOVA test using Statistical Package for Social Sciences (SPSS)
version 18.0.
The present study was helpful to determine the effect of bacteriophage in reduction of
Salmonella in commercial broilers sector. Antibiotic resistant Salmonella infection is difficult
to control using conventional ways of antibiotic therapy and is responsible for huge economic
losses in poultry. Chicken groups that were used for invivo study showed that bacteriophage
was proved very effective in reduction of Salmonella either gives it in combination with
Salmonella or separately to poultry. It is predicted that bacteriophage therapy is better than
the conventional ways for reduction of Salmonella infection in poultry sector. It is essential
that the research should be continue to study the effect of bacteriophage in reduction of
specie specific Salmonella, and to determine the effect of different physicochemical factor on
their activity. Availability: Items available for loan: UVAS Library [Call number: 2511-T] (1).
8.
Microbiological Quality Assessment Of Human And Veterinary Drugs
by Sadaf Riaz (2007-va-277) | Dr. Ali Ahmad Sheikh | Dr. Fareeha Akhtar | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The continued increase in population results in increasing demand for pharmaceutical products. According to Pakistan pharmaceutical manufacturers association, Pakistan has approx. 400 pharmaceutical industries including 25 multinational industries. The Pakistan pharmaceutical industry meets 70% of country’s demand of pharmaceutical drugs.
Medicines are chemical compound that are administrated to human or animal as an aid to diagnosis, treatment or prevention of disease (Lecca, 1978).
Microbiological quality assessment is very important point of pharmaceutical manufacturing. The term quality in its wide sense means “Safety “. Microbial contamination means, presence of undesired micro-organisms or their metabolic products (Uba, 1990).
Pharmaceutical drugs are used in many ways in treatment of diseases (Mugoyela et al. 2010). Pharmaceuticals of different forms are susceptible to contamination by different microbes (Aulton, 2002). Contamination of medicines with micro-organisms can bring about changes in their physiochemical properties, results in product degradation before their expiry date (Shaikh et al. 2000). Microbial contamination rang from true pathogen like Clostridium tetani, to opportunistic pathogens, such as Pseudomonas aeruginosa (Mwambete, 2009). Microbial infections not only due to physical presence of micro-organisms, but also their metabolites/toxins can be very harmful even in low quantities (Shukla et al. 2004). Medicines used in treatments cannot afford to have microbial contamination. Patients who take medicines have very weak immune system which makes them vulnerable to wide range of infections. Microbial contamination of drugs may contribute to secondary microbial infections in immunologically weak patients (Adeshina et al. 2009). Contaminated pharmaceuticals may lead to medication complicacy in patients.
Drugs can be divided into two types on the basis of microbiology; sterile pharmaceutical products and non-sterile pharmaceutical products (Clement et al. 2013). Sterile pharmaceutical products should not contain any viable micro-organisms and non-sterile pharmaceutical products may contain viable micro-organisms but it should be within official limit and all drugs should not have any pathogenic bacteria. Most of drugs can be contaminated during storage and handling (Takon and Antai, 2006). Antimicrobial agents are added to medicines to minimize microbial growth but should not be added to mask poor manufacturing process (Gad et al. 2011).
The level of microbial contamination in medicines depends on availability of nutrients, water, presence of other micro-organisms, osmotic pressure and oxygen etc. The factors determining the results of drug-borne infections may include the type and amount of microbes, patient’s immune system and route of administration (Baird, 2004).
In recent years manufacturing of drugs have improved. The occurrence of microbes in drugs has been well documented. There have been many reports of infections caused by contaminated drugs (Ibezim, 2002; Coker, 2005). But majority of cases of infections caused by medicines are not recognized and reported as such (Denyer and Baird, 2006). Previous studies have showed microbiological quality concern related to commercially available drugs (Denyer and Baird, 2006).
Contamination of drugs results in loss of faith of a consumer in pharmaceutical industry and sales would go down (Mugoyela and Mwambete, 2010).
Although pharmaceutical industries are one of the growing and expanding sector in Pakistan, but the quality of medicines varies as they are retail oriented. Unfortunate situation is present in sales of drugs. A number of unlicensed drugs stores selling poorly manufactured drugs
2
Hence the microbiological quality of pharmaceuticals, assessment of number and type of bacteria and fungi within drugs, is essential to ensure consumer safety (Urmi et al. 2014; Tamalli et al. 2013)
This study to relate to national need and reflects its importance. New international trend focus on the quality of pharmaceutical products, unfortunately few studies has been carried out in this prospective which is not proven to be sufficient in predicting the quality of drugs. This has compelled me to do research on this topic in order to assess the level of such microbial contaminants in pharmaceuticals given to patients in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2275-T] (1).
