​​Screening Cowpea Genotypes for Resistance to Cowpea Aphid Borne Mosaic Virus (CABMV) and Cowpea Severe Mosaic Virus (CPSMV) in Paraguay

Background: Cowpea [Vigna unguiculata (L) Walp.] is one of the Paraguayan rural families’ main crops, serving as an essential protein and carbohydrate source. Cowpea aphid borne mosaic virus (CABMV) and cowpea severe mosaic virus (CPSMV) were identified infecting cowpea plants. Disease control caused by both viruses is difficult because there is no information about local cowpea resistant cultivars and vector control is not practical. Methods: In the present work, sixteen cowpea genotypes/cultivars were mechanically inoculated with local isolates of CABMV and CPSMV to identify resistant genotypes/cultivars that can be used in breeding programs. Virus infections were determined by symptoms expression and confirmed by PTA-ELISA. Result: Genotypes Arroz rojo (V. angularis), TVu 379, TVu 382, TE94-256-2E and TE97-309G-9 were resistant to CABMV. Genotypes Arroz rojo (V. angularis), CNCX-698-128F, TVu 379, TVu 382, TVu-3961, TE97-309G-9 and TE97-309G-3 were resistant to CPSMV. Overall, this study showed that local cowpea cultivars do not offer any resistance to virus infection and the need for resistant germplasms for cowpea breeding programs in the country.

↵Peptide based Indirect ELISA for Detection of Antibodies against Peste des petits ruminants virus

Background: Infectious virus antigen is not recommended for disease monitoring in global Peste des petits ruminants eradication strategies. Native virus antigens are gradually being replaced with recombinant or synthetic peptide antigens. The focus of the present study is to optimize and develop peptide-based immunoassay for the detection of antibodies to PPRV N and H proteins. Methods: Epitopes of PPRV H and N proteins were selected based on prediction with bioinformatics tools and from previous studies. Two peptides each were synthesized for N and H proteins and peptide ELISA developed. The peptide ELISA’s sensitivity and specificity were tested with sera samples collected at different time intervals of vaccination (goat =73, sheep= 62) and 88 random serum samples (goat =47, sheep=41). The collected sera were screened using cELISA before proceeding to peptide ELISA. Result: In competitive ELISA, 106 goat serum samples and 96 sheep serum samples were found to be positive. Fourteen goat serum samples and seven sheep serum sample were shown to be negative. Among120 goat serum samples tested, 114 were found to be positive by peptide ELISA. Similarly, out of 103 sheep serum samples analyzed, 96 were found to be positive with peptide ELISA. The peptide ELISA based on the highly conserved and antigenic N and H epitope detected antibodies to PPRV in precise manner. This study demonstrated the effective use of synthetic peptides as an antigen in the detection of antibodies to PPRV.

Increased interleukin-6 levels in neuron-derived plasma small extracellular vesicles of subarachnoid haemorrhage patients

Background: Aneurismal subarachnoid hemorrhage (aSAH) is a serious type of stroke with high mortality and disability. Identifying circulating biomarkers is helpful to improve theranostics of aSAH. In this study, we are for the first time to report circulating interleukin-6(IL-6) in neuron-derived small extracellular vesicles(NDSEVs) were identified to be the potential biomarkers in prognosis of aSAH. Methods: We extracted small extracellular vesicles from the plasma of aSAH patients and healthy controls and were enriched by sequential precipitation and anti-L1CAM antibody immunoabsorption. Subsequently,we determined IL-6 levels by an enzyme-linked immunosorbent assay (ELISA). Result: Plasma IL-6 NDSEVs showed distinct pattern differences between aSAH patients and healthy controls. The IL-6 NDSEVs levels were increased and positively associated with disease monitoring and prognosis of aSAH patients. These data suggest an elevated neuroinflammatory cascade in aSAH patients. Conclusion: The IL-6 NDSEVs maybe prospective biomarkers to indicate its progression, and thus may own great potential in applications such as prognostic evaluation of aSAH in the near future.

Presence of Antibodies against Bluetongue Virus (BTV) in Sheep 5 to 7.5 Years after Vaccination with Inactivated BTV-8 Vaccines

Thirty-six female sheep, previously vaccinated against Bluetongue virus serotype 8 (BTV-8) using inactivated vaccines, were included in this field study. In Germany, vaccination was compulsory in 2008 and 2009, voluntary in 2010 and early 2011, and later, was prohibited in 2011. Due to their age, eighteen sheep had been vaccinated for two or more consecutive years, while a further eighteen animals had only been vaccinated once or not at all. The sheep were blood sampled five (n = 31) to 7.5 years (n = 5) after their last vaccination. All serum samples (n = 36) were tested for BTV group-specific antibodies by an ELISA (IDScreen® Bluetongue Competition assay, ID Vet). In five of the animals, the BTV-8 serotype-specific antibody titers were measured by serum neutralization (SN). The majority of sheep that were vaccinated annually for two or more years showed a positive ELISA (14/18 sheep) and a SN (two of two sheep) result 5 years after their last vaccination. Most of the sheep vaccinated fewer than twice showed a negative ELISA result 5 to 7.5 years after their last vaccination (13/18 animals). The three animals in this group tested by SN showed one negative and two positive results. This short communication is the first to describe the presence of BTV antibodies in sheep 5 to 7.5 years after vaccination with inactivated BTV-8 vaccines.

Deteksi Johne’s Disease pada Sapi Potong di Kabupaten Kebumen Berdasarkan Uji Elisa

Kebumen Regency have been set as the source of the beef cattle breeds by the Ministry of Agriculture of the Republic of Indonesia. As the parent stock of beef cattle, these should be free from any infectious disease. Base on Balai Besar Veterinary Wates survey at 2015, some beef cattle in some districts at Kebumen indicated Johne's Disease. This research was carried out with the epidemiologic approach to detect Johne's Disease. About 173 serum specimens were taken from 173 heads of cows who have clinical symptoms of Johne's Disease. Sera were analyzed by ELISA. Base on ELISA result, 38 from 173 serum were seropositive. These cows sample come from local ancestor and there were not Johne’s disease vaccination program in Kebumen. This research proved that Johne's Disease have occurred in this populations which dedicated for parent stock of beef cattle in the District of Kebumen. Disease control program for Johne’s disease should be undertaken in this area to prevent the disease transmition.

Penggunaan Imunostik sebagai Uji Serologi untuk Deteksi Brucella abortus pada Sapi (APPLICATION IMMUNOSTICK ASSAY FOR SEROLOGICAL TEST BRUCELLA ABORTUS IN BOVINE)

Serological test is one of diagnostic method to detect pathogenicity brucella. Several methods are being improving such as Rose Bengal Test (RBT), Complement Fixation Test (CFT) dan Enzyme-linked Immunosorbent Assay (ELISA). Immunostick has an accuracy equivalent to ELISA and is easy to apply in the field so it is possible to be applied as a rapid test for brucellosis detection. The study aim was to know sensitivity and specificity of immunostick that were used to detecte antibody Brucella abortus using commercial antigens of B. abortus Strain 19 (S19) and B. abortus Strain 99 (S99). The test have compared with ELISA. The tests were conducted in two stages, namely (i) immunostick ability to detect antibodies in seropositive and seronegative serum, and (ii) the immunostick result were compared with ELISA result in serum grup that were be know and unknown status. A total of 250 serums were examined and result indicated that immunostick can be detect B. abortus antibodies in cattle serum with sensitivity 100%. Immunostick specifity were 45,45% for B. abortus S99(1) antigen; 78,79% for B. abortus S99(2) antigen and 51,52% for B. abortus S19 antigen. When the test compared with ELISA, the sensitivity 82,86% and the spesifity were 52,31% for B. abortus S99(1) antigen; 93,54% and 79,71 for B. abortus S99(2) antigen and 82,86% and 58,46% for B. abortus S19 antigen.

Rotavirus Diarrhoea among Children under Five Years in a Tertiary Level Government of Rajasthan

Introduction: Rotavirus is the leading cause of severe, dehydrating diarrhoea in young children globally. Studies indicate that rotavirus causes approximately 40 percent of childhood diarrhoea hospitalization worldwide and around 39 percent in India in less than 5 years of age. This study aimed to estimate the prevalence of rotavirus diarrhea among hospitalized children aged under five years.Materials and Method: Stool samples were collected from children who fall within the age range of 0-5 years with acute diarrhea and samples are tested for rotavirus by the enzyme linked immunosorbent assay (ELISA).Result: Out of 349 samples, 104(29.8%) cases were positive for rotavirus by ELISA. Therefore the prevalence of rotavirus infection among hospitalized patient under this study was 29.8%.Conclusion: Rotavirus is an important cause of diarrhea in hospitalized children.J Nepal Paediatr Soc 2016;36(3):273-276

The collection of lymphatic fluid from the bovine udder and its use for the detection of Mycobacterium avium subsp. paratuberculosis in the cow

The objective of the current study was to evaluate the feasibility of lymph collection from the bovine udder and to investigate if the lymphatic fluid might be of diagnostic value in cows infected with Mycobacterium avium subsp. paratuberculosis (MAP), the etiologic agent of paratuberculosis. Lymph fluid collection was attempted from 58 cows, and the reactions of the cows as well as the level of difficulty of the procedure were recorded in 56 animals. Lymph samples (51 in total) were tested for the presence of MAP by nested polymerase chain reaction. Collection of the lymphatic fluid caused no or mild signs of discomfort in 94.6% of the cows; in 51.8% of cows, lymphatic fluid was attained on the first attempt, while sample collection was unsuccessful in 12.1%. Mycobacterium avium subsp. paratuberculosis was detected in 43.1% of all lymph samples. The bacterium was present in 66.7% of cows with clinical Johne’s disease, in 42.8% of asymptomatic cows with a positive or suspicious enzyme-linked immunosorbent assay (ELISA) result in blood, and in 38.7% of cows with a negative ELISA result in blood. The present study shows that the procedure was well tolerated by most cows and can easily be performed on farm. The current report of the isolation of MAP from lymph fluid suggests that the present approach could be used for the early detection of Johne’s disease in cattle.