Amazon tropical fishes of commercial interest show human-cell contamination but no SARS-CoV-2 in a real-life scenario.

BACKGROUND: Amazonas was one of the most impacted Brazilian states by the COVID-19 pandemic. Mortality rates were high, and the health systems collapsed. It is important to identify possible intermediate reservoirs to avoid animal-to-human contamination. Several tropical fish are of commercial interest and are sold in large open-air markets in the region, representing a large economic and dietary importance. OBJECTIVES: This study aimed to verify if fish species of commercial importance, aerosols, and fish wastewater in local open-air markets, at a major capital city in the western Brazilian Amazon, are contaminated by SARS-CoV-2. METHODS: 488 fish, 50 aerosol, and 45 wastewater samples were analyzed for the presence of SARS-CoV-2. The samples were subjected to extraction using the BIOGENE Viral DNA/RNA Extraction kit, and the molecular diagnosis was tested for SARS-CoV-2 using the Bio-Manguinhos SARS-CoV-2 (EDx) Molecular Kit. RESULTS: It was not possible to detect the virus (Ct≤40, for Gene E) in these samples, however, in 181 samples of fish it was possible to detect the human RP gene (Ct≤35, for the RP Gene), indicating human contact. There was a high number of COVID-19 diagnoses in all city districts in which the samples were collected, showing that SARS-CoV-2 was circulating. CONCLUSION: This study indicates that fish of local commercial importance do not carry SARS-CoV-2 viral particles, despite circulation of SARS-CoV-2, and are not an important source of animal-to-human contamination. Despite these results, the human RP gene was found detectable in fish, air, and fish wastewater, showing that such places may carry human pathogens.

Incidence of hepatitis A and hepatitis E viruses and norovirus and rotavirus in fish and shrimp samples caught from the Persian Gulf / [Incidência de vírus da hepatite A e hepatite E e norovírus e rotavírus em amostras de peixes e camarões capturados no Golfo Pérsico]

Foodborne viruses including hepatitis A virus (HAV), norovirus (NoV), rotavirus (RoV) and hepatitis E virus (HEV) are easily transmitted through contaminated seafoods. The current research was done to assess the incidence of RoV, NoV GI and GII,hAV and hEV in fish and shrimp samples caught from the Persian Gulf, Iran. Three-hundred and twenty fish and shrimp samples were collected. The presence of foodborne viruses were assessed by the real-time PCR. Forty-nine out of 320 (15.31%) fish and shrimp samples were positive for foodborne viruses. Distribution of hAV, NoV GI and NoV GII amongst all studied samples were 0.93%, 5.93% and 8.43%, respectively. hEV and RoV viruses were not found in studied samples. Parastromateus niger and Scomberomorus commerson fish and Penaeus monodon shrimp were the most frequently contaminated samples. Simultaneous incidence of hAV and NoV GI and hAV and NoV GII were 0.31% and 0.93%, respectively. Distribution of foodborne viruses in samples collected through spring, summer, autumn and winter seasons were 14.28%, 9.33%, 11.76% and 24.44%, respectively. Findings revealed that the incidence of foodborne viruses was significantly associated with seafood species and also season of sampling.(AU)

Vírus transmitidos por alimentos, incluindo hepatite A (HAV), norovírus (NoV), rotavírus (RoV) e hepatite E (HEV) são facilmente transmitidos através de frutos do mar contaminados. Esta pesquisa foi realizada para avaliar a incidência de RoV, NoV GI e GII, hAV e hEV em amostras de peixes e camarões capturadas no Golfo Pérsico, Irã. Foram coletadas 300 amostras de peixes e camarões. A presença de vírus transmitidos por alimentos foi avaliada por PCR em tempo real. Quarenta e nove das 320 amostras de peixes e camarões (15,31%) foram positivas para vírus transmitidos por alimentos. A distribuição de hAV, NoV GI e NoV GII entre as amostras estudadas foi 0,93%, 5,93% e 8,43%, respectivamente. Os vírus hEV e RoV não foram encontrados nas amostras estudadas. Os peixes Parastromateus niger e Scomberomorus commerson e o camarão Penaeus monodon foram as amostras mais frequentemente contaminadas. A incidência simultânea de hAV e NoV GI, e hAV e NoV GII foi de 0,31% e 0,93%, respectivamente. A distribuição dos vírus transmitidos por alimentos nas amostras coletadas na primavera, verão, outono e inverno foi de 14,28%, 9,33%, 11,76% e 24,44%, respectivamente. Os resultados demonstram que a incidência de vírus transmitidos por alimentos foi significativamente associada às espécies de frutos do mar e também à época da amostragem.(AU)

Detection and molecular characterization of Lymphocystivirus in Brazilian ornamental fish.

The aim of this study is to report the occurrence of Lymphocystivirus in Brazilian ornamental fish. From 25 ornamental fish species submitted for molecular diagnosis, only one sample (Pomacanthus imperator) was positive, and its viral nucleotide sequence obtained clustered with sequences of genotype VII. To our knowledge, this is the first report on the genetic characterization of Lymphocystivirus in Brazil.

Genome analysis of Ranavirus frog virus 3 isolated from American Bullfrog (Lithobates catesbeianus) in South America.

Ranaviruses (family Iridoviridae) cause important diseases in cold-blooded vertebrates. In addition, some occurrences indicate that, in this genus, the same virus can infect animals from different taxonomic groups. A strain isolated from a Ranavirus outbreak (2012) in the state of Sao Paulo, Brazil, had its genome sequenced and presented 99.26% and 36.85% identity with samples of Frog virus 3 (FV3) and Singapore grouper iridovirus (SGIV) ranaviruses, respectively. Eight potential recombination events among the analyzed sample and reference FV3 samples were identified, including a recombination with Bohle iridovirus (BIV) sample from Oceania. The analyzed sample presented several rearrangements compared to FV3 reference samples from North America and European continent. We report for the first time the complete genome of Ranavirus FV3 isolated from South America, these results contribute to a greater knowledge related to evolutionary events of potentially lethal infectious agent for cold-blooded animals.

Infectious diseases in aquarium ornamental pet fish: prevention and control measures / Doenças infeciosas em peixes ornamentais de aquário: medidas de prevenção e controle

Aquarium ornamental pet fish constitute a major segment in the pet industry, with the United States, Europe, and Japan dominating the market. There are approximately 1,500 marine fish species and over 4,500 freshwater fish species commercialized as aquarium ornamental pet fish. Fish are the fourth most common pet present in Brazilian homes. In Brazil, aquarium ornamental pet fish can be marketed and distributed from different parts of the Brazilian territory and the world. Commercialization and circulation of living animals without the use of adequate prophylactic management procedures enables dissemination of a number of agents responsible for infectious diseases. Aquarium pet fish can also carry pathogenic agents, of bacterial, viral, fungal, or parasitic etiology, that may have a zoonotic feature endangering the persons handling the animals. This review presents the main pathogenic infectious agents of bacterial, viral, and fungal etiology that affect aquarium pet fish, as well as the prevention and control measures to ensure sanitary excellence in this segment.(AU)

Peixes ornamentais de aquário representam um grande segmento no mercado de animais de estimação, no qual Estados Unidos, Europa e Japão dominam. São aproximadamente 1.500 espécies de peixes marinhos e em torno de 4.500 de espécies de água doce comercializados com fins ornamentais. O peixe é a quarta espécie mais comum nos domicílios brasileiros. Peixes ornamentais de aquário são comercializados e distribuídos em diferentes partes do território nacional e do mundo. O comércio e circulação de animais vivos sem o uso de procedimentos de manejo profilático adequados possibilita a disseminação de inúmeros agentes patogênicos. Peixes ornamentais de aquário carreiam consigo agentes patogênicos de etiologia bacteriana, viral, fúngica e parasitária, sendo alguns de caráter zoonótico colocando em risco pessoas que os manipulam. O objetivo desta revisão é apresentar os principais agentes infeciosos patogênicos - de natureza bacteriana, viral e fúngica - que acometem peixes ornamentais de aquário, bem como os métodos de prevenção e controle que permitam excelência no segmento.(AU)

The Geographic Structure of Viruses in the Cuatro Ciénegas Basin, a Unique Oasis in Northern Mexico, Reveals a Highly Diverse Population on a Small Geographic Scale.

The Cuatro Ciénegas Basin (CCB) is located in the Chihuahuan desert in the Mexican state of Coahuila; it has been characterized as a site with high biological diversity despite its extreme oligotrophic conditions. It has the greatest number of endemic species in North America, containing abundant living microbialites (including stromatolites and microbial mats) and diverse microbial communities. With the hypothesis that this high biodiversity and the geographic structure should be reflected in the virome, the viral communities in 11 different locations of three drainage systems, Churince, La Becerra, and Pozas Rojas, and in the intestinal contents of 3 different fish species, were analyzed for both eukaryotic and prokaryotic RNA and DNA viruses using next-generation sequencing methods. Double-stranded DNA (dsDNA) virus families were the most abundant (72.5% of reads), followed by single-stranded DNA (ssDNA) viruses (2.9%) and ssRNA and dsRNA virus families (0.5%). Thirteen families had dsDNA genomes, five had ssDNA, three had dsRNA, and 16 had ssRNA. A highly diverse viral community was found, with an ample range of hosts and a strong geographical structure, with very even distributions and signals of endemicity in the phylogenetic trees from several different virus families. The majority of viruses found were bacteriophages but eukaryotic viruses were also frequent, and the large diversity of viruses related to algae were a surprise, since algae are not evident in the previously analyzed aquatic systems of this ecosystem. Animal viruses were also frequently found, showing the large diversity of aquatic animals in this oasis, where plants, protozoa, and archaea are rare.IMPORTANCE In this study, we tested whether the high biodiversity and geographic structure of CCB is reflected in its virome. CCB is an extraordinarily biodiverse oasis in the Chihuahuan desert, where a previous virome study suggested that viruses had followed the marine ancestry of the marine bacteria and, as a result of their long isolation, became endemic to the site. In this study, which includes a larger sequencing coverage and water samples from other sites within the valley, we confirmed the high virus biodiversity and uniqueness as well as the strong biogeographical diversification of the CCB. In addition, we also analyzed fish intestinal contents, finding that each fish species eats different prey and, as a result, presents different viral compositions even if they coexist in the same pond. These facts highlight the high and novel virus diversity of CCB and its "lost world" status.

Molecular detection and phylogenetic analysis of megalocytivirus in Brazilian ornamental fish.

Megalocytiviruses have a worldwide distribution, causing serious economic loss to the global aquaculture industry. They also present a threat to ornamental fish trade because megalocytiviral infections have unspecified symptoms, making early diagnosis difficult. In this study, 100 ornamental fish from 24 different species were tested by PCR for megalocytivirus, with a 47% positive rate being identified. Phylogenetic reconstruction, based on the major capsid protein (MCP) gene, clustered all Brazilian samples into a single clade, showing identity values ranging from 99% to 100% when compared to each other. This is the first report of megalocytivirus infection in some ornamental fish species in Brazil.

Lymphocystis virus in ornamental fish imported in Brazil / Lymphocystis vírus em peixes ornamentais importados para o Brasil

Lymphocystis is a chronic viral disease found in both fresh and marine water fishes, characterized by nodules that typically appear on the skin or fins. Lymphocystis is the most common viral infection of ornamental fish. This study aimed to identify fish showing clinical signs of Lymphocystis virus in an ornamental fish importation company and to highlight the importance of involving veterinary doctors to assure the health of ornamental fish, a relatively unexplored practice in Brazil. Seventeen ornamental fishes studied during quarantine presented with clinical signs of the disease, confirmed after microscopic analysis. Every single fish presented with enlarged fibroblastic cells typical of the Lymphocystis virus. In this case, lesions were removed with a scalpel blade and the fish were treated with an antibiotic in a hospital aquarium for seven days. Thirty days later, the fish were healthy and ready for sale. The study also reinforces the fact that it is extremely important to provide the staff with examples of good practices in handling fishes, provide the technical team with professional training, and avoid situations of stress on fishes in order to restrain opportunistic diseases. The enablement of veterinary doctors in establishments that trade aquatic ornamental fishes is also essential to allow them to perform their work in prevention and disease control.

Linfocitose é uma doença viral crônica encontrada em peixes de água doce e marinha, caracterizada por nódulos principalmente na pele e/ou nadadeiras. É a doença viral mais comum nos peixes de aquário. O objetivo do presente estudo de caso foi identificar os animais com sintomatologia clínica condizente com infecção por Lymphocystis vírus, em um estabelecimento importador de organismos aquáticos ornamentais, além de alertar para a importância de médicos veterinários capacitados na área de sanidade de animais aquáticos, relativamente a um agente pouco explorado nessa área no Brasil. Um total de dezessete peixes ornamentais foram estudados durante o período de quarentena, no qual apresentaram sinais clínicos da doença, confirmada por análise microscópica. Todos os peixes apresentaram células fibroblásticas de tamanho aumentado, típicas de Lymphocystis vírus. As lesões foram removidas com lâmina de bisturi e os peixes foram tratados com antibiótico em um aquário hospital por sete dias para prevenir contra infecções bacterianas secundárias. Após trinta dias, os peixes estavam completamente curados e disponíveis para venda. Este relato reforça a importância de hábitos de boas práticas de higiene e manipulação, além do fornecimento de treinamentos aos profissionais que manipulam diretamente os animais comercializados, a fim de diminuir situações desnecessárias de estresse

Lymphocystis virus in ornamental fish imported in Brazil / Lymphocystis vírus em peixes ornamentais importados para o Brasil

Lymphocystis is a chronic viral disease found in both fresh and marine water fishes, characterized by nodules that typically appear on the skin or fins. Lymphocystis is the most common viral infection of ornamental fish. This study aimed to identify fish showing clinical signs of Lymphocystis virus in an ornamental fish importation company and to highlight the importance of involving veterinary doctors to assure the health of ornamental fish, a relatively unexplored practice in Brazil. Seventeen ornamental fishes studied during quarantine presented with clinical signs of the disease, confirmed after microscopic analysis. Every single fish presented with enlarged fibroblastic cells typical of the Lymphocystis virus. In this case, lesions were removed with a scalpel blade and the fish were treated with an antibiotic in a hospital aquarium for seven days. Thirty days later, the fish were healthy and ready for sale. The study also reinforces the fact that it is extremely important to provide the staff with examples of good practices in handling fishes, provide the technical team with professional training, and avoid situations of stress on fishes in order to restrain opportunistic diseases. The enablement of veterinary doctors in establishments that trade aquatic ornamental fishes is also essential to allow them to perform their work in prevention and disease control.(AU)

Linfocitose é uma doença viral crônica encontrada em peixes de água doce e marinha, caracterizada por nódulos principalmente na pele e/ou nadadeiras. É a doença viral mais comum nos peixes de aquário. O objetivo do presente estudo de caso foi identificar os animais com sintomatologia clínica condizente com infecção por Lymphocystis vírus, em um estabelecimento importador de organismos aquáticos ornamentais, além de alertar para a importância de médicos veterinários capacitados na área de sanidade de animais aquáticos, relativamente a um agente pouco explorado nessa área no Brasil. Um total de dezessete peixes ornamentais foram estudados durante o período de quarentena, no qual apresentaram sinais clínicos da doença, confirmada por análise microscópica. Todos os peixes apresentaram células fibroblásticas de tamanho aumentado, típicas de Lymphocystis vírus. As lesões foram removidas com lâmina de bisturi e os peixes foram tratados com antibiótico em um aquário hospital por sete dias para prevenir contra infecções bacterianas secundárias. Após trinta dias, os peixes estavam completamente curados e disponíveis para venda. Este relato reforça a importância de hábitos de boas práticas de higiene e manipulação, além do fornecimento de treinamentos aos profissionais que manipulam diretamente os animais comercializados, a fim de diminuir situações desnecessárias de estresse(AU)

Isolamento de Aeromonas hydrophila em Bettas plendens – alterações histopatológicas nas brânquias / Isolation of Aeromonas hydrophila in Bettas plendens - histopathologic changes in gills / Aislamiento de Aeromonas hydrophila en Bettas plendens - alteraciones histopatológicas en las branquias

O objetivo deste estudo foi isolar e caracterizar Aeromonas hydrophila, causadora de mortalidade em peixes descrever as alterações histopatológicas provocadas nas brânquias de machos de beta Bettas plendens. Foram utilizados dez peixes que apresentavam característica de alteração de comportamento como anorexia, letalidade e apatia. Amostras de água, muco e brânquias foram inoculadas em meio de cultura Agar sangue a 25ºC por 48 horas. Foi caracterizado A. hidrófila como principal causa do surto de mortalidade. Foram observadas alterações histopatológicas nas brânquias desde fusão lamelar secundária e necrose do tecido. A. hidrophila prejudica o órgão respiratório de forma irreversível, comprometendo as trocas gasosas levando o peixe ao óbito.

The objective of this study was to isolate and characterize Aeromonas hydrophila, a cause of mortality, and to describe the histopathological changes caused in the gills of Bettas plendens. Ten fish presenting change behavior such as anorexia, lethality and apathy were used in this study. Water, mucus and gill samples were inoculated in blood agar culture medium at 25°C for 48 hours. Aeromonas hidrophila was characterized as the main cause of mortality outbreak. Histopathological changes were observed in the gills, from secondary lamellar fusion to tissue necrosis. A. hidrophila irreversibly harms the respiratory organ, compromising the gaseous exchanges, which leads to the death of the fish.

El objetivo de este estudio fue aislar y caracterizar Aeromonas hydrophila, causante de mortalidad en peces, y describir las alteraciones histopatológicas causadas en las branquias de machos de beta Bettas plendens. Se utilizó 10 peces que presentaban características de cambio de comportamiento como anorexia, mortalidad y apatía. Las muestras de agua, moco y branquias fueron inoculadas en medio de cultivo Agar sangre a 25° C durante 48 horas. Fue caracterizado A. hidrófila como principal causa del surto de mortalidad. Se ha observado cambios histopatológicos en las branquias, desde fusión lamelar secundaria y necrosis de los tejidos. A. hidrophila compromete el órgano respiratorio de forma irreversible, comprometiendo los cambios gaseosos y llevando los peces a la muerte.

Isolamento de Aeromonas hydrophila em Bettas plendens alterações histopatológicas nas brânquias / Isolation of Aeromonas hydrophila in Bettas plendens - histopathologic changes in gills / Aislamiento de Aeromonas hydrophila en Bettas plendens - alteraciones histopatológicas en las branquias

O objetivo deste estudo foi isolar e caracterizar Aeromonas hydrophila, causadora de mortalidade em peixes descrever as alterações histopatológicas provocadas nas brânquias de machos de beta Bettas plendens. Foram utilizados dez peixes que apresentavam característica de alteração de comportamento como anorexia, letalidade e apatia. Amostras de água, muco e brânquias foram inoculadas em meio de cultura Agar sangue a 25ºC por 48 horas. Foi caracterizado A. hidrófila como principal causa do surto de mortalidade. Foram observadas alterações histopatológicas nas brânquias desde fusão lamelar secundária e necrose do tecido. A. hidrophila prejudica o órgão respiratório de forma irreversível, comprometendo as trocas gasosas levando o peixe ao óbito.(AU)

The objective of this study was to isolate and characterize Aeromonas hydrophila, a cause of mortality, and to describe the histopathological changes caused in the gills of Bettas plendens. Ten fish presenting change behavior such as anorexia, lethality and apathy were used in this study. Water, mucus and gill samples were inoculated in blood agar culture medium at 25°C for 48 hours. Aeromonas hidrophila was characterized as the main cause of mortality outbreak. Histopathological changes were observed in the gills, from secondary lamellar fusion to tissue necrosis. A. hidrophila irreversibly harms the respiratory organ, compromising the gaseous exchanges, which leads to the death of the fish.(AU)

El objetivo de este estudio fue aislar y caracterizar Aeromonas hydrophila, causante de mortalidad en peces, y describir las alteraciones histopatológicas causadas en las branquias de machos de beta Bettas plendens. Se utilizó 10 peces que presentaban características de cambio de comportamiento como anorexia, mortalidad y apatía. Las muestras de agua, moco y branquias fueron inoculadas en medio de cultivo Agar sangre a 25° C durante 48 horas. Fue caracterizado A. hidrófila como principal causa del surto de mortalidad. Se ha observado cambios histopatológicos en las branquias, desde fusión lamelar secundaria y necrosis de los tejidos. A. hidrophila compromete el órgano respiratorio de forma irreversible, comprometiendo los cambios gaseosos y llevando los peces a la muerte.(AU)

Risk of introducing viral hemorrhagic septicemia virus (VHSV) to the Chilean South Pacific via sardine imports from Europe.

Chile imports from Spain 100s of metric tons of frozen sardine Sardina pilchardus fished in European oceans, which, with several other clupeids, are presumed susceptible to infection with viral hemorrhagic septicemia virus (VHSV). The frozen sardines are directly introduced into the sea as bait to catch southern hake Merluccius australis in the same areas where wild and pen-raised salmonids are present. A simulation model was therefore developed to evaluate the potential risk of infection of wild Chilean southern hake with VHSV from imported bait. The model indicated that VHSV-susceptible fish species present in Chilean waters, like southern hake, are not at immediate risk of infection. However, sensitivity analyses showed that infectious doses at lower concentrations of VHSV combined with higher VHSV-prevalence import scenarios could likely result in VHSV infections of a moderate number of indigenous southern hake (> or =54 fish yr(-1)).

Estomatite Vesicular Alagoas (VSA) am tilápia nilótica (Oreochromis niloticus). Possível implicaçäo epidemiológica / Vesicular Stomatitis Alagoas (VSA) in tilapia nilotica (Oreochromis niloticus). Possible epidemiological concern

Foram realizadas inoculaçöes experimentais do vírus da Estomatite Vesicular Alagoas (VSA) em tilápia nilótica (Oreochromis niloticus), pela via intraperitoneal e por imersäo em suspensäo viral, com o objetivo de verificar se o mesmo se replicava neste sistema hospedeiro e se o vírus era eliminado na água, o que poderia representar um importante papel no ciclo epidemiológico da virose. Os peixes foram dispostos em grupos testemunhos e inoculados com suspensäo de vírus VSA, tanto pela via intraperitoneal quanto por imersäo. Foram observadas alteraçöes anatomopatológicas em órgäos da cavidade abdominal e cérebro. Esses órgäos e a água do aquário foram inoculados em células BHK21 e observados a presença de efeito citopático característico dos vírus pertencentes a este gênero. A segunda passagem do vírus em peixes confirmou as lesöes encontradas anteriormente e o isolamento em novas inoculaçöes em cultivo celular. Os resultados sugerem que a tilápia é sensível ao VSA nas duas vias de inoculaçäo, elimina formas infectantes do vírus na água, podendo representar um importante elo na cadeia epidemiológica da estomatite vesicular.