RESUMO: A mastite bovina é a doença mais impactante na indústria de laticínios e é caracterizada por uma complexidade de agentes causais, os quais têm revelado uma variação geográfica entre regiões e países. Os patógenos relacionados à mastite têm sido tradicionalmente classificados como contagiosos ou ambientais, com base na fonte de infecção ou origem e nas vias de transmissão. Além disso, a gravidade das infecções mamárias tem sido associada à virulência dos patógenos, e ao estado imunológico e nutricional dos animais. Considerando esse cenário, investigamos os agentes etiológicos, os escores de gravidade clínica e os dados de dias em lactação (DEL) em 4.273 casos clínicos de mastite bovina em dez fazendas de grande porte, quatro localizadas no sudoeste do estado de São Paulo e seis no sul do Estado de Minas Gerais, Brasil. Streptococcus dysgalactiae (283/4.273 = 6,6%), Escherichia coli (190/4.273 = 4,4%), Prototheca spp. (112/4.273 = 2,6%) e Streptococcus uberis (95/4.273 = 2,2%) foram os patógenos predominantes isolados. Entre os 4.273 casos clínicos, a pontuação de gravidade clínica estava disponível em 43,8% (1.871/4.273) dos animais. Desses, 69,8% (1.306/1.871), 27,3% (510/1.871) e 2,9% (55/1.871) foram classificados como leves, moderados e graves, respectivamente. Os patógenos foram observados principalmente em amostras obtidas durante os primeiros 100 dias em lactação, e a gravidade clínica foi principalmente classificada como leve. Nossos resultados contribuem para a identificação etiológica, a pontuação de gravidade clínica e os aspectos de ordenha de vacas com mastite em fazendas leiteiras com histórico de infecções mamárias clínicas. RESUMO causais países ambientais transmissão disso animais cenário etiológicos DEL (DEL 4273 4 273 4.27 porte Gerais Brasil 283/4.273 2834273 283 (283/4.27 6,6%, 66 6,6% , 6 6,6%) 190/4.273 1904273 190 (190/4.27 4,4%, 44 4,4% 4,4%) spp 112/4.273 1124273 112 (112/4.27 2,6% 26 2 95/4.273 954273 95 (95/4.27 2,2% 22 isolados 438 43 8 43,8 1.871/4.273 18714273 1 871 (1.871/4.273 Desses 698 69 69,8 1.306/1.871, 13061871 1.306/1.871 306 (1.306/1.871) 27 3 27,3 510/1.871 5101871 510 (510/1.871 29 9 2,9 55/1.871 551871 55 (55/1.871 leves graves respectivamente 10 leve etiológica clínicas 427 4.2 283/4.27 283427 28 (283/4.2 6,6 190/4.27 190427 19 (190/4.2 4,4 112/4.27 112427 11 (112/4.2 2,6 95/4.27 95427 (95/4.2 2,2 43, 1.871/4.27 1871427 87 (1.871/4.27 69, 1306187 1.306/1.87 30 (1.306/1.871 27, 510/1.87 510187 51 (510/1.87 2, 55/1.87 55187 5 (55/1.87 42 4. 283/4.2 28342 (283/4. 6, 190/4.2 19042 (190/4. 4, 112/4.2 11242 (112/4. 95/4.2 9542 (95/4. 1.871/4.2 187142 (1.871/4.2 130618 1.306/1.8 (1.306/1.87 510/1.8 51018 (510/1.8 55/1.8 5518 (55/1.8 283/4. 2834 (283/4 190/4. 1904 (190/4 112/4. 1124 (112/4 95/4. 954 (95/4 1.871/4. 18714 (1.871/4. 13061 1.306/1. (1.306/1.8 510/1. 5101 (510/1. 55/1. 551 (55/1. 283/4 (283/ 190/4 (190/ 112/4 (112/ 95/4 (95/ 1.871/4 1871 (1.871/4 1306 1.306/1 (1.306/1. 510/1 (510/1 55/1 (55/1 283/ (283 190/ (190 112/ (112 95/ (95 1.871/ 187 (1.871/ 130 1.306/ (1.306/1 510/ (510/ 55/ (55/ (28 (19 (11 (9 1.871 18 (1.871 13 1.306 (1.306/ (510 (55 (2 (1 ( 1.87 (1.87 1.30 (1.306 (51 (5 1.8 (1.8 1.3 (1.30 1. (1. (1.3

ABSTRACT: Bovine mastitis is the most impacting disease of dairy industry, and it is characterized by a complexity of causal agents, which have revealed a geographical variation among regions and countries. The mastitis-related pathogens have been traditionally classified as contagious or environmental, based on habits of the microorganisms and transmission routes. Besides, the severity of mammary infections has been associated with the virulence of the pathogens, and immune and nutritional status of the hosts. Considering this scenario, we investigated the etiological pattern, clinical severity scores, and days in milk (DIM) data in 4,273 clinical cases of bovine mastitis among ten large-dairy farms located in Brazil. Streptococcus dysgalactiae (283/4,273 = 6.6%), Escherichia coli (190/4,273 = 4.4%), Prototheca spp. (112/4,273 = 2.6%), and Streptococcus uberis (95/4,273 = 2.2%) were the predominant pathogens isolated. Among 4,273 clinical cases, clinical severity score was available in 43.8% (1,871/4,273) animals. From these, 69.8% (1,306/1,871), 27.3% (510/1,871), and 2.9% (55/1,871) were scored as mild, moderate, and severe, respectively. Pathogens were observed mainly in samples obtained during the first 100 days in milk, and clinical severity scored mainly as mild. Our results contributed to the etiological identification, clinical severity scoring, and milking aspects of mastitic cows in dairy farms with a history of clinical mammary infections. ABSTRACT industry agents countries mastitisrelated related environmental routes Besides hosts scenario pattern scores DIM (DIM 4273 4 273 4,27 largedairy large Brazil 283/4,273 2834273 283 (283/4,27 6.6%, 66 6.6% , 6 6.6%) 190/4,273 1904273 190 (190/4,27 4.4%, 44 4.4% 4.4%) spp 112/4,273 1124273 112 (112/4,27 2.6%, 26 2.6% 2 2.6%) 95/4,273 954273 95 (95/4,27 2.2% 22 isolated 438 43 8 43.8 1,871/4,273 18714273 1 871 (1,871/4,273 animals these 698 69 69.8 1,306/1,871, 13061871 1,306/1,871 306 (1,306/1,871) 27 3 27.3 510/1,871, 5101871 510/1,871 510 (510/1,871) 29 9 2.9 55/1,871 551871 55 (55/1,871 mild moderate severe respectively 10 identification scoring 427 4,2 283/4,27 283427 28 (283/4,2 6.6 190/4,27 190427 19 (190/4,2 4.4 112/4,27 112427 11 (112/4,2 2.6 95/4,27 95427 (95/4,2 2.2 43. 1,871/4,27 1871427 87 (1,871/4,27 69. 1306187 1,306/1,87 30 (1,306/1,871 27. 510187 510/1,87 51 (510/1,871 2. 55/1,87 55187 5 (55/1,87 42 4, 283/4,2 28342 (283/4, 6. 190/4,2 19042 (190/4, 4. 112/4,2 11242 (112/4, 95/4,2 9542 (95/4, 1,871/4,2 187142 (1,871/4,2 130618 1,306/1,8 (1,306/1,87 51018 510/1,8 (510/1,87 55/1,8 5518 (55/1,8 283/4, 2834 (283/4 190/4, 1904 (190/4 112/4, 1124 (112/4 95/4, 954 (95/4 1,871/4, 18714 (1,871/4, 13061 1,306/1, (1,306/1,8 5101 510/1, (510/1,8 55/1, 551 (55/1, 283/4 (283/ 190/4 (190/ 112/4 (112/ 95/4 (95/ 1,871/4 1871 (1,871/4 1306 1,306/1 (1,306/1, 510/1 (510/1, 55/1 (55/1 283/ (283 190/ (190 112/ (112 95/ (95 1,871/ 187 (1,871/ 130 1,306/ (1,306/1 510/ (510/1 55/ (55/ (28 (19 (11 (9 1,871 18 (1,871 13 1,306 (1,306/ (510/ (55 (2 (1 ( 1,87 (1,87 1,30 (1,306 (510 (5 1,8 (1,8 1,3 (1,30 (51 1, (1, (1,3

RESUMO: A raiva é uma encefalite viral que acomete os mamíferos, incluindo humanos. O diagnóstico laboratorial rápido e eficaz do vírus da raiva é importante para a saúde pública. Este estudo teve como objetivo avaliar a viabilidade operacional, técnica e financeira da RT-qPCR em substituição à inoculação em camundongos para o diagnóstico da raiva na rotina laboratorial. Foram analisadas 316 amostras de cérebros de mamíferos, 121 positivas e 195 negativas, previamente diagnosticadas por imunofluorescência direta (IFD) e isolamento viral em camundongos, por inoculação intracerebral em camundongos. As amostras foram submetidas à técnica duplex TaqMan RT-qPCR. A precisão, sensibilidade e especificidade da RT-qPCR foram analisadas. Os custos para realizar a técnica de RT-qPCR foram analisados e comparados com o custo do isolamento viral em camundongos. Os resultados demostraram 99,37% de acurácia, 99,17% de sensibilidade e 99,49% de especificidade pela RT-qPCR quando relacionada aos resultados de IFD e isolamento viral em camundongos, que se mostrou uma técnica robusta e com repetibilidade. O tempo mínimo para diagnóstico positivo foi reduzido na RT-qPCR (1 dia) quando comparado ao isolamento viral em camundongos (9,64 dias), e com redução de 17,7% no custo pela técnica molecular. O presente estudo demonstrou que a técnica de biologia molecular é uma ferramenta eficiente para o diagnóstico da raiva na rotina laboratorial, podendo substituir o isolamento viral em camundongos. RESUMO mamíferos humanos pública operacional RTqPCR RT qPCR 31 12 19 negativas (IFD RTqPCR. qPCR. precisão 9937 99 37 99,37 acurácia 9917 17 99,17 9949 49 99,49 repetibilidade 1 ( dia 9,64 964 9 64 (9,6 dias, dias , dias) 177 7 17,7 3 993 99,3 991 99,1 994 4 99,4 9,6 96 6 (9, 17, 99, 9, (9

ABSTRACT: Rabies is a viral encephalitis that affects mammals, including humans. Rapid and effective laboratory diagnosis of the rabies virus is critical for public health. This study evaluated the operational, technical, and financial viability of the RT-qPCR in replacement of inoculation in mice for diagnosing rabies in the laboratory routine. A total of 316 samples of mammalian brains were analyzed, 121 positives and 195 negatives, previously diagnosed by direct immunofluorescence (DIF) and mouse inoculation test (MIT). The samples were submitted to the duplex TaqMan RT-qPCR technique. The accuracy, sensitivity, and specificity of RT-qPCR were analyzed. We analyzed the costs for performing the RT-qPCR technique and compared it with the cost of MIT. The results showed 99.37% accuracy, 99.17% sensitivity, and 99.49% specificity by RT-qPCR when related to DIF and MIT results, which proved to be a robust and repeatable technique. The minimum time for a positive diagnosis was reduced in RT-qPCR (1 day) if compared to MIT (9.64 days), with a 17.7% reduction in the cost of the molecular technique. The present study demonstrated that the molecular biology technique is an efficient tool to diagnose rabies in the laboratory routine, being able to replace MIT. ABSTRACT mammals humans health operational technical RTqPCR RT qPCR routine 31 12 19 negatives (DIF . (MIT) accuracy sensitivity 9937 99 37 99.37 9917 17 99.17 9949 49 99.49 1 ( day 9.64 964 9 64 (9.6 days, days , days) 177 7 17.7 3 (MIT 993 99.3 991 99.1 994 4 99.4 9.6 96 6 (9. 17. 99. 9. (9

RESUMO: Escherichia coli é reconhecida como um dos principais microrganismos responsáveis pelo desencadeamento da mastite clínica, doença que causa perdas econômicas consideráveis na indústria de laticínios. Neste contexto, este estudo teve como objetivo principal a identificação de isolados de E. coli presentes em amostras individuais de leite coletadas de vacas com diagnóstico de mastite clínica, de diversas regiões do Brasil. Adicionalmente, por reação em cadeia da polimerase (PCR), foi investigada a presença dos genes de virulência eae, bfpB, escN, aatA, aggR, ipaH, stx1, stx2, est e eltA, todos associados aos patótipos de E. coli diarreiogênica (DEC). Como parte integrante do estudo, foi realizada uma avaliação abrangente do perfil de sensibilidade dos isolados a 11 antimicrobianos diferentes amplamente utilizados no tratamento da mastite. Foram coletadas 198 amostras de leite de vacas com diagnóstico de mastite clínica. Dentre essas amostras, 12 isolados (6,07%) demonstraram crescimento bacteriano superior a três Unidades Formadoras de Colônia (UFC) quando cultivadas em meio ágar MacConkey e características morfológicas de E. coli. Para avaliar a suscetibilidade desses isolados aos antimicrobianos foi utilizado o teste de disco-difusão, sendo observada a resistência mais predominante em relação à estreptomicina e à tetraciclina, afetando 16,67% das cepas analisadas. É relevante ressaltar que todos os isolados investigados não demonstraram a presença dos genes eae, aatA, aggR, ipaH, stx1, stx2, est e eltA. Estes resultados indicam que estes isolados não se enquadram nos patótipos conhecidos como Escherichia coli diarreiogénica (DEC). Porém, um dos isolados testados apresentou positividade para os genes bfpB e escN. A detecção de E. coli resistente associada à mastite clínica aponta para possíveis lacunas no tratamento da doença. Além disso, a presença de genes de resistência em estirpes de E. coli indica a capacidade potencial de transmitir estes genes entre animais e talvez ao longo da cadeia alimentar. RESUMO laticínios contexto E Brasil Adicionalmente PCR, PCR , (PCR) eae escN aatA aggR ipaH stx1 stx stx2 eltA DEC. DEC . (DEC) 1 19 6,07% 607 6 07 (6,07% UFC (UFC discodifusão, discodifusão disco difusão, difusão disco-difusão tetraciclina 1667 16 67 16,67 analisadas Porém disso alimentar (PCR (DEC 6,07 60 0 (6,07 166 16,6 6,0 (6,0 16, 6, (6, (6 (

ABSTRACT: Escherichia coli is recognized as one of the main microorganisms responsible for triggering clinical mastitis, a disease that causes considerable economic losses in the dairy industry. In this context, this study aimed to identify E. coli isolates present in individual milk samples collected from cows diagnosed with clinical mastitis from various regions of Brazil. Additionally, through polymerase chain reaction (PCR), the presence of virulence genes eae, bfpB, escN, aatA, aggR, ipaH, stx1, stx2, est, and eltA was investigated; all associated with the pathotypes of diarrheagenic Escherichia coli (DEC). As an integral part of the study, a comprehensive assessment of the sensitivity profile of the isolates to 11 different antimicrobials widely used in mastitis treatment was also conducted. A total of 198 milk samples were collected from cows diagnosed with clinical mastitis. Among these samples, 12 isolates (6.07%) demonstrated bacterial growth greater than three Colony-Forming Units (CFU) when grown on MacConkey agar medium and morphological characteristics of E. coli. The disc-diffusion test was used to evaluate the susceptibility of these isolates to antimicrobials, and the most predominant resistance was observed concerning streptomycin and tetracycline, affecting 16.67% of the strains analyzed. Notably, all isolates investigated did not demonstrate the presence of the genes eae, aatA, aggR, ipaH, stx1, stx2, est, and eltA. These results indicate that these isolates do not fit the pathotypes known as diarrheagenic Escherichia coli (DEC). However, one of the isolates tested was positive for the bfpB and escN genes. The detection of resistant E. coli associated with clinical mastitis points to possible gaps in the treatment of the disease. Additionally, the presence of resistance genes in E. coli strains indicates the potential to transmit these genes between animals and, perhaps, along the food chain. ABSTRACT industry context E Brazil Additionally PCR, PCR , (PCR) eae aatA aggR ipaH stx1 stx stx2 est DEC. DEC . (DEC) 1 conducted 19 6.07% 607 6 07 (6.07% ColonyForming Colony Forming CFU (CFU discdiffusion disc diffusion tetracycline 1667 16 67 16.67 analyzed Notably However perhaps (PCR (DEC 6.07 60 0 (6.07 166 16.6 6.0 (6.0 16. 6. (6. (6 (

ABSTRACT Background: Leptospirosis represents a One Health issue, affecting humans and animals. This study investigated pathogenic leptospires in small wild rodents in São Paulo, Brazil. Methods: Kidney samples from 164 rodents underwent qPCR testing, targeting pathogenic Leptospira spp. Results: Thirty-five animals (21.34%) tested positive, including five rodent species: Akodon montensis (2/21; 9.5%), Necromys lasiurus (1/4; 25%), Oligoryzomys nigripes (24/92; 26.1%), Oligoryzomys flavescens (5/26; 19.2%), and Sooretamys angouya (3/14; 21.4%). Botucatu municipality exhibited the highest prevalence, with 42.5% (20/47) of the animals testing positive. Conclusions: The presence of Leptospira spp. in wild rodents suggests they may be chronic carriers, contaminating the environment. Background issue Paulo Brazil Methods 16 spp Results Thirtyfive Thirty 21.34% 2134 21 34 (21.34% positive species 2/21 221 2 (2/21 9.5%, 95 9.5% , 9 5 9.5%) 1/4 14 1 4 (1/4 25%, 25 25% 25%) 24/92 2492 24 92 (24/92 26.1%, 261 26.1% 26 26.1%) 5/26 526 (5/26 19.2%, 192 19.2% 19 19.2%) 3/14 314 3 (3/14 21.4%. 214 21.4% . 21.4%) prevalence 425 42 42.5 20/47 2047 20 47 (20/47 Conclusions carriers environment 21.34 213 (21.34 2/2 22 (2/2 9.5 1/ (1/ 24/9 249 (24/9 26.1 5/2 52 (5/2 19.2 3/1 31 (3/1 21.4 42. 20/4 204 (20/4 21.3 (21.3 2/ (2/ 9. (1 24/ (24/ 26. 5/ (5/ 19. 3/ (3/ 21. 20/ (20/ (21. (2 ( (24 (5 (3 (20 (21

ABSTRACT This review aims to provide current information about Q fever, elucidating the etiological, epidemiological, pathogenic, clinical, diagnostic, therapeutic, and prophylactic aspects of the disease for the medical community. We discuss the main forms of presentation of the agent, its ability to persist in the body, the infinite possibilities of susceptible hosts, the main known forms of transmission, its importance in populations at occupational risk, and the role of arthropods in the natural history of the disease. Focusing on Brazil, we present the cases already described and studies developed since its first report, and how there is still much to unravel. We are aware of the possibilities of the persistence of the agent and the development of severe clinical pictures and the specific treatments currently instituted. We also wish to raise awareness about the future, the new genotypes that are emerging, the need to study the effects of vaccines, and the impact of Q fever on the population. Q fever is a poorly understood disease in Latin America, and recent studies, especially in Brazil, have revealed the importance of developing new studies. etiological epidemiological pathogenic diagnostic therapeutic community body hosts transmission risk Brazil report unravel instituted future emerging vaccines population America

Resumo Objetivo: investigar o efeito do uso de diferentes agentes (hialuronidase tópica, fotobiomodulação e da associação da fotobiomodulação com a hialuronidase tópica) na prevenção de formação de lesões causadas por extravasamento de doxorrubicina bem como na diminuição de lesões formadas pelo extravasamento desta droga. Método: estudo experimental com 60 ratos Wistar, randomizados em quatro grupos de 15 animais. Grupo 1 (Controle); Grupo 2 (Hialuronidase); Grupo 3 (Fotobiomodulação) e Grupo 4 (Hialuronidase + Fotobiomodulação). Induziu-se ferida aplicando 1 mg de doxorrubicina no subcutâneo do dorso dos animais. A concentração da hialuronidase tópica foi de 65 unidades de turbidez/g, a energia empregada foi de 1 joule de laser vermelho 100 mW por centímetro quadrado. Com avaliação macroscópica a cada dois dias por 28 dias, observou-se as variáveis: integridade da pele, presença de flictema, hiperemia, exsudato, sangramento, edema, crosta, descamação e tecido de granulação. Resultados: os animais dos grupos com fotobiomodulação obtiveram melhores resultados na avaliação das variáveis: sangramento, hiperemia, exsudato, pele íntegra e edema. Conclusão: evidenciou-se que a associação da fotobiomodulação com a hialuronidase tópica foi eficaz na diminuição dos efeitos locais e auxiliou no processo de cicatrização da ferida e que a FBM isolada foi capaz de prevenir o aparecimento de lesões.

Abstract Objective: to investigate the effect of using different agents (topical hyaluronidase, photobiomodulation, and the association of photobiomodulation with topical hyaluronidase) in preventing the formation of lesions caused by doxorubicin extravasation, as well as in the reduction of lesions formed by extravasation of this drug. Method: a quasi-experimental study conducted with 60 Wistar rats, randomized into four groups with 15 animals each. Group 1 (Control); Group 2 (Hyaluronidase); Group 3 (Photobiomodulation); and Group 4 (Hyaluronidase + Photobiomodulation). A wound was induced by applying 1 mg of doxorubicin to the subcutaneous tissue of the back of the animals. The concentration of topical hyaluronidase was 65 turbidity units/g and the energy employed was 1 joule of 100 mW red laser per square centimeter. With macroscopic evaluation every two days for 28 days, the following variables were observed: skin integrity, presence of blisters, hyperemia, exudate, bleeding, edema, crust, peeling and granulation tissue. Results: the animals from the groups subjected to photobiomodulation obtained better results in the assessment of the following variables: bleeding, hyperemia, exudate, intact skin and edema. Conclusion: it was evidenced that the association of photobiomodulation with topical hyaluronidase was effective in reducing the local effects and assisted in the wound healing process, and that PBM alone was able to prevent appearance of lesions.

Resumen Objetivo: investigar el efecto del uso de diferentes agentes (hialuronidasa tópica, fotobiomodulación y la combinación de fotobiomodulación y hialuronidasa tópica) en la prevención de la formación de lesiones causadas por la extravasación de doxorrubicina y en la reducción de las lesiones formadas por la extravasación de ese fármaco. Método: estudio experimental con 60 ratas Wistar, distribuidos aleatoriamente en cuatro grupos de 15 animales. Grupo 1 (Control); Grupo 2 (Hialuronidasa); Grupo 3 (Fotobiomodulación) y Grupo 4 (Hialuronidasa + Fotobiomodulación). La herida se indujo aplicando 1 mg de doxorrubicina por vía subcutánea en el lomo de los animales. La concentración de hialuronidasa tópica fue de 65 unidades de turbidez/g, la energía utilizada fue de 1 joule de láser rojo de 100 mW por centímetro cuadrado. En la evaluación macroscópica cada dos días durante 28 días se observaron las siguientes variables: piel intacta, presencia de flictena, hiperemia, exudado, sangrado, edema, costra, descamación y tejido de granulación. Resultados: los animales de los grupos con fotobiomodulación obtuvieron mejores resultados en la evaluación de las variables: sangrado, hiperemia, exudado, piel intacta y edema. Conclusión: se demostró que la combinación de fotobiomodulación y hialuronidasa tópica fue eficaz para reducir los efectos locales y ayudó en el proceso de cicatrización de heridas y que la FBM por sí sola previno la aparición de lesiones.

Resumo Toxoplasma gondii é um parasito zoonótico de distribuição mundial que pode infectar várias espécies de animais homeotérmicos. Poucos estudos avaliaram a exposição de animais silvestres em cativeiro ao T. gondii. Este estudo envolveu uma pesquisa sorológica de anticorpos anti-T. gondii em mamíferos mantidos no Zoológico Nacional de Cuba (PZN) e no Zoológico do Rio de Janeiro (RIOZOO) no Brasil. O estudo consistiu em um total de 231 amostras de soro de mamíferos, sendo 108 do PZN e 123 do RIOZOO. Todas as amostras foram submetidas à pesquisa de IgG anti-T. gondii pelos métodos de ELISA de inibição (PZN) e teste de aglutinação modificado (RIOZOO). Anticorpos de T. gondii foram detectados em 85,2% das amostras do PZN e 32,5% das amostras do RIOZOO. No PZN, Perissodactyla (92,3%) foi a ordem com maior taxa de prevalência sorológica, enquanto no RIOZOO a ordem Primatas (46,7%) se destacou (p <0,05). Além dessa associação, a origem dos animais PZN também foi associada à infecção por T. gondii. Esse achado demonstra a necessidade de monitoramento veterinário constante de mamíferos silvestres em cativeiro, a fim de vincular o diagnóstico sorológico a alterações clínicas indicativas de toxoplasmose.

Abstract Toxoplasma gondii is a zoonotic parasite of worldwide distribution that can infect several species of homeothermic animals. Few studies have evaluated the exposure of captive wild animals to T. gondii. This study involved a serological survey of anti-T. gondii antibodies in mammals kept in Cuba’s National Zoo (PZN) and in the Rio de Janeiro Zoo (RIOZOO) in Brazil. The study consisted of a total of 231 serum samples from mammals, 108 from PZN and 123 from RIOZOO. All the samples were subjected to IgG anti-T. gondii testing by means of the inhibition ELISA method and the modified agglutination test, respectively. T. gondii antibodies were detected in 85.2% samples from PZN and 32.5% samples from RIOZOO. At the PZN, Perissodactyla (92.3%) was the order with the highest serological prevalence rate, whereas at the RIOZOO, the order Primates (46.7%) stood out (p<0.05). In addition to this association, the origin of the PZN animals was also associated with T. gondii infection. This finding demonstrates the need for constant veterinary monitoring of captive wild mammals in order to link the serological diagnosis with clinical alterations indicative of toxoplasmosis.

Resumo A toxoplasmose está mundialmente distribuída e causa perdas na produção animal e problemas de saúde pública. Objetivou-se detectar Toxoplasma gondii e Sarcocystis spp. em 141 produtos cárneos de origem suína (49), bovina (48) e de quibe cru (44), comercializados em mercados de Botucatu, SP, Brazil. Realizou-se bioensaio das amostras em camundongos para isolamento do parasita, e detecção do DNA pela reação em cadeia pela polimerase, tendo como alvo a região do elemento repetitivo de 529 pares de bases (PCR-529-bp). Todas as amostras foram negativas ao bioensaio e 9 (6,38%) positivas à PCR, sendo 5/48 bovinas, 3/49 suínas e 1/44 quibe. Determinou-se a genotipagem das amostras positivas pela multiplex-, nested- e RFLP-PCR com 11 marcadores (SAG1, 5’-3’SAG2, alt.SAG2, SAG3, B-TUB, GRA6, L358, c22-8, c29-6, PK1, Apico). Obteve-se genótipo completo em uma amostra, semelhante a outros já identificados (MAS, TgCkBr89 e TgCkBr147). Nested- e RFLP-PCR do gene 18S rRNA das amostras positivas à PCR foram realizadas, e os produtos da nested-PCR, sequenciados e alinhados com dados do GenBank no NCBI. Quatro apresentaram 100% de homologia com T. gondii (L37415.1), duas Sarcocystis hominis (AF006471.1), duas Sarcocystis cruzi (AF176934.1), uma Sarcocystis hirsuta (AF006469.1), indicando a circulação de T. gondii e Sarcocystis spp.

Abstract Toxoplasmosis occurs worldwide causing economic losses to the animal production and problems to the public health. The study aimed to detect Toxoplasma gondii and Sarcocystis spp.in 141 meat products from commercial meat cuts of pork, beef, and kibbeh sold in commercial markets from Botucatu, SP, Brazil. Samples were bioassayed in mice to isolate the parasite, and the parasite DNA detected by PCR targeting the 529 base pairs repeat element region (PCR-529-bp). All samples resulted negative on bioassay, whereas PCR positive for 9 (6,38%), distributed as 5/48 beef, 3/49 pork, and 1/44 kibbeh. PCR-positive were investigated for the the parasite genotype using multiplex-, nested-, and RFLP-PCR for 11 markers (SAG1, 5’-3’SAG2, alt.SAG2, SAG3, B-TUB, GRA6, L358, c22-8, c29-6, PK1, Apico). Complete genotype was determined on just one PCR-positive sample that matched MAS, TgCkBr89 and TgCkBr147 isolates already identified. In addition, nested- and RFLP-PCR targeting 18S rRNA was run for all PCR-positive samples and, the products, sequenced and aligned to the GenBank at NCBI website. Four samples showed 100% homology with T. gondii (GenBank #L37415.1), three with Sarcocystis hominis (GenBank #AF006471.1), two Sarcocystis cruzi (GenBank #AF176934.1), and one Sarcocystis hirsuta (GenBank #AF006469.1), indicating the circulation of T. gondii and Sarcocystis spp.

Abstract INTRODUCTION: Leptospirosis and brucellosis cause immunosuppression that worsens the clinical condition of people living with HIV/AIDS (PLWHA). We investigated the serological profile and risk factors of PLWHA. METHODS: Serum samples (n=238) were researched for Brucella spp. antibodies using Rose Bengal and tube agglutination tests and Leptospira spp. antibodies using the microscopic agglutination test. RESULTS: All samples were negative for Brucella spp. For leptospirosis, four samples (1.69%) were positive, and Andamana was the prevalent serovar. CONCLUSIONS: Low or no detection of these zoonoses does not reduce their importance in PLWHA. Vigilant, educational, and preventive measures should be adopted.

Abstract INTRODUCTION: Visceral leishmaniasis (VL) is an important zoonosis in Brazil. Previous identification of parasitized dogs can also help prevent the disease in humans, even in non-endemic areas of the country. The Brazilian Ministry of Health recommends diagnosis in dogs using a DPP® (rapid test) as a screening test and an immunoenzymatic assay (ELISA) as a confirmatory test (DPP®+ELISA), and culling infected dogs as a legal control measure. However, the accuracy of these serological tests has been questioned. METHODS: VL in dogs was investigated in a non-endemic area of the São Paulo state for three consecutive years, and the performances of different diagnostic tests were compared. RESULTS: A total of 331 dog samples were collected in 2015, 373 in 2016, and 347 in 2017. The seroprevalence by DPP®+ELISA was 3.3, 3.2, and 0.3%, respectively, and by indirect immunofluorescence assay (IFA), it was 3.0, 5.6, and 5.5%, respectively. ELISA confirmed 18.4% of DPP® positive samples. The concordance between the IFA and DPP® was 83.9%. The concordance between IFA and DPP®+ELISA was 92.9%. A molecular diagnostic test (PCR) was performed in 63.2% of the seropositive samples, all of which were negative. CONCLUSIONS: In non-endemic areas, diagnostic tests in dogs should be carefully evaluated to avoid false results.

RESUMO Objetivo: Analisar a contração de feridas e padrão histomorfométrico das lesões em ratos Wistar submetidos ao extravasamento de doxorrubicina. Método: Utilizados 60 ratos fêmeas adultos, divididos em quatro grupos de quinze animais: Grupo 1 (Controle, sem antídoto); Grupo 2 (Hialuronidase); Grupo 3 (Fotobiomodulação) e Grupo 4 (Hialuronidase + Fotobiomodulação). Aplicado 1 mg (0,5 ml) de doxorrubicina no subcutâneo do dorso dos animais, induzindo a ferida. Realizada avaliação macroscópica e morfométrica das lesões a cada dois dias por 28 dias. No 30º dia foi realizada a eutanásia e coleta do material para avaliação histológica. Resultados: Os animais dos grupos fotobiomodulação e fotobiomodulação+Hialuronidase apresentaram tecido de regeneração com neovascularização e inflamação aguda, com melhora na cicatrização das feridas, o que não ocorreu nos outros grupos. As taxas de contração foram melhores nos tratados com fotobiomodulação e fotobiomodulação + Hialuronidase, com percentual de cicatrização de 76,6% e 72,1%, respectivamente. Conclusão: A combinação da fotobiomodulação (660 nm–1 J) com hialuronidase tópica (65 UTR) mostrou-se eficaz no processo de cicatrização de feridas por extravasamento de doxorrubicina, podendo ser incorporada à pratica da oncologia clínica.

ABSTRACT Objective: To analyze wound contraction and histomorphometric pattern of lesions in Wistar rats undergoing doxorubicin extravasation. Method: Sixty adult female rats were used, divided into four groups of fifteen animals: Group 1 (Control, without antidote); Group 2 (Hyaluronidase); Group 3 (Photobiomodulation), and Group 4 (Hyaluronidase + Photobiomodulation). Doxorubicin 1mg (0.5 ml) was applied subcutaneously on the animals’ back, inducing the wound. Macroscopic and morphometric evaluation of the lesions was performed every two days for 28 days. On the 30th day, euthanasia was performed and the material was collected for histological evaluation. Results: The animals in the photobiomodulation and photobiomodulation + Hyaluronidase groups presented regeneration tissue with neovascularization and acute inflammation, with improvement in wound healing, which did not occur in the other groups. The contraction rates were better in those treated with photobiomodulation and photobiomodulation + Hyaluronidase, with healing percentages of 76.6% and 72.1%, respectively. Conclusion: The combination of photobiomodulation (660 nm–1 J) with topical hyaluronidase (65 UTR) proved to be effective in the process of wound healing due to extravasation of doxorubicin, and can be incorporated into the practice of clinical oncology.

RESUMEN Objetivo: Analizar la contracción de heridas y patrón histomorfométrico de las lesiones en ratones Wistar sometidos a la extravasación de doxorrubicina. Método: Fueron utilizadas 60 ratas adultas, divididas en cuatro grupos de quince animales: Grupo 1 (Control sin antídoto); Grupo 2 (Hialuronidasa); Grupo 3 (Fotobiomodulación) y Grupo 4 (Hialuronidasa + Fotobiomodulación). Se aplicó 1 mg (0,5 ml) de doxorrubicina en el subcutáneo del dorso de los animales, induciendo la herida. Realizada evaluación macroscópica y morfométrica de las lesiones cada dos días por 28 días. En el 30° día se realizó la eutanasia y recolección del material para evaluación histológica. Resultados: Los animales de los grupos fotobiomodulación y fotobiomodulación+ Hialuronidasa presentaron tejido de regeneración con neovascularización e inflamación aguda, con mejoría de la cicatrización de las heridas, lo que no ocurrió en otros grupos. Las tasas de contracción fueron mejores en los tratados con fotobiomodulación y fotobiomodulación + Hialuronidasa, con porcentaje de cicatrización de 76,6% y 72,1%, respectivamente. Conclusión: la combinación de la fotobiomodulación (660nm–1J) con hialuronidasa tópica (65 UTR) demostró ser más eficaz en el proceso de cicatrización de heridas por extravasación de doxorrubicina, lo que posibilitó adoptarla en la práctica de la oncología clínica.

Abstract Background: The early symptoms of leptospirosis and dengue fever are difficult to distinguish and can cause diagnostic confusion. Due to the large dengue epidemics that has occurred in Brazil in recent years, it is possible that cases of leptospirosis were unreported. Therefore, we performed a retrospective study to detect leptospirosis in patients who were tested for dengue, but whose laboratory diagnoses were negative. Methods: Sera samples from 2,017 patients from 48 cities located in the central region of São Paulo state, Brazil, were studied. All samples were subjected to the microscopic agglutination test (MAT), 305 of which were taken from patients five days or less since the onset of symptoms, and were additionally subjected to real-time polymerase chain reaction (PCR). Results: The overall prevalence of leptospirosis cases was 21 (1.04%), with 20 through MAT (18 for Icterohaemorrhagiae and two for the Cynopteri serogroup) and one through PCR (amplicon sequencing compatible with Leptospira interrogans). According to previously established criteria, eight cases of leptospirosis were classified as “confirmed” and 13 as “probable”. The Brazilian notification system for health surveillance had no records for 16 patients positive for leptospirosis and, thus, they were considered unreported cases. Statistical analyses revealed that the prevalence of leptospirosis was higher in men (1.56%) than in women (0.56%), and the mean age was higher in positive patients (43.7 years) than in negative ones (32.3 years). Conclusion: The results indicated that patients suspected of dengue fever had evidence of leptospirosis or Leptospira infection, and most of these cases were unreported in the Brazilian notification system. The high burden of dengue may contribute to the misdiagnosis of leptospirosis, and health professionals should increase their awareness of leptospirosis as an important differential diagnosis of patients with suspicion of dengue.

RESUMO: O gênero Mycoplasma inclui mais de 200 espécies que causam doenças nos animais. É responsável por quadros de mastite em bovinos, podendo também estar relacionado à outras manifestações como artrite e pneumonia em bezerros e novilhas. O presente estudo objetivou a detecção de Mycoplasma bovis isolados a partir de amostras de leite de mastite clínica bovina, bem como, a comparação da taxa de isolamento em dois meios de cultura: Hayflick e SP4. Para efeito de triagem amostral, foram avaliadas quanto à presença da classe Mollicutes 1166 amostras de leite de casos de mastite clínica pela técnica de PCR convencional. Das 1166 amostras de leite avaliadas, 8,6% (100/1166) foram positivas à classe. Nas análises moleculares, obteve-se 1,1% (13/1166) de positividade para Mycoplasma bovis na PCR convencional e 0,9% (11/1166) na PCR em tempo real. Os resultados do cultivo microbiológico das 100 amostras triadas previamente demonstraram 6% (6/100) de crescimento de colônias ao se utilizar o meio Hayflick e 11% (11/100) ao se utilizar o meio SP4 (incluindo as positivas ao primeiro). A partir dos 11 isolados obtidos no cultivo microbiológico, a PCR convencional confirmou Mycoplasma bovis em nove deles, e dois foram negativos para o agente. Na análise filogenética dos isolados, todos agruparam no cluster Mycoplasma bovis e Mycoplasma agalactiae. Frente aos resultados, ressalta-se a importância da presença de Mycoplasma bovis na etiologia da mastite clínica bovina e reforça a necessidade de estudos mais aprofundados para a elucidação de outras espécies de micoplasmas que possam estar envolvidas na mastite bovina.

ABSTRACT: The genus Mycoplasma includes more than 200 bacterial species that cause disease in animals. It is responsible for causing mastitis in bovines and may be related to other manifestations, such as arthritis and pneumonia in calves and heifers. The present study aimed to detect Mycoplasma bovis isolated from milk samples of bovine clinical mastitis, and to compare the isolation rates in two culture media: Hayflick and SP4. An initial screening was performed in order to detect the presence of the class Mollicutes in 1166 milk samples from clinical mastitis by the conventional Polymerase Chain Reaction (PCR) technique. According to the 1166 milk samples evaluated, 8.6% (100/1166) were positive to class Mollicutes. Regarding molecular analyses, 1.1% (13/1166) of conventional PCR for positive M. bovis was obtained and 0.9% (11/1166) in real-time PCR. The results of the microbiological culture of the 100 samples previously screened demonstrated that 6% (6/100) of colony growth have been developed when using the Hayflick medium, and 11% (11/100) when using the SP4 medium (including the positive on Hayflick medium). Concerning the 11 isolates obtained in the microbiological culture, conventional PCR confirmed M. bovis in nine of them, and two cultures were negative. In the phylogenetic analysis of the isolates, all of them were grouped in M. bovis and M. agalactiae clusters. The results confirmed the importance of the presence of M. bovis in the etiology of bovine clinical mastitis and reinforced the need for further studies to elucidate other Mycoplasma species that may be involved in bovine clinical mastitis in Brazil.

Resumo Toxoplasma gondii é um dos parasitas mais importantes mundialmente. Embora muitos estudos de soroprevalência tenham sido realizados em espécies domésticas e selvagens, dados sobre a incidência cumulativa e distribuição espacial de T. gondii em animais são extremamente escassos. No presente estudo, cães do município de Botucatu, estado de São Paulo, foram acompanhados por um ano, e suas amostras de sangue foram colhidas em três momentos: dias 1, 180 e 360. O soro foi submetido ao teste de imunofluorescência indireta (RIFI), para detectar anticorpos IgG para T. gondii. A idade e o sexo foram comparados com os resultados da RIFI por meio de testes estatísticos. Dos 350 cães soronegativos no dia, 1,53 se tornaram soropositivos nas amostragens subsequentes; portanto, a incidência cumulativa foi de 15,1% de cães expostos/ano. A idade e o sexo não tiveram associação com os resultados da sorologia. A análise espacial demonstrou que os cães soropositivos se distribuíram em todas as áreas compreendidas pelo estudo, com uma concentração significativa em uma região com condições sanitárias precárias e baixos níveis socioeconômicos. T. gondii é frequente e amplamente distribuído na área urbana de Botucatu, e regiões empobrecidas podem estar associadas a maiores níveis de contaminação ambiental por esse parasita.

Abstract Toxoplasma gondii is one of the most important protozoa parasites worldwide. Although many seroprevalence studies have been performed in domestic and wild species, data on the cumulative incidence and the spatial distribution of T. gondii in animals are extremely scarce. In the present study, dogs from Botucatu municipality, São Paulo state, were followed for one year and their blood samples were collected on three moments: days 1, 180, and 360. The sera were submitted to the immunofluorescence antibody test (IFAT) to detect IgG antibodies to T. gondii. Age and sex were compared with IFAT results through statistical tests. Spatial analysis was used to detect clusters of seropositive dogs. Among the 350 dogs that were seronegative on day 1, 53 became seropositive in subsequent samplings; thus, cumulative incidence was 15.1% exposed dogs/year. Age and sex were not associated with serological results. The spatial analysis revealed that seropositive dogs were distributed in all the studied areas, with a significant cluster in a zone with poor sanitary conditions and low socioeconomic status. T. gondii is frequent and widely distributed in the urban area of Botucatu, and impoverished areas are possibly associated with high levels of environmental contamination by this parasite.