An ELISA test using a circulating Mycobacterium bovis peptide for detecting bovine tuberculosis in dairy cattle.

This study aimed to determine the sensitivity (Se) and specificity (Sp) of a circulating pathogen-specific biomarker (polyketide synthetase 5, Pks5)-based enzyme-linked immunosorbent assay (ELISA) independently or in conjunction with a caudal fold tuberculin (CFT) test for bovine tuberculosis (bTB) screening in dairy cattle. We enrolled 987 dairy cows from 34 herds in Chiang Mai province, Thailand. A conditionally independent Bayesian model with a single population was inferred from the test results. The percentage of positive results for the Pks5-ELISA using 0.4 OD cutoff test and CFT test were 9.0% (89/987) and 10.5% (104/987), respectively. The median of posterior estimates of Se for the Pks5-ELISA test was 90.2% (95% posterior probability interval [PPI] = 76.6-97.4%), while the estimated Sp was slightly higher (median = 92.9, 95% PPI = 91.0-94.5%). The median estimated Se of the CFT test was 85.9% (95% PPI = 72.4-94.6%), while the estimated Sp was higher, with a median of 90.7% (95% PPI = 88.7-92.5%). The posterior estimate for true disease prevalence was 2.4% (95% PPI = 1.2-3.9%). The Pks5-ELISA test yielded characteristics at or above the acceptable standards for bTB detection. Therefore, the pathogen-specific biomarker, Pks5, is a potential detection system for bTB screening and may be applied as an ancillary test together with the currently applied standard method (CFT test) to reinforce the bTB control and eradication programs.

Clinical Evidence and Risk Factors for Reproductive Disorders Caused by Bacterial Infections in Meat Goats in Northeastern Thailand.

The objective of this study was to identify risk factors related to reproductive disorders caused by bacterial infections in goats in northeastern Thailand. Two hundred twenty farms were investigated, and 49 herds were found to have clinical reproductive disorders. Moreover, 96% (47/49) of herds showing clinical reproductive failure preferred to circulate bucks between herds. A total of 118 sera, including 85 clinical reproductive disorder cases such as abortion (n = 70), abortion with arthritis (n = 1), orchitis (n = 3), repeat breeder (n = 6), sterile (n = 1), and weak kids (n = 4), and 33 bucks' circulations were serologically tested for bacterial infections caused by Coxiella burnetii, Chlamydophila abortus, and Brucella spp. Results showed 69% (81/118 cases) were seropositive for Q fever (n = 55; 46.61%), brucellosis (n = 8; 6.78), and chlamydiosis (n = 18; 15.25%), respectively; 82% of herds (40/49 herds) were infected with at least one of those diseases. Moreover, 40% of infected herds (16/40) had coinfection among the three of those diseases. Approximately 60% (20/33) of buck circulation showed seropositivity to at least one of the diseases, and 85% of infected bucks were seropositive for Q fever (17/20). Buck circulation between herds is a risk factor for diseases on farms (p=0.001); odds ratio (OR = 109.29; 95% confidence interval (CI) = 6.61-1,807.38). Moreover, the annual brucellosis test is a protective factor against reproductive failure cases on farms (p=0.022; OR = 0.45; 95% CI = 0.23-0.89). Reproductive disorder cases can be caused by sexual transmission, so buck circulation can yield Q fever, brucellosis, and chlamydiosis in communities. This investigation is the first report of chlamydiosis infection in our area. Concerning Q fever, chlamydiosis, and brucellosis are zoonotic diseases that impact animal health and production losses. Control and prevention measures related to risk factors together with active surveillance programs should be incorporated into client education.

Prevalence and Demographic Risk Factors of Mycobacterium tuberculosis Infections in Captive Asian Elephants (Elephas maximus) Based on Serological Assays.

To address putative TB statuses of elephants and to identify and quantify potential demographic risk factors for TB, three ELISAs specific for different mycobacterial antigens (ESAT6, CFP10, MPB83) and the TB Stat-Pak assay were used as surrogate serological markers for TB infection in elephants. In view of the low number of animals of which the infected status could be confirmed (4 out of 708) Latent Class Analyses of TB serology test outcomes was used to predict the putative TB status of each of 708 elephants as positive (17.3%), inconclusive (48.7%), or negative (34%) when assessed on a population basis. Correlation between test performance of the individual assays was high between the ELISAs, but low with that of the TB Stat-Pak assay. Risk factors, assessed based on cut off values for each of the ELISAs determined by ROC analysis, included sex, BCS, age, working time, feed type, management system, camp size and region. Old age elephants were more likely to show a positive TB serology test outcome, than younger ones. Elephants working 7 h per day and the ones in good condition BCS (7-11) were less likely to be positive in TB serology testing. In addition, fewer animals in the large camp size (31-50 elephants) were found to be positive in ELISA tests, compared to elephants in the other camp sizes. In this study, the North region had the lowest percentages of elephants with positive TB test outcome, the West region and to a lesser extend the other regions showed clearly higher percentages of positive animals. Even though assays used in the present study have not been validated yet, results obtained showed promise as diagnostic or screening tests. For the diagnosis of animals suspected to be infected, the ELISA tests, once further optimized for the individual antigens, can be used in parallel. For screening of complete camps for presence or absence of infection, a single optimized ELISA test can be utilized.

Detection of virus-neutralising antibodies and associated factors against rabies in the vaccinated household dogs of Kathmandu Valley, Nepal.

BACKGROUND: Rabies is a vaccine-preventable neglected tropical viral zoonosis. It occurs worldwide, creating a very heavy burden in many developing countries, including Nepal. Dogs are the principle vector for the transmission of this disease in urban areas. Vaccination is the most important preventive measure in areas where dogs are the principle source of infection. This study was conducted with the aim of detecting virus-neutralising antibodies and associated factors against rabies in vaccinated household dogs of Kathmandu valley. METHODS: Blood samples were collected from 110 vaccinated pet dogs in Kathmandu, Bhaktapur, and Lalitpur districts of Nepal. The samples were taken to the laboratory of the National Zoonosis and Food Hygiene Research Center where serum was separated. An indirect immune-enzymatic assay (PlateliaTM Rabies II kit ad usum Veterinarium, Biorad, China) was used for the detection of rabies virus anti-glycoprotein antibodies in the dog serum samples following the manufacturer's recommendations and instructions. Optical density values for unknown samples were compared with the positive sera titers in quantification tests obtained after a direct reading on the standard curve. Results were expressed as equivalent units per ml (EU/ml). FINDINGS: Of the total samples, 89.09% exceeded the required seroconversion level (≥ 0.5 EU/ml); another 9.09% did not reach the seroconversion level (0.125-0.5 EU/ml); and 1.81% had undetectable seroconversion levels (<0.125 EU/ml) suggesting that the animal had not seroconverted according to the PLATELIA™ RABIES II test. Only one factor, the condition under which the dog was kept, was significantly associated with the antibody titer level. No association was found for any of the other factors included in the study. INTERPRETATION: Vaccination is the most effective measure for prevention and control of rabies. The locally manufactured brand of vaccine, which is available in Nepal, is potent enough to generate a sufficient amount of protective antibodies, equal to international brands.

Mycobacterium tuberculosis infection of domesticated Asian elephants, Thailand.

Four Asian elephants were confirmed to be infected with Mycobacterium tuberculosis by bacterial culture, other diagnostic procedures, and sequencing of 16S-23S rDNA internal transcribed spacer region, 16S rRNA, and gyrase B gene sequences. Genotyping showed that the infectious agents originated from 4 sources in Thailand. To identify infections, a combination of diagnostic assays is essential.

Identification of virulence-associated antigens and plasmids in Rhodococcus equi from patients with acquired immune deficiency syndrome and prevalence of virulent R. equi in soil collected from domestic animal farms in Chiang Mai, Thailand.

The prevalence of virulent Rhodococcus equi in soil collected from 17 domestic animal farms (from 12 cattle, 1 pig, and 4 horse farms) and in 6 clinical specimens from patients with acquired immune deficiency syndrome (AIDS) in Chiang Mai, Thailand, was investigated. The isolates were tested for the presence of 15-17-kDa antigens (VapA) and a 20-kDa antigen (VapB) by immunoblotting and for the presence of virulence plasmid DNA. Rhodococcus equi was isolated from most soil samples (68 of 80) obtained from the 17 farms, with 2.0 x 10(2) to 6.0 x 10(5) colony-forming units per gram of soil. We detected VapA in none of the 537 isolates from the soil samples. In one isolate from a pig farm, both VapB and virulence plasmid DNA were detected. Of the 6 clinical isolates from patients with AIDS, however, 4 isolates contained both VapB and virulence plasmid DNA. The remaining 2 isolates were avirulent.

Seroprevalence of melioidosis in dairy cattle in Chiang Mai Province, northern Thailand.

The seroprevalence of melioidosis in dairy cattle in Chiang Mai Province was investigated using of the indirect hemagglutination antibody (IHA) method. Two hundred and fifty-three samples were tested for serum antibodies to Burkholderia pseudomallei. The samples were from a total population of 8,688 dairy cattle in the province; random sampling, stratified by the location of cattle, was used. The seroprevalence was determined as 2% at 1:40 cut-off value, which was estimated to equate to 0.3% to 3.7% (95% CI). This report of relatively low disease prevalence in the animal population corresponds to other prevalence studies of the agent in the environment and the human population in the region. The prevalence is markedly different to that reported from northeastern Thailand, where the disease is highly endemic.