Luciano Bastos Lopes

Parameter estimation and use of gamma interferon assay for the diagnosis of bovine tuberculosis in Brazil

This study aimed to evaluate the interference of tuberculin test on the gamma-interferon (INFg) assay, to estimate the sensitivity and specificity of the INFg assay in Brazilian conditions, and to simulate multiple testing using the comparative tuberculin test and the INFg assay. Three hundred-fifty cattle from two TB-free and two TB-infected herds were submitted to the comparative tuberculin test and the INFg assay. The comparative tuberculin test was performed using avian and bovine PPD. The INFg assay was performed by the BovigamTM kit (CSL Veterinary, Australia), according to the manufacturers specifications. Sensitivity and specificity of the INFg assay were assessed by a Bayesian latent class model. These diagnostic parameters were also estimate for multiple testing. The results of INFg assay on D0 and D3 after the comparative tuberculin test were compared by the McNemars test and kappa statistics. Results of mean optical density from INFg assay on both days were similar. Sensitivity and specificity of the INFg assay showed results varying (95% confidence intervals) from 72 to 100% and 74 to 100% respectively. Sensitivity of parallel testing was over 97.5%, while specificity of serial testing was over 99.7%. The INFg assay proved to be a very useful diagnostic method.

Epidemiological investigation of Borrelia burgdorferi in horses in the municipality of Sinop—MT, Brazil

Borrelia burgdorferi sensu stricto is the main etiological agent of Lyme disease (LD) in the USA. In Brazil, it is believed that a similar spirochete is the causal agent of the Baggio–Yoshinari syndrome (BYS), a zoonosis also transmitted by ticks, whose clinical manifestations are similar to those of LD. Despite the epidemiological importance, there are no studies reporting the presence and the prevalence of B. burgdorferi among horses in Mato Grosso State. The aim of this study was to detect and measure the frequency of IgG antibodies anti-B. burgdorferi American strain G39/40 in horses in the municipality of Sinop, MT—Brazil, using the indirect enzyme-linked immunosorbent assay (ELISA) for serological diagnosis. Blood samples from 367 horses were collected in 81 farms. An epidemiological questionnaire was applied during the visits to obtain information related to the animals and the farms. From the 367 horses, 214 were positive for B. burgdorferi sensu stricto according to the results of the ELISA test, representing an apparent prevalence of 54.04% [CI = 0.4548051–0.6237234]. Concomitantly, 89 blood samples were taken for molecular analysis by nested polymerase chain reaction (PCR). According to the PCR test results, none of the samples were reactive, although 53 of these samples were reactive according to ELISA. Seventy five farms (92.59%) had at least one reactive horse for B. burgdorferi. Our results support the hypothesis of the presence of anti-Borrelia spp. antibodies in horses in Mato Grosso, reaching a high animal prevalence. Besides that, leisure/sport purposes proved to be a risk factor, with an odds ratio of 3.16. These findings clearly indicate the need of borreliosis control in Sinop and make a significant contribution to the knowledge of the disease in Mato Grosso.

Diagnosis of Leptospira spp. Infection in Sheep Flocks in the State of Mato Grosso, Brazil.

Background: Leptospirosis has a worldwide distribution with high social and economic impact. It is caused by a variety of Leptospira spp. serovars, particularly in tropical regions. Various species of animals can be affected by Leptospira spp. including cattle, horses, sheep, goats and swine, and can act as a reservoir for human infection. In Brazil, sheep leptospirosis has been diagnosed in a variety of States, however, little is known about sheep flocks of Mato Grosso. Considering the marked growth of sheep industry in Mato Grosso State in past recent years, this study aimed to evaluate the frequency of serologically positive sheep for Leptospira spp. serovars in the State of Mato Grosso (Brazil). Materials, Methods & Results: Blood samples of 346 sheep, which belonged to 16 sheep flocks of the medium northern region of Mato Grosso was obtained by jugular vein puncture. Serum was obtained by blood centrifugation and stored at -20°C until the test to be performed. The diagnosis was performed using the microscopic agglutination test. Serovars Australis, Autumnalis, Bataviae, Bolivia, Castelonis, Celledoni, Grippotyphosa, Hardjo, Hardjobovis, Hebdomadis, Icterohaemorrhagiae, Javanica, Lagoa, Norma, Panama, Pomona, Sejroe, Shermani, Szwajizak, and Wolffi were tested. Reactions were considered positive with minimum titration of 1:100. Considering all serovars, 54.9% of sheep were positive, and the serovar more frequently detected was Celledoni (22.8%), followed by Javanica (19.4%), Castellonis (16.5%), and Norma (15.3%), with predominance of titration of 100 for these serovars, while only one sheep had a titration of 1:800 detected for serovar Hardjo. All flocks (100%) were positive (at least one positive animal), with the frequency of animals positive to Leptospira spp. varying of 31.3% to 79.2% in flocks. Discussion: Infection for Leptospira spp. in humans has a large social and sanitary impact. Under an animal health perspective, beyond sanitary impact, leptospirosis have a large economic impact and can act as important reservoirs for Leptospira spp. for human infection, especially in tropical conditions, which favor the persistence of agent in the environment. The occurrence of sheep leptospirosis has been diagnosed in several States of Brazil as Rio Grande do Sul, Paraiba, Rio de Janeiro, Sao Paulo and Maranhao, with frequency of herds positive for Leptospira spp. varying of 5.4% to 47.4%, however, the frequency of positive sheep in this study was higher than these previous reports in other Brazilian States. The most frequent serovar in sheep flocks have a great importance for prophylactic control. However, there were identified as main frequent serovar Celledoni and Javanica, which are considered unusual in sheep, and indicates a peculiar profile of ovine serovars for the region. Considering de high frequency of positive farms (100%) in Mato Grosso and the lack of studies about the occurrence of leptospirosis in this region, our results suggest the occurrence of risk factors that can favor the dispersion and survival of agent. Contact of the sheep evaluated in this study with other animal species such as cattle, dogs, and horses that can serve as a source for sheep infection or may become contaminated from sheep. Then, the profile of sheep infection by Leptospira spp. in the State of Mato Grosso has a peculiar profile of serovars, with high frequency of infection of animals and herds.