Uterine Microbial Flora Of Sahiwal Cattle During Oestrus And Its Relayionship With Pregnancy Rate
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation 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.
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Microbiological Quality Assessment Of Human And Veterinary Drugs
Material type: Book ; Literary form:
Publisher: 2015 Dissertation 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
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.
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Assessment Of Microbial Load, Protease Activity And Aflatoxin M1 In Raw And Uht Milk Procured From Local Markets Of Lahore
Material type: Book ; Literary form:
Publisher: 2015 Dissertation 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.
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.
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Seroprevalence And Risk Factor Analysis Of Bluetongue Virus In Lahore And Faisalabad Districts Of Punjab Province, Pakistan
Material type: Book ; Literary form:
Publisher: 2016 Dissertation 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,
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.
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Evaluation Of Antimicrobial Activity Of Essential Oil And Extracts Of Nigella Sativa Against Antibiotic Resistant Salmonella Enterica Isolates Of Human And Poultry Origin
Material type: Book ; Literary form:
Publisher: 2017 Dissertation 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
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.
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