Linfoma intraocular em cães - achados da ultrassonografia contrastada e elastografia ARFI / Intraocular lymphoma in dogs - findings of contrast enhanced ultrasound and ARFI elastography

Background: Ocular lymphoma can affect the iris, conjunctiva, choroid, and retina and is mostly associated with multicentric disease. Elastography is an ultrasound technique that provides noninvasive, pain-free assessment of tissue stiffness. It has the ability to assess subtle changes throughout the organ as well as focal lesions. Microbubble contrast ultrasound enables the detection of incipient vascular flows, which are difficult to detect using traditional ultrasound methods. This study aimed to describe acoustic radiation force impulse (ARFI) elastography and microbubble contrast ultrasound findings in the eyes of two dogs diagnosed with intraocular T-cell lymphoma. Cases: Case 1. Physical examination revealed an exophytic mass in the left eye. Schirmer test revealed a secretion of 22 mm/min. Negative threat reflex, glare, direct pupillary light reflex, and consensual response were also noted. Biomicroscopy revealed hyperplasia of the third eyelid, overlapping with the affected eye. When the membrane was removed, moderate conjunctival hyperemia, mucoid secretion, and buphthalmia were observed. In addition, significant corneal edema was present, making it impossible to visualize the anterior chamber and perform fundus examination. The intraocular pressure, as measured with a rebound tonometer, was 39 mmHg. B-mode ultrasonography identified amorphous, hyperechoic, and heterogeneous structures throughout the anterior chamber, iris, and ciliary body. The elastogram showed that the mass had greenish tones and intermediate stiffness, and the mean SWV of the ciliary body and iris was 2 m/s. Contrast-enhanced ultrasound (CEUS) revealed vascularization of the neoformation region, with wash-in, peak, and wash-out values of 9.89 s, 24.56 s, and 107.87 s, respectively. Case 2. On physical examination, a change in the shape of the right pupil was observed. Schirmer test revealed a secretion of 20 mm/min, with negative threat, glare, and pupillary reflexes to direct and consensual light. Biomicroscopy revealed neoformation from 7 am to 11 am in the sclera, retina, and choroid complex, concomitant with dyscoria and conjunctival hyperemia. The intraocular pressure, as measured by rebound tonometry, was 33 mmHg. Fundoscopy revealed a mass in the temporal region and focal retinal detachment. No changes were observed in the contralateral eye. B-mode ultrasound revealed an increase in volume in the temporal region of the iris, ciliary body, and choroid with diffuse heterogeneity and partial retinal detachment. Elastographic examination revealed shades of green and yellow compatible with increased tissue stiffness. On quantitative examination, the mean SWVs of the ciliary body and iris were 3.14 m/s. On CEUS, the neoformation region presented wash-in, peak, and wash-out values of 8.67 s, 22.33 s, and 80.20 s. Discussion: B-mode ultrasonography established the tumor extent and evaluated echogenicity, verifying the involved ocular structures. The examination played an important role in the diagnosis as well as clinical management. ARFI elastography can detect small tissue changes, helping to define nodules and masses more reliably, in addition to allowing the verification of tissue stiffness. In both dogs, it was possible to verify that the masses presented greater rigidity than the adjacent tissues both qualitatively and quantitatively. In previous studies, it was found that cutaneous and breast lymphomas in humans were more rigid than adjacent tissues on elastography. This increase in rigidity and heterogeneity observed on elastograms can be explained by the extramedullary interactions of the matrix in T-cell lymphomas. Tumor growth is dependent on the blood supply, which was evaluated using CEUS in these cases. Furthermore, the ciliary body contrast filling times were longer than those described in normal dogs.

Sampling techniques and environmental variables influence the distribution of pseudoscorpions in urban forest fragments in the central Amazon / Métodos de amostragem e variáveis ambientais influenciam a distribuição de pseudoescorpiões em fragmentos florestais urbanos na Amazônia central

The growth of urban landscapes has genarally reduced biodiversity worldwide. Invertebrates have explored different environments, and it usually takes different sampling techniques to get a representative sample of the species assemblage in a given location. Some studies have sought to determine the minimum necessary number of sampling techniques, including ecological relationships or costs to guide the sampling protocol. In the Amazon, the effect of soil characteristics on invertebrate distribution is well known. We evaluated if sampling techniques have a complementary effect on the detection of pseudoscorpion assemblages and tested whether environmental variables affect the distribution of pseudoscorpion species. The study sites were two urban forest fragments in the city of Manaus, in the central Amazon. In each fragment, we sampled 20 palm trees using the beating technique, and installed transects with 12 sampling points for collection of soil and litter samples for extraction of arthropods in a Berlese funnel and a Winkler extractor, respectively. We collected 267 individuals of 11 species of pseudoscorpions. Most records were obtained through the Winkler extraction in both fragments. The assemblage from the palm trees was different from that in the edaphic samples. Pseudoscorpion species composition also differed significantly between soil and litter, and was influenced by potassium concentration. The number of species in the fragments and the environmental effect on the distribution of pseudoscorpions was similar to that recorded in environmental protection areas, evidencing that urban forest fragments can serve as an efficient repository of Amazonian pseudoscorpion biodiversity.(AU)

O crescimento das paisagens urbanas geralmente reduziu a biodiversidade em todo o mundo. Os invertebrados exploram diferentes ambientes, e geralmente são necessárias diferentes técnicas de amostragem para obter uma amostra representativa da assembleia de espécies em um determinado local. Alguns estudos têm buscado determinar o número mínimo necessário de técnicas de amostragem, incluindo relações ecológicas ou custos para orientar o protocolo de amostragem. Na Amazônia, o efeito das características do solo na distribuição dos invertebrados é bem conhecido. Nós avaliamos se as técnicas de amostragem têm um efeito complementar na detecção de assembleias de pseudoescorpiões e testamos se as variáveis ambientais afetam a distribuição das espécies de pseudoescorpiões. Os locais de estudo foram dois fragmentos florestais urbanos na cidade de Manaus, Amazônia central. Em cada fragmento, amostramos 20 palmeiras com guarda-chuva entomológico e instalamos transectos com 12 pontos amostrais para coleta de solo e serrapilheira para extração de artrópodes em funil de Berlese e extrator de Winkler, respectivamente. Coletamos 267 indivíduos de 11 espécies de pseudoescorpiões. A maioria dos registros foi obtida com Winkler em ambos fragmentos. A assembleia em palmeiras foi diferente das amostras edáficas. A composição de espécies de pseudoescorpiões também diferiu significativamente entre solo e serapilheira, e foi influenciada pela concentração de potássio. O número de espécies nos fragmentos e o efeito ambiental na distribuição de pseudoescorpiões foram semelhantes aos registrados em áreas de proteção ambiental, evidenciando que fragmentos florestais urbanos podem servir como um eficiente repositório da biodiversidade de pseudoescorpiões amazônicos.(AU)

Serological and molecular detection of Theileria equi in horses from Sinop, Mato Grosso state, Brazil / Detecção sorológica e molecular de Theileria equi em equinos oriundos de Sinop, Mato Grosso, Brasil

Equine piroplasmosis is the most important tick-borne disease to affect horses in Brazil. Theileria equi is one of the causative agents of equine piroplasmosis. Chronic cases are expected, in which the animals show no apparent signs of infection and remain asymptomatic but constitute a source of the infectious agent that ticks can spread. This study was conducted across 81 ranches located in the municipality of Sinop, State of Mato Grosso, Brazil. A sample calculation was performed to estimate the apparent prevalence of T. equi among horses. A total of 1,853 animals were included in the sampling analysis based on the information available from the Institute of Agricultural and Livestock Defense of Mato Grosso State. The serological analysis of 367 serum samples using an indirect enzyme-linked immunosorbent assay (ELISA) to detect anti-T. equi antibodies revealed that 337 animals were positive, representing a frequency of 90.70%. The molecular analysis to amplify the EMA-1 gene showed positivity in 20 of 89 tested samples. The fragments of four samples were sequenced and analyzed to determine their similarities to sequences from other species, based on sequences deposited at GenBank. All showed 100% similarity with T. equi. Our study represents the first report of T. equi antibodies among the equids in north-central region of Mato Grosso, revealing the widespread distribution of seropositive animals.

A piroplasmose equina é a doença transmitida por carrapatos mais importante em cavalos no Brasil. Theileria equi é um dos agentes causadores da piroplasmose equina. São esperados casos crônicos, nos quais os animais não apresentam sinais aparentes de infecção e permanecem assintomáticos, mas constituem uma fonte de infecção e disseminação por carrapatos. Este estudo foi realizado em 81 fazendas localizadas no município de Sinop, Estado de Mato Grosso, Brasil. Um cálculo amostral foi realizado para estimar a prevalência aparente de T. equi entre cavalos. No total, 1.853 animais foram incluídos na análise amostral com base nas informações disponíveis no Instituto de Defesa Agropecuária do Estado de Mato Grosso. A análise sorológica de 367 amostras de soro por meio de ensaio imunoenzimático indireto (ELISA) para detecção de anticorpos anti-T. equi revelou que 337 animais eram positivos, representando uma frequência de 90,70%. A análise molecular para o gene EMA-1 mostrou positividade em 20 das 89 amostras testadas. Os fragmentos de quatro amostras foram sequenciados e analisados para determinar suas semelhanças com sequências de outras espécies, a partir das sequências depositadas no GenBank. Todos mostraram 100% de similaridade com T. equi. Nosso estudo representa o primeiro relato de anticorpos contra T. equi entre os equídeos na região centro norte de Mato Grosso, revelando a ampla distribuição de animais soropositivos.

High level of infection by Toxoplasma gondii in pigs slaughtered in the city of São Luís, Maranhão / Alto nível de infecção por Toxoplasma gondii em suínos abatidos na cidade de São Luís, Maranhão

Abstract A serological, molecular and histopathological study was carried out in order to investigate occurrences of Toxoplasma gondii in pigs slaughtered with and without inspection service. Serum samples were collected from 60 pigs to detect anti-T. gondii antibody by indirect fluorescent antibody (IFAT). Tongue, masseter and diaphragm fragments were also collected for parasite DNA detection by means of the polymerase chain reaction (PCR) and histopathological analysis. The serological results showed that 77% (44/60) of the pigs were positive. Regarding PCR, 66.67% (40/60) were positive for T. gondii. Among the tissues evaluated, the diaphragm was the one with the highest frequency of positivity (40%; 24/60), followed by the masseter (38.33%; 23/60) and tongue (33.3%; 20/60). Histopathological changes were only observed in the diaphragm, which presented inflammatory infiltrates of lymphohistiocytic and neutrophilic types. These results not only show the potential threat of T. gondii to human health, but also demonstrate the dynamic epidemiological situation of toxoplasmosis in pigs in the city of São Luís, providing support for food security regarding pigs and for T. gondii control programs in Brazil.

Resumo Realizou-se um estudo sorológico, molecular e histopatológico com o objetivo de verificar a ocorrência de Toxoplasma gondii em suínos abatidos com e sem serviço de inspeção. Foram coletados soros de 60 suínos para a pesquisa de anticorpos anti-T. gondii pela reação de imunofluorescência indireta (RIFI). Também foram coletados fragmentos de língua, masseter e diafragma para a detecção do DNA do parasito por meio da reação em cadeia da polimerase (PCR) e análise histopatológica. A análise sorológica demonstrou que 77% (44/60) dos suínos apresentaram anticorpos anti-T. gondii. Com relação ao PCR, 66,67% (40/60) foram positivos para T. gondii. Dentre os tecidos avaliados, o diafragma foi o que obteve maior frequência de positividade (40%; 24/60), seguidos de masseter (38,33%; 23/60) e língua (33,3%; 20/60). Alterações histopatológicas foram observadas apenas no diafragma, que apresentou infiltrado inflamatório do tipo linfohistiocitário e neutrofílico. Esses resultados não evidenciam apenas a ameaça potencial de T. gondii à saúde humana, mas também demonstram a dinâmica situação epidemiológica da toxoplasmose em suínos na região da cidade de São Luís, fornecendo suporte para a segurança alimentar de suínos e programas de controle de T. gondii no país.

Toxoplasmose encefálica em um gambá-de-orelha-branca (Didelphis albiventris) / Encephalic toxoplasm in a white-eared possum (Didelphis albiventris)

Background: Toxoplasmosis is caused by Toxoplasma gondii, an obligate intracellular protozoan that belongs to the Apicomplexa phylum, coccidian subclass, and affects all warm-blooded animals. The role of opossums in the epidemiology of toxoplasmosis in Brazil is not fully understood, and there are very few descriptions of toxoplasmosis lesions in these animals. This report describes the anatomopathological, molecular and immunohistochemical findings of a case of encephalic toxoplasmosis in free-living white-eared possum (Didelphis albiventris). Case: A young male opossum (D. albiventris), was treated at the Veterinary Hospital of Wild Animals of the University of Brasília, Federal District. The animal was apathetic, uncoordinated, reluctant to move, and had an exposed proximal fracture in the left radius and ulna with laceration of muscles and adjacent tendinous structures. Amputation on the left thoracic limb was performed followed by analgesia and antibiotic therapy. The environment is frequented by other wild animals, and stray cats have access to the patio of the building. Twenty-five days after arriving at the hospital, the animal was found dead in its cage. After death, a necropsy was performed. Organ fragments from the abdominal cavity, thoracic and central nervous system were collected, processed routinely for histology and stained with hematoxylin and eosin. Macroscopic lesions in the central nervous system were not observed. On microscopy, the brain showed moderate random glial nodules throughout the neuropil associated with the presence of spherical to elongated parasitic cysts of about 20 µm, with a thin wall and with its interior full of bradyzoites, consistent with Toxoplasma gondii. There was also moderate fibrinoid necrosis and moderate multifocal lymphoplasmacytic infiltrate surrounding the blood vessels (perivascular cuffs) To investigate the etiology of the brain injury, brain sections were subjected to immunohistochemistry (IHC) and real-time polymerase chain reaction (qPCR) technique for detection of T. gondii and Neospora caninum. Immunostaining for T. gondii in the cyst wall and in bradyzoites and negative immunostaining for N. caninum. qPCR was positive for T. gondii and negative for N. caninum. Discussion: Diagnosis of encephalic toxoplasmosis in a Didelphis albiventris was possible based on histopathological, immunohistochemical and molecular findings. The morphological classification of the brain lesion was important for the diagnosis. Brain toxoplasmosis in opossums usually results in focal areas of malacia on macroscopy and focally extensive necrosis on microscopy, neutrophil infiltrate, calcified necrotic material, and perivascular cuffs of lymphocytes and plasma cells. In the present case, similar histopathological lesions were noted, but no significant macroscopic changes were observed. The etiology here was defined by immunohistochemistry and qPCR, techniques proven to be useful and with good specificity for diagnosing toxoplasmosis in mammals. It is believed that the positive immunohistochemical and molecular result for Toxoplasma gondii together with the negative result for Neospora caninum were conclusive for the diagnosis. Thus, we demonstrate here a post mortem diagnosis of toxoplasmosis in a free-living synanthropic opossum and the use of anatomopathology, immunohistochemistry and real-time polymerase chain reaction as a diagnostic option for this disease in opossums.

Evaluation of quantitative polymerase chain reaction for the detection of Toxoplasma gondii oocysts shed by cats / Avaliação da reação em cadeia da polimerase quantitativa para a detecção de oocistos de Toxoplasma gondii eliminados por gatos

Abstract Felines are definitive hosts of Toxoplasma gondii and can shed oocysts in their feces, contaminating the environment. Sporulated oocysts are highly resistant to the environment and have higher infectivity, which are attributed to many toxoplasmosis outbreaks. The aim of the present study was to evaluate a quantitative polymerase chain reaction (qPCR) technique for the detection of T. gondii oocysts shed by cats. Twelve cats from a previous vaccine experiment were challenged orally with 600 cysts of the TgDoveBr8 strain on day 72. Fecal samples were collected daily using the centrifugal flotation technique, with microscopic examination (Sheather technique) and qPCR for 20 days after the challenge. Cats from all groups shed oocysts in their feces. Five negative cats in the Sheather were positive according to qPCR on the 3rd day post-inoculation (dpi). Oocysts were detected on the 4th dpi using the Sheather; however, there was no statistical difference between the two methods (p=0.1116). In addition, there was no statistically significant difference in oocyst shedding between the groups according to the Sheather technique (p=0.6534) and qPCR (p=0.9670). In conclusion, these results demonstrate that qPCR can be used as an alternative to the Sheather to detect and quantify T. gondii oocysts.

Resumo Felinos são hospedeiros definitivos do Toxoplasma gondii e podem eliminar oocistos nas fezes, contaminando o meio ambiente. Oocistos esporulados são altamente resistentes ao meio ambiente com elevada infectividade, sendo atribuído a muitos surtos de toxoplasmose. O objetivo do estudo foi avaliar a reação em cadeia da polimerase quantitativa (qPCR) para a detecção de oocistos de T. gondii eliminados por gatos. Doze gatos de um experimento prévio de vacina foram desafiados por via oral com 600 cistos da cepa TgDoveBr8 no dia 72. Amostras fecais foram coletadas diariamente pela técnica de centrifugo-flutuação seguida de exame microscópico (técnica de Sheather) e qPCR por 20 dias após desafio. Gatos de todos os grupos eliminam oocistos nas fezes. Cinco gatos negativos na técnica Sheather foram positivos de acordo com a qPCR no 3º dia pós-inoculação (dpi). Oocistos foram detectados no 4º dpi no Sheather; entretanto, não houve diferença estatística entre os dois métodos (p=0,1116). Não houve diferença estatisticamente significativa na eliminação de oocistos entre os grupos de acordo com a técnica de Sheather (p = 0,6534) e qPCR (p = 0,9670). Em conclusão, esses resultados demonstram que qPCR pode ser usada como uma alternativa ao Sheather para detectar e quantificar oocistos de T. gondii.

Macroscopic, histological, and molecular aspects of Sarcocystis spp. infection in tissues of cattle / Aspecto Macroscópico, histológico e molecular de Sarcocystis spp. em tecidos de bovinos e ovinos

The macroscopic, histological, and molecular aspects of Sarcocystis spp. were examined in the tissues of two cattle and four sheep, 16 and eight fragments analyzed respectively, condemned in the slaughterhouse. All 24 samples were collected and analyzed for detecting macrocysts and macroscopic lesions. Subsequently, subdivided for direct examination, polymerase chain reaction and histopathological examination. All sheep tissues samples had grossly white round to oval tissue cysts, ranging from 0.3 to 1 cm in diameter. In contrast, cattle tissues did not present grossly visible cysts but had randomly distributed white-yellow foci with irregular contours. All samples from cattle and sheep had microscopic cysts. In the histological examination of sheep tissues, circular to elongated, encapsulated, basophilic structures ranging from 30 to 3,000 µm in length and 20 to 1,000 µm in width were observed within the skeletal muscle fibers. In cattle tissues, all cardiac muscle four fragments analyzed contained circular to elongated basophilic structures inside cardiomyocytes and in some Purkinje fibers. PCR were performed using the primers: 2L and 3H. In conclusion, all 24 tissues were infected with Sarcocystis spp., and S. gigantea (in sheep) and S. cruzi (in cattle). were the identified species by sequencing.(AU)

Os aspectos macroscópicos, histológicos e moleculares de Sarcocystis spp. foram examinados nos tecidos de dois bovinos e quatro ovinos, 16 e oito fragmentos analisados, respectivamente, condenados no matadouro. Todas as 24 amostras foram coletadas e analisadas para detecção de macrocistos e lesões macroscópicas. Posteriormente, subdivididas para exame direto, reação em cadeia da polimerase e exame histopatológico. Todas as amostras de tecidos de ovelha apresentavam cistos grosseiramente visíveis, caracterizados como brancos, de redondos a ovais e estruturas variando de 0,3 a 1 cm de diâmetro. Em contraste, os tecidos de bovinos não apresentavam cistos grosseiramente visíveis, mas tinham focos branco-amarelos com contornos irregulares, distribuídos aleatoriamente. Todas as amostras de bovinos e ovinos apresentavam cistos microscópicos. No exame histológico de tecidos ovinos foram observadas estruturas basofílicas circulares a alongadas, encapsuladas, variando de 30 a 3.000 µm de comprimento e 20 a 1.000 µm de largura dentro das fibras do músculo esquelético. Nos tecidos de bovinos, todos os quatro fragmentos de músculo cardíaco analisados continham estruturas basofílicas circulares a alongadas, dentro dos cardiomiócitos e em algumas fibras de Purkinje. PCRs foram realizadas utilizando-se os "primers" 2L e 3H. Em conclusão, todos os 24 tecidos estavam infectados com Sarcocystis spp., sendo S. gigantea (em ovinos) e S. cruzi (em bovinos) as espécies identificadas por sequenciamento.(AU)

Diagnóstico de cinomose canina por meio de teste imunocromatográfico e sua correlação com achados clínicos e hematológicos no semiárido da Paraíba / Diagnosis of canine distemper by immunochromatographic test and its correlation with clinical and hematological findings in the semiarid region of Paraíba

Objetivou-se através deste trabalho, determinar a prevalência de cinomose canina no semiárido da Paraíba, através de testes rápidos imunocromatográficos, correlacionando-a com os principais achados clínicos e hematológicos. Foram analisadas 67 fichas de animais em que foram realizados testes rápidos para pesquisa de antígeno em amostras nasais e oculares no período de janeiro a dezembro de 2019. Observou-se que 47% (32/67) dos cães analisados foram positivos para cinomose canina. As variáveis que apresentaram diferença estatística significativa (p<0,05) para a infecção foram animais sem raça definida 60% (21/35), animais não vacinados 70% (29/42), e período seco do ano, sendo o mês de agosto (40%; 13/32), com maior ocorrência. Os principais sistemas afetados foram o respiratório 61% (17/28), oftalmológico 70% (22/31), nervoso 69% (13/19), dermatológico 45% (9/20), e gastrintestinal 42% (6/14). As principais alterações hematológicas foram anemia 66% (23/32), leucopenia 76% (19/25) e linfopenia 48% (15/31). Concluiu-se que foi elevada a ocorrência de cinomose canina em animais com suspeita clínica no Semiárido Paraibano, e animais sem raça definida, não vacinados, no período seco do ano foram mais diagnosticados com a enfermidade.

The objective of this study was to determine the prevalence of distemper canine distemper vírus (CDV) infection in the semi-arid region of Paraíba, using rapid immunochromatographic tests, correlating it with the main clinical and hematological findings. 67 records of animals were analyzed in which rapid tests were performed for antigen research in nasal and ocular from January to December 2019. It was observed that 47% (32/67) of compulsory dogs were positive for canine distemper. The variables that defined difference difference (p <0.05) for infection were mixed breed animals 60% (21/35), unvaccinated animals 70% (29/42), and dry period of the year, being the August (40%; 13/32), with greater occurrence. The main affected systems were the respiratory 61% (17/28), ophthalmological 70% (22/31), nervous 69% (13/19), dermatological 45% (9/20), and gastrointestinal 42% (6/14 )) The main changes were hematological, anemia 66% (23/32), leukopenia 76% (19/25) and lymphopenia 48% (15/31). It was concluded that the occurrence of canine distemper in animals with clinical suspicion in the Semiarid Paraibano was high, and non-vaccinated mixed-breed animals in the dry period of the year were more diagnosed with the disease.

Técnicas reversíveis de rufião em ovinos (Ovis aires) / Reversible techniques of ruffian in sheep (Ovis aires)

Background: The use of teaser rams is an essential practice for detecting estrus in ewes as well as for accelerating puberty and synchronizing ovulation in the animal. There are several methods for preparing teasers, and the method used should be based on an assessment of the producer’s requirements. The ideal technique should be low cost and safe, ensuring the non-fertilization of ewes. This study evaluated the feasibility and effectiveness of two reversible teaser preparation techniques using a reversible plastic clamp. The techniques were compared in terms of functionality, possible post-surgical complications, and hematological changes of the rams as well as durability and reversibility. Materials, Methods & Results: Twelve healthy rams, aged 14-20 months, were divided into two groups (G1 and G2). Blood samples were collected through the jugular vein to perform the following analyses: blood count, total plasma protein, and fibrinogen. Following local infiltrative anesthesia with 5.0 mL 2% lidocaine without vasoconstrictor, the procedure was performed as follows: in G1, the preputial ostium was partially closed, and in G2, sigmoid flexure was performed in the cranial region, approximately 5-8 cm immediately caudal to the scrotal sac. In the postoperative period, 20 mg/kg oxytetracycline and 2.2 mg/kg flunixinmeglumine were intramuscularly administered as a single dose. The wounds were dressed, sprayed with repellent, and allowed to heal for seven days. The procedures in both groups were simple to perform, low cost, and low risk; caused minimal tissue injury; enabled rapid recovery; promoted little or no stress to the animals; are reversible; and left no complications. The animals of both groups satisfactorily identified the females in estrus during the three-month experimental period, maintained libido, and failed to mate with any female. The blood count levels...

Evaluation of teaser buffaloes subjected to surgical technique for fixation of the caudal curvature of the sigmoid flexure with myectomy of the penis retractor muscle / Avaliação de rufiões bubalinos submetidos à técnica cirúrgica de fixação da curvatura caudal da flexura sigmóide com miectomia do músculo retrator do pênis

The objective of this work was to evaluate the efficiency technique of fixation of the caudal curvature of the sigmoid flexure with myectomy of the penis retractor muscle and the performance of teaser bulls after surgery, through the detection of females in estrus, without performing the copulation. 12 teaser buffaloes were used without defined breed, with an average age of 20 months, weighing on average 200 kg. The animals were subjected to food and water fasting for 12 hours to provide tranquilization in advance. The surgical procedure consisted of the exteriorization of the sigmoid flexure, myectomy of  penis retractor muscle and fixation of the caudal curvature of the penis sigmoid flexure. Of the 12 evaluated teaser buffaloes in the libido test, 100% performed  the  flehmen reflex, mount attempt, incomplete and complete jump  with total inability of  the penis exposure.    It was concluded that this technique can be used safely, having  simplicity in its execution, effective results and mainly low cost, not interfering in the animals libido. The animals when tested as teaser buffaloes kept the libido during the experimental period with total inability of exposure of the penis.

O objetivo do presente trabalho foi avaliar a técnica de fixação da curvatura caudal da flexura sigmóide com miectomia do músculo retratordo pênis e o desempenho de rufiões bubalinos após cirurgia, através da detecção de fêmeas em estro, sem executar a cópula. Foram utilizados 12 bubalinos sem raça definida, com idade média de 20 meses, pesando em média 200 kg. Os animais foram submetidos ajejum alimentar e hídrico de 12 horas para tranquilização prévia. O procedimento cirúrgico constou naexteriorização da flexura sigmóide, miectomia do músculo retrator do pênis e fixação da curvatura caudal da flexura sigmóide do pênis. Dos 12 rufiões avaliados no teste de libido, 100% realizaram reflexode flehmen, tentativa de monta, salto incompleto e completo com total incapacidade de exposição do pênis. Conclui-se que esta técnica pode ser utilizada com segurança, apresentando simplicidade na sua execução, eficácia nos resultados e principalmente baixo custo, não interferindo na libidodos animais. Os animais quando testados como rufiões mantiveram a libido durante o período experimental com total incapacidade de exposição do pênis.

Evaluation of teaser buffaloes subjected to surgical technique for fixation of the caudal curvature of the sigmoid flexure with myectomy of the penis retractor muscle / Avaliação de rufiões bubalinos submetidos à técnica cirúrgica de fixação da curvatura caudal da flexura sigmóide com miectomia do músculo retrator do pênis

The objective of this work was to evaluate the efficiency technique of fixation of the caudal curvature of the sigmoid flexure with myectomy of the penis retractor muscle and the performance of teaser bulls after surgery, through the detection of females in estrus, without performing the copulation. 12 teaser buffaloes were used without defined breed, with an average age of 20 months, weighing on average 200 kg. The animals were subjected to food and water fasting for 12 hours to provide tranquilization in advance. The surgical procedure consisted of the exteriorization of the sigmoid flexure, myectomy of  penis retractor muscle and fixation of the caudal curvature of the penis sigmoid flexure. Of the 12 evaluated teaser buffaloes in the libido test, 100% performed  the  flehmen reflex, mount attempt, incomplete and complete jump  with total inability of  the penis exposure.    It was concluded that this technique can be used safely, having  simplicity in its execution, effective results and mainly low cost, not interfering in the animals libido. The animals when tested as teaser buffaloes kept the libido during the experimental period with total inability of exposure of the penis.(AU)

O objetivo do presente trabalho foi avaliar a técnica de fixação da curvatura caudal da flexura sigmóide com miectomia do músculo retratordo pênis e o desempenho de rufiões bubalinos após cirurgia, através da detecção de fêmeas em estro, sem executar a cópula. Foram utilizados 12 bubalinos sem raça definida, com idade média de 20 meses, pesando em média 200 kg. Os animais foram submetidos ajejum alimentar e hídrico de 12 horas para tranquilização prévia. O procedimento cirúrgico constou naexteriorização da flexura sigmóide, miectomia do músculo retrator do pênis e fixação da curvatura caudal da flexura sigmóide do pênis. Dos 12 rufiões avaliados no teste de libido, 100% realizaram reflexode flehmen, tentativa de monta, salto incompleto e completo com total incapacidade de exposição do pênis. Conclui-se que esta técnica pode ser utilizada com segurança, apresentando simplicidade na sua execução, eficácia nos resultados e principalmente baixo custo, não interferindo na libidodos animais. Os animais quando testados como rufiões mantiveram a libido durante o período experimental com total incapacidade de exposição do pênis.(AU)

Detection of Neospora caninum and Toxoplasma gondii in semen of naturally infected rams

Background: Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle. Moreover, abortion cases attributed to N. caninum and T. gondiiinfection have been occasionally reported in sheep. Due to the relatively scarce information on the molecular detection ofN. caninum in the semen of naturally infected rams, this study aimed to detect parasitic DNA in fresh semen samples andin frozen extended semen straws from male sheep from artificial inseminations centers in Southern Brazil.Materials, Methods & Results: Semen samples of 38 rams from artificial insemination centers were evaluated. Eleven ramswere naturally infected (seropositive for anti-N. caninum and/or anti-T. gondii IgG) and were selected for fresh semen collection.We tested all the samples for the closely related protozoan T. gondii to detect a possible cross-reaction and co-infection, due tothe close similarity with N. caninum. The indirect fluorescent antibody test was used to detect IgG antibodies in the 11 serumsamples from rams. Fresh semen samples were collected from 11 rams on days 1, 50, 55, and 58 using an artificial vaginaand ewe in estrus. Other 27 rams had their frozen extended semen straws analyzed. A total of 20 fresh semen samples and27 frozen extended semen straws samples were used to detect the presence of N. caninum and T. gondii DNA by polymerasechain reaction (PCR). Nc-5 and B1 genes were used as target regions to detect N. caninum and T. gondii DNA, respectively.The presence of N. caninum DNA was confirmed in the third collection of a fresh semen sample of one seropositive ram. T.gondii DNA was detected in a fresh semen sample of one seropositive ram. The DNA sequences of 186 bp from N. caninum(GenBank accession: MH806393) and 492 bp from T. gondii (GenBank accession: MH793503)...

Detection of Neospora caninum and Toxoplasma gondii in semen of naturally infected rams

Background: Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle. Moreover, abortion cases attributed to N. caninum and T. gondiiinfection have been occasionally reported in sheep. Due to the relatively scarce information on the molecular detection ofN. caninum in the semen of naturally infected rams, this study aimed to detect parasitic DNA in fresh semen samples andin frozen extended semen straws from male sheep from artificial inseminations centers in Southern Brazil.Materials, Methods & Results: Semen samples of 38 rams from artificial insemination centers were evaluated. Eleven ramswere naturally infected (seropositive for anti-N. caninum and/or anti-T. gondii IgG) and were selected for fresh semen collection.We tested all the samples for the closely related protozoan T. gondii to detect a possible cross-reaction and co-infection, due tothe close similarity with N. caninum. The indirect fluorescent antibody test was used to detect IgG antibodies in the 11 serumsamples from rams. Fresh semen samples were collected from 11 rams on days 1, 50, 55, and 58 using an artificial vaginaand ewe in estrus. Other 27 rams had their frozen extended semen straws analyzed. A total of 20 fresh semen samples and27 frozen extended semen straws samples were used to detect the presence of N. caninum and T. gondii DNA by polymerasechain reaction (PCR). Nc-5 and B1 genes were used as target regions to detect N. caninum and T. gondii DNA, respectively.The presence of N. caninum DNA was confirmed in the third collection of a fresh semen sample of one seropositive ram. T.gondii DNA was detected in a fresh semen sample of one seropositive ram. The DNA sequences of 186 bp from N. caninum(GenBank accession: MH806393) and 492 bp from T. gondii (GenBank accession: MH793503)...(AU)

Comparison of three protocols to preserve Leptospira spp. in cat urine for efficient DNA extraction and PCR amplification

Background: The pathogenic leptospira infection in mammalian species can cause a range of acute or chronic manifestations and may result in a carrier state. Previous studies have suggested that cats were resistant to acute leptospirosis however, the description of some clinical cases suggests that Leptospira spp. may also be pathogenic to this species. Recentstudies have shown that leptospires may be shed in the urine of infected cats. Endogenous substances present in urine mayinhibit PCR and allow leptospires to evade detection. This study aims to compare three protocols for sample processingto optimize the detection of pathogenic leptospires in cat urine.Materials, Methods & Results: Three protocols to optimize the detection of pathogenic leptospires in cat urine were tested.Aliquots of standard concentration of L. interrogans serovar Canicola culture were added to urine samples to achieveconcentrations of 1×105 to 1×102 leptospires/mL for each protocol. In protocols A and B the urine was neutralized by theaddition of phosphate-buffered saline (PBS), pH 7.4, in a proportion of 1 PBS: 2.5 urine (v/v). In protocol A, PBS wasadded to neutralize the urine pH for the leptospiral organisms immediately after addition of leptospires. In protocol B,PBS was added just before DNA extraction. In protocol C, no PBS was added. DNA extraction was performed at 4, 24and 48 h after addition of the leptospires using a modified protocol. Samples were incubated at 37ºC for 10 min. Sampleswere then centrifuged (850 g) for 15 min, at 25ºC. The supernatants were transferred to another tube, and the pellets werediscarded. The supernatants were centrifuged (16060 g) for 20 min at 4ºC. The supernatants were then discarded, and thepellets resuspended and washed with 1000 µL of PBS. All the samples were centrifuged at 16060 g for an additional 20min at 25ºC. The supernatants were discarded and the pellets were resuspended in 100 µL of PBS and incubated at 94ºCfor...(AU)

Comparison of three protocols to preserve Leptospira spp. in cat urine for efficient DNA extraction and PCR amplification

Background: The pathogenic leptospira infection in mammalian species can cause a range of acute or chronic manifestations and may result in a carrier state. Previous studies have suggested that cats were resistant to acute leptospirosis however, the description of some clinical cases suggests that Leptospira spp. may also be pathogenic to this species. Recentstudies have shown that leptospires may be shed in the urine of infected cats. Endogenous substances present in urine mayinhibit PCR and allow leptospires to evade detection. This study aims to compare three protocols for sample processingto optimize the detection of pathogenic leptospires in cat urine.Materials, Methods & Results: Three protocols to optimize the detection of pathogenic leptospires in cat urine were tested.Aliquots of standard concentration of L. interrogans serovar Canicola culture were added to urine samples to achieveconcentrations of 1×105 to 1×102 leptospires/mL for each protocol. In protocols A and B the urine was neutralized by theaddition of phosphate-buffered saline (PBS), pH 7.4, in a proportion of 1 PBS: 2.5 urine (v/v). In protocol A, PBS wasadded to neutralize the urine pH for the leptospiral organisms immediately after addition of leptospires. In protocol B,PBS was added just before DNA extraction. In protocol C, no PBS was added. DNA extraction was performed at 4, 24and 48 h after addition of the leptospires using a modified protocol. Samples were incubated at 37ºC for 10 min. Sampleswere then centrifuged (850 g) for 15 min, at 25ºC. The supernatants were transferred to another tube, and the pellets werediscarded. The supernatants were centrifuged (16060 g) for 20 min at 4ºC. The supernatants were then discarded, and thepellets resuspended and washed with 1000 µL of PBS. All the samples were centrifuged at 16060 g for an additional 20min at 25ºC. The supernatants were discarded and the pellets were resuspended in 100 µL of PBS and incubated at 94ºCfor...

Evolução e perspectivas da inseminação artificial em bovinos / Evolution and perspectives of timed artificial insemination in cattle

A inseminação artificial (IA) em bovinos tem sido aplicada em todo o mundo com o intuito de melhorar o ganho genético e a eficiência reprodutiva dos rebanhos. Com o objetivo de facilitar a utilização da IA foram desenvolvidos protocolos de inseminação artificial em tempo fixo (IATF) que promovem o controle do crescimento folicular e da ovulação e permitem a aplicação da IA em dias predeterminados, sem a necessidade de detecção de estro e com elevadas taxas de prenhez. Nos últimos 20 anos, vários protocolos de sincronização para IATF foram estudados para atender diferentes realidades de manejo, raças e categorias de animais. Atualmente, 86% das inseminações no Brasil estão sendo realizadas por IATF (13,6 milhões de IATF de um total de 15,4 milhões de doses de sêmen comercializadas em 2018). Com a colaboração dessa tecnologia, verificou-se que o percentual de fêmeas em idade reprodutiva inseminadas artificialmente passou de 5,8% em 2002 para 13,1% em 2018. O aumento de uso da IATF representa considerável incremento de produtividade quando comparado com a monta natural, maximizando o retorno econômico para as fazendas de corte e de leite.(AU)

Artificial insemination in cattle has been used worldwide to improve the genetic gain and reproductive efficiency of the herds. In order to facilitate the use of AI, several protocols for timed artificial insemination (TAI) have been developed, which promote the control of follicular growth and ovulation, allowing the use of AI in predetermined days, without the necessity of estrus detection and with satisfactory pregnancy rates. In the last 20 years, a wide variety of protocols have been studied to address different management realities, breeds and animal categories. Currently, 86% of artificial inseminations in Brazil are performed using TAI (13.6 million TAI of 15.4 million batches of semen commercialized in 2018). The percentage of females in reproductive age inseminated in Brazil went from 5.8% in 2002 to 13.1% in 2018. Reproductive programs that use TAI present increase in productivity when compared to natural breeding, maximizing the economic gain for beef and dairy properties.(AU)

Evolução e perspectivas da inseminação artificial em bovinos / Evolution and perspectives of timed artificial insemination in cattle

A inseminação artificial (IA) em bovinos tem sido aplicada em todo o mundo com o intuito de melhorar o ganho genético e a eficiência reprodutiva dos rebanhos. Com o objetivo de facilitar a utilização da IA foram desenvolvidos protocolos de inseminação artificial em tempo fixo (IATF) que promovem o controle do crescimento folicular e da ovulação e permitem a aplicação da IA em dias predeterminados, sem a necessidade de detecção de estro e com elevadas taxas de prenhez. Nos últimos 20 anos, vários protocolos de sincronização para IATF foram estudados para atender diferentes realidades de manejo, raças e categorias de animais. Atualmente, 86% das inseminações no Brasil estão sendo realizadas por IATF (13,6 milhões de IATF de um total de 15,4 milhões de doses de sêmen comercializadas em 2018). Com a colaboração dessa tecnologia, verificou-se que o percentual de fêmeas em idade reprodutiva inseminadas artificialmente passou de 5,8% em 2002 para 13,1% em 2018. O aumento de uso da IATF representa considerável incremento de produtividade quando comparado com a monta natural, maximizando o retorno econômico para as fazendas de corte e de leite.

Artificial insemination in cattle has been used worldwide to improve the genetic gain and reproductive efficiency of the herds. In order to facilitate the use of AI, several protocols for timed artificial insemination (TAI) have been developed, which promote the control of follicular growth and ovulation, allowing the use of AI in predetermined days, without the necessity of estrus detection and with satisfactory pregnancy rates. In the last 20 years, a wide variety of protocols have been studied to address different management realities, breeds and animal categories. Currently, 86% of artificial inseminations in Brazil are performed using TAI (13.6 million TAI of 15.4 million batches of semen commercialized in 2018). The percentage of females in reproductive age inseminated in Brazil went from 5.8% in 2002 to 13.1% in 2018. Reproductive programs that use TAI present increase in productivity when compared to natural breeding, maximizing the economic gain for beef and dairy properties.

Occurrence of anti-Neospora caninum antibodies in cattle in the dairy farming region of the state of Piauí, Brazil / Ocorrência de anticorpos anti- Neospora caninum em vacas na bacia leiteira do estado do Piauí, Brasil

This study focused on the detection of anti-Neospora caninum IgG antibodies in cows in the dairy farming region of the state of Piauí, Brazil. To this end, serum samples were collected from 255 dairy cows on 17 farms located in the dairy farming region of the municipality of Parnaíba, Piauí, Brazil. The indirect fluorescence antibody test (IFAT) was employed to detect anti-N. caninum IgG antibodies, using anti-bovine IgG (Sigma®) conjugated to fluorescein isothiocyanate (FITC) and a cutoff point of 1:200. Of the 255 samples analyzed, 69 (27.06%) were positive for anti- N. caninum IgG antibodies, the relative frequency found by property was: 1 (20.00%), 2 (53.33%), 3 (46.66%), 4 (53.33%), 5 (26.66%), 6 (6.66%), 7 (6.66%), 8 (20.00%), 9 (26.66%), 10 (26.66%), 11 (20.00%), 12 (20.00%), 13 (46.66%), 14 (26.66%), 15 (26.66%), 16 (20.00%) and 17 (13.33%). with titers of 200 (15.94%), 400 (20.30%), 800 (24.63%), 1600 (23.18%) and 3200 (15.94%), being the highest frequency for the titer of 800. This study demonstrates for the first time that cows from dairy herds of Parnaíba municipality, state of Piauí, are exposed to N. caninum.(AU)

Objetivou-se estudar a ocorrência de anticorpos IgG anti- Neospora caninum em vacas, na bacia leiteira do estado do Piauí, Brasil. Foram obtidas amostras séricas de 255 vacas leiteiras provenientes de 17 propriedades localizadas na bacia leiteira do município de Parnaíba, Piauí, Brasil. Para a pesquisa de anticorpos IgG anti- N. caninum foi empregada a técnica de Reação de Imunofluorescência Indireta, utilizando-se conjugado anti-IgG bovine (Sigma®) conjugado ao isotiocianato de fluoresceína e ponto de corte 1:200. Das 255 amostras analisadas, 69 (27,06%) foram positivas para anticorpos IgG anti- N. caninum, a frequência relativa encontrada por propriedade foi de: 1 (20,00%), 2 (53,33%), 3 (46,66%), 4 (53,33%), 5 (26,66%), 6 (6,66%), 7 (6,66%), 8 (20,00%), 9 (26,66%) 10 (26,66%), 11 (20,00%), 12 (20,00%), 13 (46,66%), 14 (26,66%), 15 (26,66%), 16 (20,00%) e 17 (13,33%) com títulos de 200 (15,94%), 400 (20,30%), 800 (24,63%), 1600 (23,18%) e 3200 (15,94%), sendo a maior frequência para o título de 800. Este estudo demonstra pela primeira vez que, vacas de rebanhos leiteiros do município de Parnaíba, estado do Piauí, estão expostas ao N. caninum.(AU)

Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods

This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.(AU)

Este trabalho teve como objetivos determinar a ocorrência e realizar a caracterização molecular de Cryptosporidium spp. em 498 amostras fecais de canários (Serinus canaria) criados em cativeiro, utilizando três métodos de diagnóstico: análise microscópica pela coloração negativa com verde malaquita, nested PCR seguida de sequenciamento dos fragmentos amplificados e PCR duplex em tempo real específica para detecção de Cryptosporidium galli e Cryptosporidium genótipo III de aves. A positividade total para Cryptosporidium spp. (total de amostras positivas em pelo menos um método de diagnóstico) obtida pela análise microscópica, nested PCR e PCR duplex em tempo real foi de 13,3% (66/498). As taxas de positividade para Cryptosporidium spp. foram 2,0% (10/498) e 4,6% (23/498) por microscopia e nested PCR, respectivamente. O sequenciamento de 20 amostras amplificadas pela nested PCR identificou C. galli (3,0%; 15/498), Cryptosporidium genótipo I de aves (0,8%; 4/498) e Cryptosporidium avium (0,2%; 1/498). A PCR duplex em tempo real revelou positividade de 7,8% (39/498) para C. galli e 2,4% (12/498) para Cryptosporidium genótipo III de aves. A análise microscópica diferiu significativamente da nested PCR para detecção de Cryptosporidium spp. A PCR duplex em tempo real apresentou maior sensibilidade que a nested PCR/sequenciamento para detectar as espécies/genótipos gástricos de Cryptosporidium.(AU)

Immunohistochemical detection of viral etiopathogenesis in lambs and goat kids with neonatal diarrhea

Background: Neonatal enteritis is an important disease that causes deaths of animals before 3 weeks of age, and results in significant economic losses. Viral agents can predispose the young animals to secondary infections in the gastrointestinal tract, especially in lambs and goat kids younger than 21 days. Although the neonatal diarrhea is common in calves, there is still little knowledge about pathology, pathogenesis and immunohistochemical localization of viral agents that cause neonatal enteritis in lambs and goat kids. In this study, we carried out investigations with the aim of detecting adenovirus, rotavirus, coronavirus and herpes virus in the guts of goat kids and lambs with viral enteritis.Materials, Methods & Results: Adenovirus, rotavirus, coronavirus and herpes virus antisera were applied to paraffinembedded intestinal tissue from neonatal lambs and kids that had died from enteritis. In addition, viral agents in the gut cells were detected and evaluated by electron microscopy. The study material consisted of 15 lambs and 15 goat kids that were presented for diagnosis. Viral agents were detected by immunohistochemically in 20 out of 30 animals. Rotavirus was diagnosed in 10 animals, adenovirus in five, herpes virus in three and coronavirus in two animals; and these results were supported by the electron microscopy. This study showed that viral agents play an important role in neonatal enteritis in lambs and kids.Discussion: Bacteria, viruses and protozoa may have a role in the etiology of neonatal enteritis and identifying the etiological agents is not always possible without laboratory studies. In addition, the immune system of the animal and environmental factors are important factors for to occurrence of the disease.[...]