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.

Serious lesions in Green turtles (Chelonia mydas) afflicted by fatal Spirorchiidiasis found stranded in south and southeastern Brazil.

Several diseases have been reported as affecting endangered wild sea turtle population worldwide, including spirorchiidiasis. This parasitic infection results in serious circulatory disorders in sea turtles, as well as tissue damage due to the presence of spirorchiids eggs. However, few reports of organs severely affected by tissue replacement caused by granulomatous inflammatory processes due to spirorchiidiasis in sea turtles are available. In this regard, this study describes massive lesions in 16 juvenile green turtles from southeastern Brazil presenting no other detectable diseases or injuries, associated to parasitic compression of air spaces, parasitic thyroid atrophy, parasitic encephalic compression and parasitic splenic lymphoid depletion. These rare injuries were categorized as extremely severe, affecting most spirorchiidiasis-infected organs. Spirorchiidiasis was, thus, noted herein as capable of causing a variety of lethal injuries to vital or extremely important organs in sea turtles. Spirorchiidiasis should, therefore, also be considered a potential cause of death in stranded green sea turtle monitoring efforts.

Pathological changes by spirorchiid eggs in hawksbill sea turtle (Eretmochelys imbricata) stranded off brazilian coast.

Eretmochelys imbricata (hawksbill turtle) is classified as a critically endangered species at world level; in Brazil, it is listed among the threatened species. Spirorchiids are parasites of the circulatory system of chelonians which may cause serious lesions in the various tissues of the host due to deposition of eggs in the bloodstream. In this context, the aim of the present study was to describe the pathology caused by spirorchiid eggs in E. imbricata from the Brazilian over a five year period. A total of 29 animals were analyzed, of which nine (31%) presented lesions associated to spirorchiids eggs. The lesions were: granulomatous enteritis in six (66.66%), granulomatous splenitis in five (55.55%), granulomatous hepatitis in three (33.33%), granulomatous pneumonia in three (33.33%), granulomatous pancreatitis in two (22.22%), and granulomatous adrenalitis in one specimen (11.11%). Concluded the main lesion in Eretmochelys imbricata was giant-cell granulomatous inflammatory reaction to the parasite's eggs. Animals exhibiting poor physical health were more susceptible to presenting such lesions.

A new haemosporidian parasite from the Red-legged Seriema Cariama cristata (Cariamiformes, Cariamidae).

Haemoproteids (Haemosporida, Haemoproteidae) are a diverse group of avian blood parasites that are transmitted by hematophagous dipterans. In this study, we describe Haemoproteus pulcher sp. nov. from a Red-legged Seriema (Cariama cristata) in southeast Brazil. Analysis of the mitochondrial cytb gene indicates this parasite is closely related to Haemoproteus catharti (from Turkey Vulture, Cathartes aura) and the unidentified haemosporidian lineages PSOOCH01 (from Pale-winged Trumpeter, Psophia leucoptera) and MYCAME08 (from Wood Stork, Mycteria americana). This group of parasites appears to represent an evolutionary lineage that is distinct from other Haemoproteus spp., being instead more closely related to Haemocystidium spp. (from reptiles), Plasmodium spp. (from reptiles, birds, and mammals) and other mammal-infecting haemosporidians (Nycteria, Polychromophilus, and Hepatocystis). Current evidence suggests that parasites of this newly discovered evolutionary lineage may be endemic to the Americas, but further studies are necessary to clarify their taxonomy, life cycle, vectors, hosts, geographic distribution and host health effects. Additionally, it should be borne in mind that some PCR protocols targeting the cytb gene might not reliably detect H. pulcher due to low primer affinity.

Investigation of Leishmania (Viannia) braziliensis Infection in Wild Mammals in Brazil.

PURPOSE: Leishmaniasis are infectious and zoonotic diseases and present in cutaneous and visceral forms. Cutaneous leishmaniasis is endemic and widely distributed throughout the state of Espírito Santo, Brazil. Several cases of cutaneous leishmaniasis in humans and dogs associated with Leishmania (Viannia) braziliensis have been reported in the state. This study aimed to investigate the occurrence of natural infection by Leishmania(Viannia) braziliensis in wild mammals found dead (by trampling or natural death) in the Sooretama Biological Reserve, Espírito Santo State. METHODS: From January 2018 to December 2019, 60 animals were collected. Of these, 47 animals from 12 different species were analyzed. The results were demonstrated using descriptive analysis of the observations to calculate the absolute and relative frequencies of the data. RESULTS: In the PCR, using specific primers for the genus Leishmania (D1, D2, and D3) and the species Leishmania (Viannia) braziliensis (ISVB/ISVC), 4 positive animals (8.5%) were detected: 1 Cuniculus paca (paca) (25%) and 3 Callithrix geoffroyi (white-faced marmoset) (25%). In the histopathological analysis, the parasitic amastigote form was not observed. CONCLUSIONS: The natural infection, detected by PCR, by Leishmania (Viannia) braziliensis in Cuniculus paca (paca) and Callithrix geoffroyi (white-faced marmoset) constitutes the first report of infection of this rodent and primate species in the literature. Despite the confirmation of Leishmania (Viannia) braziliensis infection in rodents and primates, the role of these species in the transmission of this zoonosis still needs further observational studies to identify their seasonal variation, transmissibility, infection stability, and the effects of a given parasite on the population and/or individual.

(R)-(+)-Lasiodiplodin isolated from the endophytic fungus Sordaria tamaensis exhibits potent antimycobacterial and anti-inflammatory activities in vitro and in vivo: a dual approach for the treatment of severe pulmonary tuberculosis.

OBJECTIVES: This study aimed to evaluate endophytic fungi isolated from Tocoyena bullata and Humiria balsamifera plant species for their antimycobacterial and anti-inflammatory activities, focusing on severe pulmonary tuberculosis cases which are often associated with exacerbated inflammation. METHODS: Mycobacterium suspensions were incubated with the samples for 5 days. RAW 264.7 macrophages stimulated with LPS were also incubated with them for 24 h to assess the inhibition of inflammatory mediator production and cytotoxicity. C57BL/6 mice were infected with Mtb M299 and treated for 15 days with lasiodiplodin (Lasio). KEY FINDINGS: Endophytic fungus Sordaria tamaensis, obtained from T. bullata, was the most promising. Its ethanolic extract impaired mycobacterial growth with MIC50 (µg/ml): 1.5 ± 0.6 (BCG), 66.8 ± 0.1 (H37Rv) and 80.0 ± 0.1 (M299). (R)-(+)-Lasio showed MIC50 92.2 ± 1.8 µg/ml (M299). In addition, Lasio was able to inhibit NO, IL-1ß and TNF-α production and was not cytotoxic for macrophages. M. tuberculosis-infected C57BL/6 animals treated by Lasio reduced the number of acid-fast bacilli, lung pathology, leucocyte influx and proinflammatory cytokine production in the lungs. The class IIa fructose 1,6-bisphosphate aldolase was the predicted hypothetical target of Lasio. CONCLUSIONS: (R)-(+)-Lasio stood out as a promising anti-TB compound, exhibiting anti-inflammatory and antimycobacterial effects, as well as low cytotoxicity.

A Murine Model of Mycobacterium kansasii Infection Reproducing Necrotic Lung Pathology Reveals Considerable Heterogeneity in Virulence of Clinical Isolates.

Among non-tuberculous mycobacteria, Mycobacterium kansasii is one of the most pathogenic, able to cause pulmonary disease indistinguishable from tuberculosis in immunocompetent susceptible adults. The lack of animal models that reproduce human-like lung disease, associated with the necrotic lung pathology, impairs studies of M. kansasii virulence and pathogenicity. In this study, we examined the ability of the C57BL/6 mice, intratracheally infected with highly virulent M. kansasii strains, to produce a chronic infection and necrotic lung pathology. As a first approach, we evaluated ten M. kansasii strains isolated from Brazilian patients with pulmonary disease and the reference strain M. kansasii ATCC 12478 for virulence-associated features in macrophages infected in vitro; five of these strains differing in virulence were selected for in vivo analysis. Highly virulent isolates induced progressive lung disease in mice, forming large encapsulated caseous granulomas in later stages (120-150 days post-infection), while the low-virulent strain was cleared from the lungs by day 40. Two strains demonstrated increased virulence, causing premature death in the infected animals. These data demonstrate that C57BL/6 mice are an excellent candidate to investigate the virulence of M. kansasii isolates. We observed considerable heterogeneity in the virulence profile of these strains, in which the presence of highly virulent strains allowed us to establish a clinically relevant animal model. Comparing public genomic data between Brazilian isolates and isolates from other geographic regions worldwide demonstrated that at least some of the highly pathogenic strains isolated in Brazil display remarkable genomic similarities with the ATCC strain 12478 isolated in the United States 70 years ago (less than 100 SNPs of difference), as well as with some recent European clinical isolates. These data suggest that few pathogenic clones have been widely spread within M. kansasii population around the world.

Brown booby (Sula leucosgaster) envenomation and death caused by toadfish (Porichthys porosissimus) ingestion in Southeastern Brazil / Envenenamento e morte de atobás-marrons (Sula leucogaster) causados pela ingestão de magangá-liso (Porichthys porosissimus) no sudeste do Brasil

Abstract A case report of six brown booby (Sula leucogaster) envenomation following the ingestion of toadfish (Porichthys porosissimus) specimens in Rio de Janeiro, Southeastern Brazil is discussed herein. Several macro- and microscopic pathological alterations were observed in the examined brown booby specimens, and rapid envenomation (<24 h) is suggested due to the digestion stage of the fish found in their gastrointestinal tracts. This is the first report to mention the death of adult and juvenile brown boobies due to envenomation by the toadfish P. porosissimus. Further studies assessing the role of discarded fish as a driving force of negative effects on seabirds, such as envenomations, for example, along the southeastern coast of Brazil are recommended.

Resumo Um relato de caso sobre o envenenamento de seis indivíduos de atobás-marrons (Sula leucogaster) após a ingestão de espécimes de mangagá-liso (Porichthys porosissimus) no Rio de Janeiro, sudeste do Brasil, é discutido neste estudo. Diversas alterações patológicas macro e microscópicas foram observadas nos espécimes de atobá-marrom examinados, e um envenenamento rápido (<24 h) é sugerido devido ao estágio de digestão dos peixes encontrados em seus tubos digestórios. Este é o primeiro registro que menciona a morte de indivíduos juvenis e adultos de atobás marrons por envenenamento devido a ingestão do magangá-liso P. porosissimus. Novos estudos avaliando o papel dos peixes descartados como uma força motriz dos efeitos negativos sobre as aves marinhas como, por exemplo, envenenamentos, ao longo da costa sudeste do Brasil são recomendados.

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.

Causes of Fatal Cyathostomiasis in Brown Booby (Sula leucogaster) from Brazil: Identification of Pathogen and Implications for Management.

Fatal infection by Cyathostoma (Cyathostoma) phenisci (Nematoda: Syngamidae), was identified in 2 of 52 brown boobies (Sula leucogaster) collected on beaches in the state of Rio de Janeiro, Brazil, and admitted to the veterinary clinic for rehabilitation. Both infected birds were in poor physical condition, with atrophied pectoral muscles, and died soon after starting treatment. The parasitological and pathological examination of the carcasses revealed the presence of C. (C.) phenisci in the trachea, resulting in tracheitis, as well as severe parasitic granulomatous bronchopneumonia caused by eggs deposited in the lungs. In our opinion, these serious pathological changes were the primary cause of chronic respiratory illness. This is the first description of fatal cyathostomiasis in a fish-eating avian host caused by infection by a member of the subgenus Cyathostoma (Cyathostoma). Therefore, it is reasonable to consider C. (C.) phenisci to be a real threat to a wide range of their definitive hosts, and cyathostomiasis should be considered in the differential diagnosis for fish-eating marine birds, even in cases without respiratory signs. This is also the first record of the genus Cyathostoma in Brazil.

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.

Atrophic Proventriculitis Associated with Gravid Females of Tetrameres sp. (Nematoda: Tetrameridae) in the Magellanic Penguin (Spheniscus magellanicus) on the Coast of Brazil.

The present study offers the first description of proventriculitis associated with the presence of gravid female nematodes of the genus Tetrameres (Nematoda: Tetrameridae) in 3 juvenile Magellanic penguins (Spheniscus magellanicus) found dead on the coast of Brazil. This study broadens knowledge on parasites associated with these hosts and the real impact of this association.

Spontaneous Primary Right Tibiotarsal Osteosarcoma with Pulmonary Metastasis in a Free-Ranging Cory's Shearwater (Calonectris diomedea).

We report the pathologic features of a primary right tibiotarsal productive osteoblastic osteosarcoma with pulmonary metastasis, a unique finding in a Cory's Shearwater (Calonectris diomedea).

Fatal Rameshwarotrema uterocrescens infection with ulcerative esophagitis and intravascular dissemination in green turtles.

The trematode Rameshwarotrema uterocrescens (Digenea: Pronocephalidae) parasitizes the glands of the caudal esophagus of Chelonia mydas. In the present study, 741 C. mydas were examined, 85 animals had adult specimens of R. uterocrescens associated with necrotizing ulcerous esophagitis, of these 85, 21 presented invasion of the esophageal mural vessels in the caudal esophagus of juvenile green turtles (Chelonia mydas). Necrotizing granulomatous splenitis, hepatitis, and nephritis were associated with the presence of parasites. The eggs from R. uterocrescens are birefringent under plane-polarized light, which distinguishes them from those of spirorchiid trematodes. This study contributes novel data on R. uterocrescens, methods for detecting this parasite, and demonstrates the fatal potential of parasitism in C. mydas.

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.

Myrtenal-induced V-ATPase inhibition - A toxicity mechanism behind tumor cell death and suppressed migration and invasion in melanoma.

BACKGROUND: Metastatic tumor cells have acidic extracellular pH and differential electrochemical H+ gradients generated across their cell membranes by V-type H+-ATPases. This study shows that inhibition of the V-ATPases by the plant-derived monoterpene Myrtenal results in tumor cell death and decreased metastatic dissemination in mice. METHODS: The Myrtenal anticancer toxicity was evaluated in vitro using murine (B16F0 and B16F10) and human (SkMel-5) melanoma cell lines, and in in vivo mouse metastatic dissemination model. Proton flux and extracellular acidification were directly evaluated at the surface of living cells using a non-invasive selective ion electrode approach. RESULTS: The inhibition of V-ATPases by 100 µM Myrtenal disrupted the electrochemical H+ gradient across the cell membranes, strongly induced cell death (4-5 fold), and decreased tumor cells migration and invasion in vitro. Myrtenal (15 mg/kg) also significantly reduced metastasis induced by B16F10 in vivo, further reinforcing that V-ATPase is a molecular target to halt the progression of cancers. CONCLUSIONS: These data revealed the therapeutic potential of Myrtenal as inhibitor of melanoma progression proposing a mechanism of action by which once inhibited by this monoterpene the proton pumps fail to activate cancer-related differential electrochemical gradients and H+ fluxes across the tumor cell membranes, disrupting pH signatures inherent in tumor progression, resulting in reprogrammed cell death and metastasis inhibition. GENERAL SIGNIFICANCE: The work represents a new mechanistic strategy for contention of melanoma, the most aggressive and deadly form of cutaneous neoplasm, and highlights Myrtenal, other related monoterpenes and derivatives as promising proton pump inhibitors with high chemotherapeutic potential.

Diagnostic methods to detect uterus illnesses in mares.

Uterine illnesses are the major problem in horse reproductive programs. The diagnosis of these pathologies is often neglected or does not provide enough information about the cause. The aim of this study was to evaluate different diagnoses to provide more accurate information about endometritis and the real endometrial profile at the beginning of a breeding season. Fifty-one mares with reproductive failure were evaluated. Two different uterine cytology diagnoses, culture of uterine flush material and biopsy assay, were performed. The data were analyzed by the Chi-squared test to compare different diagnostic methods for the same sample from each mare, and sensitivity and specificity tests were performed. Most of evaluations showed no association between cytology and other diagnostic methods, except with bacterial culture. The most common bacterium found was Escherichia colli, but it was not associated with neutrophil presence in cytology. The presence of ß-hemolitic streptococcus was well characterized by positive cytology and cloudy lavage fluid aspect. Endometrial profile was affected by age and whether the mare was a donor or recipient. The most sensitive diagnostic method was gross aspect of lavage fluid. Endometritis should be diagnosed by at least two methods to increase accuracy. Endometritis is not the only uterine disease that causes embryo loss, since endometrial fibrosis can also be a cause.

Chronic Cystitis Associated with Plesiochorus cymbiformis ( Rudolphi, 1819 ) Looss, 1901 (Digenea: Gorgoderidae) in a Loggerhead Turtle Caretta caretta (Linnaeus 1758) (Testudines, Cheloniidae) from Brazil: A Case Report.

This paper reports the first occurrence of Plesiochorus cymbiformis (Digenea: Gorgoderidae) in a loggerhead turtle ( Caretta caretta [Testudines, Cheloniidae]) in Brazil and describes the histological findings of related injuries to the urinary bladder.

Correction: Hypervirulent Mycobacterium tuberculosis strain triggers necrotic lung pathology associated with enhanced recruitment of neutrophils in resistant C57BL/6 mice.

[This corrects the article DOI: 10.1371/journal.pone.0173715.].