Brucellosis: A Threat to Human Population of District Rawalpindi

Objective: This study was conducted to detect cases of brucellosis among shepherds and to identify risk factors associated with human brucellosis. Study Design: Case control study Methodology:  A descriptive study followed by case control study was conducted during the month of August, 2016 at Village Hassar tehsil Taxilla, district Rawalpindi. A case was defined as “intermittent fever, profuse night sweats, headache and positive brucella antibodies on ELISA in a resident of Hassar from August 21-25, 2016. Epidemiological information was recorded on structured questionnaire. Cases and controls were matched by age and sex (1:4). Blood samples were collected from sheep/goat handlers (n=30) and small ruminants (n=144). Rose Bengal plate test (RBPT) and indirect Enzyme linked immunosorbent assay (I-ELISA) was used for testing of  serum samples. Frequencies were calculated, odd ratios were determined at 95% confidence interval with p value less than 0.05. Results: A total of six cases of brucellosis were identified. Among cases 42% were having direct  contact with small ruminants and 60% were raw milk consumers. Animal handler (OR =12 CL=1.19-123.6: p<.026) were likely to have brucellosis as compared to those who were not directly involved in animal handling. Persons consuming raw milk are more likely to have brucellosis (OR=11: Cl= 1.3-95: p<0.04) as compared to those consuming pasteurized milk. Among small ruminants tested, 52%  were found positive for brucellosis. Conclusion: Animal handlers/shepherds of district Rawalpindi were infected with brucellosis. Animal handler and raw milk consumer were more likely to get brucella infection. Infected small ruminant are potential source of infection for human. Presence of brucella infection in animal handlers/shepherd of Rawalpindi is suggestive of brucella infection all across the country. &nbsp

Comparative analysis of BTS-34 and Vero-76 cell lines for isolation of Peste des Petits Ruminants (PPR) virus

International audiencePeste de petits ruminant (PPR) is a highly contagious viral disease affecting predominantly sheep and goats. The virus belongs to the genus Morbillivirus accounting for causing immunosuppression and severe lymphopenia in host lymphoid cells principally due to the presence of a glycoprotein known as Signaling Lymphocyte Activation Molecule (SLAM) acting as cellular receptor. Limitations regarding the efficiency of PPR virus isolation using a variety of cell lines paved the way to the use of a new cell line (BTS-34) expressing these receptors. In the present study, a total of 18 PPRV confirmed strains were inoculated onto low passage Vero 76 and BTS-34 cell lines simultaneously to compare the susceptibilities of Vero and BTS cells for PPR virus isolation. The recovered viruses were then re-confirmed by Ic-ELISA and RT-PCR. In order to investigate the differences in titers of virus strains, serial tenfold dilutions were made separately using both cells at a density of 0.01x10(6) and tissue culture infective dose fifty (TCID50) was calculated. The results revealed that the characteristic pattern of the CPEs was more obvious in BTS-34 cell line marked by giant cells ultimately transforming into distinguished syncytia in 2 to 3 days. The average incubation time for virus isolation on Vero cells was about 10 days and syncytia formation were less marked. The growth curve indicated a one log increase in virus titer in case of BTS-34 cell line as compared to Vero-76. Statistically, there was significant difference between two types of cell lines in terms of number of days taken for PPR virus isolation as determined by single factor ANOVA (P<0.001). The study showed that BTS cells produced high virus titer with clearly distinct CPEs in short time due to the presence of SLAM receptors as compared to Vero cells suggesting the BTS cells to be more efficient and sensitive for PPRV isolation

Filter paper is a simple and cost-effective transport medium for serological diagnosis of Peste des petits ruminants

Peste des petits ruminants (PPR) is a highly contagious disease caused by peste-des-petits-ruminants virus. Following the successful eradication of the related rinderpest virus, a program to control and eradicate PPR was launched by the FAO and OIE. PPR is today present in many tropical countries where maintaining the cold chain for sample transportation is one of the major barriers for timely processing. Transport of samples on filter paper is a simple and cost-effective method, however validation and optimization is required to fully adapt this approach. The objective of this study was to evaluate and validate the use of filter paper in serological diagnosis of PPR. Blood samples (serum and filter paper) were collected from sheep and goats in both Tanzania and Pakistan and analysed using a PPRV-specific cELISA. The positive proportion was 10.7% in Tanzania and 80% in Pakistan when performing the analysis on serum. These results were then considered as reference and compared to the results from the filter papers analysed by the same cELISA. According to the statistical analysis the cut-off for a positive results for samples stored on filter paper was adjusted from &lt; 50 % competition percentage to &lt; 84% in Tanzania and to &lt; 69% in Pakistan.These results demonstrate that filter papers are an acceptable and cost-effective transport method of whole blood samples for later use in serological analysis

Supplementary Material.docx

Details of samples collection of Peste des Petits Ruminants Virus (PPRV) from different regions of Pakistan