Ivan Roque de Barros Filho

Mycoplasma ovis in captive cervids: prevalence, molecular characterization and phylogeny.

Hemotrophic mycoplasmas (hemoplasmas) are bacteria that attach to red blood cells of mammals, leading to acute and/or subclinical disease in infected animals. It has been suggested that Mycoplasma ovis, a hemoplasma that infects sheep and goats worldwide, may also infect deer. The aim of this study was to evaluate whether South American deer are infected with M. ovis. EDTA-anticoagulated blood samples from a herd of 32 captive South American deer were collected. DNA extraction of blood samples was performed followed by PCR amplification of the 16S and 23S rRNA genes, and sequencing of products. Using M. ovis PCR, 27/31 (87%) were positive, including 21/22 Mazama nana; 2/3 Mazama americana and 4/6 Blastocerus dichotomus. Sequencing of the nearly entire 16S rRNA gene of 26/27 positive samples showed 98.2-98.8% identity to M. ovis of sheep (GenBank, AF338268) and 98.6-99.4% identity to M. ovis-like of a fawn (FJ824847); the 23S rRNA gene from one of these isolates and the fawns had 97.6% identity. The remaining isolate had just 94.9% identity to the 16S rRNA gene of M. ovis and only 89.4% identity to the 23S rRNA gene of the fawns M. ovis. This is the first report of M. ovis in captive South American deer, revealing a high prevalence of hemoplasma infection in these animals.

Detection of a novel hemoplasma based on 16S rRNA gene DNA in captive and free-ranging capybaras (Hydrochaeris hydrochaeris).

Two different species of hemoplasmas, Mycoplasma coccoides and M. haemomuris, are known to infect small rodents such as mice and rats. However, there are no previous reports of hemoplasma infection in capybara (Hydrochaeris hydrochaeris). The aim of our study was to determine whether these hemoplasmas might infect capybaras from Southern Brazil. Blood samples from 31 animals: 10 captive and 21 free-ranging capybaras were collected and packed cell volume and total plasma protein were measured. DNA was extracted and PCR assays for M. coccoides and M. haemomuris were performed. Using the M. coccoides-PCR assay 64% of the capybaras were positive, 80% free-ranging and 30% from captive animals. The prevalence of infection between the groups was significantly different (p=0.001). Sequencing of the nearly entire 16S rRNA gene from the positive samples suggested a novel hemoplasma isolate with identity of 92% with M. coccoides and 86% with M. haemomuris. All capybara samples were negative for M. haemomuris infection. DNA of a housekeeping gene was successfully amplified from all samples. This is the first evidence of a hemoplasma infection in capybaras.

Detection of Neospora sp. antibodies in cart horses from urban areas of Curitiba, Southern Brazil

Neospora caninum is a protozoan parasite which affects dogs as definitive hosts and several mammalian species as intermediate hosts mainly causing abortions and central nervous system disorders. The reemerging population of cart horses for carrying recycling material in urban areas of major cities in Brazil may have an impact on disease spreading, and these animals may be used as sentinels for environmental surveillance. Thus, the present study investigated the frequency of Neospora sp. antibodies in cart horses from Curitiba and surrounding areas, Paraná State, Southern Brazil. IgG antibodies against Neospora sp. were detected using indirect fluorescence antibody test (IFAT), and titers equal to or higher than 1:50 were considered reactive. Of all samples, 14/97 (14.4%) were positive: 2/29 (6.9%) were younger than 5; 5/26 (19.2%) between 6 and 9; and 6/31 (19.4%) older than 10 years of age. One of the 11 animals with unknown age was positive (9.1%). Cart horses are likely to be more exposed to dog feces and to Neospora sp. oocyst contamination in urban settings and a lower frequency of disease in dogs may have a negative impact on horse infection risk in these areas.

Prevalence and Molecular Characterization of Mycoplasma ovis in Selected Free-Ranging Brazilian Deer Populations

Mycoplasma ovis is a hemoplasma that may cause anemia and mortality in small ruminants. Our aim was to determine whether M. ovis infects populations of free-ranging deer in Brazil. Buffy coat samples from 64 Blastocerus dichotomus from Porto Primavera, 18 Ozotocerus bezoarticus from Pantanal, and 21 O. bezoarticus from Emas National Park were tested. Using a M. ovis PCR protocol to amplify extracted DNA, 46/64 (72%) of deer from Porto Primavera, 10/18 (56%) from Pantanal, and 4/21 (19%) from Emas National Park were positive, giving an overall positive rate of 58% for hemoplasma in these wild deer. Sequencing and phylogenetic analysis of the 16S rRNA gene revealed 3 genetically distinct hemoplasmas including M. ovis, ‘Candidatus Mycoplasma erythrocervae’, and a hemoplasma most closely related to M. ovis. Phylogenetic analysis of the 23S rRNA gene from selected sequences confirmed these relationships.

Detection of anti-Toxoplasma gondii antibodies in carthorses in the metropolitan region of Curitiba, Paraná, Brazil.

Toxoplasma gondii, the agent for toxoplasmosis, has worldwide distribution. Horses normally play a secondary role in its life cycle, but movement around urban areas, feeding on grass and the increasing use of carthorses for gathering recyclable material in some urban areas of Brazil may increase their exposure to T. gondii infection. The aim of the present study was to investigate the frequency of anti-T. gondii antibodies in carthorses in the metropolitan region of Curitiba, PR. IgG antibodies against T. gondii were detected using the indirect fluorescence antibody test (IFAT) (titers ≥ 64). Seventeen (17.0%) of the 100 horses sampled were seropositive. There were no statistical differences in relation to sex (p = 0.28) or age (p = 0.15). Our findings suggest that carthorses are exposed to T. gondii infections and that no associations with age or sex exist.

Hemotropic mycoplasma in a free-ranging black howler monkey (Alouatta caraya) in Brazil

Hemotropic mycoplasmas are bacteria that infect erythrocytes and cause sub-clinical infections to life-threatening disease. We describe hemotropic mycoplasma infection in a free-ranging black howler monkey (Alouatta caraya). This is the first molecular detection of a hemotropic mycoplasma in a nonhuman primate from Brazil.

DETECTION OF RDRIO STRAIN OF MYCOBACTERIUM TUBERCULOSIS IN TAPIRS (TAPIRUS TERRESTRIS) FROM A ZOO IN BRAZIL

Tuberculosis is a chronic infection caused by strains of the Mycobacterium tuberculosis complex and occurs in both animal and human populations. The death of a tapir showing purulent material and a hard mass in the lungs at necropsy raised suspicion of a potential disease caused by mycobacteria species in a Brazilian zoo. Later, two other tapirs with similar signs died and were further investigated. Polymerase chain reaction (PCR) from bronco-alveolar lavages was performed, and both animals tested positive for the RD(Rio) strain of M. tuberculosis, which is a recently discovered Latin American-Mediterranean sublineage and the main cause of human tuberculosis in Rio de Janeiro, Brazil. To investigate the possibility of human infection and the source of transmission, all 50 zoo employees underwent tuberculin skin testing; four were reactive, but radiographic exams and direct sample staining did not suggest tuberculosis. Thus, direct human to animal transmission was not proven. However, the presence of RD(Rio) M. tuberculosis in tapirs highlights the lack of attention to diseases that human beings may transmit to wildlife.

SEROLOGICAL AND MOLECULAR SURVEY OF Leptospira spp. AMONG CART HORSES FROM AN ENDEMIC AREA OF HUMAN LEPTOSPIROSIS IN CURITIBA, SOUTHERN BRAZIL

Introduction: Cart horses are a re-emerging population employed to carry recyclable material in cities. Methods: Sixty-two horses were sampled in an endemic area of human leptospirosis. The microscopic agglutination test (MAT) and real-time polymerase chain reaction (qPCR) were performed. Results: A seropositivity of 75.8% with serovar Icterohaemorrhagiae in 80.8% of the horses was observed. Blood and urine were qPCR negative. MAT showed positive correlations with rainfall (p = 0.02) and flooding (p = 0.03). Conclusions: Although horses may be constantly exposed to Leptospira spp. in the environment mostly because of rainfall and flooding, no leptospiremia or leptospiruria were observed in this study.

Treponemes Detected in Digital Dermatitis Lesions in Brazilian Dairy Cattle and Possible Host Reservoirs of Infection

ABSTRACT The main pathogenic treponemes causing bovine digital dermatitis were identified from 17 infected herds in southern Brazil for the first time in this study using PCR. We did not find a relationship between treponeme phylogroup composition and clinical classification. Treponema phagedenis was present in all lesions. Rumen fluid was implicated as a reservoir location for these pathogens.

Fatores etários no leucograma de fêmeas zebuínas sadias da raça Nelore (Bos indicus)

In order to evaluate the influence of the age on the white blood cell counts of Nelore (Zebu) cattle, 158 blood samples from seven groups of different ages (group I-up to three months; group II-three to six months; group III-six to 12 months; group IV-12 to 24 months; group V-24 to 48 months; group VI-48 to 72 months and group VII-older than 72 months) were studied. The highest and lowest mean values observed were total white blood cell/mm3-16992±4104 (group III) and 10353±2397 (group VI); neutrophils/mm3-3931±1578 (group I) and 2416 ±1118 (group III); eosinophils/ mm3-999±499 (group V) and 265±276 (group II); basophils/mm3-67±88 (group VII) and 39±78 (group III); typical lymphocytes/ mm3-12758±3608 (group III) and 5906±1883 (group VI); atypical lymphocytes/ mm3-1310±603 (group II) and 760±419 (group VI) ; lymphocytes total/ mm3-14079±4027 (group III) and 6666±2059 (group VI); monocytes/mm3-27±62 (group I) and 0 (group III). The differences (p>0.05) observed among the groups in relation to total white blood cell, lymphocytes, neutrophils and eosinophils can be related to the influence of the age on leukogram of Nelore (Zebu) cattle raised in Sao Paulo- Brazil.Professores Assistentes da Escola de Medicina Veterinaria da Universidade Federal da Bahia. Professor Associado, Departamento de Clinica Medica da Faculdade de Medicina Veterinaria e Zootecnia da Universidade de Sao Paulo (USP), Av. Professor Doutor Orlando Marques de Paiva, 87, CUASO. Sao Paulo, SP, 05508-000. E-mail: borges@usp.br. Autor para correspondencia. Professor Titular, Departamento de Clinica Medica da Faculdade de Medicina Veterinaria e Zootecnia da USP. Professor Associado, Departamento de Clinica Medica da Faculdade de Medicina Veterinaria e Zootecnia da USP. Professor Assistente da Faculdade de Medicina Veterinaria da Universidade Federal do Parana. RESUMO