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.

Chickens treated with a nitric oxide inhibitor became more resistant to Plasmodium gallinaceum infection due to reduced anemia, thrombocytopenia and inflammation.

Malaria is a serious infectious disease caused by parasites of the Plasmodium genus that affect different vertebrate hosts. Severe malaria leads to host death and involves different pathophysiological phenomena such as anemia, thrombocytopenia and inflammation. Nitric oxide (NO) is an important effector molecule in this disease, but little is known about its role in avian malaria models. Plasmodium gallinaceum-infected chickens were treated with aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase, to observe the role of NO in the pathogenesis of this avian model. AG increased the survival of chickens, but also induced higher parasitemia. Treated chickens demonstrated reduced anemia and thrombocytopenia. Moreover, erythrocytes at different stages of maturation, heterophils, monocytes and thrombocytes were infected by Plasmodium gallinaceum and animals presented a generalized leucopenia. Activated leukocytes and thrombocytes with elongated double nuclei were observed in chickens with higher parasitemia; however, eosinophils were not involved in the infection. AG reduced levels of hemozoin in the spleen and liver, indicating lower inflammation. Taken together, the results suggest that AG reduced anemia, thrombocytopenia and inflammation, explaining the greater survival rate of the treated chickens.

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.

(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.

Mixed infection with Libyostrongylus dentatus and Libyostrongylus douglassii induces a heterophilic inflammatory infiltrate in the proventriculus of ostriches.

Libyostrongylus dentatus and Libyostrongylus douglassii are haematophagous nematodes found in the proventriculus and the ventriculus of ostriches. Pathological damage leading to bird death has been attributed to L. douglassii. However, histopathology of the mixed infection has not been reported. The aim of the present work was to characterize the cellular inflammatory infiltrate found in the proventriculus of ostriches with a mixed infection. Analysis of the collected nematodes confirmed a mixed infection in the proventriculus of examined birds. Histopathological examination of the proventriculus showed an inflammatory infiltrate composed of granular cells in close proximity to the nematodes. The granulocyte infiltrate was composed mainly of heterophils identified by the lack of peroxidase and presence of fusiform granules.

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).

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.

Pathologies of the digestive system caused by marine debris in Chelonia mydas.

The growth of human population and deficient pollution control measures pose significant challenge to the environment. Despite conservation efforts, all sea turtle species are at some risk of extinction. The present study investigated the effect of marine debris on the gastrointestinal tract of green turtles in southeastern Brazil. Of the 777 animals evaluated, 290 showed marine debris in one segment of the gastrointestinal tract. The presence of these materials in the gastrointestinal tract may be harmful, independent of the segment involved, and increases the risk of impaction. Marine debris has become a significant hazard to Chelonia mydas in the region surveyed, causing perforation, rupture, or fecal impaction that, when not treated, is potentially fatal, exposing the intestine to bacterial infection.

Disseminated pythiosis in three horses.

Three cases of equine subcutaneous pythiosis with dissemination to the internal organs were investigated. The subcutaneous lesions were observed on the mammary gland, nostrils and limbs of the infected horses. Histopathological analysis of the infected tissues revealed a strong eosinophilic reaction, with macrophages, mast cells and giant cells. Sparsely septated hyphal filaments of 4-6 microm diameter were identified in the center of the eosinophilic areas. Specific fluorescent antibody against Pythium insidiosum confirmed the hyphae in the infected tissues in three examined horses. In one of the three cases, the DNA sequences amplified from the infected subcutaneous tissues and internal organs, revealed that P. insidiosum's 18S SSU rDNA amplicons shared 100% identity with those sequences deposit in GenBank. This is the first report confirming by immunochemical and genetic techniques that P. insidiosum can disseminated from superficial to deep structures.

Differential immunohistochemical expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in cow uteri with adenomyosis during follicular phase.

The present investigation was intended to show a different immunohistochemical profile of matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2 in bovine uteri with adenomyosis during follicular phase. Uterine samples of 32 cows in reproductive age were taken from the medial third of one of the uterine horns and grouped according to the adenomyosis degree (superficial and deep). Tissue sections (4 µm) were incubated overnight at 4°C with monoclonal antibody for matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2. Staining intensities were evaluated in the luminal epithelium, ectopic and dystopic endometrial tissue (stroma, capillaries and glands), endometrial-myometrial border, myometrium, myometrial vessels (middle tunic and endothelium). The matrix metalloproteinase-2 expression was higher for deep adenomyosis samples, showing a differential mean reactivity in superficial endometrium, myometrial vessels, myometrium adjacent to adenomyotic focus and endometrial-myometrial border (P < 0.05). Moreover, matrix metalloproteinase-2 expression was higher in deep adenomyosis samples than that of Tissue inhibitor metalloproteinase-2 in almost all uterine structures analyzed (except for the endometrial and myometrial vessels and endometrial-myometrial border). The opposite was observed in the follicular phase, for both normal specimens and with superficial adenomyosis, where Tissue inhibitor metalloproteinase-2 expression was higher than that of matrix metalloproteinase-2. In conclusion, a differential pattern of matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2 was observed in cow uteri with adenomyosis.

Difference in virulence of Mycobacterium avium isolates sharing indistinguishable DNA fingerprint determined in murine model of lung infection.

BACKGROUND: Opportunistic Mycobacterium avium typically causes disease in immunocompromised patients and in some groups of apparently healthy individuals. The high virulence of some bacterial lineages increases the disease risk. High-resolution molecular genotyping studies of M. avium clinical isolates demonstrated that some genotype patterns were more prevalent than others, suggesting that close genetic relatedness of these successful isolates sharing a similar genotype could determine similar biological properties associated with high virulence. METHODS AND FINDINGS: In this study, we aimed to compare the virulence and pathogenic properties of two epidemiologically unrelated M. avium isolates sharing an indistinguishable DNA fingerprint in a well-characterized model of pulmonary infection in mice, resistant or susceptible to mycobacteria. The mice, C57BL/6 wild- type or IFN-gamma gene disrupted (GKO), respectively, were intratracheally infected with two isolates, H27 (human blood isolate) and P104 (pig lymph node isolate), and the lungs were examined for bacterial loads, histopathology and cytokine gene expression. The obtained data demonstrated significant differences in the virulence properties of these strains. Although the H27 strain grew significantly faster than P104 in the early stage of infection, this bacterium induced protective immunity that started to reduce bacterial numbers in the wild-type mice, whereas the P104 strain established a chronic infection. In the GKO mice, both strains were capable of causing a chronic infection, associated with higher bacterial burdens and severe lung pathology, in a similar manner. CONCLUSIONS/SIGNIFICANCE: The results demonstrated that the studied isolates differed in the pathogenic properties although were indistinguishable by actually widely used genotyping techniques demonstrating that the genotype similarity does not predict similarity in virulence of M. avium isolates.

First description of corpus luteum with central cavity during pregnancy in Sotaia guianensis (Van Bénéden, 1864) - Case report / Primeira descrição de corpo lúteo cavitário gestacional em Sotalia guianensis (Van Bénéden, 1864) - Relato de Caso

O objetivo foi descrever pela primeira vez o corpo lúteo cavitário gestacional em Sotalia guianensis, espécie de golfinhocosteiro que se distribui ao longo da costa Atlântica Ocidental. Os ovários de um espécime com 196cm de comprimentocorporal, com 9,2 meses de gestação, capturado acidentalmente durante atividade pesqueira na costa norte do estado doRio de Janeiro. O material foi avaliado morfologicamente e processado para cortes em parafina, corado pela hematoxilina eeosina para microscopia. O ovário direito apresentava forma amendoada, com sulcos em uma lateral, três folículos e umcorpo lúteo polar em involução. O ovário esquerdo pesava sete vezes mais que o direito. A superfície era enrugada, com umfolículo, seis corpos lúteos e quatro albicans. O maior corpo lúteo era protruso, com diâmetro médio de 3cm e uma cavidadecentral de 1cm de diâmetro, revestida por paredes regulares e espessas, sugerindo ser um corpo lúteo em desenvolvimento.Os outros corpos lúteos eram maciços e estavam em regressão. A microscopia do corpo lúteo não demonstrou envolvimentopatológico. Esta descrição, ímpar na espécie, representa uma contribuição ao conhecimento da biologia (reprodutiva) da S.guianensis, que consta como "Dados Deficientes" na lista vermelha da União Internacional para Conservação da Natureza.

This is the first study to describe the corpus luteum with central cavity during pregnancy in Sotalia guianensis, a coastal dolphin species distributed along the Atlantic Ocean coast. The material analyzed was a pair of ovaries of a 196-cm-long female was 9.2 months pregnant, accidentally captured on the north coast of the state of Rio de Janeiro, Brazil. Both ovaries were measured and inspected for anatomic structures. After, ovaries were process in paraffin-embedded and stained with hematoxylin and eosin for microscopy. The left ovary had a wrinkled outside, one follicle, six corpora lutea and four corpora albicans. The largest corpus luteum was protruded, 3 cm long in mean diameter with one central cavity. It was lined by uniform thick walls and peripheral vascularization, which suggests that the corpus luteum is under development. The left ovary was deformed due to the presence of corpus luteum, which accounted for the seven-fold higher weight as compared to the right ovary. The histology of the corpus luteum did not evince any pathologic status of the material. This description, the first ever made for the species, offers a contribution to the existing knowledge about the reproduction biology of S. guianensis, which is a member of the "Insufficient Data" Red List prepared by the World Conservation Union.