Molecular detection of porcine circovirus (PCV2 and PCV3), torque teno swine virus 1 and 2 (TTSuV1 and TTSuVk2), and histopathological findings in swine organs submitted to regular slaughter in Southeast, Brazil.

Porcine circovirus 2 and 3 (PCV2 and PCV3) and torque teno sus virus 1 and 2 (TTSuV1 and TTSuVk2) are important pathogens in pig associated with post-weaning mortality, different clinical syndromes in adults (PCVAD), and a decrease of average daily weight gain (PCV2-SI) but little is known about the infection on asymptomatic pigs. The aim of this study was to evaluate the presence of PCV2, PCV3, TTSuV1, and TTSuVk2 in swine organ samples from asymptomatic pigs slaughtered in Espírito Santo State, South-eastern Brazil, through molecular detection and histopathological analysis. Nested PCR showed the presence of PCV2 DNA in 10% (14/140), PCV3 in 13.6% (19/140), TTSuV1 in 12.9% (18/140), and TTSuVk2 in 30% (42/140) of the tissue samples. All four viruses were detected in the lung, kidney, lymph node, and liver. TTSuVk2 was detecded in 30% (42/140), PCV3 in 13.6% (19/140), TTSuV1 in 12.9% (18/140), and PCV2 in 10% (14/140) of the samples. Single infections were observed in 30.7% (43/140), while co-detections in the same tissue occurred in 15.7% (22/140). The most frequent combinations were TTSuV1/TTSuVk2 in 31.8% (7/22), PCV2/TTSuVk2 in 18.1% (4/22), and PCV2/PCV3/TTSuVk2 in 13.6% (3/22). Lymphocyte depletion was associated with TTSuVk2 infection (p = 0.0041) suggesting that TTSuVK2 plays an induction of PMWS-like lymphoid lesions in pigs. The data obtained in this study show that PCV2, PCV3, TTSuV1, and TTSuVk2 are related to infection in asymptomatic animals with different tissue lesions, and the molecular diagnosis for these pathogens should be considered in the sanitary monitoring of herds.

O circovírus suíno 2 e 3 (PCV2 e PCV3) e os Torque Teno vírus suínos 1 e 2 (TTSuV1 e TTSuVk2) são patógenos importantes na suinocultura associados a diferentes síndromes clínicas e morte de leitões pós desmame (PCVAD) e redução no ganho diário de peso (PCV2-SI). Entretanto, pouco se sabe sobre a circulação desses agentes e o impacto da infecção em porcos assintomáticos. O objetivo deste estudo foi avaliar a presença de PCV2, PCV3, TTSuV1 e TTSuVk2 em amostras de órgãos de suínos assintomáticos abatidos no estado do Espírito Santo, região sudeste do Brasil, por meio de detecção molecular e análise histopatológica. A análise tecidual por nested PCR mostrou a presença de DNA de PCV2 em 14 (10%), PCV3 em 19 (13,6%), TTSuV1 em 18 (12,9%) e de TTSuVk2 em 42 (30%) das amostras. Todos os quatro vírus foram detectados no pulmão, rim, nódulo linfático e fígado TTSuVk2 foi detectado em 30% das amostras teciduais (42/140), PCV3 em 13.6% (19/140), TTSuV1em 12.9% (18/140), e o PCV2 em 10% (14/140. Mono infecções foram observadas em 30.7% (43/140) das amostras enquanto infecções múltiplas observadas em 15.7% (22/140 das amostras de tecido). As combinações mais frequentes foram TTSuV1/TTSuVk2 em 31.8% (7/22), PCV2/TTSuVk2 em 18.1% (4/22), e PCV2/PCV3/TTSuVk2 em 13.6% (3/22). A depleção de linfócitos foi associada à infecção por TTSuVk2 (p = 0,0041) e esses achados sugerem que TTSuKV2 desempenha uma indução de lesões linfoides semelhantes a PMWS em porcos. Os dados obtidos neste estudo mostram que PCV2, PCV3, TTSuV1 e TTSuVk2 estão relacionados à infecção em animais assintomáticos com lesões teciduais diversas, e sugerem que o diagnóstico molecular para esses patógenos deve ser considerado no monitoramento sanitário dos rebanhos.

Helicobacter spp. and gastric lesions association in minipigs / Associação entre Helicobacter spp. e lesões gástricas em minipigs

ABSTRACT: Due to their similarity to humans, studies regarding gastric ulcers in pigs have become of great interest. It is known that Helicobacter spp. is related to the occurrence of these ulcers, as they possess a high prevalence in pigs. This study aimed to associate gastric lesions findings naturally occurring in minipigs related to or not related to the presence of Helicobacter spp., through the ultra-rapid urease test and immunohistochemical analysis, reaffirming the disease in swine as a natural biomedical model for human cases. For this, samples of formalin-fixed paraffin-embedded gastric tissues from 40 minipigs were obtained at the Laboratório de Morfologia e Patologia Animal (LMPA) in the Universidade Estadual do Norte Fluminense (UENF). Regarding the ultra-rapid urease test, no animal was positive in all regions. However, 18 were positive in at least one: six (15%) were positive in the aglandular region, eight (20%) in the antrum region, 13 (32.5%) in the region of the cardiac gland, and one (2.5%) in the region of the fundic gland. Regarding immunohistochemical analysis, only one animal was positive in all regions, and 32 animals were positive in at least one: four (10%) were positive in the aglandular region, 10 (25%) in the antrum region, 23 (57.5%) in the region of the cardiac gland, and three (7.5%) in the region of the fundic gland. The gastric lesion findings showed a close relationship with Helicobacter spp., enriching the laboratory animal pathologies list. The immunostaining of the bacteria not associated with gastric lesions in certain regions demonstrated the saprophytic and opportunisitic nature of Helicobacter.

RESUMO: Por sua semelhança com a da espécie humana, a úlcera gástrica nos suínos tem despertado muito interesse. Sabe-se que o Helicobacter spp. está relacionado à ocorrência dessas úlceras, apresentando alta prevalência. Este estudo teve como objetivo associar as lesões gástricas, de ocorrência natural em miniporcos, à presença, ou não, de Helicobacter spp., por meio do teste ultra-rápido da urease e da análise imunohistoquímica, reafirmando a enfermidade em suínos como modelo biomédico natural para os casos em humanos. Para este fim, foram utilizados histossetes de tecido gástrico de 40 minipigs obtidos no Laboratório de Morfologia e Patologia Animal (LMPA) da Universidade Estadual do Norte Fluminense (UENF). Em relação ao teste ultra-rápido da urease, nenhum animal foi positivo em todas as regiões, entretanto, 18 foram positivos em pelo menos uma: seis foram positivos na região aglandular, oito na região do antro, 13 na região das glândulas cárdicas e um na região das glândulas fúndicas. Em relação à análise imuno-histoquímica, somente um animal foi positivo em todas as regiões e 32 animais foram positivos em pelo menos uma: quatro foram positivos na região aglandular, 10 na região do antro, 23 na região das glândulas cárdicas e três na região das glândulas fúndicas. As lesões gástricas demonstraram estreita relação com o Helicobacter spp. em miniporcos, enriquecendo a lista de patologias de animais de laboratório. A imunomarcação da bactéria não associada à lesão, em certas regiões gástricas, demonstra seu caráter saprofítico e oportunista.

Associação de alterações anatomopatológicas com o Helicobacter spp. por meio de análise imuno-histoquímica em estômagos de leitões / Association of anatomopathological changes with the Helicobacter spp. through immunohistochemical analysis in piglets stomachs

O objetivo deste trabalho foi relacionar os achados anatomopatológicos das lesões gástricas subclínicas de ocorrência natural em leitões com a presença, ou não, de Helicobacter spp. por meio da imuno-histoquímica. Foram utilizados 48 leitões de linhagem genética comercial. Os animais foram adquiridos em uma granja comercial, com peso médio de 34 Kg e idade média de 79 dias; após o abate, seus estômagos foram coletados e avaliados. Avaliações histopatológicas e imuno-histoquímicas foram realizadas em amostras das regiões anatômicas aglandular e glandular. Macroscopicamente, 34 (70,83%) leitões apresentaram lesões na região aglandular, enquanto que em 14 animais (29,17%) não foram encontradas alterações nesta região. Dos estômagos com lesão, 14 foram classificados como grau 1, seis como grau 2 e 14 como grau 3. Microscopicamente, 44 amostras (91,66%) apresentaram paraqueratose. Deste total, 22 apresentaram a forma discreta, 20 a moderada e dois a acentuada. Na avaliação macroscópica da porção glandular, 41 (85,4%) animais apresentaram alteração em pelo menos uma das três regiões, e em somente sete (14,6%) não foram encontradas lesões em nenhuma delas. Em 14 deles, houve aumento da atividade mucípara, em dois, houve erosão e, em cinco, hiperemia. As lesões na região glandular do estômago foram mais extensas no antro e no cárdia, seguidas do fundo. Em relação à análise imuno-histoquímica, 21 (43,8%) amostras tiveram resultados negativos em todas as regiões, e 24 (50%) foram positivas em pelo menos uma delas, sendo que nenhuma foi positiva em todas. Os achados anatomopatológicos demonstraram relação estatística com a bactéria e, sua imunomarcação não associada à lesão em certas regiões gástricas, demonstra seu caráter saprofítico e oportunista.

The aim of this study was to relate the anatomopathological findings of naturally occurring subclinical gastric lesions in piglets, with or without Helicobacter spp. through immunohistochemistry. Forty-eight piglets of commercial genetic lineage were used. The animals were acquired in a commercial farm, with an average weight of 34 kg and an average age of 79 days, and after slaughter, their stomachs were collected and evaluated. Samples from the glandular and aglandular anatomical regions were evaluated. Macroscopically, 34 (70.83%) samples had lesions on aglandular region, while 14 (29.17%) nothing had. Of the injured stomachs, 14 were classified as grade 1, six as grade 2 and 14 as grade 3. Microscopically, 44 samples (91.66%) showed parakeratosis. Of these, 22 showed a discreet manner, 20 moderate and two severe. In the glandular region, in 41 (85.4%) samples there was a change in at least one of the three regions, and only seven animals (14.6%) showed no change in any of the three. Fourteen samples showed increased muciparous activity, two showed erosion and five hyperemia. The lesions were higher in antral regions and cardic, followed the fundus. In relation to immunohistochemistry, 21(43.8%) samples were negative in all areas, 24 (50%) were positive in at least one, and none were positive in all. The anatomopathological findings showed a statistical relationship with the bacteria, and its immunostaining, not associated with gastric lesions in certain regions, demonstrates its saprophytic and opportunistic character.

Associação de alterações anatomopatológicas com o Helicobacter spp. por meio de análise imuno-histoquímica em estômagos de leitões

O objetivo deste trabalho foi relacionar os achados anatomopatológicos das lesões gástricas subclínicas de ocorrência natural em leitões com a presença, ou não, de Helicobacter spp. por meio da imuno-histoquímica. Foram utilizados 48 leitões de linhagem genética comercial. Os animais foram adquiridos em uma granja comercial, com peso médio de 34 Kg e idade média de 79 dias; após o abate, seus estômagos foram coletados e avaliados. Avaliações histopatológicas e imuno-histoquímicas foram realizadas em amostras das regiões anatômicas aglandular e glandular. Macroscopicamente, 34 (70,83%) leitões apresentaram lesões na região aglandular, enquanto que em 14 animais (29,17%) não foram encontradas alterações nesta região. Dos estômagos com lesão, 14 foram classificados como grau 1, seis como grau 2 e 14 como grau 3. Microscopicamente, 44 amostras (91,66%) apresentaram paraqueratose. Deste total, 22 apresentaram a forma discreta, 20 a moderada e dois a acentuada. Na avaliação macroscópica da porção glandular, 41 (85,4%) animais apresentaram alteração em pelo menos uma das três regiões, e em somente sete (14,6%) não foram encontradas lesões em nenhuma delas. Em 14 deles, houve aumento da atividade mucípara, em dois, houve erosão e, em cinco, hiperemia.  As lesões na região glandular do estômago foram mais extensas no antro e no cárdia, seguidas do fundo. Em relação à análise imuno

The aim of this study was to relate the anatomopathological findings of naturally occurring subclinical gastric lesions in piglets, with or without Helicobacter spp. through immunohistochemistry. Forty-eight piglets of commercial genetic lineage were used. The animals were acquired in a commercial farm, with an average weight of 34 kg and an average age of 79 days, and after slaughter, their stomachs were collected and evaluated. Samples from the glandular and aglandular anatomical regions were evaluated. Macroscopically, 34 (70.83%) samples had lesions on aglandular region, while 14 (29.17%) nothing had. Of the injured stomachs, 14 were classified as grade 1, six as grade 2 and 14 as grade 3. Microscopically, 44 samples (91.66%) showed parakeratosis. Of these, 22 showed a discreet manner, 20 moderate and two severe. In the glandular region, in 41 (85.4%) samples there was a change in at least one of the three regions, and only seven animals (14.6%) showed no change in any of the three. Fourteen samples showed increased muciparous activity, two showed erosion and five hyperemia. The lesions were higher in antral regions and cardic, followed the fundus. In relation to immunohistochemistry, 21(43.8%) samples were negative in all areas, 24 (50%) were positive in at least one, and none were positive in all. The anatomopathological findings showed a statistical relationship with the bacteria, and its immunostaining, not associated with gastric lesions in certain regions, demonstrates its saprophytic and opportunistic character.

Anatomopathological changes, quantification and distribution of Libyostrongylus spp. in regions of the proventriculus and ventriculus of naturally- and experimentally-infected ostriches.

Nematodes of the genus Libyostrongylus parasitize ostriches, causing high mortality rates. These nematodes are found in the proventriculus and ventriculus of ostriches, but little is known about their distribution and the possible anatomopathological changes they cause in the various regions of these organs. This paper describes the distribution and quantification of Libyostrongylus and pathological changes found in regions of the proventriculus and ventriculus of ostriches with high and low levels of both natural and experimental infection. Ostriches were necropsied and tissue samples from the distinct regions of both organs were analysed based on nematode counts and histopathology after staining with haematoxylin and eosin, Masson's trichrome or Alcian blue/PAS. The cranial and glandular regions of the proventriculus were the most parasitized. The ventriculus contained more nematodes in the caudal region. No macro- or microscopic pathological changes were observed in either of these organs of experimentally-infected birds. However, naturally-infected birds with high levels of infection presented proventriculus with macroscopic lesions and heterophilic infiltrates surrounding nematodes. In the glandular region of this organ, nematodes were located in the adenomeres of the secretory ducts, causing altered architecture and erosions and ulcerative lesions with damaged epithelium. Nematode eggs were found in the koilin layer of the middle and caudal regions of the ventriculus only of these birds. The pH of the regions assessed by Alcian blue/PAS staining changed from acidic in the proventriculus to more alkaline in the caudal region of the ventriculus. These data add knowledge to the biology of Libyostrongylus. RESEARCH HIGHLIGHTS The most parasitized areas were the cranial and glandular regions of the proventriculus. Naturally-infected birds with high levels of infection presented macro lesions in the proventriculus and damaged epithelium. Nematode eggs were found in the ventriculus. The proventriculus had an acidic pH, which turned alkaline towards the ventriculus.

Comparison among bone marrow mesenchymal stem and mononuclear cells to promote functional recovery after spinal cord injury in rabbits

Abstract Purpose: To investigate the efficacy of allogeneic mesenchymal stem-cells and autologous mononuclear cells to promote sensorimotor recovery and tissue rescue. Methods: Female rabbits were submitted to the epidural balloon inflation method and the intravenous cells administrations were made after 8 hours or seven days after injury induction. Sensorimotor evaluation of the hindlimbs was performed, and the euthanasia was made thirty days after the treatment. Spinal cords were stained with hematoxylin and eosin. Results: Both therapies given 8 hours after the injury promoted the sensorimotor recovery after a week. Only the group treated after a week with mononuclear cells showed no significant recovery at post-injury day 14. In the days 21 and 28, all treatments promoted significant recovery. Histopathological analysis showed no difference among the experimental groups. Our results showed that both bone marrow-derived cell types promoted significant sensorimotor recovery after injury, and the treatment made at least a week after injury is efficient. Conclusion: The possibilities of therapy with bone marrow-derived cells are large, increasing the therapeutic arsenal for the treatment of spinal cord injury.

Comparison among bone marrow mesenchymal stem and mononuclear cells to promote functional recovery after spinal cord injury in rabbits.

PURPOSE: To investigate the efficacy of allogeneic mesenchymal stem-cells and autologous mononuclear cells to promote sensorimotor recovery and tissue rescue. METHODS: Female rabbits were submitted to the epidural balloon inflation method and the intravenous cells administrations were made after 8 hours or seven days after injury induction. Sensorimotor evaluation of the hindlimbs was performed, and the euthanasia was made thirty days after the treatment. Spinal cords were stained with hematoxylin and eosin. RESULTS: Both therapies given 8 hours after the injury promoted the sensorimotor recovery after a week. Only the group treated after a week with mononuclear cells showed no significant recovery at post-injury day 14. In the days 21 and 28, all treatments promoted significant recovery. Histopathological analysis showed no difference among the experimental groups. Our results showed that both bone marrow-derived cell types promoted significant sensorimotor recovery after injury, and the treatment made at least a week after injury is efficient. CONCLUSION: The possibilities of therapy with bone marrow-derived cells are large, increasing the therapeutic arsenal for the treatment of spinal cord injury.

Lesões pré-ulcerativas gastroesofágicas em miniporcos / Gastroesophageal ulcer research in mini pigs

A úlcera gastroesofágica figura como uma das principais causas de morte súbita de suínos em fase de reprodução. O objetivo deste trabalho foi avaliar os achados anatomopatológicos das lesões gástricas subclínicas em miniporcos. Quarenta animais, com peso médio de 39kg e idade de 17 meses, foram mantidos em biotério, em condições de temperatura e umidade constantes, alimentados duas vezes ao dia, sendo depois eutanasiados e necropsiados. Os estômagos foram colhidos e avaliados. Macroscopicamente, 29 animais apresentaram alterações na região aglandular, sendo que 37,5% foram classificadas no grau 1; 22,5%, no grau 2; e 12,5%, no grau 3. Microscopicamente, 29 (72,5%) miniporcos apresentaram paraqueratose. Destes, 35% tinham a forma moderada, 25% a discreta e 12,5% a acentuada. As lesões glandulares foram mais extensas na região cárdica, seguida da antral e fúndica. Trinta e seis (90%) animais apresentaram lesão em pelo menos uma das três regiões. Em 21 (52,5%) mini- porcos observou-se hiperatividade mucípara com catarro; em 17 (42,5%), aumento da atividade mucípara; em dez (25%), erosão. Os achados revelaram alta prevalência de lesões pré-ulcerativas nos estômagos dos miniporcos. A presença destas relacionou-se à gravidade macroscópica: quanto mais graves, mais intensa a paraqueratose.

Gastro-esophageal ulcers are one of the principal causes of reproduction pigs sudden death. This work aimed to evaluate the pathological findings of subclinical gastric lesions naturally occurring in mini pigs. Forty mini pigs with a mean weight of 39.0 kg and 17 months of age kept in animal facility, under constant temperature and humidity, and fed twice a day, were euthanized and necropsied. Their stomachs were collected and evaluated. Macroscopic lesions in aglandular portion were observed in 29 animals, and 37.5% of them were classified as grade 1, 22.5% as grade 2, and 12.5% as grade 3. In 29 (72.5%) animals parakeratosis was microscopically seen: 35% had the moderate manner, 25% discreet and 12.5% severe one. Glandular lesions were more extensive in the cardia region, followed by antral and fundic. Thirty-six (90%) animals had lesions in at least one of three regions. In 21 (52.5%) mini pigs, it was observed mucipara hyperactivity with phlegm; in 17 (42.5%), increased mucipara activity; and in ten (25%), erosion. Findings revealed high prevalence of pre-ulcerative lesions in the stomachs of mini pigs. The presence of these was related to the macroscopic gravity: the more serious, more intense was the parakeratosis.