Comparison Of Two Imported Live Attenuated PPR Vaccines In Local Sheep In Pakistan
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Peste des petits ruminants (PPR) also famous as goat plaque is of viral origin and is extremely contagious disease of sheep and goat (Dhar et al. 2002; Asim et al. 2009). PPR can cause high mortality about 50 – 80 % in non-immunized sheep and goat population. Due to its similarity with other diseases, Peste des petits ruminants (PPR) is being devalued but at the same time it is said to be one of the major constraints to successful small ruminant farming in tropics (Sen et al. 2010). PPR virus is paramyxovirus, enveloped and belongs to the genus morbillivirus. These viruses comprise of 16Kb long, single stranded RNA showing negative polarity (Barrett et al. 2005).
The various vaccines like homologous and recombinant vaccines have been manufactured for the management of Peste des petits ruminants (PPR), as no accurate treatment is available for its control. For the immunity of animals against this disease, the tissue culture based, attenuated rinderpest vaccine (TCRV) had been accustomed over a extensive period because of the antigenic association among RPV and PPRV (Diallo et al. 1989).With the help of fresh freeze-drying methods and stabilizing agents the thermostability of the present PPR homologous vaccine has been enhanced significantly (Worrwall et al. 2001).
In Pakistan, PPR vaccine was manufactured with the help of PPRV Nigerian 75/I (PPR 75/1 LK 6 Vero 75) for the sheep and goat immunization (Asim et al. 2009). India had manufactured numerous live attenuated vaccines like the PPRV Sungri/96 that has been regularized for use (Hegde et al. 2008). ). The Peste des Petits Ruminants (PPRV-Sungri/96 ) vaccine is being manufactured on small and large scale for prevention of Peste des Petits Ruminants (PPR) outbreaks in India (Singh et al. 2004).
The current study was designed to study the immunogenicity of two imported live attenuated PPR vaccines in local sheep. A total of sixty (60) animals were selected and further separated into two groups, viz. Group-A and Group-B, having thirty (30) animals each. Group-A was further sub-divided into A1 comprising 10 sheep to which Raksha PPR vaccine (Sungri 96) was administered, A2 comprising of 10 sheep to which PPR vaccine (Nigeria 75/1) was administered and A3 comprising of 10 non-vaccinated sheep which served as control. Group B was separated into two sub-groups i.e B1 and B2 having fifteen (15) animals each. The Group-B1 was sub-divided into B1a having 05 sheep to which Raksha PPR vaccine (Sungri 96) was only administered, B1b having 05 sheep to which along with Raksha PPR vaccine (Sungri 96), Vitamin AD3E was administered and B1c having 05 unvaccinated sheep which served as control.
Similarly the Group-B2 was sub-divided into B2a having 05 sheep to which PPR vaccine (Nigeria 75/1) was only administered, B2b having 05 sheep to which along with PPR vaccine (Nigeria 75/1), Vitamin AD3E was administered and B2c having 05 non-vaccinated sheep and served as control group respectively. The serum samples were collected and mean antibody titer was calculated by complement fixation test (CFT) at zero day, 7th day, 14th day, 28th day and 48th day post-vaccination.
The live attenuated, Raksha PPR (Sungri 96) vaccine induced the mean antibody titers of 0 ±0.00, 4.7±0.48, 4.7±0.48, 4.9±0.31 and 4.9±0.31 which was significantly higher than the mean antibody titers shown by the PPR (Nigeria 75/1) vaccinated animals i.e. 0±0.00, 3.3±0.51, 3.4±0.51, 4±1.15 and 4.1±1.19 at zero, 7th, 14th, 28th, 48th day post-vaccination respectively. Similarly the mean antibody titers shown by the PPR (Nigeria 75/1) vaccinated animals were 0 ±0.00, 10.4± 3.86, 11.2±4.13, 20±11.31 and 21.6±11.80 at zero, 7th,14th, 28th and 48th day post vaccination respectively. Result of present study demonstrated
that the mean antibody titer values of animals vaccinated with Raksha PPR (Sungri 96) was significantly higher than animals vaccinated with PPR (Nigeria 75/1) at zero, 7th,14th, 28th and 48th day post vaccination respectively. The study also concluded that the mean antibody titer of animals receiving vaccination along with vitamin supplementation was significantly higher than animals receiving only vaccination. While performing the statistical analysis of data, it was revealed that the results were significant (p<0.05).
The present study summarized and concluded that the mean antibody titer values of Raksha PPR (Sungri 96) was significantly higher than PPR vaccine (Nigeria 75/1). As both India and Pakistan are two neighbouring countries, so PPR among them also falls in trans-boundary disease category. It signifies that both being part of Asia subcontinent and PPRV strain of lineage IV prevails in both regions. Keeping these factors under consideration proper vaccination strategy should be followed for the immunization of animals. In past, Nigeria 75/1 strain of PPRV vaccine had been used in Pakistan but the results were not reliable in terms of desired immune response and protection. Although titer was shown by this vaccine but protection is not reliable for proper health care of small ruminants. There was an immense need to come up with the authentic research on PPRV vaccine Raksha PPR (Sungri 96) in Pakistan which is already being used in India with desirable results.
The results of present research project were mostly similar with the findings of other scientists. The results of this study were analyzed through Independent t-test for independent samples.
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Evaluation Of Risk Factors And Molecular Diagnosis Of Dermatophytosis In Dogs
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Publisher: 2016 Dissertation note: Dogs are most kept and beloved pets in Pakistani society. Dermatophytosis is among the common disease of the pets. Many predisposing factors are involved in development of clinical cases of dermatophytosis including climatic conditions, housing condition of dogs and physical attributes such as coat hair size. Dermatophytosis is not only of concern as being infection of pets but also of its zoonotic importance hence it is very crucial to diagnose dermatophytic infection well in time. Dermatophytosis is caused by Dermatophytes,Microsporum, Trichophyton and Epidermophyton, the fungal species. It is difficult to diagnose the Dermatophytosis from other skin infections by routine tests in most of the cases especially subclinical. Polymerase Chain Reaction (PCR) is advanced and the most reliable technique to detect genome of Dermatophytes even in minute quantities specifically and can efficiently detect the presence of any Dermatophyte specie on the skin of dog. The current study was planned to develop and validate a diagnostic assay which could be able to detect and distinguish tree important dermatophytes species including Microsporum, Trichophyton and Epidermophytonby a uniplex PCR reaction. Analysis of involvement of certain predisposing factors in dermatophytosis was second goal to be worked on in this study. Samples of suspected pet dogs (n=50) were collected by scraping the skin at affected areas over skin. DNA was extracted from the skin scraping samples by organic Phenol Chloroform Isoamyle Alcohol method. Primers, specific to the 18-S ribosomal RNA region of genomes of the Dermatophytes, were designed after alignment of available sequences of Microsporum,Trichophyton and Epidermophyton at NCBI. Annealing temperature and recipe of PCR reaction was optimized by gradient PCR in BIO-Rad thermal cycler. Amplification reaction of all samples collected was carried out as per optimized reaction conditions, afterwards. Amplified products obtained were subjected to genotyping by agarose gel electrophoresis for size based separation of the amplified products. The specific amplified bands of desired genomic region of dermatophytes were seen in UV light transilluminator. The data of results of predisposing factors involved in dermatophytosis wasanalysedby using Pearson’s chi squared test with the help of Statistical Package for the Social Science (SPSS) Program.
Genome specific product sizes of Microsporum and Trichophyton i.e. 366 bp and 351 bp in respective positive samples were observed. Out of 50 suspected samples 46 samples were positive for dermatophytosis out of which 38 samples (82.6%) were positive for Microsporum, 6 samples (13%) for Trichophyton and 2 samples (4.4%) were positive for both Microsporumand Trichophyton.
This study will help to validate a diagnostic technique for Dermatophytosis with greater efficacy and reliability. Moreover, this investigation may become basis for the future research activities in this field in Pakistan.
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Epidemiology Of Major Bacterial And Parasitic Causes Of Foal Diarrhea
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Publisher: 2017 Dissertation note: Present study was carried out in District Lahore and District Sargodha, Punjab province of Pakistan, from January, 2016 to December, 2016. The study was conducted to study the prevalence of Diarrhea in foals and to identify the major viral, bacterial and parasitic causes of diarrhea in foals in these districts. The foals that passed lose feces a least 4 to 5 times a day were considered diarrheic. The results showed that the prevalence of diarrhea was 72.8% in the foals. District wise prevalence showed that the prevalence of diarrhea in foals were 73.7% in district Sargodha and were 72% in District Lahore. According to the results the prevalence of diarrhea in male foals was 74% and in female foal were 72%. The diarrhea was more prevalent in donkeys at is 76.6% as compaired to horses which was 74.5%.
The viral (rotavirus), bacterial (Salmonella, Clostridium perfirengens and E. coli) and parasitic causes of diarrhea were identified by appropriate technique. The viral causes were diagnosed using ELISA technique. The bacteria were isolated by culturing and were confirmed by polymerase Chain Reaction (PCR). The parasitic causes studied using microscopic examination. To identify the cause of diarrhea 400 samples (200 from each district) were collected and processed for viral, bacterial and parasitic detection.
The results showed that 91.1% of the samples were positive for one or more infectious agents. District wise results showed that the prevalence of more or more than infectious agents were higher in district Lahore (95.5%) as compared to district Sargodha which was 87.5%. The isolation of one or more than one infectious agents were higher in males it is 92.7% while were low in females which was 90.5%. The results showed that the prevalence of one or more than one infectious agents were higher in horses (92.4%) in comparision with donkey which was 87.8%.
Experiment No. I: Investigation of Parasitic causes of Foal Diarrhea
Fecal samples were preserved in 10% formalin and transported to the laboratory for diagnosis of parasites. The fecal samples from foals suffering from diarrhea were processed by using following parasitological examination.
4. Direct microscopic examination
The sample negative with direct microscopic examination was examined using simple floatation examination.
5. Simple floatation examination
The sample negative with Simple floatation examination was examined by using sedimentation floatation technique.
6. Sedimentation floatation Technique
The sample negative by using Sedimentation technique was recorded as negative for parasites.
The results show that 340 (85%) out of 400 samples were positive for one or more than one endo-parasites. The prevalence of endo-parasites was higher in district Sargodha it is 87.5% as compared to district Lahore, which was 82.5% (Table No.7). Gastrodiscus Spp were the higher prevalent endo-parasite and 308 (77%) (Table No. 10) of the samples were positive for Gastrodiscus Spp while the lowest prevalent endo-parasite was Anoplocephala spp with (3) 0.75% prevalence (Table No. 12). other helmenth such as Dictyocaulus Spp. (22.5%), Oxyuris Spp. (15.75%), Strongyloides Spp. (15.75%), Ascaris equorum (4.75), Tridontophorus Spp. (2%), Trichomena spp. (1.5%) Strongylus spp. (1.5%), and Paranoplocephala Spp. (5%)
Experiment No. II: Molecular Diagnosis of Bacteria Causes of Foal Diarrhea
The samples were culture for Salmonella, E.coli and Clostridium perfirengins on respective selective media and DNA was extracted from the culture. DNA was amplified by PCR and the bacteria were confirm using PCR. To diagnose Lasonia the DNA was extracted directly from fecal sample and were processed for lawsonia. The result show that 55% of the samples were positive for one or more than one type of bacteria. Maximum prevalence were observe of E. coli 48.75% and none of the sample were positive for lawsonia. The other isolated bacteria were Salmonella 18.24% and Clostridium perfiengens 18%.
Experiment No. III: Investigation of Viral causes of Foal Diarrhea
Foal suffering from diarrhea were screened and analyzed for presence of rotavirus by using commercially available ELISA kit
The result of detection of rotavirus shows that rotavirus was detected in (70) 17.5% of the sample processed for the diagnosis of rotavirus.
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