Clonality test by PCR - PARR in real time of canine lymphomas

Lymphoma is a neoplasm of hematopoietic origin that affects canines. The proper establishment of prognosis and rapid institution of treatment are essential for a better quality of life, and immunophenotyping is one of the tools used for this purpose. The objective of this study was to perform a clonality test for immunophenotypic characterization of canine lymphomas using the polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) technique in real-time from samples fixed in formalin and embedded in paraffin. The 23 analyzed samples were fixed in formalin and embedded in paraffin canine lymphoma from the collection Laboratory of Histopathology of the Animal Pathology Area of the Departament of Veterinary Medicine - Federal Rural University of Pernambuco (UFRPE). Samples were processed, their DNA was extracted, quantified, diluted, and standardized at a concentration of 50 ng/µL. After extraction, all samples were subjected to conventional PCR for endogenous control (detection of the IgM target region), in which the extracted DNA was amplified in a final volume of 25 µL. The 128 bp amplified product was detected by 1.5% agarose gel electrophoresis. Of the 23 samples analyzed for the detection of the conserved region referring to the endogenous gene, 91.30% (21/23) amplified the conserved region Cµ by conventional PCR, and two samples 8.70% (2/23) were negative. Endogenous control positive samples were subjected to real-time PCR-PARR for detection of IgH Major and IgH Minor for B lymphocytes (LB), and TCRy for lymphocytes T (LT) target regions. All reactions were performed in duplicate to reduce the risk of false-positive or false-negative results due to technical errors. Samples previously confirmed by immunohistochemistry were used as positive controls for T cell and B cell lymphoma, and MilliQ water was used as a negative reaction control. After amplification, the melting curve gradually increased the temperature by 1o C/5 s to 95o C during continuous fluorescence monitoring. Of the 21 samples analyzed, 100.00% (21/21) demonstrated clonal amplification. Of these, 57.15% (12/21) were positive for phenotype B, and 42.85% (9/21) were positive for phenotype T. Due to the importance of researching and confirming samples from files fixed and embedded in paraffin samples in laboratories, PCR-PARR is a good tool for this purpose. In the present study, real-time PCR analysis demonstrated greater sensitivity in the characterization of the immunophenotype of lymphomas from old samples fixed in formalin and embedded in paraffin. The temperature of melting curve analysis may vary depending on the amount of DNA and its quality. In the present study, it was found that the average melting temperature in the samples varied between ± 3o C when compared to that in the control sample for LB and LT, 83.5o C and 80o C, respectively: in the literature, there is a relative difference in this temperature, which may vary up to 4o C. Real-time PCR-PARR was satisfactory in the characterization of the immunophenotype of canine lymphomas from formalin-fixed and paraffin-embedded samples; therefore, its use is recommended for both retrospective studies. The use of PCR-PARR associated with histopathological and/or cytopathological examination in cases of canine lymphomas strongly helps pathologists, provide a safe establishment of the immunophenotype, minimize errors, and optimize the diagnosis, thus directly contributing to the establishment of the prognosis.(AU)

Sexual diagnosis through morphometric evaluation of the proximal femur / El diagnóstico sexual a través de la evaluación morfométrica del fémur proximal

The determination of variables such as age, height, ethnicity and sex are extremely important to the identification of biological findings, especially in accidents with fatal victims. The diagnosis of sex can be 100 % certain for cases in which the skeleton is complete and in a good state of conservation, the individual is an adult and the morphometric variables of the population to which the individual belongs are known.: The aim of the present study was to perform a morphometric evaluation of the proximal femur and compare measurements between males and females. Sixty pairs of femurs were acquired from the Department of Animal Morphology and Physiology of the Universidade Federal Rural de Pernambuco. The bones were measured with the aid of the ImageJ software program and the following variables were compared between bones from male and female cadavers: DFH - diameter of femoral head on the craniocaudal and sagittal axes; DFN - diameter of femoral neck on the craniocaudal and sagittal axes; FNL - femoral neck length; ILL - intertrochanteric line length. The morphometric variables had the following mean values in the male bones: DFH-craniocaudal axis - 45.10 ± 0.35 mm; DFH-sagittal axis - 48.27 ± 0.35 mm; DFN-craniocaudal axis - 33.21 ± 0.40 mm; DFN-sagittal axis - 29.96 ± 0.05 mm; FNL - 31.71 ± 0.05 mm; ILL - 66.47 ± 0.59 mm. The mean values for the female bones were as follows: DFH-craniocaudal axis - 40.68 ± 0.20 mm; DFH-sagittal axis - 42.61 ± 0.20 mm; DFN-craniocaudal axis - 29.11 ± 0.03 mm; DFN-sagittal axis - 26.05 ± 0.04 mm; FNL - 31.10 ± 0.04 mm; ILL - 60.80 ± 0.41 mm. With the exception of the femur neck length, all variables measurements were significantly larger (p < 0.0001) on the male bones. The present findings demonstrate that the femur bone, particularly the proximal portion, exhibits important sexual dimorphism and has high potential for forensic purposes.

La determinación de variables como la edad, la altura, el origen étnico y el sexo son extremadamente importantes para la identificación de los hallazgos biológicos, especialmente en accidentes con víctimas fatales. El diagnóstico de sexo puede entregar una coincidencia del 100 % para los casos en los que el esqueleto está completo y en un buen estado de conservación, el individuo es un adulto y se conocen las variables morfométricas de la población a la que pertenece el individuo. El objetivo del presente estudio consistió en realizar una evaluación morfométrica del fémur proximal y comparar las mediciones entre hombres y mujeres. Se adquirieron sesenta pares de fémures del Departamento de Morfología y Fisiología Animal de la Universidade Federal Rural de Pernambuco. Los huesos se midieron con la ayuda del programa de software ImageJ y se compararon las siguientes variables entre los huesos de cadáveres masculinos y femeninos: DCF: diámetro de la cabeza femoral en los ejes craneocaudal y sagital; DCF: diámetro del cuello femoral en los ejes craneocaudal y sagital; LCF - longitud del cuello femoral; LLI - longitud de línea intertrocantérea. Las variables morfométricas tenían los siguientes valores medios en los huesos masculinos: Eje craniocaudal DCF - 45,10 ±0,35 mm; DCF-eje sagital - 48,27±0,35 mm; DCF-eje craneocaudal - 33,21 ± 0,40 mm; DFN-eje sagital - 29,96 ± 0,05 mm; LCF - 31,71 ± 0,05 mm; LLI - 66,47 ± 0,59 mm. Los valores medios para los huesos femeninos fueron los siguientes: DCFeje craneocaudal - 40,68 ± 0,20 mm; DCF-eje sagital - 42,61 ± 0,20 mm; DCF-eje craneocaudal - 29,11 ± 0,03 mm; DCF-eje sagital - 26,05 ± 0,04 mm; LCF - 31,10 ± 0,04 mm; LLI - 60,80 ± 0,41 mm. Con la excepción de la longitud del cuello del fémur, todas las medidas variables fueron significativamente más grandes (p <0,0001) en los huesos masculinos. Los presentes hallazgos demuestran que el hueso del fémur, particularmente la porción proximal, exhibe un importante dimorfismo sexual y tiene un alto potencial para fines forenses.

Low prevalence of infection by Sarcocystis neurona in horses from the State of Alagoas, Brazil / Baixa prevalência da infecção por Sarcocystis neurona em cavalos do estado de Alagoas, Brasil

Abstract The aim of this study was to determine the prevalence of infection by Sarcocystis neurona in horses and identify potential risk factors. Were analyzed 427 samples from 36 farms in 21 municipalities in the Alagoas State, Brazil. Presence of anti-S. neurona antibodies was diagnosed by indirect immunofluorescence antibody test (IFAT) and was confirmed using the immunoblot test. Risk factors were assessed through investigative questionnaires on animal management on the farms. The prevalence of anti-S.neurona antibodies was 2.8% (confidence interval, CI: 1.5-4.9%) from IFAT and 1.6% (CI:0.8-3.34%) from immunoblot, and there were positive horses on 16.6% of the studied farms. None of the variables studied presented associations with serological status for S. neurona. This is the first report on infection by S. neurona in horses reared in Alagoas, Brazil showing a low exposure to S. neurona in this region, but with significant numbers of foci.

Resumo Objetivou-se neste estudo determinar a prevalência e os fatores de risco associados à infecção por Sarcocystis neurona em equinos. Foram analisadas 427 amostras de 36 propriedades localizadas em 21 municípios do estado de Alagoas. O diagnóstico de anticorpos anti-S. neurona foi realizado pela técnica de Imunofluorescência Indireta (IFI) e confirmada por immunoblot. O estudo dos fatores de risco foi realizado a partir de questionários investigativos sobre o manejo dos animais nas propriedades. A prevalência de anticorpos anti-S. neurona foi de 2,8% (I.C. 1,5-4,9%) na IFI e de 1,6% (I.C. 0,8-3,34%) no immunoblot com equinos positivos em 16,6% das propriedades estudadas. Nenhuma variável estudada apresentou associação com o status sorológico para S. neurona. Este é o primeiro relato da infecção por S. neurona em equinos criados no Estado de Alagoas, Brasil, confirmando que os animais desta região têm baixa exposição a S. neurona, mas com significativo número de focos.

Determination of sex based on the morphometric evaluation of the proximal tibia / Determinación del sexo basada en la evaluación morfométrica de la tibia proximal

SUMMARY: Anatomic differences are found between human males and females. These characteristics of sexual dimorphism vary across populations and have important implications for the study of human evolution. Researchers have employed anthropological tools for the characterization of human cadavers in different states of decomposition using complete skeletons, isolated bones or even bone fragments. The aim of the present study was to differentiate males and females based on morphometric aspects of human tibias from the Brazilian population. Sixty pairs of adult human tibias were obtained from bone collection of the Anatomy Sector of the Department of Animal Morphology and Physiology of the Rural Federal University of Pernambuco, Brazil. Ten measurements were made on the joint face of the tibial plateau: Anteroposterior diameter of the joint surface of the medial condyle (APM), Transverse diameter of the joint surface of the medial condyle (TM), Anteroposterior diameter of the joint surface of the lateral condyle (APL), Transverse diameter of the joint surface of the lateral condyle (TL), Anterior transverse measure of inter-condyle area (ATI), Posterior transverse measure of inter-condyle area (PTI), Middle transverse measure of inter-condyle area (MTI), Anteroposterior measure of inter-condyle area (API), Anterior measure of intercondyle area (AI) and Posterior measure of inter-condyle area (PI). The median morphometric values for the males' proximal tibia were: APM - 4.707 ± 0.406; TM - 3.024 ± 0.307; APL -4.023 ± 0.414; TL - 3.405 ± 0.323; ATI - 3.365 ± 0.425; PTIc - 1.788 ± 0.307; MTI - 1.167 ± 0.279; API - 5.140 ± 0.377; AI - 2.865 ± 0.249 and PI - 2.297 ± 0.344. The median morphometric numbers for the female's proximal tibia were: APM - 4.334 ± 0.317; TM - 2.702 ± 0.298; APL - 3.549 ± 0.382; TL - 2.991 ± 0.281; ATI - 2.818 ± 0.307; PTI - 1.555 ± 0.222; MTI - 1.054 ± 0.262; API - 4.596 ± 0.303; AI - 2.565 ± 0.21; PI - 2.031 ± 0.225. All parameters measured showed values significantly higher (p < 0.05) for the male's proximal tibia. The findings demonstrate sexual dimorphism in this leg bone, which can be used with considerable reliability for scientific and forensic purposes. The results of these studies can be useful in cases where other methods are not applicable, increasing the range of options for forensic investigation.

RESUMEN: Se encuentran diferencias anatómicas entre hombres y mujeres. Estas características del dimorfismo sexual varían entre las poblaciones y tienen implicaciones importantes para el estudio de la evolución humana. Los investigadores han empleado herramientas antropológicas para la caracterización de cadáveres humanos en diferentes estados de descomposición utilizando esqueletos completos, huesos aislados o incluso fragmentos óseos. El objetivo del presente estudio fue diferenciar a hombres y mujeres basándose en aspectos morfométricos de las tibias humanas de la población brasileña. Se obtuvieron sesenta pares de tibias adultas en la recolección ósea del Sector de Anatomía del Departamento de Morfología Animal y Fisiología de la Universidad Federal Rural de Pernambuco, Brasil. Se realizaron diez mediciones en la cara articular de la meseta tibial: Diámetro anteroposterior de la superficie articular del cóndilo medial (APM), diámetro transversal de la superficie articular del cóndilo medial (TM), diámetro anteroposterior de la superficie articular del códilo lateral (APL), diámetro transversal de la superficie articular del cóndilo lateral (TL), medida transversa anterior del área intercondílea (ATI), medida transversal posterior del área intercondílea (PTI), medida anteroposterior del área intercondílea (API), medida anterior del área intercondílea (AI) y medida posterior del área intercondílea (PI). Los valores morfométricos medianos para la tibia proximal de los varones fueron: APM - 4.707 ± 0.406; TM - 3,024 ± 0,307; APL 4,023 ± 0,414; TL - 3,405 ± 0,323; ATI - 3.365 ± 0,425; PTI - 1,788 ± 0,307; MTI - 1,167 ± 0,279; API - 5.140 ± 0.377; AI - 2,865 ± 0,249 y PI - 2,297 ± 0,344. Los números morfométricos medianos para la tíbia proximal de la hembra fueron: APM - 4.334 ± 0.317; TM - 2,702 ± 0,298; APL - 3,549 ± 0,382; TL - 2,991 ± 0,281; ATI - 2,818 ± 0,307; PTI - 1.555 ± 0.222; MTI 1,054 ± 0,262; API - 4.596 ± 0.303; AI - 2,565 ± 0,21; PI 2.031 ± 0,225. Todos los parámetros medidos mostraron valores significativamente más altos (p <0,05) para la tibia proximal del varón. Los hallazgos demuestran dimorfismo sexual en este hueso de la pierna, que se puede utilizar con una fiabilidad considerable para fines científicos y forenses. Los resultados de estos estudios pueden ser útiles en casos donde otros métodos no son aplicables, aumentando el rango de opciones para la investigación forense.

Co-Infection by Sarcoptes scabiei and Microsporum gypseum in Free-Ranging Crab-Eating Fox, Cerdocyon thous (Linnaeus, 1766)

ABSTRACT The aim of the present study was to describe the clinical manifestation, treatment and outcome of a case of co- infection by Sarcoptes scabiei and Microsporum gypseum in Cerdocyon thous (crab-eating fox) from Northeastern Brazil.

Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil

Abstract The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples.

Morphometric analysis of the scapula and their differences between females and males / Análisis morfométrico de la escápula y sus diferencias entre hombres y mujeres

Sexual dimorphism is a key role for forensic anthropology. It allows ascertaining the sex of certain biological discoveries. By postmortem studies it is possible to establish a profile of individuals. This profile allows the identification in cases where bodies are skeletonized. Several bones present dimorphism and have been studied to increase approach for forensic identification. This study morphometrically evaluated the scapula bone and compared the measurements between scapula bone of female and male cadavers. Sixty parts were utilized in this experiment. All cadavers belonged to the Department of Animal Morphology and Physiology of the Federal Rural University of Pernambuco. The scapula bones were photographed and measured through software ImageJ®. Eight parameters of measurements were considered: scapula height (SH), distance between basis and spine (DBSp), scapula width (SW), spine width (SpW), acromion-coracoid width (ACW) and scapula body width at 2 (L2), 4 (L4) and 6 (L6) cm of the lower angle. The median morphometric values for the male scapulas were: SH - 15,143±1,144; DBSp - 7,94±0,61; SW - 10,24±0,65; SpW - 14,01±0,81; ACW - 9,06±0,58; L2 - 4,40±0, 54; L4 - 5, 51 ± 0, 67; L6 - 6, 70±0, 71. The median morphometric numbers for the female scapulas were: SH - 13, 26±0, 79; DBSp - 6, 96±0,47; SW-9, 08±0,50; SW - 12,43±0,69; ACW-7, 79±0,55/L2 - 3,83±0,39; L4 4,97±0,41; L6 - 6,29±0,45. All parameters measured showed values significantly higher (p < 0.05) for the male's scapula. The scapula is a bone sexually dimorphic and can potentially be used in forensic investigations. Consequently this data presents the need of more studies utilizing this bone for personal identification in other populations as well. Thus the results of these studies can be useful in cases where other methods are not applicable, increasing the range of options in forensic investigation.

El dimorfismo sexual es una función clave para la antropología forense. Permite determinar el sexo de ciertos descubrimientos biológicos. Para estudios post mortem, es posible establecer el perfil de los individuos. Este perfil permite la identificación de los cuerpos en el caso que sean solo esqueletos. Varios huesos presentaron dimorfismo y fueron estudiados para aumentar el enfoque en la identificación forense. Este estudio evaluó morfométricamente la escápula y se compararon las mediciones entre la escápula de cadáveres femeninos y masculinos. Se utilizaron 60 piezas en esta investigación. Todos los cadáveres pertenecían al Departamento de Morfología y Fisiología Animal de la Universidad Federal Rural de Pernambuco. Los huesos fueron fotografiados y se midieron a través del software ImageJ®. Se consideraron ocho parámetros de las mediciones: altura de la escápula (SH), distancia entre la columna vertebral y la cavidad glenoidea (DBSP), ancho de la escápula (SW), ancho de la columna vertebral (SpW), distancia entre acromion y proceso coracoides (ACW) y ancho del cuerpo de la escápula a 2 (L2), 4 (L4) y 6 (L6) cm del ángulo inferior de la escápula. Los valores morfométricos de la mediana para las escápulas de los hombres fueron: SH - 15.143 ± 1.144; DBSP - 7,94 ± 0,61; SW - 10,24 ± 0,65; SpW - 14,01 ± 0,81; ACW - 9,06 ± 0,58; L2 - 4,40 ± 0,54; L4 - 5,51 ± 0,67; L6 - 6,70 ± 0,71. La mediana de las características morfométricas de las escápulas de las mujeres fueron: SH - 13,26 ± 0,79; DBSP - 6,96 ± 0,47; SW - 9,08 ± 0,50; SW - 12,43 ± 0,69; ACW - 7,79 ± 0,55 / L2 - 3,83 ± 0,39; L4 4,97 ± 0,41; L6 - 6,29 ± 0,45. Todos los parámetros medidos mostraron valores significativamente mayores (p <0,05) para la escápula de los hombres. La escápula es un hueso que presenta dimorfismo sexual y puede ser utilizado en investigaciones forenses. En consecuencia, estos datos sugieren la necesidad de realizar más estudios de este hueso para la identificación en otras poblaciones. Los resultados de este estudio pueden ser útiles en aquellos casos en los que otros métodos no puedan aplicarse, debido a que presentan mayores opciones para la investigación forense.

Occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chicken breeding in Northeast, Brazil / Ocorrência de anticorpos anti-Toxoplasma gondii e DNA do parasita em galinhas de criações domésticas no Nordeste, Brasil

Abstract The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chickens bred in the metropolitan area of Recife, Brazil. In total, 212 serum samples were collected from 16 properties, and 12 backyard chickens were collected in the six sanitary districts of Recife. An indirect immunofluorescence assay (IFA) was used to investigate the occurrence of anti-Toxoplasma gondii antibodies. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in brain, heart, liver and lung specimens. Of the samples analyzed by serology, 86/212 (40.56%) were positive; of the samples analyzed by PCR, 2/12 (16.7%) were positive, with both samples positive by both tests (serological and molecular). The presence of antibody anti-T. gondii and parasite DNA in tissues of these animals are worrying aspects for public health because there is a risk of transmission of the parasite to humans through eating undercooked or raw meat. Based on the results, the adoption of preventive measures to prevent the cats access to the chickens creations should be encouraged, since these animals were identified in most of the studied properties.

Resumo O objetivo do presente estudo foi investigar a ocorrência de anticorpos anti-Toxoplasma gondii e de DNA do parasito em galinhas de criações domésticas, na região metropolitana de Recife, Brasil. No total, 212 amostras de soro foram coletadas de aves de 16 estabelecimentos e de 12 galinhas de criações domésticas nos seis distritos sanitários de Recife. Para a pesquisa de anticorpos anti-Toxoplasma gondii foi utilizada a Reação de Imunofluorescência Indireta (RIFI). A Reação em Cadeia da Polimerase (PCR) foi utilizada para detectar o DNA de T. gondii em fragmentos de cérebro, coração, fígado e pulmão. Das amostras analisadas por sorologia, 86/212 (40,56%) foram positivas. Das amostras analisadas por PCR, 2/212 (16,7%) foram positivas, em ambos os testes (sorológicos e moleculares). A presença de anticorpos anti-T. gondii e de DNA parasitário nos tecidos desses animais são aspectos preocupantes para saúde pública, porque há o risco de transmissão do parasita para humanos através da ingestão de carne mal cozida ou crua. Com base nos resultados obtidos, a adoção de medidas preventivas que evitem o acesso de gatos às criações de galinhas deve ser incentivada, uma vez que esses animais foram identificados na maioria das propriedades estudadas.

Paratuberculose em bovinos de corte na região Sul do Rio Grande do Sul / Paratuberculosis in beef cattle in Southern Brazil

Descrevem-se os aspectos epidemiológicos e clínico-patológicos de paratuberculose diagnosticada no sul do Rio Grande do Sul em uma propriedade de bovinos de corte. Dois bovinos criados extensivamente que apresentavam emagrecimento progressivo e diarreia crônica foram necropsiados. Os linfonodos mesentéricos estavam aumentados e edematosos. A mucosa do intestino estava espessada e enrugada com aspecto cerebroide principalmente na porção final do íleo, válvula íleo-cecal e ceco. Fragmentos dos órgãos foram fixados em formalina 10%, incluídos em parafina, cortados e corados pela técnica de hematoxilina e eosina (HE) e Ziehl-Neelsen (ZN). Fezes foram encaminhadas ao Departamento de Medicina Veterinária, Área de Medicina Veterinária Preventiva da Universidade Federal Rural de Pernambuco para o cultivo de Mycobacterium aviumsubsp.paratuberculosis em meio Lowenstein Jensen com micobactina e para realização da PCR. Histologicamente, havia enterite granulomatosa no jejuno, íleo, ceco e reto, afetando multifocalmente, também, o duodeno e o cólon. Havia, ainda, linfangite e adenite granulomatosa. Pela coloração de ZN foram observados numerosos bacilos álcool-ácido resistentes (BAAR) no interior de macrófagos, células gigantes de Langhans e nos linfonodos mesentéricos no jejuno, íleo ceco e reto. Não houve crescimento bacteriano nas amostras de fezes e cinco amostras amplificaram a sequência genética IS900 específica do Mycobacterium aviumsubesp. paratuberculosis. Pelo presente trabalho pode-se concluir que a paratuberculose apesar dos poucos relatos ocorre também em bovinos de corte criados extensivamente no sul do Rio Grande do Sul e pode ter uma prevalência maior do que se supõe na região. Alerta-se para a necessidade do diagnóstico e da tomada de medidas efetivas de controle para esta doença que, por muitos, ainda é considerada uma doença exótica no Brasil.

The epidemiological, clinical and pathological aspects of paratuberculosis diagnosed in southern Rio Grande do Sul in a beef cattle property are described. Two cattle raised extensively and that presented progressive weight loss and chronic diarrhea were necropsied. The mesenteric lymph nodes were enlarged and edematous. The intestinal mucosa was thickened and wrinkled with cerebroid aspect, especially in the final portion of the ileum, ileocecal valve and cecum. Fragments of the organs were fixed in 10% formalin, embedded in paraffin, sectioned and stained with hematoxylin and eosin (HE) and Ziehl-Neelsen (ZN). Feces samples were referred to Departamento de Medicina Veterinária, Área de Medicina Veterinária Preventiva da Universidade Federal Rural de Pernambuco, for the cultivation of Mycobacterium avium subsp. paratuberculosis with Lõwenstein Jensen for mycobacterial and PCR analysis. Histologically, granulomatous enteritis was observed in the jejunum, ileum, cecum and rectum, and in multifocal areas the duodenum and colon. Lymphangitis and granulomatous adenitis was also observed. In ZN staining numerous resistant acid-fast bacilli (AFB) within macrophages, giant cells of Langhans and mesenteric lymph nodes in the jejunum, ileum cecum and rectum were observed. There was no bacterial growth in stool samples. Five samples amplified the gene sequence IS900 specific for Mycobacterium aviumsubsp. paratuberculosis. It can be concluded that paratuberculosis, despite the few reports, occurs in beef cattle raised extensively in southern Rio Grande do Sul, Paratuberculosis may have a higher prevalence in the region than it is assumed. There is need for diagnosis of this disease and to assume effective measures for its control, as for many it is still considered an exotic malady in Brazil.

Detection of Ureaplasma spp. in semen samples from sheep in Brazil

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

Seroprevalence of antibodies to Neospora caninum in dogs in the state of Alagoas, Brazil / Soroprevalência de anticorpos anti-Neospora caninum em cães no estado de Alagoas, Brasil

This study investigated occurrences of antibodies to anti-Neospora caninum in dogs in the state of Alagoas, Brazil, and the risk factors associated with the infection. To investigate antibodies against N. caninum, an indirect immunofluorescence antibody test (IFAT) was used. To identify risk factors associated with infection by N. caninum, univariate analyses on variables of interest were conducted. The study analyzed 237 samples from dogs of both sexes and different breeds and ages, both from rural and from urban areas. Ten animals (4.2%) were seropositive: five (4.8%) from rural areas and five (3.8%) from urban areas. Regarding sex, five (4.0%) were male and five (4.5%) were female. In relation to breed, four animals of undefined breed and six of defined breed were positive. The ages of the seropositive animals ranged from two months to eleven years. Results show there are no significant associations between seropositivity and the variables analyzed. Results indicate neosporosis must be included in differential diagnosis of dog neurological disorders.

Investigou-se a ocorrência de anticorpos anti-Neospora caninum em cães no Estado de Alagoas, Brasil e os fatores de risco associados à infecção. Para a pesquisa de anticorpos contra N. caninum empregou-se a Reação de Imunofluorescênica Indireta (RIFI). Para identificar os fatores de risco associados à infecção pelo N. caninum foi realizada análise univariada das variáveis de interesse. Foram analisadas 237 amostras de cães de ambos os sexos, diferentes raças e idades e procedentes de áreas urbanas e rurais. Obteve-se 10 (4,2%) animais soropositivos, sendo cinco (4,8%) de áreas rurais e cinco (3,8%) de áreas urbanas. Com relação ao sexo, cinco (4,0%) eram machos e cinco (4,5%) fêmeas. Quanto às raças foram positivos quatro animais de raças indefinidas e seis de raças definidas. As idades dos animais soropositivos variaram de dois meses a 11 anos. Os resultados da análise estatística revelaram não haver associação significativa entre a soropositividade e as variáveis analisadas. Os resultados indicam a necessidade de incluir a neosporose no diagnóstico diferencial das desordens neurológicas caninas.

Occurrence and risk factors associated with infection by Toxoplasma gondii in goats in the State of Alagoas, Brazil / Ocorrência e fatores de risco associados à infecção por Toxoplasma gondii em caprinos do Estado de Alagoas

INTRODUCTION: Goats are considered very susceptible to infection by Toxoplasma gondii and when this occurs during pregnancy, it may cause fetal death with subsequent fetal resorption, abortion, mummification, stillborn or the birth of weak goats. The objective of this work was to determine the occurrence of and identify risk factors for T. gondii infection in goats in different mesoregions in the State of Alagoas. METHODS: The research was conducted on 24 goat breeding farms in 10 municipalities. A total of 454 blood samples were examined for anti-T. gondii antibodies by indirect immunofluorescence antibody test. To evaluate the risk factors of toxoplasmosis in goats, questionnaires were applied analyzing the farm's production system and nutritional, reproductive and sanitary management. RESULTS: Disease occurrence was 39 percent with 95.8 percent of farms presenting seropositive animals. Significant associations were observed for mesoregion (OR = 0.23; 95 percentCI = 0.09 - 0.57), age (OR = 0.36; 95 percentCI = 0.20 - 0.64), semi-intensive herd management (OR = 8.70; 95 percentCI = 1.87 - 40.43), access of cats to water provided for goats (OR = 3.38; 95 percentCI = 1.89-6.02) and cats feeding on placental remnants (OR = 2.73; 95 percentCI = 1.38 - 5.40). CONCLUSIONS: Toxoplasma gondii infection is disseminated in the State of Alagoas and the adoption of a program that monitors infection foci is required to reduce the risk factors identified in the present study.

INTRODUÇÃO: Caprinos são considerados susceptíveis à infecção por Toxoplasma gondii que quando ocorre durante a gestação pode causar morte fetal com subsequente reabsorção, aborto, mumificação, natimortos ou nascimento de cabritos fracos. Objetivou-se com este estudo determinar a ocorrência e identificar os fatores de risco associados à infecção por T. gondii em caprinos nas diferentes mesorregiões do Estado de Alagoas. MÉTODOS: A pesquisa foi realizada em 10 municípios, sendo analisadas 24 propriedades de produção caprina com aptidão mista. Foram coletadas amostras sanguíneas de 454 animais para realização da pesquisa de anticorpos anti-Toxoplasma gondii através da prova sorológica de imunofluorescência indireta. Para o estudo dos fatores de risco, foram aplicados questionários com questões referentes ao sistema de produção e manejos nutricional, reprodutivo e sanitário. RESULTADOS: A ocorrência encontrada foi de 39 por cento com 95,8 por cento das propriedades apresentando animais positivos. Foi observada associação significativa para as variáveis: mesorregião (OR = 0,23; IC 95 por cento = 0,09 - 0,57), idade (OR = 0,36; IC 95 por cento = 0,20 - 0,64), sistema de criação semi-intensivo (OR = 8,70; IC 95 por cento = 1,87 - 40,43), acesso dos gatos à água fornecida aos animais (OR = 3,38; IC 95 por cento = 1,89 - 6,02) e gatos se alimentando de restos placentários (OR = 2,73; IC 95 por cento = 1,38 - 5,40). CONCLUSÕES: A infecção por T. gondii está disseminada no Estado de Alagoas, sendo necessária a realização de um programa de monitoramento dos focos da infecção no Estado com objetivo de reduzir os fatores de riscos identificados no presente estudo.

Perfil da contagem de células somáticas na infecção intramamária em búfalas na região Nordeste do Brasil / Somatic cell count profile in intramammary infection of buffaloes from the Brazilian Northeast

Objetivou-se com esse estudo avaliar o perfil de células somáticas na mastite subclínica em búfalas leiteiras no Nordeste do Brasil. Foram analisadas 1896 amostras de leite de 474 búfalas procedentes de quatro propriedades de exploração leiteira nos Estados de Pernambuco, Alagoas, Bahia e Ceará. A secreção láctea de cada teto foi submetida ao Califórnia Mastitis Test (CMT) e as amostras positivas, a partir de duas cruzes foram coletadas para realização da contagem de células somáticas (CCS) e exame microbiológico. Observou-se que as amostras positivas no exame microbiológico apresentaram CCS entre 280.000 a 401.000 cel/mL com mediana de 328.000 cel/mL. Conclui-se que valores de CCS acima de 280.000 cel/mL é um indicativo de infecção da glândula mamária, contudo o exame microbiológico do leite é o melhor método para diagnóstico da mastite subclínica em búfalas. A observação dos Staphylococcus coagulase negativa exercendo influência na elevação da CCS deve ser objeto de outros estudos para elucidar a patogenicidade desse grupo de microrganismos na mastite de búfalas.

The aim of this study was to evaluate the somatic cell profile in the subclinical mastitis in dairy buffaloes in the Brazilian Northeastern. One thousand eight hundred ninety six milk samples from 474 buffaloes proceeding from four dairy farms located in the states of Pernambuco, Alagoas, Bahia and Ceará were analyzed. The lactic secretion was submitted to the California Mastitis Test (CMT) and the positive samples, from two crosses on were collected for the somatic cell count (SCC) and microbiologic exam. It was observed that the positive samples at the microbiologic exam presented SCC between 280,000 and 401,000 cells/mL with median of 328,000 cells/mL. It was concluded that SCC values above 280,000 cells/mL is an indication of mammary gland infection, however, the microbiologic exam of the milk is the best method for the diagnostic of subclinical mastitis in buffaloes. The observation of Staphylococcus coagulase-negative exerting influence in the SCC elevation must be subject of further studies in order to elucidate the pathogenicity of this group of microorganisms in mastitis of buffaloes.