1.
Uterine Microbial Flora Of Sahiwal Cattle During Oestrus And Its Relayionship With Pregnancy Rate
by Habib- Ur- Rehman | Prof. Dr. Masood Rabbani | Dr. Ali Ahmad Sheikh | Prof. Dr. Nasim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2011Dissertation note: In the present study uterine microbial flora of Sahiwal cattle during oestrus and its relationship with pregnancy rate was determined. According to the results a total of 11 bacterial species were isolated from 50 uterine samples of estrus Sahiwal cattle, maintained at Livestock Production Research Institute (LPRI), Bahardur Nagar, district Okara, Punjab province, Pakistan. The isolates include E. coli, Micrococcus spp., Bacillus spp., Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp., Pseudomonas spp., Citrobacter diversus, Salmonella spp., Proteus spp. and Corynebacterium spp. Tabulation of results showed that prevalence of these isolates was different among pregnant and non-pregnant animals. Moreover, E .coli, Micrococcus spp., Bacillus spp., Staphylococcus aureus and Citrobacter diversus are found to be thriving in uterus as normal microbial flora, whereas, Streptococcus spp. isolate as abnormal microbial flora appearing to be having some role in decreasing pregnancy rate. While, Pseudomonas spp., Corynebacterium spp. Staphylococcus epidermidis, Salmonella spp., and Proteus spp. Isolates could not be differentiable as normal and abnormal uterine microbial flora due to insignificant available data. Furthermore, complete blood counts of 50 blood samples of these same animals indicated that those animals harboring isolates like Streptococcus spp., Pseudomonas spp., and Corynebacterium spp. in their uterus, had more likelihood of abnormally increased value of Mean Corpuscular Volume (MCV) than to presence of any other bacteria. But due to lower data of Pseudomonas spp., and Corynebacterium spp isolated from total samples, only Streptococcus spp. seemed to be ranked as abnormal in Pakistani Sahiwal cattle cows. Interestingly all those animals from where Corynebacterium spp. was isolated, were showing increased values both of MCV and HCT (Hematocrit) which is indicative of their pathogenic role in causing uterine infections.
On the basis of this study it can be modestly concluded that uterine microbial flora identification may serve as a better tool in assessing and foretelling the reproductive health status of the breeding animals. After necessary assessment, presence of any harmful microbial flora or pathogen can be effectively treated through either selecting an appropriate antibiotic by using culture sensitivity testing or by using any suitable bactericidal agent thereby help in boosting conception and pregnancy rates.
Availability: Items available for loan: UVAS Library [Call number: 1293,T] (1).
2.
Characterization Of Antimicrobial Resistance In Escherichia Coli Recovered From Retail Chicken In Lahore
by Fayyaz Yasin | Dr. Ali AHmad Sheikh | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Escherichia coli is normal inhabitant of lower intestinal tract of all warm blooded animals. The background of drug resistant studies on E. coli represent that extensive and irrational use of antimicrobials in human and veterinary sector for treatment, prophylaxis and feed additive made this organism resistant to commonly used antimicrobials.
It is assumed that E. coli have the ability to transfer resistant genes via bacterial conjugation, transduction and transformation. As a result pathogenic organisms are also becoming resistant to commonly used antibiotics. And there is a need to check the extent of resistance to ensure the efficacy of antimicrobials used in public health.
The purpose of current study was to estimate the prevalence of antimicrobial resistant E. coli in retailed chicken meat samples of Lahore city. In current study hundred chicken meat samples were collected from local market in various areas of Lahore.These samples were processed for isolation of generic E. coli. Initial confirmation of generic E. coliwas made using standard culturing and biochemical reactions. The prevalence rate of E. coli was found 85%. The antimicrobial resistance against in the E. coli isolates was determined by disk diffusion method. All the E. coli isolates were found resistant to at least seven antimicrobials and 34 different antimicrobial patterns were found.
Availability: Items available for loan: UVAS Library [Call number: 1477,T] (1).
3.
Antibacterial Activity Of Herbal Extracts Against Multi-Drug Resistant Escherichia Coli Recovered Form Retail
by Arfat | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Escherichia coli is normally present in lower part of intestinal tract of all warm blooded animals. The background of drug resistant studies on E. coli represents that extensive and irrational use of antimicrobials in human and veterinary sector for treatment, prophylaxis and feed additive made this organism resistant to commonly used antimicrobials. Retail chicken meat is one of the major sources of spread of MDR E. coli infections in humans.
The purpose of present study was to evaluate the prevalence of multi drug resistant E. coli recovered from retail chicken meat samples collected from various areas of Lahore city. Also study the effect of antibacterial activity of selected herbal extracts against isolated MDR E. coli. In current study 100 E. coli isolates were processed for isolation of generic E. coli. Identification of generic E. coli was made using standard culturing, biochemical reactions and confirmed through PCR. The recovery rate of E. coli was found 80% and multidrug resistant pattern in the E. coli isolates was determined using disk diffusion method. A total 73.86% of E. coli isolates were found resistant with at least three antimicrobials related to different groups.
Current study revealed the effectiveness of herbal extracts against MDR E. coli. Clove, cinnamon and mint have good antibacterial activity as compared to coriander, kalonji and garlic. Hence, these herbal extracts can be used as promising alternatives of antimicrobials against multiple drug resistant E. coli species.
Availability: Items available for loan: UVAS Library [Call number: 1606,T] (1).
4.
Isolation, Identification And Control Of Vancomycin Resistant Staphylococcus Aureus
by Fakhra Liaqat | Dr. Ali Ahmad Sheikh | Dr. Jawad Nazir | Dr. Tanveer Hussain.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Antimicrobial resistance (AMR) is an emerging issue throughout the world. Vancomycin resistant Staphylococcus aureus has been recently reported from many countries including Asian region. The failure of antibiotics diverts the focus of modern science towards the discovery and application of new alternative antimicrobial agents. Herbal plants not only known for their antimicrobial potential but are being used since centuries for the treatment of infections. This study has been conducted to isolate Vancomycin Resistant Staphylococcus aureus from wounds of hospitalized patients and these isolates were not only tested against Linezolid, Moxifloxacin, and Clindamycin antibiotics but also against the ethanolic extracts of Garlic, Mint, Coriander, Turmeric, Kalonji, Cinnamon and Cloves for their antibacterial activity.
Methicillin resistant Staphylococcus aureus were isolated from wounds and resistance to vancomycin was confirmed according to Clinical and Laboratory Standards Institute recommended method. Minimum inhibitory concentrations of selected antibiotics and selected plant extract was determined by broth microdilution method followed by the measurement of minimum bactericidal concentration by culturing on agar plates.
In current study 104 S. aureus isolates were recovered from 150 wound exudates samples. Resistance to methicillin was shown by 49.04% isolates. Final results yielded, 22 vancomycin intermediate and 5 vancomycin resistant S. aureus strains. Vancomycin resistant isolates were tested for susceptibility against selected antibiotics and ethanolic extracts of plants. Almost all the isolates displayed susceptibility to all three antibiotics and the plant extracts. The data was analyzed statistically by chi square test and one way ANOVA using Statistical Package for Social Science (SPSS) 18.0 software. This study was helpful to find out the effective antibiotics against Vancomycin Resistant Staphylococcus aureus. Plant extracts which were found effective are the best alternate to the conventional antibiotics without having any drawback of antibiotic resistance development.
Availability: Items available for loan: UVAS Library [Call number: 1615,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.
Correlation Of Deifferent Managment Systems And Facilities Of Retail Milk Shops With That Of Microbial Load In Raw And Pasteurized Milk
by Faria kanwal | Prof. Dr. Masood rabbani | Dr. Ali ahmad sheikh | Dr.Wasim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1953,T] (1).
7.
Characterization Of Beta-Lactamase Producing Genes From The Chicken Meatborne Antimicrobial Resistant
by Ayesha tabassum | Dr. Ali ahmad sheikh | Prof. Dr.Masood rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1955,T] (1).
8.
Detection Of Gram Positive Foodborne Pathogen From Retail Quail Meat Through Optimized Multiplex Pcr
by Iqra Safdar | DR. Ali ahmad sheikh | Dr.Tanveeer | Ms.Fareeha akhtar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1956,T] (1).
9.
Comparative Study On Prevalence And Antimicrobial Susceptibility Pattern Of Extendedspectrum B- Lactamase
by Karam rasool | Prof. Dr. Masood rabbani | Dr. Ali ahmad sheikh | Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1992,T] (1).
10.
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).
11.
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).
12.
Assessment Of Microbial Load, Protease Activity And Aflatoxin M1 In Raw And Uht Milk Procured From Local Markets Of Lahore
by Sadaf Almas (2007-VA-250) | Dr. Imran Altaf | Dr. Ali Ahmad Sheikh | Dr. Muhammad Nasir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is among the largest milk producing countries. The requirement of milk is
increasing day by day. It has rapidly increasing demand and competition in national and
international markets. Milk consumers in Pakistan often face low-quality, lack of hygiene and
absence of cold chains as primary contributors to this low quality. Milk and dairy products also
become contaminated during manufacturing and packaging processes. The consumption of low
quality milk may cause milk borne diseases. Not only the bacteria but the presence of their
enzymes also can cause deterioration of the milk quality. The heat stable enzymes can cause the
spoilage of commercial UHT products without presence of any viable count. Aflatoxin (M1) is
metabolite of AFB1 in milk. It causes chronic diseases and immunosuppression in children. It is
found carcinogenic and cytotoxic in nature.
In this project microbial load, free amino acid estimation to predict any protease activity
and Aflatoxin M1 were studied in both UHT and Raw milk samples (n=15) procured from local
markets of Lahore. Three UHT brands A, B and C were purchased. The UHT milk was studied
for microbial growth and protease activity at purchase and at expiry of the products. The
microbial load was evaluated by testing of milk for Total viable count, Coliforms, Yeast and
Molds, Anaerobic Clostridia and Bacillus cereus in both raw and UHT milk. Protease activity
was estimated by assessing the free amino acid by using ninhydrin assay while the Aflatoxin M1
was detected through High performance liquid chromatography.
SUMMARY
80
CONCLUSION:
It was found that the locally available raw milk quality was poor for consumption and dairy
processing for safe and stable milk products. UHT milk quality was found better with low
microbial load. Protease activity with reference to free amino acid was detected in raw milk
which is indication of the poor milk storage conditions, cold chain maintenance and
unavailability of fresh milk. Protease activity was also found in UHT milk and an increase in free
amino acid which could be due to heat stable proteases active during shelf life of the milk
brands. Aflatoxin M1 was found in majority of raw milk sample which showed the poor animal
feed storage and monitoring system. Aflatoxin M1 was also found in some samples of UHT
brands with high concentration which depicted that AFM1 was heat stable and it retained in the
commercial UHT products as well. Availability: Items available for loan: UVAS Library [Call number: 2354-T] (1).
13.
Antimicrobial Effect Of Various Herbs On Sore Throat Causing Antibiotic Resistant Bacterial Isolates
by Abida Mushtaque (2010-VA-298) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Muhammad Nasir .
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Sore throat or pharyngitis in childrenis mainly caused by group A (β-hemolytic) Streptococcus pyogenes. The bacterium transferred from infected person to susceptible one through coughing, sneezing and by direct contact. Irrational use of antibiotics to control sore throat infection results in development of antibiotics resistance. To overcome the problem of antibiotic resistance, various herbal extracts(Liquorice, ginger, cinnamon and red dates)are used which have known antimicrobial activity against these bacterial isolates.
In current study 70 throat swab samples were collected from Children Hospital, Lahore and processed to isolate and identify antibiotic resistant sore throat bacteria by swabbing on blood agar andTryptic soya agar.Initial identification was done using conventional biochemical tests and confirmation was done through API 20 Strep. Antibiotic resistance pattern in isolated bacteria was done by modified Kirby Bauer disc diffusionas per CLSI,2014 criteria.Extracts (Ethanolic, boiling and distilled water) of herbs (Liquorice, ginger, cinnamon, red dates and kalonji) was obtained and tested for antimicrobial activity against resistant isolates. Efficacy of the herbal extracts was evaluated through Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentrations. Ethanolic extracts of Liquorice, ginger, cinnamon and red dates have good antimicrobial activity against resistant sore throat bacterial isolates than boiling and distilled water extracts.
The present study demonstrated that S. pyogenes and Staphylococcus are major bacteria responsible for sore throat infection in children. These isolates have variety of different antibiotic resistance patterns. In order to minimize emergence of antibiotic resistant isolates, herbal extracts can be used.Ethanolic extracts of liquorice, ginger, cinnamon, red dates have good antibacterial activity as compared to kalonji. Statistical analysis showed that ethanolic extracts of Liquorice has a significant activity with ginger, cinnamon and red dates.The degree of antibacterial activity of plant extracts testedcan be graded in the following order: Liquorice> ginger>Cinnamon>red dates>kalonji. Itis estimated that by using natural products will reduce the resistance in microorganism.
Statistical analysis:
The data collected in MS Excel 2016 and analyzed statistically by one way Analysis of Variance using Statistical Package for Social Science (SPSS) 18.0 software.
Availability: Items available for loan: UVAS Library [Call number: 2488-T] (1).
14.
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).
15.
Seroprevalence And Risk Factor Analysis Of Bluetongue Virus In Lahore And Faisalabad Districts Of Punjab Province, Pakistan
by Syeda Marriam Maqbool (2014-VA-522) | Dr. Muhammad Zubair Shabbir | Dr. Ali Ahmad Sheikh | Dr. Maryam Javed.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Domestic animals play an important role in the rural and agricultural economies of developing countries. Therefore, animal diseases pose a threat to country’s economy, animal welfare, the environmental and public health. One of the important animal diseases is Bluetongue, listed as notifiable disease by OIE. Causative agent is Bluetongue virus (BTV) an arbovirus that belongs to genus Orbivirus with in family Reoviridae. The main route of transmission is through the bite of Culicoides biting midges. Disease is enzootic and widely distributed in areas where susceptible animals and vector species are prevalent. It has been reported worldwide including the neighboring countries of Pakistan. BTV is also considered an endemic in Pakistan but little information is available on its epidemiology in this area. Serological tests can detect antibodies produced against infection and helpful to analyze the prevalence of a pathogen in circumstances when there lacks vaccination practice to ruminants in a given geographical area.
Competitive ELISA was used to identify antibodies to BTV in the sera samples of animals in Faisalabad and Lahore districts. Blood samples were collected from randomly selected villages of both districts and processed for serum separation by using gel/clot activator tubes. Separated serum was analyzed by competitive ELISA. Further, statistical analysis was done by OpenEpi to check the association between BTV seroprevalence and potential risk factors. Later the BTV prevalence has been mapped in relation to different villages of both districts.
Results of present study revealed that Bluetongue virus is prevalent in Faisalabad and Lahore districts with high seropositivity observed for Faisalabad district. Antibodies to BTV were detected in all studied animals irrespective of their age, sex, parity and breed. Risk factor analysis is implicating the association of BTV seroprevalence with breed, sex and age for sheep,
SUMMARY
44
cattle and buffalo respectively. Further studies should be conducted to expand the geographical area for the assessment of Bluetongue prevalence and to explore the genetic diversity of Bluetongue virus. Availability: Items available for loan: UVAS Library [Call number: 2497-T] (1).
16.
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).
17.
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).
18.
Effects Of Storage Temperature On The Microbiological Quality Of Fish Meat
by Rida Haroon Durrani (2010-VA-307) | Dr. Fareeha Akhtar | Dr. Ali Ahmad Sheikh | Dr. Mati- ur- Rahman.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Fish is an affluent resource of various significant nutritional constituents such as high- quality protein, vitamins, minerals and n-3 rich polyunsaturated fatty acids. Sea foods are categorized as one of the briskly perishable food items than other muscle foods, due to their high water activity (Aw), existence of self-degrading enzymes and neutral pH value which is the cause of quick production of unwanted whiff and flavors. Present research was designed to study the effect of storage temperature and associated spoilage of fish meat through various pathogens of bacterial nature. Samples were procured from two different managemental systems to observe the effect of storage place on bacterial load. Effect of variations in seasonal temperature were also observed on the bacterial load by collecting samples in three different seasons of the year i.e. winter, spring and summer season. Total viable count was performed along with the selective culturing of bacterial species capable of causing gastrointestinal disorders. Statistical analysis reveals that a significant difference (p< 0.05) exists between seasonal variations and patterns of bacterial types. With respect to storage up to 8 hours a significant difference (p< 0.05) is observed with the existing bacterial types. A significant difference (p< 0.05) lies between bacterial types and the source from which samples were procured. A significant difference (p< 0.05) was observed between sampling season and storage duration. While a non-significant difference (p> 0.05) was recorded between seasons and source from where samples were collected. With respect to storage time a significant difference (p< 0.05) was recorded with the sources of samples. While comparing the patterns of bacterial types in samples collected in different seasons of year a significant difference (p<
Summary
90
0.05) exists with respect to storage time. While a significant difference (p< 0.05) was observed
between growth patterns of bacterial types with respect to seasonal variations and source. With
respect to the storage time there lies a significant difference (p< 0.05) to the bacterial types
observed in samples procured from two different management systems. With respect to season
a significant difference (p< 0.05) was noticed with storage time and source of sample. While
significant difference (p< 0.05) was present between the bacterial types isolated during
different seasons of the year and subjecting them to different storage time and the source of
samples. Availability: Items available for loan: UVAS Library [Call number: 2672-T] (1).
19.
In Vitro Biological Control Of Avian Pathogenic Escherichia Coli
by Aleena kokab (2011-VA-418) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Waseem Shehzad.
Material type: Publisher: 2017Dissertation note: Avian pathogenic E. coli (APEC) has meticulous virulence properties making it a
potential pathogen causing insidious infections in poultry, termed as colibacillosis either
as primary pathogen or as a secondary pathogen. All over the world, there are major
economic losses in poultry industry due to this disease. To prevent APEC infection,
strategies include improving hygienic methods, vaccination, antibiotic treatment and
introduction of novel immunopotentiators but all these measures had limited success.
Moreover, because of extensive use of antibiotics for treatment purposes, antibiotic
resistance is major growing concern. Bacteriophage therapy is promising new alternative
to antibiotics. There are many benefits of phages over antibiotics and are now used in
research for the treatment of enteric and respiratory problems in poultry and are proved to
be effective in reducing E.coli infection.
In present study bacteriophages was used to minimize and control the number of
E.coli causing colibacillosis in poultry. For this, E.coli (n = 10) from liver of infected
commercial poultry birds and E.coli (n = 5) from intestines of apparently healthy birds
was isolated and characterized using biochemical tests. Final confirmation of E.coli
isolates was done by PCR. To differentiate between pathogenic and non-pathogenic
E.coli, Congo Red Dye binding test (CR test) was performed. Antibiotic resistance
profiling of the isolates was also determined against 10 commonly used antibiotics
through Kirby-Bauer disk diffusion method.
For the isolation of bacteriophages, sewage or poultry sludge sample was
processed according to literature method and purified bacteriophages (n = 5) were
Summary
63
assessed for their stability at various temperature and pH. Bacteriophages were also
assessed for their In-Vitro Lytic Activity of against E. coli isolates.
Five numbers of lytic bacteriophages were isolated and purified against avian
pathogenic E.coli, showing resistance against Ampicillin, Doxycycline, Ciprofloxacin,
Neomycin, Trimethoprim-sulphamethoxazole, Nalidixic acid and Ceftriaxone. Isolates
were also classified into pathogenic and non-pathogenic on the basis of their ability to
bind Congo red dye on Congo red media. Phages were highly lytic against pathogenic
isolates while there was no lytic activity against non-pathogenic isolates thus making
them unique for in vivo trials. Phages were found to be stable at temperature from 25oC
to 56oC with highest stability at 37oC while on the basis of pH, phages were stable
between 5-9 pH with highest stability at pH 7. Maximum lytic activity observed was for
12 hours and then there was emergence of resistant bacteria and this problem may be
tackled by increasing MOI or by using combinations of phages against resistant bacteria.
Present study helped to reveal the effectiveness of bacteriophages against antibiotic
resistant E.coli and proved that bacteriophages can be used as promising alternatives to
antibiotics for reducing E.coli infection in poultry. Availability: Items available for loan: UVAS Library [Call number: 2829-T] (1).
20.
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).
