Detection and diversity of maize yellow mosaic virus infecting maize in Nigeria Abstract Maize yellow mosaic virus (MaYMV; genus Polerovirus; family Luteoviridae) was recently characterized from maize in China and subsequently detected in mixed infection with sugarcane mosaic virus (genus Potyvirus; family Potyviridae) in sugarcane and itch grass in Nigeria. This study was conducted to understand the status and genetic diversity of MaYMV in maize fields in the northern guinea savannah region of Nigeria. A survey was conducted in 2017 and maize (n = 90) and itch grass (n = 10) plants suspected of virus infection based on appearance of mosaic and/or yellowing symptoms were sampled in Kaduna (n = 65) and Katsina (n = 35) states. The samples were screened individually by reverse transcription polymerase chain reaction using the genus-specific primers targeting poleroviruses and potyviruses Pol-G-F and Pol-G-R primers encompassing the partial P1–P2 fusion protein and coat protein genes of poleroviruses and primer pair CI-For & CI-Rev encompassing the partial cylindrical inclusion proteins of most potyviruses. A subset of amplified DNA fragments was cloned, Sanger-sequenced, and the obtained sequences were bioinformatically analyzed along with corresponding sequences from GenBank. The ~ 1.1 Kb polerovirus fragment was detected in 32.2% (29/90) of the maize samples while all 10 itch grass samples tested negative. BLASTN analysis of sequences derived from six polerovirus samples confirmed the virus identity as MaYMV. In pairwise comparisons, the MaYMV sequences derived in this study shared 97–99% nucleotide identity with sequences of other MaYMV isolates available in the NCBI GenBank. Phylogenetic analysis revealed the segregation of global MaYMV sequences into three host-independent clusters with pattern of geographic structuring.

Scorecard method for assessing the severity of peste des petits ruminants in sheep and goats Abstract A methodology to assess the clinical severity of peste des petits ruminants (PPR) in sheep and goats in the field condition was developed using a scorecard by considering five specific cardinal clinical signs (pyrexia, oculo-nasal discharge, oral lesions, respiratory signs, and diarrhoea) of disease. The scores were assigned for the signs based on the severity of the disease that ranged from 1 (low) to 4 (high). The assigned weightage for signs, morbidity, and mortality was 0.75, 0.05 and 0.2, respectively summing up to unity. The scoring and weightages and guidelines were devised by Delphi technique based on the field investigation, field veterinarian’s assessment and specific inputs from PPR experts. The estimated Weighted Score Index (WSI) was considered to classify the severity into mild (WSI < 40) or moderate (WSI 41–60) or severe (WSI > 60) form. This scorecard will help preliminarily to the extent for the identification of the suspected flocks with a required case definition at the first instance, before making decisions on what merits further field investigation. This is first of its kind of methodology to assess the disease pattern in small ruminants and could be used as a disease severity assessment tool in different geographical areas in endemic settings.

Occurrence and variability of begomoviruses associated with bhendi yellow vein mosaic and okra enation leaf curl diseases in south-western India Abstract Bhendi yellow vein mosaic disease (BYVMD) and Okra enation leaf curl disease (OELCuD) are common diseases of okra/bhendi [Abelmoschus esculentus (L.) Moench] affecting both pod yield and quality in the Indian subcontinent. BYVMD is caused by the infection of a begomovirus and associated betasatellite. In this study, we have made an attempt to investigate the diversity of begomoviral and the satellite sequences in okra samples showing BYVMD and OELCuD, by using a rapid PCR-based approach on 46 samples collected from 23 locations of Southern and Western India. We have also analyzed nine RCA-generated full-length begomoviral clones, some generated from the above samples displaying BYVMD and some OELCuD. By the PCR approach, we find the presence of begomovirus okra enation leaf curl virus (OELCuV) in most samples, irrespective of the disease being displayed (BYVMD or OELCuD). The nine apparently full-length sequences also show high identities with OELCuV and show instances of both intra-specific as well as intra-strainal recombination. We have also analyzed the begomoviral sequences associated with BYVMD and OELCuD from publicly available nucleotide sequence databases and show much higher sequence diversity amongst BYVMV, as compared to OELCuV. This is the first study which comprehensively demonstrates the presence of OELCuV in okra samples showing BYVMD and those showing OELCuD.

Expression and characterization of the non-structural protein V of small ruminant morbillivirus Abstract Peste-des-petits ruminants is a transboundary viral disease of small ruminants caused by small ruminant morbillivirus (SRMV). In the present study, the full-length V gene of SRMV was constructed through site-directed mutagenesis from the P gene transcripts of the vaccine virus (Sungri/96 India) and expressed in a prokaryotic expression system. In animals, the seroconversion against this protein occurs from 14-days and is getting produced from 48 h in cell culture. An indirect ELISA developed using this protein has a relative sensitivity and relative specificity of 77.73% and 73.775%, respectively as compared to c-ELISA. In this ELISA, it was observed that most of the convalescent animals elicited higher level of antibodies than vaccinated animals.

Protective immunity against hepatitis B virus infection in a group of vaccinated Sri Lankan military service men following a complete course of vaccination Abstract Vaccination is the appropriate measure to protect military personnel against the hepatitis B virus (HBV) infection. Testing the military personnel for anti-HBs levels after vaccination is vital in re-vaccinating those that have not developed protective immunity. The aim of the current study was to determine the immunity in a group of vaccinated Sri Lankan military personnel (n = 150; age = 26–44 years) following a complete course of hepatitis B virus surface antigen (HBsAg) vaccination by assessing the antibodies against HBsAg (anti-HBs) levels. Three months after the last dose of the vaccination, blood samples were collected from the study population and tested for anti-HBs levels using a commercially available ELISA. Of the 150 military service men tested, 139 (92.67%) had anti-HBs levels higher than 10 mIU/mL, WHO approved levels for protective immunity against HBV infection. Of the 139 that had sufficient anti-HBs levels, 24% (36/150) had anti-HBs levels between 10 and 100 mIU/mL and 68.67% (103/150) had anti-HBs levels > 100 mIU/mL. Overall, 7.33% (11/150) participants had anti-HBs levels < 10 mIU/mL. Sero-conversion to > 10 mIU/mL anti-HBs was more than 90% in those that were less than 40 years of age and it was less than 90% in those that were more than 40 years of age.

Occurrence of pepper mottle virus on tomato in India Abstract Tomato plants exhibiting mosaic and mottling symptoms were collected from the research field of Punjab Agricultural University during 2016-17. DAS-ELISA test using pepper mottle virus (PepMoV) antiserum showed positive reaction to 5 of 25 tomato samples collected. Further molecular characterization of virus was carried out by using potyvirus specific primer pairs corresponding to most conserved and variable genes viz., Nuclear inclusion b (NIb) and P1 respectively. RT-PCR assay confirmed the association of PepMoV with tomato crop under field conditions in India.

Emerging horizon for bat borne viral zoonoses Abstract Bats are the only flying placental mammals that constitute the second largest order of mammals and present all around the world except in Arctic, Antarctica and a few oceanic islands. Sixty percent of emerging infectious diseases originating from animals are zoonotic and more than two-thirds of them originate in wildlife. Bats were evolved as a super-mammal for harboring many of the newly identified deadly diseases without any signs and lesions. Their unique ability to fly, particular diet, roosting behavior, long life span, ability to echolocate and critical susceptibility to pathogens make them suitable host to harbor numerous zoonotic pathogens like virus, bacteria and parasite. Many factors are responsible for the emergence of bat borne zoonoses but the most precipitating factor is human intrusions. Deforestation declined the natural habitat and forced the bats and other wild life to move out of their niche. These stressed bats, having lost foraging and behavioral pattern invade in proximity of human habitation. Either directly or indirectly they transmit the viruses to humans and animals. Development of fast detection modern techniques for viruses from the diseased and environmental samples and the lessons learned in the past helped in preventing the severity during the latest outbreaks.

Clinical evaluation, serological response and lesions generated by the A/Mexico/La Gloria-3/2009/H1N1 and A/swine/New Jersey/11/1976/H1N1 influenza viruses in colostrated and non-colostrated pigs Abstract Influenza A viruses cause respiratory disease in piglets, and maternal immunity plays an important role in protecting against Influenza virus infection. Nevertheless, in the presence of high levels of maternal antibodies against influenza, an adequate immune response is not developed. In this study, the effect of maternal antibodies against the swine influenza A/swine/New Jersey/11/1976/H1N1 virus (swH1N1) on clinical presentation, serological response, and lesions produced in colostrated and non-colostrated pigs was evaluated in pigs infected with the human influenza A/Mexico/La Gloria-3/2009/ H1N1 (pH1N1) and swH1N1 viruses. Our results indicated that between 2 and 4 days post-challenge, sneezing and mild nasal discharge were observed in all pigs. Body temperature in pigs from all treatment groups ranged between 39.2 and 39.3 °C. Pigs inoculated with the pH1N1 virus (421 g) exhibited a significantly lower daily weight gain than those inoculated with the swH1N1 virus (524 g). HI antibody titers against the pH1N1 virus were significantly different between colostrated (1.62) and non-colostrated (0.43) pigs. Significant differences in antibody titers were detected between pigs inoculated with the pH1N1 (1.28) or the swH1N1 virus (0.77) (P < 0.05). The highest percentage of pulmonary lesions was observed in non-colostrated/pH1N1 pigs (11.88%) at 6 days post-challenge. Cross reactivity was observed between the pH1N1 and swH1N1 viruses, as the maternal antibodies against the swH1N1 virus successfully neutralized the pH1N1 virus infection.

Persistence of antibody response in chikungunya Abstract Chikungunya is a mosquito-borne viral illness associated with chronic arthritic symptoms that persist for months. The IgM antibody appears within a week post any infection and declines at 2–3 months. The present study was aimed to demonstrate the presence of specific IgM antibody among chikungunya confirmed cases. Blood samples were collected from chikungunya PCR positive patients at the time of diagnosis, at 1-week, 1, 8, 10 and 12 months post infection. All acute and follow-up serum samples were evaluated for chikungunya virus-specific IgM antibodies using ELISA technique. Our findings indicate the persistence of anti-chikungunya IgM up to 10-months post-infection in a majority of chikungunya virus infected persons. Interpretation of results should be carefully done as only IgM ELISA is used to diagnose acute infection, especially post chikungunya outbreak. The presence of IgM antibody does not rule out the absence of any other diagnosis due to its persistence. Thus, we hypothesize that real-time PCR is more reliable for the detection of acute chikungunya cases in endemic areas while IgM detection may be useful in identifying exposure to this disease.

Viral hepatitis among acute hepatitis patients attending tertiary care hospital in central India Abstract Viral hepatitis is a considerable public health burden affecting millions of people throughout the world. The incidence of viral hepatitis varies greatly depending upon geographic locations, age and gender. Exploring the etiological spectrum and clinic-epidemiological profile of acute viral hepatitis (AVH) becomes essential for strategizing the preventive measures to control the diseases. An epidemiological data depicting AVH situation and its etiologies is missing from central India. With the aim of fulfilling this lacuna, the present analysis was done on samples tested over a period of 2 years from July 2015 to June 2017. Of the 1901 hepatitis cases, 597 individuals (31.4%) were positive for AVH infection and HEV was the predominant cause followed by HBV, HAV and HCV. Co-infections of hepatitis viruses were detected in 42 cases. Co-infection of HEV with HBV was the commonest pattern. Male preponderance was observed among AVH positive cases and the age group of 26–45 years was the most susceptible to the viral hepatitis infections, except hepatitis A, which was the most frequent among children. Two hundred patients (33.45%) required hospitalization and 51 deaths were attributed to AVH infections. The analysis for the first time reports intricacies and viral etiologies of AVH in central India. Regular diagnosis of AVH etiology and monitoring of cases will help in patient management and assist disease control programs to take policy decisions.