RESUMO Este estudo teve como objetivo avaliar a situação demográfica e de saúde de cães e gatos pertencentes a famílias assistidas pela Estratégia de Saúde da Família (ESF), de Santa Maria/RS, Brasil. Esta pesquisa foi um estudo transversal baseado na população adscrita, desenvolvido por meio da aplicação de questionário aos residentes das 16 áreas de ESF da cidade. Este foi o primeiro estudo que abordou as condições dos animais de estimação na área de ESF. Foram estudados 414 domicílios, e 88,5% deles tinham animais de estimação (cães e/ou gatos), com uma média de 2,2 cães e 0,8 gatos por domicílio. Apenas 18,4% (228/1,241) dos animais foram esterilizados (cães, 15,1% [135/891]; gatos, 26,7% [93/348]). Ao considerar o número de cães, os lares com um residente tinham menos cães que os lares com dois ou mais residentes (p=0,006). O nível de educação e a renda familiar não estavam associados ao número de animais (p>0,001). Entretanto, níveis mais altos de educação e renda familiar foram associados com a esterilização de cães, monitoramento veterinário, vacinação e tratamento de ectoparasitas em cães e gatos (p<0,0001). Adicionalmente, a maior renda familiar foi associada a uma maior freqüência de tratamento endoparasitário (p<0,05). O estudo mostra um alto número médio de animais de estimação por domicílio nas áreas de ESF em comparação com a população domiciliar média, bem como uma falta de cuidados veterinários, tornando essencial promover a custódia responsável.

ABSTRACT This study aimed to evaluate the demographic and healthcare situation of dogs and cats owned by families assisted by the Family Health Strategy (FHS), from Santa Maria/RS, Brazil. This research was a cross-sectional and population-based study developed by applying a questionnaire to residents in the 16 FHS areas of the city. This was the first study addressing pet animal conditions in the FHS area. A total of 414 households were studied, and 88.5% of them had pets (dogs and/or cats), with an average of 2.2 dogs and 0.8 cats per household. Only 18.4% (228/1.241) of the animals were sterilized (dogs, 15.1% [135/891]; cats, 26.7% [93/348]). When considering the number of dogs, households with one resident had fewer dogs than households with two or more residents (p=0.006). The level of education and family income were not associated with the number of animals (p>0.001). However, higher levels of education and family income were associated with the sterilization of dogs, veterinary monitoring, vaccination, and treatment of ectoparasites in dogs and cats (p<0.0001). Additionally, the higher family income was associated with a higher frequency of endoparasite treatment (p<0.05). The study shows a high average number of pets per household in FSH areas compared to the average household population as well as a lack of veterinary care, making it essential to promote responsible custody.

RESUMO A mastite é considerada uma das principais doenças que afetam rebanhos leiteiros ao redor do mundo e é causada principalmente por Staphylococcus aureus e Streptococcus spp. ambientais. Eventualmente, patógenos não convencionais, como Mycobacterium de crescimento rápido (RGM), podem também causar mastite crônica, a qual não respondera ao tratamento com antimicrobianos. O diagnóstico de mastite causada por RGM é uma tarefa difícil, e a maioria das vezes esse agente pode ser subdiagnosticado. Neste relato, descreveu-se um caso de mastite clínica em uma vaca, no sul do Brasil, causada por RGM Mycobacteroides abscessus, confirmado por caracterização microbiológica e molecular. Os resultados reforçam a necessidade de um diagnóstico laboratorial detalhado para a identificação do agente e a inclusão deste como diagnóstico diferencial no reconhecimento de mastite crônica não responsiva ao tratamento.

ABSTRACT Mastitis is considered the main disease that affects dairy cattle worldwide, and it is caused mainly by Staphylococcus aureus and environmental Streptococcus spp. Eventually, nonconventional pathogens, as rapidly growing mycobacteria(RGM), may also cause chronic mastitis, which will not be responsive to antibiotic treatments. Diagnosis of mastitis caused by RGM is a difficult task, and most of time this agent may be misdiagnosed. Here we describe a case of clinical mastitis caused by the RGM Mycobacteroides abscessus in a cow from Southern Brazil, confirmed by microbiological and molecular characterization. Our results reinforce the necessity of a detailed laboratorial identification of the agent and to include this agent in differential diagnosis of chronical clinical mastitis nonresponsive to treatment.

RESUMO Neospora caninum é o principal agente etiológico da neosporose em animais domésticos, e sua patogênese compreende duas fases características: aguda e crônica. Roedores são usados como modelos experimentais para simular neosporose bovina aguda e crônica. Neste estudo, foi inoculado um total de 27 gerbilos, fêmeas, com diferentes doses de taquizoítos de N. caninum, visando induzir doença crônica. O DNA foi extraído de diferentes órgãos de cada animal após a morte espontânea ou a eutanásia. Os tecidos encefálicos foram submetidos à PCR em tempo real de alta sensibilidade para detecção de animais com infecção crônica. Todas as outras amostras foram submetidas à PCR padrão. Um total de 11 gerbilos morreu devido à neosporose aguda, como confirmado pela detecção de DNA de N. caninum nos órgãos. A dosagem de antígeno que pode induzir infecção crônica foi de 5x103 taquizoítos/mL de N. caninum. Em seções do encéfalo de alguns animais, que apresentaram sinais clínicos de infecção persistente, encontraram-se 70% de positividade para a seção do encéfalo anterior, sugerindo essa área como preferencial para a formação de cisto. Assim, foram determinadas,, em gerbilos, as dosagens de taquizoítos capazes de induzir infecção crônica ou aguda, bem como foram detectados tecidos positivos, preferencialmente, em órgãos sistêmicos, na fase aguda, e no encéfalo, na crônica.

ABSTRACT Neospora caninum is the main etiologic agent of neosporosis in domestic animals and its pathogenesis comprises two characteristic phases: acute and chronic. Rodents are used as experimental models to mimic acute and chronic bovine neosporosis. In this study, we inoculated a total of 27 female gerbils, with different doses of N. caninum tachyzoites aiming to induce chronic disease. DNA was extracted from different organs of each animal after spontaneous death or euthanasia. Encephalic tissues were submitted to a highly sensitive real time PCR aiming to detect chronically infected animals. All the other samples were submitted to standard PCR. A total of 11 gerbils died due to acute neosporosis, as confirmed by N. caninum DNA detection in organs. 5x103 tachyzoites/mL of N. caninum was the dosage of antigen that can induce chronic infection in gerbils. In the encephalon sections of some animals that showed clinical signs of persistent infection, we found 70% positive for the anterior encephalon section, suggesting this area as preferential for cyst formation. Therefore, we determined the doses of tachyzoites that cause acute or chronic infection and detection of positive tissues, preferably, systemic organs during acute and encephalon in chronic phases.

RESUMO Galinhas criadas ao ar livre podem ingerir oocistos de T. gondii presentes no ambiente e, com isso, albergar cepas virulentas desse parasita em diferentes tecidos, sem sinais clínicos. O isolamento de T. gondii por meio de bioensaios em camundongos e gatos, a partir de tecidos de galinhas naturalmente infectadas, tem sido descrito em vários países. Isso demonstra a importância das galinhas caipiras na epidemiologia desse parasita. O objetivo deste trabalho foi caracterizar genotipicamente isolados de T. gondii obtidos de galinhas caipiras naturalmente infectadas em uma área rural do município de Santa Maria, estado do Rio Grande do Sul, Brasil. Fragmentos de cérebro e de coração, de 12 galinhas soropositivas para T. gondii, foram processados pela técnica de digestão péptica para isolamento do parasita. Das 12 amostras submetidas a bioensaio com camundongos, nove isolados foram obtidos. A caracterização genotípica por RFLP-PCR foi realizada utilizando-se 11 marcadores genéticos: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 e Apico e revelou a presença de cinco genótipos atípicos de acordo com o ToxoDB (# 11, # 55, # 64, # 140 e # 163). Os resultados mostraram uma ampla diversidade genética de T. gondii em galinhas caipiras nessa região.

ABSTRACT Free-range chickens may ingest oocysts of T. gondii present in the environment and consequently harbor virulent strains of this parasite in different tissues, without any clinical signs. Isolation of T. gondii through bioassays on mice and cats from naturally infected chicken tissues has been described in several countries, demonstrating the importance of free-range chickens in the transmission of this parasite. The aim of this study was the genotypic characterization of T. gondii isolates obtained from naturally infected free-range chickens in a rural area of the state of Rio Grande do Sul, Brazil. Brain and heart tissue from 12 chickens seropositive for T. gondii were processed using peptic digestion technique for parasite isolation. From 12 samples subjected to mouse bioassay, nine isolates were obtained. RFLP-PCR genotypic characterization was performed using 11 genetic markers: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genetic characterization of the isolates revealed the presence of five atypical genotypes according to ToxoDB (# 11, # 55, # 64, # 140 and # 163). Our results showed a wide genetic diversity of T. gondii in free-range chickens in this region.

O objetivo deste estudo foi avaliar a exposição intrauterina ao Sarcocystis spp., para determinar o número de potros que possuem concentrações detectáveis de anticorpos contra esses agentes no soro, antes da ingestão do colostro, por meio da coleta de dados sobre a exposição a esses protozoários nos equinos. Amostras de soro foram coletadas de 195 éguas puro-sangue e seus respectivos potros recém-nascidos, em duas fazendas localizadas na região Sul do Brasil. Os testes utilizados na detecção de anticorpos específicos para o Sarcocystis foram a reação de imunofluorescência indireta (RIFI) e análise por meio de immunoblot. Pela RIFI, em 84,1% (159/189) das éguas e em 7,4% (14/189) dos potros foram detectados anticorpos anti-Sarcocystis spp. Todas as amostras soropositivas dos potros também foram positivas para suas respectivas mães. As amostras de soro dos potros soropositivos na RIFI, não apresentaram reatividade no immunoblot, tendo como antígenos merozoítos de S. neurona. Em conclusão, foi demonstrada a exposição intrauterina de Sarcocystis spp. em equinos, com ocorrência em éguas, porém, em seus respectivos potros, antes da ingestão de colostro a ocorrência foi reduzida.

The aim of this study was to examine the intra-uterine exposure to Sarcocystis spp. antigens, determining the number of foals with detectable concentrations of antibodies against these agents in the serum, before colostrum ingestion and collect data about exposure of horses to the parasite. Serum samples were collected from 195 thoroughbred mares and their newborns in two farms from southern Brazil. Parasite specific antibody responses to Sarcocystis antigens were detected using the indirect immunofluorescent antibody test (IFAT) and immunoblot analysis. In 84.1% (159/189) of the pregnant mares and in 7.4% (14/189) of foals we detected antibodies anti-Sarcocystis spp. by IFAT. All samples seropositive from foals were also positive in their respective mares. Serum samples of seropositive foals by IFAT, showed no reactivity on the immunoblot, having as antigens S. neurona merozoites. In conclusion, the intra-uterine exposure to Sarcocystis spp. antigens in horses was demonstrated, with occurrence not only in mares, but also in their foals, before colostrum ingestion these occurrences were reduced.

ABSTRACTSarcocystis neurona is the primary agent for Equine Protozoal Myeloencephalitis (EPM), important neurological disease characterized by behavior or muscular changes, that impairs animal performance and husbandry. Sarcocystis cruzi is a pathogen related to myositis in cattle. Although related the life cycles of the parasites are distinct. S. neurona has opossums (Didelphis spp.) and S. cruzi, dogs as definitive hosts. However, S. neurona and S. cruzi may undergo cross-reactivity in serological tests, interfering on results of EPM ante-mortem diagnostic tests. In the present study, serology of 189 mares was performed by indirect immunofluorescence antibody test, using antigens of S. neurona and S. cruzi in order to assess the exposure degree of animals to antigens. Analyzing the results, it was observed that most of the animals (84.13%) reacted with at least one protozoal species and the number of animals which showed antibodies against S. cruzi was greater than S. neurona (80.42% and 33.86%, respectively) and a third of seropositive animals reacted to antigens of both species.

O uso indiscriminado de produtos químicos no controle do carrapato bovino constitui a principal causa do gradativo aumento do número de cepas resistentes deste parasita às bases disponíveis no mercado. A utilização de óleos essenciais e extratos vegetais é uma prática antiga no controle de carrapatos, porém só recentemente tem recebido a devida atenção dos pesquisadores. O objetivo deste experimento foi avaliar a eficácia in vitro do óleo de capim limão (Cymbopogon citratus) sobre fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus através do exame de biocarrapaticidograma. Foram testadas seis diluições do óleo de C. citratus (1; 5; 10; 25; 50 e 100%) em uma população de carrapatos resistentes a amidínicos e piretróides sintéticos. A inibição de postura foi de 3; 23; 46; 66; 46 e 46%, a eclosão larval foi de 83; 58; 31; 0; 38 e 25% e a eficácia do tratamento foi de 32; 64; 83; 100; 88 e 82%, respectivamente. O óleo de C. citratus apresentou controle parcial do carrapato R. microplus in vitro, mesmo frente a populações resistentes a produtos químicos.

The indiscriminate use of chemical products to control the cattle tick is the main cause of the gradual increase in the number of strains of this parasite that are resistant to the bases currently available in the market. The use of essential oils and plant extracts is an ancient practice for tick control; however, only recently has it received due attention by researchers. The aim of this experiment was to evaluate the in vitro efficacy of lemongrass (Cymbopogon citratus) essential oil on engorged females of Rhipicephalus (Boophilus) microplus through immersion test. Six concentrations of Cymbopogon citratus oil (1; 5; 10; 25; 50 and 100%) were tested against a tick population resistant to synthetic formamidines and pyrethroids. The inhibition of egg-laying was 3; 23; 46; 66; 46 and 46%, the hatching was 83; 58; 31; 0; 38 and 25%, and the treatment efficacy was 32; 64; 83; 100; 88 and 82%, respectively. C. citratus oil showed partial control of the tick R. microplus in vitro, even against populations resistant to chemical products.

Calves born persistently infected with non-cytopathic bovine viral diarrhea virus (ncpBVDV) frequently develop a fatal gastroenteric illness called mucosal disease. Both the original virus (ncpBVDV) and an antigenically identical but cytopathic virus (cpBVDV) can be isolated from animals affected by mucosal disease. Cytopathic BVDVs originate from their ncp counterparts by diverse genetic mechanisms, all leading to the expression of the non-structural polypeptide NS3 as a discrete protein. In contrast, ncpBVDVs express only the large precursor polypeptide, NS2-3, which contains the NS3 sequence within its carboxy-terminal half. We report here the investigation of the mechanism leading to NS3 expression in 41 cpBVDV isolates. An RT-PCR strategy was employed to detect RNA insertions within the NS2-3 gene and/or duplication of the NS3 gene, two common mechanisms of NS3 expression. RT-PCR amplification revealed insertions in the NS2-3 gene of three cp isolates, with the inserts being similar in size to that present in the cpBVDV NADL strain. Sequencing of one such insert revealed a 296-nucleotide sequence with a central core of 270 nucleotides coding for an amino acid sequence highly homologous (98%) to the NADL insert, a sequence corresponding to part of the cellular J-Domain gene. One cpBVDV isolate contained a duplication of the NS3 gene downstream from the original locus. In contrast, no detectable NS2-3 insertions or NS3 gene duplications were observed in the genome of 37 cp isolates. These results demonstrate that processing of NS2-3 without bulk mRNA insertions or NS3 gene duplications seems to be a frequent mechanism leading to NS3 expression and BVDV cytopathology.

Bovine herpesvirus type 5 (BHV-5) is a major agent of meningoencephalitis in cattle and establishes latent infections mainly in sensory nerve ganglia. The distribution of latent BHV-5 DNA in the brain of rabbits prior to and after virus reactivation was studied using a nested PCR. Fifteen rabbits inoculated intranasally with BHV-5 were euthanized 60 days post-inoculation (group A, N = 8) or submitted to dexamethasone treatment (2.6 mg kg-1 day-1, im, for 5 days) and euthanized 60 days later (group B, N = 7) for tissue examination. Two groups of BHV-1-infected rabbits (C, N = 3 and D, N = 3) submitted to each treatment were used as controls. Viral DNA of group A rabbits was consistently detected in trigeminal ganglia (8/8), frequently in cerebellum (5/8), anterior cerebral cortex and pons-medulla (3/8) and occasionally in dorsolateral (2/8), ventrolateral and posterior cerebral cortices, midbrain and thalamus (1/8). Viral DNA of group B rabbits showed a broader distribution, being detected at higher frequency in ventrolateral (6/7) and posterior cerebral cortices (5/7), pons-medulla (6/7), thalamus (4/7), and midbrain (3/7). In contrast, rabbits inoculated with BHV-1 harbored viral DNA almost completely restricted to trigeminal ganglia and the distribution did not change post-reactivation. These results demonstrate that latency by BHV-5 is established in several areas of the rabbit's brain and that virus reactivation leads to a broader distribution of latent viral DNA. Spread of virus from trigeminal ganglia and other areas of the brain likely contributes to this dissemination and may contribute to the recrudescence of neurological disease frequently observed upon BHV-5 reactivation.