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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: book Book; 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: book Book; 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: book Book; 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: book Book; 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: book Book; 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: book Book; 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: book Book; 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: book Book; 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: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1992,T] (1).

10. Study Of Molecular Diagnosis, Associated Risk Factors And Treatment Of Anaplasmosis In Bovines

by Muhammah Hassan Haider (2008-VA-252) | Dr. Jawaria Ali Khan | Dr. Muhammad Avais | Dr. Ali Ahmad Sheikh.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: According to the economic survey of Pakistan 2013-14, livestock sector contributed about 55.9 and 11.8% to the agriculture and national GDP, respectively.Total cattle population in the year 2013-14 is 39.7 million. Total milk and meat production contributed by cattle is 18,027000 tons and 18,87000 tons respectively (Anonymous 2013-14) . In Pakistan, the chiefhindrance in performance and health of animals are parasitic diseases including tick born. Ticks have been easily grown and reproduce in the optimal climate of tropical weather in Pakistan. In Pakistan, there are huge number of species and genera of tick fauna (Durrani and Shakoori, 2009). Tick-borne diseases include babesiosis, theileriosis, anaplasmosis, lymedisease . Among these tick-borne diseases anaplasmosis is one of the vital diseases which is responsible for the substantial economic losses in term of high morbidity, mortality and production losses that are decreased in milk, meat and other important livestock productions. Throughout the world, the “tick-borne diseases” are extensivelyspread in sub-tropic and tropicareasand Pakistan also includes in these areas(Khan et al. 2004). Moreover, anaplsamosis is non-contagious disease but also known to be infectious, transmission of which occur through mechanical means that includes the flies or tick bites. As well as different equipment that are using during tattooing or castration, in dehorning and needles also causes transmission (Aubry & Geale. 2011). Anaplasma marginale (A. marginale) is the major cause of Bovine anaplasmosis and this speciesbelongs to the genus Anaplasma (“Rickettsiales, Anaplasmataceae”). The most important 20 ticks species which cause transmission are Argaspersicus, Boophiludecoloratus, B. microplus, B. annulatus, Dermacentoroccidentalis, D. andersoni, D. variabilis, D.albipictus, Hyalommaexcavatum, Ixodesricinus, Ornithodoroslahorensis, Rhipicephalussimus, R. saniguineus, and R. bursa(Marchette & Stiller. 1982), on the other hand, Boophilusmicroplus found as a major contributor in cause of anaplasmosis (Tick Fever Research Centre, 1996). The family anaplasmatacease was classified in 1957 of the order Rickettsiale. In the genera Anaplasma, Aegyptianella, Haemobartonella and Eperythrozoon, this family was rationalized (Ristic&Kreier. 1984), and projected a combination based on analysis of genes sequences for protein on surface and ribosomal RNA (16S)(Dumler et al. 2001; Walker & Dumler. 1996). Anaplasma is a rickettsial, obligate intracellular bacteria found in red blood cells. Disease is characterized by fever, anemia and jaundice. Infection is transmitted by ticks or mechanically by biting insects or contaminated hypodermic needles or surgical instruments (Tylor et al. 2007).The most significant parasite is Anaplasma speciesthat is transmitted by at least twenty different species of ticks. But among those all mostly Bmicroplus causes Anaplasmosis (Rajput et al.2005). A. marginale is etiological agent of bovine anaplasmosis (Bram, 1957; Dumler et al., 2001; Kocan et al., 2000). A. marginale persistently infect both cattle and ticks and help as infection reservoirs(Kocan et al. 2003). Bovine erythrocyte membrane bound parasitophorous vacuoles is the site for replication of this obligate intracellular organisms. Dogs, humans, wild and domesticated ruminants are the species effected by A. marginale. This anaplasma genus is an obligate intracellular parasite of vertebrate hosts. In cattle, theA. marginal, A. phagocytophilum, A. central, and A. bovisare major pathogenic species producing diseases (Inokuma et al. 2007). Researchers suggestedthat in cattle, some unidentified Ehrlichia or Anaplasma species are present (Awadia et al. 2006). For this pathogen, which cause the persistent infection, the reservoirs are generally present in different host like tick or mammalian (Kocan et al. 2004). The wide range of different ruminant can be affected by Anaplasmabecause the species of anaplasma are not considered as strictly specific for any particular host specific(Kuttler. 1984). The A. marginalecan only be developed inside the RBCs of bovine (Richey et al. 1981). After invading the erythrocyte the tick multiplies and form almost eight initial bodies, they then get enlarge in the outer membrane and give it an appearance of a large dot. The organism gets mature in the outer membrane of RBC and force the infected RBC to get rupture due to infection. By rupturing the outer membrane where it previously multiplied come into the blood stream to infect the other RBCs. On the progression of progresses, more number of erythrocytes is infected by this organism and afterward destruction of erythrocytes occur (Stewart et al. 1981). Upon the clinical signs and symptoms are appear when the 15 percent of the RBCsare affected by the parasite(Radostits et al. 2006). The symptoms of the disease are pyrexia, anemia, icterus, hematuria, anorexia, muscular tremors, dyspnea, depression, lethargy, constipation and yellow colour of mucous membrane (Bram et al. 1983), low milk production, miscarriage and sometime mortality (Alderink and Dietrich. 1981). The mature RBCs are the site for multiplication after the invading by A.marginale. In the acute case of Anaplasmosis the higher number of RBCs are affected by the parasite for example more the 109 red blood cells per ml and after that the disease is categorized by death, abortion, loss of weight and anemia, and the survival cattle that are infected act as reservoirs for transmission of the disease in the herd level(De Echaide et al. 1998).The haematological profile can be used as useful diagnostic tool in anaplasma infection(Van Wyk et al. 2013).There is significant decrease in packed cell volume (PCV), Hemoglobin (Hb) and erythrocytes count.Serum biochemical analysis showed increased total protein,bilirubin and alanine aminotransferase enzyme (Sharma et al. 2013). A diagnosis ofcattle anaplasmosis may be made tentatively based on geographical location, seasonal variation and presenting clinical signs or necropsy finding in infected animals (Kocan et al. 2010). A.marginale,can affect throughout the life of cattle but disease severity is dependent on the age of the cattle. The clinical disease is less occurs in the calves and disease is very rare under the age of 6 month. The mild type of the disease can occur in the animal, which are between the ages of six month to one year. The acute type of the disease occurs in the animals between the ages ofone to twoyears but this disease is never fatal for the animal between these age groups.Whereas if the acute type of the disease occur in the adult animal that are more than two year aged can face the serious consequences and this disease can cause the death of the cattle with higher percentage (29 to 49%) of mortality (Kocan et al. 2003; Richey. 1991). Beside the animal age at the infection time, onceA. marginale infect animals, the animals become carrier for the infection throughout of animal life, with or without the clinical signs of the infection (Richey et al. 1991). The animal with the strong immune system are recovered from the acute type of anaplasmosis (Palmer et al. 1989). The regular cycle of ten to fourteen days are present in the carrier animals and in this cycle there are more or less number of RBCs are infected with the pathogen(Kocan et al. 2003;Kieser et al. 1990; Viseshakul et al. 2000). For the proper diagnosis of the disease and quality control in livestock, the most important aspect is the preciseor correct detection of pathogen that is spread by the tick.In 1990, with the beginning of the diagnosis with the techniques on the molecular bases the researcher produced more accurate and sensitive techniquesfor pathogenic species determination and these techniques are still used till to date (Ahantarig et al. 2008). The Giemsa staining method is used for the diagnosis of anaplasmosis in bovinesthat can be achievedby finding ofA.marginalein smear of blood from animals that are affected clinically. For the carrier animals or pre symptomatic animals thistechnique is not reliable. In these types of conditions, the serological tests are used for general diagnosis of infection by detection of antibodies. This is confirmed by molecular detection methods. A.marginaleappears as “dark staining blue purple bodies” measuring about 0.3 to 1 µm in length. A. centrale inclusion bodies are well differentiated by their location. Serological diagnosis can be done for the identification of infected cattle, DNA based test can be used for the molecular diagnosis(Stuen et al. 2011). The conventional parasitological technique like Giemsa staining always remained gold standard for diagnosis of Bovine anaplasmosis. Light microscopy of thin blood smears stained with giemsa stain may facilitate demonstration of A. marginale organisms in the erythrocytes (Kocan et al. 2004) .As giemsa staining method is not applicable for the detection of subclinical infection. Therefore PCR can also be used as a diagnostic tool for the detection of A.marginale in this technique primer pairs are used which resulted in the amplification of only their target DNA i.e. A.marginale(Munderloh et al. 2004). In the United states, for the treatment of anaplasmosis, the specific approved compound is the tetracyclines that include chlortetracycline or oxytetracycline(Kuttler. 1980). Oxytetracycline, Chlortetracycline and Tetracycline can be used to treat anaplasma infection in cattle .Among these Oxytetracycline is the most used drug at the dose rate of 10mg/kg body weight. While single dose of imidocarb dipropionte can be used at the dose rate 3mg/kg body weight (De Waal 2000).Imidocarb dipropionate with the dose of 3 milligram/kilogram body weight repidly treat A. marginale(anaplsmosis), on the other hand, same drug with double dose (6 milligram/kilogram body weight) and each dose has gape of two weeks,fail to cure A. marginalecarrier heifers(McHardy & Simpson. 1974). The animals recovered completely with imidocarb dipropionate (Akhter et al. 2010).The E elephantina and Aloemarlothiileaf extracts demonstrated good activity against rickettsia. These 2 plants along with Usanguineaand Rtridentata in certain concoctions are believed to be effective against anaplasmosis (Naidoo 2004). Availability: Items available for loan: UVAS Library [Call number: 2228-T] (1).

11. 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: book Book; 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).

12. 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: book Book; 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).

13. 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: book Book; 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).

14. Control Of Gastrointestinal Nematodes By Anthelmintics In Buffalo Heifers And Its Effect On Weight Gain

by Tanveer Saroia (2008-VA-062) | Prof. Dr. Khalid saeed | Dr. Nisar Ahmad | Dr. Ali Ahmad Sheikh.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Parasitic problem is a major constraint of ruminant production and causes great economic losses to dairy industry by retarded growth, low productivity and increased susceptibility of animals to other infections. The parasitic infections cause productivity losses through reduced feed intake and decreased feed efficiency. Even subclinical or chronic infections are responsible for economic losses in large ruminants. Hypothesis of the study was that administration of anthelmintics will improve weight gain in infected buffalo heifers. One hundred faecal samples of buffalo heifers of age 1-2 years were screened and 58 were found positive for gastrointestinal nematodes. The species of nematodes found prevalent in these animals were Trichostrongylus, Oesophogostomum, Haemonchus, Toxocara, Trichuris and Ostertagia. Thirty buffalo heifers with high EPG were selected and divided into 3 equals groups A, B and C. In this study Group A was a positive control group. Group B was administered Ivermactin (Bomectin) pour on at the dose rate of 1ml / 20 kg body weight. Group C was administered Oxfendazole (Systamex) orally at the dose rate of 1ml/ 10 kg body weight. The effect of these anthelmintics on EPG and weight gain were recorded after every 15 days. This study period was of 90 days (3 months) duration. There was increases in the EPG of positive control group and percent increases was 7.10%, 10.65%, 18.10%, 30.17%, 33.72%, 46.15% at days 15, 30, 45, 60, 75and 90. The percentage decrease in EPG for the group B was 98.22%, 97.04%, 91.12%, 77.51%, 60.94%, 46.74% at days 15, 30, 45, 60, 75 and 90 post treatment and the percentage decrease in EPG for the Group C was 95.80%, 92.81%, 88.62%, 71.85%, 58.68%, 26.34% at days 15, 30, 45, 60, 75 and 90 post treatment. Both anthelmintics cause significant reduction in epg medicated animals as compared to non-medicated animals. There was no difference in the two anthelmintics. The average weight gain per day for Group A (Positive Control) is 0.17 kg per day, for Group B (Ivermectin) is 0.44 kg per day and for Group C (Oxfendazol) was 0.4 kg per day. This shows significant difference between treated groups with non-treated group but there was no difference between the treated groups. It is concluded that ivermectin and oxefendazol can be used for the control of gastrointestinal nematodes. It is recommended that animals may be moved to clean facility after treatment so they are not reinfected. Repeated treatment is suggested to maintain the EPG at low level. Suggestions of the present study are 1. Monitoring of animals for gastrointestinal nematodes by modified McMaster Egg Counting technique. 2. Regular deworming 3. Improved management Availability: Items available for loan: UVAS Library [Call number: 2405-T] (1).

15. 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: book Book; 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).

16. 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: book Book; 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).

17. 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: book Book; 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).

18. 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: book Book; 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).

19. 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: book Book; 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).

20. 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: book Book; 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).

21. Prevalence And Associated Risk Factors Of Needle Stick Injuries Among Health Care Workers In Government Tertiary Care Hospitals Of Lahore

by Gurdeep Singh (2014-VA-957) | Dr. Mamoona Chaudhry | Dr. Muhammad Hassan Mushtaq | Dr. Ali Ahmad sheikh.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: In present study, seventy six percent (n=228) of respondents were involved in sharp injuries and twenty four percent (n=72) were not involved. This study concluded there was high occurrence of needle stick sharps injuries among healthcare workers in tertiary care hospitals of Lahore. There are several risks associated with handling healthcare sharps among healthcare workers which include, needle prick that could be contributed to cases of occupational infections such as HIV/AIDS. Majority of reported cases were due to following reasons , inadequate supply of barrier products ,unsafe practice such as recapping of needle In paramedical staff due to failure in standard guide lines to handle medical sharps and lack of reporting of sharps incidents and very lower ratio utilization of post exposure prophylaxis. It was observed that risks of needle stick injuries were reduced with the experience. Those who have work experience of 5 years or less had higher ratio of sharp injuries than those who had work experience five to eight years. Respondents with nursing profession and laboratory technicians were at high risk; their exposure to handle sharps is greater and due to failure in precautionary measure risk of needle stick injuries in this category is higher compared to other. While Females were at high exposure risk of sharp injuries at tertiary care hospitals than male. Respondents with education level of diploma were more exposed to needle stick injuries as compared with degree holders in paramedical staff. It was observed that shift of eight hour work is absolute and healthcare workers were normal with it. If work load increased and time was exceeded then it was difficult to manage such injuries .while training on infection control and prevention has no effect in government hospitals of Lahore to decrease the risk of sharps injury among healthcare workers. Several recommendations which need to be implemented at the hospitals were identified as discussed below. and special attention need to be given to nurses , cleaners , the critical care section and the department of medicines There is need of fully trained Summary 38 health care workers in critical care units , medical and surgical ward Momentary to developed infection control department in every government and private hospital of Lahore and give them special training about the handling of medical sharps. adequate supply of barrier products (gloves, masks, lab coats etc) Check and balance on proper use of barrier product , there is need of timely collection of used medical sharps in puncture proof containers such as safety boxes that should be ¾ full when emptying .Need of safe transportation in a covered vehicle for incarnation .Administration of hospital need to developed polices for reporting of needle stick injuries by designing a log form that capture information on demographic data of the employee. Data and time of injury, type of sharp, procedure involved, part of body involved where it occurred. So it’s important to analyzed enter data in an order to understand root cause. It’s important to conduct surveillance and prevent further injury. Infection control department should need to check post-exposure prophylaxis status in health care workers go through counseling and are followed up to enhance post exposure prophylaxis in health care workers it’s not up to individual effort it’s a team work by involving administration and health care workers.. In government hospitals of Lahore there is many factors which contribute to needle stick injuries it might be reduced by strengthening of reporting system is necessary. The responsibility of government is to strengthen the check and balance on use of barrier products (gloves, pads, lab coats etc.) and create awareness on standard guidelines on handling of medical sharps. Thus rate of needle stick injuries could be reduced to acceptable level. Availability: Items available for loan: UVAS Library [Call number: 2748-T] (1).

22. 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: book Book 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).

23. 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: book Book; 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).

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