Ocurrencia de lepas pedunculadas (Conchoderma virgatum) (Cirripedia: thoracica) en pingüinos de Magallanes (Spheniscus magellanicus)

Magellanic Penguins (Spheniscus magellanicus) breed in Argentina, Chile and the Falkland (Malvinas) Islands, and seasonally migrate towards the coasts of Uruguay, Brazil and Peru. We report the finding of pedunculate barnacles Conchoderma virgatum (Cirripedia: Thoracica) on the body surface of seven out of 62 Magellanic Penguins. These birds were beach-cast along the coast of São Paulo state (23º57’S; 46º23’W), Brazil, on winter 2008 andwinter 2010. Barnacles were attached on the surface of the flippers, feet and lower chest, and in some cases the barnacle had over 40 grouped individuals, some measuring up to 4 cm. Affected penguins had low body mass (range = 1.6 – 2.05 kg) and were in generally poor health (cachexia, anemia and dehydration). The barnacles were considered to have occurred secondarily to the poor health status of the examined penguins. KEY WORDS.- Spheniscus, Conchoderma, penguin, parasite, BrazilPingüinos de Magallanes (Spheniscus magellanicus) son nativos de Argentina, Chile y de las Islas Malvinas (Falkland), y estacionalmente migran hacía las costas de Uruguay, Brasil y Perú. En este trabajo se reporta el hallazgo de percebes pedunculados Conchoderma virgatum (Cirripedia: Thoracica) sobre la superficie del cuerpo de siete pingüinos de Magallanes. Las aves fueron encontradas en las playas a lo largo de la costa del Estado de São Paulo(23º57’S, 46º23’O), Brasil, en los inviernos de 2008 y 2010. Los crustáceos estaban en lasuperficie de las aletas, pies y abdomen, y en algunos casos, los percebes tenían más de 40 individuos agrupados, algunos midiendo hasta 4 cm. Los pingüinos afectados tenían masa corporal baja (rango = 1.6 – 2.05 kg) y se encontraban en un mal estado de salud (caquexia, anemia y deshidratación). Se considera que los percebes observados ocurrieron posteriormente y de modo secundario al mal estado de salud de los pingüinos examinados. PALABRAS CLAVE.- Spheniscus, Conchoderma, pingüino, parásito, BrasilFAPESP 2009/53956-9CNPq 301517/2006-

Epidemiology and molecular phylogeny of Babesia sp. in Little Penguins Eudyptula minor in Australia

Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia), hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium) and kinetoplastids (Trypanosoma). We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp.) and B. uriae from murres (Uria aalge). None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected

Protease-driven lysosomal activity is a core mechanism for macrophage driven collagen remodeling during liver and kidney fibrosis

Trabajo presentado en el 2nd International Workshop on Liver and Gut Fibrosis, celebrado en Valencia (España), los días 26 y 27 de octubre de 202

Novel insights on the contribution of collagen degradative macrophages to fibrosis resolution

Trabajo presentado en el EASL Congress 2023, celebrado en Viena (Austria), del 21 al 24 de junio de 2023Background and aims: Liver fibrosis is caused by an excessive accumulation of extracellular matrix (ECM) proteins. Macrophages are important effectors for ECM remodelling through recycling of the ECM within acidic compartments and can contribute to liver fibrosis resolution. Proteases, such as cathepsins, are essential for lysosomal proteolytic activity; however, their contribution to ECM remodelling within the macrophages is unknown. Thus, the aim of this study was to investigate the proteolytic and degradative signalling pathways associated to macrophages during liver fibrosis. Method: A novel macrophage-cathepsin D KO mouse strain (CtsDΔMyel) was generated by breeding LysMCre (macrophages) with CtsD-floxed mice. Fibrosis was established by chronic CCl4 administration and bile duct ligation in CtsDF/F or CtsDΔMyel mice and determined by hydroxyproline, Sirius Red, α-SMA, Col1A1 and TGF-β RT-PCR. Proteomic profile was determined by LC-MS/MS in fibrotic livers. Reversion was assessed 72 h post-challenge in a 4-week CCl4 model. Collagen degradation in liver was determined by R-CHP staining. Macrophage polarization and proteolytic secretome was assessed by RT-PCR and protease array, respectively. Collagen degradation and endocytosis was determined by FACS in Kupffer cells (KC). Single-cell RNA sequencing analysis was performed using GSE136103 dataset. Results: ScRNAseq analysis and CtsD IHP demonstrated high expression of CtsD in liver macrophages from cirrhotic patients. Next, CtsDΔMyel mouse was validated by FACS and WB in KC and dual IHP (F4/80-CtsD) in liver. CtsD deletion in macrophages enhanced liver fibrosis with enriched matrisome proteomic signatures in chronic CCl4 and BDL models. Analysis of KC isolated from 72hCCl4-treated livers demonstrated significantly lower expression of markers associated with resolutive macrophages (CD206, TREM-2 and TGF-β) and defective proteolytic secretome profile in CtsDΔMyel KC. In addition, CtsDΔMyel KC displayed defective proteolytic processing of collagen I without impairment of the Endo180 receptormediated endocytosis demonstrated by FACS. Analysis of CtsD macrophage subclusters in control and cirrhotic human livers, confirmed cirrhotic CtsD-expressing subclusters were differentially enriched in ECM degradation and organization signalling pathways. In addition, it revealed a decrease in the number of CtsD-expressing macrophage subclusters in cirrhotic livers, which could contribute to inadequate ECM recycling, perpetuating fibrosis and hampering resolution. Indeed, CtsDΔMyel mouse was unable to remodel collagen in vivo when subjected to a fibrosis reversion model determined by both percentage of HP and fluorescent intensity of collagen hybridizing peptide (CHP) binding to liver tissue. Conclusion: CtsD is essential in regulating the collagenolytic activity of macrophages during liver fibrosis and is part of a novel and currently unknown degradome landscape of restorative macrophages

Unraveling the proteolytic network controlling collagen remodeling during liver fibrosis

Resumen del trabajo presentado en el 48 Congreso anual Asociación Española para el Estudio del Hígado, celebrado en Madrid (España), del 15 al 17 de marzo de 202

Happy feet in a hostile world? The future of penguins depends on proactive management of current and expected threats

Penguins face a wide range of threats. Most observed population changes have been negative and have happened over the last 60 years. Today, populations of 11 penguin species are decreasing. Here we present a review that synthesizes details of threats faced by the world's 18 species of penguins. We discuss alterations to their environment at both breeding sites on land and at sea where they forage. The major drivers of change appear to be climate, and food web alterations by marine fisheries. In addition, we also consider other critical and/or emerging threats, namely human disturbance near nesting sites, pollution due to oil, plastics and chemicals such as mercury and persistent organic compounds. Finally, we assess the importance of emerging pathogens and diseases on the health of penguins. We suggest that in the context of climate change, habitat degradation, introduced exotic species and resource competition with fisheries, successful conservation outcomes will require new and unprecedented levels of science and advocacy. Successful conservation stories of penguin species across their geographical range have occurred where there has been concerted effort across local, national and international boundaries to implement effective conservation planning.This work was supported by the WWF-UK and PEW Foundation. SJ is supported by NSF OPP PICA #1643901

Happy feet in a hostile world? The future of penguins depends on proactive management of current and expected threats

Penguins face a wide range of threats. Most observed population changes have been negative and have happened over the last 60 years. Today, populations of 11 of the 18 penguin species are decreasing. Here we present a review that synthesizes details of threats faced by the world’s 18 species of penguins. We discuss alterations to their environment at both breeding sites on land and at sea where they forage. The major drivers of change appear to be climate, and food web alterations by marine fisheries. In addition, we also consider other critical and/or emerging threats, namely human disturbance near nesting sites, pollution due to oil, plastics and chemicals such as mercury and persistent organic compounds. Finally, we assess the importance of emerging pathogens and diseases on the health of penguins. We suggest that in the context of climate change, habitat degradation, introduced exotic species and resource competition with fisheries, successful conservation outcomes will require new and unprecedented levels of science and advocacy. Successful conservation stories of penguin species across their geographical range have occurred where there has been concerted effort across local, national and international boundaries to implement effective conservation planning

Influence of the moulting cycle on the immune and inflammatory responses in captive Magellanic penguins (Spheniscus magellanicus)

A mudança anual de penas é um componente crítico do ciclo biológico de todas as espécies de aves e pode modificar as respostas imunológicas individuais, sendo que suas consequências são pouco estudadas. Neste contexto, uma vez que são escassos os estudos acerca do perfil clínico e parâmetros imunológicos de pinguins-de-Magalhães em diferentes fases da muda, este estudo propôs avaliar determinados aspectos relacionados a esses perfis, em animais mantidos em ambiente controlado. Foram estudados 21 indivíduos adultos (dez machos e 11 fêmeas) e os parâmetros avaliados foram: teste de hipersensibilidade cutânea tardia à fitohemaglutinina (FHA) com a avaliação histopatológica de biópsias que incluíram a análise qualitativa e quantitativa dos tipos celulares presentes na reação e no controle às 6, 24 e 48 horas; parâmetros clínicos (massa corpórea e hematologia); fagocitose e burst oxidativo; fenotipagem e resposta linfoproliferativa. O teste de FHA demonstrou diferença significativa na espessura da membrana interdigital de ambas as patas inoculadas com FHA e com o controle (PBS) quando comparadas antes da inoculação e antes da biópsia às 24 e 48 horas. A análise histomorfométrica revelou não haver diferenças significativas na frequência relativa de granulócitos (heterófilos/ eosinófilos), linfócitos, macrófagos ou trombócitos entre as patas inoculadas com FHA e PBS em nenhum dos momentos analisados. Este resultado sugere que a diferença na espessura da membrana interdigital das patas inoculadas seja em decorrência do edema e não do infiltrado inflamatório. Para analisar os efeitos do ciclo da muda nos parâmetros clínicos e hematológicos, os animais foram divididos em três grupos: pré-muda (-5 a 0 dias da muda), muda (10º ao 21º dia da muda) e padrão basal (>120 dias da muda). Os resultados mostraram que em relação ao momento basal: houve aumento significativo na massa corpórea dos animais no momento da pré-muda; diminuição significativa de proteínas totais, hematócrito, eritrócitos totais e leucócitos totais na pré-muda e muda. Não houve diferença no número total de monócitos e heterófilos, contudo, houve eosinofilia e linfopenia na pré-muda e muda, respectivamente. Com relação aos parâmetros imunológicos, houve aumento no burst oxidativo gerado pelo estímulo biológico (Staphylococcus aureus) na pré-muda em relação à muda e ao padrão basal; ocorreu aumento do burst oxidativo gerado pelo estímulo químico (éster de forbol) na pré-muda em relação à muda. Em relação à fagocitose, houve diminuição desta capacidade dos leucócitos dos animais na pré-muda e muda em relação ao padrão basal quando o agente foi bacteriano; entretanto, o mesmo não foi observado para a fagocitose de leveduras (Zymosan A); houve diminuição de linfócitos T CD4+ circulantes na pré-muda e muda em relação ao padrão basal e o índice de proliferação de linfócitos aumentou durante o período de muda em relação ao basal. Esses resultados tomados em conjunto sugerem que as mudanças fisiológicas observadas durante o ciclo da muda interferem na modulação de parâmetros imunológicos e são sugestivas de serem consequência do prejuízo energético sofrido, mesmo em animais mantidos em ambientes controladosThe annual replacement of feathers is a critical component of the biological cycle of all bird species. Molt can modify their individual immune response, and the consequences of this are not well known. Within this context, and the few studies on the clinical profile and immunological parameters of Magellanic penguins at different stages of their molt, this study aimed to evaluate certain aspects related to the clinical profiles of animals kept in a controlled environment. A total of 21 adult individuals (ten males and 11 females) was analyzed and the parameters evaluated in this study were: delayed-type hypersensitivity test to phytohemagglutinin (PHA) based on the histopathological evaluation of foot web biopsies that included qualitative and quantitative analysis of cell types present in the essay and control at 6, 24 and 48 hours; clinical parameters (body mass and hematology); phagocytosis and oxidative burst; phenotyping and lymphoproliferative response. The PHA test showed significant differences in the thickness of the webbing of both feet inoculated with PHA and control (PBS), when compared before inoculation and before biopsy at 24 and 48 hours. Histomorphometric analysis revealed no significant differences in the relative frequencies of granulocytes (heterophils/ eosinophils), lymphocytes, macrophages, thrombocytes, or between the feet inoculated with PHA and with PBS in any of the samples. This result suggests that the difference in thickness of the foot webbing is due to edema and not an inflammatory infiltrate. To analyze the effects of the molting cycle on clinical and hematological parameters, the birds were divided into three groups: pre-molt (from -5 days to the start of the molt, day 0), molt (10th to 21st days of molt) and baseline (>120 days after the completion of the molt). The results showed that in comparison with baseline values: a significant increase in body mass occurred during pre-molt; and a significant decrease in total protein, hematocrit, total erythrocytes and total leukocytes occurred during pre-molt and molt. There was no difference in the total numbers of monocytes and heterophils, however, there were eosinophilia and lymphopenia during pre-molt and molt, respectively. There was an increase in the oxidative burst generated by the biological stimulus (Staphylococcus aureus) in the pre-molt compared to molt and baseline levels; and there was an increased oxidative burst generated by the chemical stimuli (phorbol esters) in the pre-molt in relation to the molt. There was a decreased capacity of phagocytosis for the leukocytes in animals during pre-molt and molt compared to baseline when the agent was bacterial; however, this was not observed for the phagocytosis of yeast (Zymosan A). There was a decrease of circulating CD4+ T lymphocytes in the pre-molt and molt compared to baseline, and the lymphocyte proliferation index increased during the molt compared to baseline. The results suggest that the physiological changes observed during the molt cycle does interfere with the modulation of immune parameters, and are suggestive of resulting as a consequence of from the high energy demands of molt, even for birds kept in controlled environment

Comparative pathology of cetaceans and pinnipeds

O conhecimento das causas de morbidade e mortalidade dos mamíferos aquáticos pertencentes à ordem Cetacea e subordem Pinnipedia no Brasil é escasso. Este trabalho teve por objetivo caracterizar as principais causas que contribuem para a morte destes animais, incluindo dentre outras as enfermidades infecciosas, parasitárias, traumáticas, metabólicas e nutricionais. Foram analisadas amostras provenientes de necropsias de 110 indivíduos oriundos de encalhes naturais e capturas acidentais em redes de pesca ocorridos na costa sul e sudeste do Brasil. Ocasionalmente, amostras de cetáceos e pinípedes provenientes de centros de reabilitação, zoológicos, aquários e oceanários também foram analisadas. As amostras consistiram de fragmentos de 1 a 2 cm3 dos órgãos principais acondicionados em formalina a 10%. As afecções foram diagnosticadas através de exames anatomopatológicos, associados, quando possível a resultados microbiológicos e parasitológicos. Dos diagnósticos finais elucidados para os cetáceos 88,6% (70/79) tiveram a morte relacionada com o sistema respiratório, 3,8% (3/79) de origem metabólica, 2,5% (2/79) de etiologia infecciosa, 1,3% (1/79) com origem traumática antropogênica e em 3,8% (3/79) foi indeterminada. Nos pinípedes as causas de morte também estiveram relacionadas principalmente com o trato respiratório (48,5% - 15/31). A morte decorrente de outras causas incluiu: 29,1% (9/31) de causas metabólicas; 3,2% (1/31) nutricional; 3,2% (1/31) digestivo; 3,2% (1/31) urinário; 3,2% (1/31) físico; 3,2% (1/31) síndrome, colapso; 3,2% (1/31) vandalismo; 3,2% (1/31) indeterminado. A análise parasitológica foi realizada em 15,4% (17/110) dos casos e a bacteriologia em 10,9% (12/110) dos casos. As interpretações anatomopatológicas foram associadas aos demais resultados, quando presentes, e então analisados conforme informações recentes de literatura relativas ao assunto. As amostras utilizadas na realização deste projeto complementam o banco de tecidos de animais selvagens mantido pelo Laboratório de Patologia Comparada de Animais Selvagens da FMVZ, formando dessa forma o primeiro acervo de tecidos de mamíferos marinhos disponível no Brasil. Os resultados alcançados neste estudo reforçam a necessidade da abordagem multidisciplinar dos animais encalhados e também dos capturados acidentalmente em redes de pesca, buscando informações que possam colaborar para uma maior compreensão dos processos patológicos que os acometem e os predispõe à morte.The known causes of morbidity and mortality of aquatic mammals belonging to the Order Cetacea and Suborder Pinnipedia in Brazil is limited. The aim of this work was to characterize the main processes that contribute to or cause the death of these animals, and included investigating infectious, parasitic, traumatic, metabolic and nutritional diseases. Samples coming from 110 fresh animals were analyzed, including individuals from strandings and incidentally caught in fishing nets along the south and southeastern coasts of Brazil. Circumstantially, carcasses coming from zoological collections, aquariums and rehabilitation centers were also included. The samples consisted of fragments of 1-2 cm3 taken of the main organs and fixed in 10% formalin. All the tissues have been processed according to routine histological procedures and analyzed under light microscopy. Diseases were diagnosed through anatomopathologic examinations, associated, whenever possible, with microbiologic and parasitologic results and then compared to related bibliography. Final results, mentioning the cause of the death, included: cetaceans: 88,6% (70/79) had lesions associated with the respiratory tract; 3,8% (3/79) of metabolic origin, 2,5% (2/79) of infectious etiology, 1,3% (1/79) were human related injuries, and 3,8% (3/79) of the cases were undetermined. For the pinnipeds the main cause of death was also related to the respiratory tract (48,5% - 15/31). The causes of death implicated in the other cases included: 29,1% (9/31) metabolical causes; 3,2% (1/31) nutritional origin; 3,2% (1/31) digestive causes; 3,2% (1/31) urinary tract; 3,2% (1/31) physical causes; 3,2% (1/31) syndrome; 3,2% (1/31) human related injuries; 3,2% (1/31) were undetermined. Parasitological analyses were performed in 15,4% (17/110) of the cases and bacteriology in 10,9% (12/110). The samples utilized in this project were stored to create a tissue bank for further studies. The results achieved in this work reinforce the need for a multidisciplinary view of stranded and by-caught animals, searching for information that can collaborate to better understand the pathologic processes affecting marine mammals and predisposing their death

Comparative pathology of cetaceans and pinnipeds

O conhecimento das causas de morbidade e mortalidade dos mamíferos aquáticos pertencentes à ordem Cetacea e subordem Pinnipedia no Brasil é escasso. Este trabalho teve por objetivo caracterizar as principais causas que contribuem para a morte destes animais, incluindo dentre outras as enfermidades infecciosas, parasitárias, traumáticas, metabólicas e nutricionais. Foram analisadas amostras provenientes de necropsias de 110 indivíduos oriundos de encalhes naturais e capturas acidentais em redes de pesca ocorridos na costa sul e sudeste do Brasil. Ocasionalmente, amostras de cetáceos e pinípedes provenientes de centros de reabilitação, zoológicos, aquários e oceanários também foram analisadas. As amostras consistiram de fragmentos de 1 a 2 cm3 dos órgãos principais acondicionados em formalina a 10%. As afecções foram diagnosticadas através de exames anatomopatológicos, associados, quando possível a resultados microbiológicos e parasitológicos. Dos diagnósticos finais elucidados para os cetáceos 88,6% (70/79) tiveram a morte relacionada com o sistema respiratório, 3,8% (3/79) de origem metabólica, 2,5% (2/79) de etiologia infecciosa, 1,3% (1/79) com origem traumática antropogênica e em 3,8% (3/79) foi indeterminada. Nos pinípedes as causas de morte também estiveram relacionadas principalmente com o trato respiratório (48,5% - 15/31). A morte decorrente de outras causas incluiu: 29,1% (9/31) de causas metabólicas; 3,2% (1/31) nutricional; 3,2% (1/31) digestivo; 3,2% (1/31) urinário; 3,2% (1/31) físico; 3,2% (1/31) síndrome, colapso; 3,2% (1/31) vandalismo; 3,2% (1/31) indeterminado. A análise parasitológica foi realizada em 15,4% (17/110) dos casos e a bacteriologia em 10,9% (12/110) dos casos. As interpretações anatomopatológicas foram associadas aos demais resultados, quando presentes, e então analisados conforme informações recentes de literatura relativas ao assunto. As amostras utilizadas na realização deste projeto complementam o banco de tecidos de animais selvagens mantido pelo Laboratório de Patologia Comparada de Animais Selvagens da FMVZ, formando dessa forma o primeiro acervo de tecidos de mamíferos marinhos disponível no Brasil. Os resultados alcançados neste estudo reforçam a necessidade da abordagem multidisciplinar dos animais encalhados e também dos capturados acidentalmente em redes de pesca, buscando informações que possam colaborar para uma maior compreensão dos processos patológicos que os acometem e os predispõe à morte.The known causes of morbidity and mortality of aquatic mammals belonging to the Order Cetacea and Suborder Pinnipedia in Brazil is limited. The aim of this work was to characterize the main processes that contribute to or cause the death of these animals, and included investigating infectious, parasitic, traumatic, metabolic and nutritional diseases. Samples coming from 110 fresh animals were analyzed, including individuals from strandings and incidentally caught in fishing nets along the south and southeastern coasts of Brazil. Circumstantially, carcasses coming from zoological collections, aquariums and rehabilitation centers were also included. The samples consisted of fragments of 1-2 cm3 taken of the main organs and fixed in 10% formalin. All the tissues have been processed according to routine histological procedures and analyzed under light microscopy. Diseases were diagnosed through anatomopathologic examinations, associated, whenever possible, with microbiologic and parasitologic results and then compared to related bibliography. Final results, mentioning the cause of the death, included: cetaceans: 88,6% (70/79) had lesions associated with the respiratory tract; 3,8% (3/79) of metabolic origin, 2,5% (2/79) of infectious etiology, 1,3% (1/79) were human related injuries, and 3,8% (3/79) of the cases were undetermined. For the pinnipeds the main cause of death was also related to the respiratory tract (48,5% - 15/31). The causes of death implicated in the other cases included: 29,1% (9/31) metabolical causes; 3,2% (1/31) nutritional origin; 3,2% (1/31) digestive causes; 3,2% (1/31) urinary tract; 3,2% (1/31) physical causes; 3,2% (1/31) syndrome; 3,2% (1/31) human related injuries; 3,2% (1/31) were undetermined. Parasitological analyses were performed in 15,4% (17/110) of the cases and bacteriology in 10,9% (12/110). The samples utilized in this project were stored to create a tissue bank for further studies. The results achieved in this work reinforce the need for a multidisciplinary view of stranded and by-caught animals, searching for information that can collaborate to better understand the pathologic processes affecting marine mammals and predisposing their death