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Peste des petits ruminant s (PPR), is a viral contagious and important transboundaiy animal disease (TAD), affecting domestic and wild small ruminants like sheep and goats, endemic in Indo-Pak subcontinent. The study was undertaken to characterize the indigenous PPRV by characterizing the fusion protein (F) gene segments by sequencing and their phylogenetic analysis for determination of genetic variation in the field viruses. Selected F gene segments of PPRV genome were amplified using RT-PCR from 110 tissues (lungs, lymph nodes and spleen) samples from dead animals and 41 swabs (nasal, occular and oral) from PPR suspected animals. The resulting amplicons were sequenced for phylogenetic analysis. The phylogenetic tree based on the F gene sequences of PPRV from different locations clustered them into lineage 4 along with the other isolates from Pakistan. Thus, genetic diversity of PPRV into the same lineages based on the F gene sequences showed the presence of lineage 4 emerged to give up the thought of molecular epidemiology for PPRV. [ABSTRACT FROM AUTHOR]

Mastitis continues to be a major economic issue for dairy producers in most parts of the world including Pakistan. Staphylococcus (S.) spp. are frequently isolated from bovine and bubaline mastitis. The indiscriminate use of antibiotics has led to the development of resistant strains of staphylococci and this factor contributed to the difficulties in its elimination from dairy herds leading to heavy economic losses to the industry. Milk samples were collected from mastitic buffaloes and cows from livestock farms around Faisalabad, Pakistan and one hundred were selected on the basis of surf field mastitis test. Isolation, identification and bacteriological examination were carried out through growth on blood agar, Grams' staining, catalase, clumping factor, coagulase and mannitol fermentation test. The purification of staph isolates was done on mannitol salt agar. Disc diffusion (oxacillin & cefoxitin) and nitrocefin tests were used to detect the production of ß-lactamase by staphylococci and hence called methicillin-resisitant staphylococci (MRS). The MRS were subjected to antimicrobial susceptibility testing through disc diffusion method against antibiotics including enrofloxacin (5(ig), norfloxacin (lOjig), amoxycillin (25|ig) and chloramphenicol (1 Ofag). Out of 100 mastitis smaples, 68 were carrying staph spp. including S. aureus (76.47%) and 5. epidermidis (23.53%). Nitroceffin test detected 30.76% as MRS. aureus (MRSA) and 25% coagulase negative methicillin resistant S. epidermidis. The disc diffusion test detected 32.69% as MRSA and 37.5% as MRSE. Nitrocefin discs applied directly to 25 milk samples, could detect -25%, while, disc diffusion test could identify 40% as MRS. Antimicrobial sensitivity test revealed that enrofloxacin was most effective followed by norfloxacin and chloramphenicol. While, amoxycillin was found resistant. The use of nitrocefin test proved better than disc diffusion method for the detection of MRS. Furthermore, aspect of methicillin-resistance must be considered while suggesting the treatment of mastitis due to Staphylococcus spp to minimize the losses. [ABSTRACT FROM AUTHOR]

Retinitis pigmentosa (RP) causes severe visual impairment early in life. The purpose of this study was to identify families with autosomal recessive RP (arRP), their exclusion analysis and identification of novel loci/genes/mutations in affected individuals. Twenty five consanguineous Pakistani families suffering from non syndromic Retinitis pigmentosa (RP) were ascertained from different areas of Pakistan to participate in this study. 85 affected and 210 normal individuals of selected families and from the healthy population took part in this research work. The clinical records of affected family members were retrospectively analyzed and ophthalmological examinations were performed in selected families. Genomic DNA was extracted from peripheral blood samples of phenotypically healthy and affected individuals. Exclusion analysis was done initially for the screening of the causative gene using highly polymorphic microsatellite markers. Genetic investigations using polymorphic microsatellite markers revealed homozygosity with CRB1 gene on chromosome 1 in affected individuals of family RP1. All other families and 210 healthy or control samples showed exclusion with the selected loci/genes. Haplotypes were constructed to appraise the results statistically. For further confirmation of linkage, sequence analysis of the linked gene (CRB1) was done. Sequence analysis revealed a novel missense mutation (?.1459?>?) in exon 6 of the CRB1 gene segregating with disease phenotype. This mutation resulted in a substitution of proline for serine at amino acid 487 (p. Ser487Pro). These findings suggest a founder effect of the p. Ser487Pro mutation in the Pakistani population and extend the spectrum of known CRB1 mutations. It was therefore concluded that the missense mutation in CRB1, leading to premature termination of the protein, is responsible forRP phenotype in the affected individuals ofRPl. [ABSTRACT FROM AUTHOR]