Frozen bovine preputial mucus as a suitable sample for the direct molecular diagnosis of Campylobacter fetus subsp. venerealis.

Long transportation times remain a challenge to the satisfactory diagnosis of Campylobacter fetus subsp. venerealis (Cfv). Here we demonstrated that samples of frozen bovine preputial mucus maintained at -20 °C for 10 days can be used as an alternative source for molecular diagnosis of Cfv. This approach will improve the analysis of this bacterium.

Adipose-derived stem cells improve full-thickness skin grafts in a rat model.

To investigate the effects of heterologous adipose-derived stem cells (ADSCs) on autologous full-thickness skin grafts, we designed a first-intention healing model using Wistar rats. We harvested and sutured two full-thickness skin grafts in the dorsal recipient beds of 15 rats, randomized into three groups. In the treatment group, 1 × 106 ADSCs resuspended in saline solution (200 µL) were administered subcutaneously to the skin graft. The control group received only saline solution subcutaneously, whereas the negative control group did not receive any treatment. Compressive dressings were maintained until postoperative day 5. The grafts were assessed by two observers, who checked for the presence of epidermolysis on day 14. Planimetry showed the relative areas of normal skin, redness, ulceration, and contraction. Graft samples were obtained on day 14 and stained with hematoxylin and eosin and Masson's trichrome. Epidermal analysis evaluated thickening, keratosis, acanthosis, hydropic degeneration, and inflammatory infiltrate. Dermal evaluation investigated the absence of hair follicles, granulation tissue formation, presence of inflammatory infiltrate, and collagen deposition. Immunohistochemistry was performed for dermal anti-VEGF and epidermal anti-Ki-67 staining. The ADSC group presented better macroscopic aspects, lower incidence of epidermolysis, and less loss of hair follicles. In addition, the ADSC group presented the lowest frequency of histopathological changes in the dermis and epidermis, as well as the largest subcutaneous and granulation tissue VEGF averages and the weakest Ki-67 staining of the epidermal basal layer. Subcutaneous administration of ADSCs may improve the integration of skin grafts, reducing the deleterious effects of ischemia and reperfusion injury.

Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I.

The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV). A domestic cat (Felis silvestris catus) presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC) evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I / Toxoplasmose fatal em um gato doméstico imunossuprimido do Brasil causada por Toxoplama gondii clonal tipo I

Abstract The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV). A domestic cat (Felis silvestris catus) presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC) evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

Resumo O objetivo deste estudo foi relatar um caso de toxoplasmose felina fatal com coinfecção com o vírus da leucemia felina (FeLV). Um gato doméstico (Felis silvestris catus) apresentou intensa dispneia e morreu três dias depois. Na necropsia, observaram-se pulmões firmes, não colabados e com múltiplas áreas brancas, além de linfoadenomegalia e esplenomegalia moderadas. No exame histopatológico, evidenciaram-se broncopneumonia intersticial necrótica acentuada e hepatite necrótica discreta associada a cistos e taquizoítas de T. gondii intralesionais positivos na imuno-histoquímica (IHC) anti-T. gondii. Evidenciou-se ainda, na medula óssea, leucemia mieloide crônica com IHC anti-FeLV positiva nas células neoplásicas. O DNA extraído dos pulmões foi positivo para T. gondii por meio da PCR-REP-529. T. gondii foi caracterizado por PCR-RFLP e pela técnica de microssatélites. Foi identificado o genótipo ToxoDB-PCR-RFLP #10, i.e., o arquétipo tipo I. A análise por microssatélites mostrou que a cepa era uma variante do tipo I, com dois alelos atípicos. Esta é a primeira vez que T. gondii clonal tipo I foi relacionado com um caso agudo de toxoplasmosis em um hospedeiro no Brasil.

Chromobacterium violaceum Infection in a Horse.

Chromobacterium violaceum is an opportunistic pathogen of mammals that produces characteristic violet pigment in bacterial culture. We report pneumonia and septicaemia caused by C. violaceum in a horse. Necropsy examination was performed on a 3-year-old Quarter Horse stallion with a history of recurrent episodes of pneumonia, fever, dyspnoea and sanguineous nasal discharge. The lungs were not collapsed, but they contained dark red foci mixed with white areas, and multiple nodules of firm consistency. Within the liver and kidney there were randomly distributed, multifocal, white pinpoint lesions (0.1-0.5 cm diameter) in the capsule and extending into the parenchyma. The brain and spinal cord contained numerous petechiae. Microscopically, the lung had severe multifocal to coalescing infiltration of degenerate neutrophils within the alveoli and parenchyma associated with extensive areas of necrosis and haemorrhage. Rod-shaped bacteria were often present in the centre of the lesions. There was intense intra-alveolar exudation of fibrin and fibrinoid degeneration of blood vessels associated with thrombosis and ischaemic necrosis adjacent to areas of severe haemorrhage. Similar lesions were present in the liver and kidneys. A pure culture of C. violaceum was obtained from samples of the lung and liver. The identity of the organism was confirmed by amplifying and sequencing the 16S rRNA gene with subsequent analysis of sequence similarity.

Clinical, pathological, immunohistochemical and molecular characterization of feline chronic gingivostomatitis.

Objectives This study presents the clinical, pathological, immunohistochemical and molecular characterization of 26 cats with feline chronic gingivostomatitis (FCG). Methods Oral mucosal biopsies, blood and swabs were collected from cats presenting with oral lesions. The tissue sections were submitted for histopathology and immunohistochemical analysis for feline calicivirus (FCV), feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV). The swabs were subjected to PCR analysis for FCV, and blood for FeLV and FIV. Results The main clinical findings were dysphagia (88.2%), halitosis (76.5%), sialorrhea (47.1%), weight loss (41.2%), intense oral discomfort (35.3%), oral hemorrhage (17.6%), and lackluster and fragile coat (11.8%). Gross inspection revealed bilateral lesions across the palatoglossal fold to the lateral tongue base. The lesions were diffuse, proliferative, intensely red and friable, and bled easily upon examination in 80.8% of cases. In 23.1% of cases, the lesions were multifocal to coalescent, at times forming multiple vesicles on a reddened, edematous palatoglossal fold. Microscopic examination showed that 15.4% of lesions had moderate (grade 2) and 84.6% had severe (grade 3) inflammation. Immunohistochemistry revealed the presence of FeLV antigens in the epithelium and the inflammatory infiltrate of 30.8% of the cats with FCG. FCV antigens were not detected in the FCG lesions. Conclusions and relevance The FCG cases analyzed could not be correlated with FCV. It is possible that FeLV plays a role as a causal agent of lesions in cases where the presence of the virus has been confirmed by immunohistochemistry in epithelial samples.

Causes of bird losses recorded in a captive-bred wild bird flock between 2011 and 2015 / Perdas registradas em um plantel de aves silvestres criadas em cativeiro, entre 2011 e 2015

ABSTRACT: This study discusses the causes of bird deaths recorded in a small aviary dedicated to the breeding of a few exotic, wild bird species. Findings from 28 birds were examined over a period of five years. About 40% of the deaths occurred in the first two weeks after hatching in the two most numerous species in the flock and such losses were mainly a result of starvation caused by inadequate nutritional management. Additionally, 28% of the cases affected recently introduced birds. Despite frequent treatment with anthelmintics; a total of 21% of the deaths in the flock could be attributed to parasitic diseases, most of them in recently acquired birds. Only three of the deaths could be associated with advanced age, all of the further cases were also attributable to management fails. Our results indicated that propagation of these and other species requires an improvement of the management of the newly hatched and newcomer birds to considerably enhance the flock’s performance. Such information may be useful in conservation initiatives and may justify, at least in part, captivity of these animals.

RESUMO: Este estudo discute as causas de morte de aves registradas em um pequeno criadouro de umas poucas espécies de aves silvestres exóticas. As mortes de 28 aves foram examinadas em um período de cinco anos. Cerca de 40% dos casos ocorreram na primeira semana de vida de filhotes das duas espécies mais numerosas no plantel e essas perdas foram consequentes à desnutrição por falhas no manejo nutricional. Outros 28% de casos ocorreram em aves, recentemente introduzidas, ou em fase de adaptação, no criadouro. Ainda que as aves do plantel sejam frequentemente dosificadas com anti-helmínticos, 21% das mortes no plantel foram secundárias a doenças parasitárias; a maioria afetou aves recentemente adquiridas. Exceto por três mortes que foram associadas com desordens atribuídas à idade avançada, todas as demais foram associadas com inadequações no manejo das aves. Esses resultados sugerem que, em sistemas dessa natureza, as aves recém-nascidas e as recém-introduzidas são categorias críticas que demandam concentração de esforços para o atendimento sistemático de suas necessidades. Tais dados podem ser aplicáveis em iniciativas de conservação de outras espécies e podem justificar, ainda que parcialmente, a manutenção desses animais, em cativeiro.

Histopathological, immunohistochemical and biomolecular diagnosis of myocarditis due to Clostridium chauvoei in a bovine / Diagnóstico anatomopatológico, imuno-histoquímico e biomolecular de um caso de miocardite por Clostridium chauvoei em um bovino

The aim of this study was to report a case of clostridial myocarditis in a bovine in Brazil with emphasis on the pathological findings, isolation and molecular identification associated with the in situ localization of C. chauvoei. The animal, a male Brangus bull with nine months of age, was found dead without prior clinical signs. Multifocal and coalescent areas of necrosis were observed in the myocardium. Rod cells in the cardiac muscle fibers were positive immunostaining for C. chauvoei, while this bacterium was also isolated and identified by polymerase chain reaction (PCR).

O objetivo deste estudo foi relatar um caso de miocardite clostridial em um bovino no Brasil, com ênfase nos achados patológicos, isolamento e identificação molecular do agente e visualização in situ de C. chauvoei. O animal, um macho da raça Brangus com nove meses de idade, foi encontrado morto sem sinais clínicos prévios. Foram observadas áreas de necrose multifocal e coalescente no miocárdio, com a presença de bastonetes imunomarcados para C. chauvoei nas fibras musculares cardíacas. Uma estirpe de C. chauvoei foi isolada do tecido e sua identidade foi confirmada por reação em cadeia da polimeras (PCR).

Rectal stenosis in pigs associated with Salmonella Typhimurium and porcine circovirus type 2 (PCV2) infection / Estenose retal em suínos associada à enterite causada por Salmonella Typhimurium em animais infectados pelo circovírus suíno tipo 2 (PCV2)

Rectal stricture is an acquired annular fibrous constriction of the rectum that results from a variety of chronic necrotizing enteric diseases. In pigs, it is in most cases a sequel of Salmonella infection. Porcine circovirus type 2 (PCV2) is a known pathogen causing immunosuppression in pigs worldwide. PCV2 infected pigs may be predisposed to salmonellosis. In this report, rectal stenosis was observed in 160 pigs from a herd that experienced an outbreak of enteric salmonellosis over a 4-month period. Distension of the abdominal wall and diarrhea were the main clinical signs observed. Five animals were analyzed showing annular cicatrization of the rectal wall 5.0-7.0 cm anterior to the anorectal junction and Salmonella-positive immunostaining in the large intestine. Salmonella Typhimurium was isolated from fragments of the large intestine. Porcine circovirus type 2 antigen was observed in the mesenteric lymph-node in 4 pigs and in the large intestine in 3 pigs.

Estenose retal é uma constrição anular fibrosa do reto que pode ser decorrente de qualquer doença entérica crônica necrotizante. Em suínos, é em muitos casos uma seqüela de infecção por Salmonella. Circovírus suíno tipo 2 é um patógeno bem conhecido que causa imunodepressão em suínos e apresenta distribuição mundial. No presente trabalho, estenose retal foi observada em 160 suínos em um rebanho que teve um surto de salmonelose entérica durante 4 meses. Distensão da parede abdominal e diarreia foram os principais sinais clínicos observados. Foram analisados cinco suínos que demonstraram cicatrização anular da parede do reto 5,0 a 7,0 cm anterior a junção anoretal e imuno-histoquímica positiva para Salmonella spp. no intestino grosso. Antígeno de Circovírus suíno tipo 2 foi observado no linfonodo mesentérico de quatro suínos e no intestino grosso de três.