Post-harvest bacterial contamination of fish, their assessment and control strategies.

Healthy fish populations lead to healthy aquatic ecosystems and it is our responsibility to be a part of the solution. Fish is one of the most favored foods and is suitable for people of all ages. Fish is an essential source of protein, vitamins, and minerals and a source of income for millions of people. Human population growth and climate change are putting a strain on our food system, demanding the development of sustainable services to enhance global food production and its security. Food safety is an intricate problem in both developed and developing countries. Fresh fish is a highly perishable food with a limited life span; as a result, it must be delivered and kept carefully to minimize deterioration and assure safety. Fish spoilage is linked to biochemical changes that occur post-harvest, such as storage and transportation. These modifications can account for fish spoilage by altering the taste, texture, and appearance. Fish harvesting, distribution, and post-harvest handling are all unhygienic, resulting in poor and unpredictable fish quality in the market. Many innovative and effective control measurements of various bacteria in fish have been proposed and evaluated. This review is a systematic approach to investigating post-harvest fish spoilage, its assessment, and control strategies.

Microbiota diversity of three Brazilian native fishes during ice and frozen storage.

This study aimed to assess the bacterial microbiota involved in the spoilage of pacu (Piaractus mesopotamics), patinga (female Piaractus mesopotamics x male Piaractus brachypomus), and tambacu (female Colossoma macropomum × male Piaractus mesopotamics) during ice and frozen storage. Changes in the microbiota of three fish species (N = 22) during storage were studied through 16S rRNA amplicon-based sequencing and correlated with volatile organic compounds (VOCs) and metabolites assessed by nuclear magnetic resonance (NMR). Storage conditions (time and temperature) affected the microbiota diversity in all fish samples. Fish microbiota comprised mainly of Pseudomonas sp., Brochothrix sp., Acinetobacter sp., Bacillus sp., Lactiplantibacillus sp., Kocuria sp., and Enterococcus sp. The relative abundance of Kocuria, P. fragi, L. plantarum, Enterococcus, and Acinetobacter was positively correlated with the metabolic pathways of ether lipid metabolism while B. thermosphacta and P. fragi were correlated with metabolic pathways involved in amino acid metabolism. P. fragi was the most prevalent spoilage bacteria in both storage conditions (ice and frozen), followed by B. thermosphacta. Moreover, the relative abundance of identified Bacillus strains in fish samples stored in ice was positively correlated with the production of VOCs (1-hexanol, nonanal, octenol, and 2-ethyl-1-hexanol) associated with off-flavors. 1H NMR analysis confirmed that amino acids, acetic acid, and ATP degradation products increase over (ice) storage, and therefore considered chemical spoilage index of fish fillets.

Potentially pathogenic bacteria in the plastisphere from water, sediments, and commercial fish in a tropical coastal lagoon: An assessment and management proposal.

Microplastics in aquatic ecosystems harbor numerous microorganisms, including pathogenic species. The ingestion of these microplastics by commercial fish poses a threat to the ecosystem and human livelihood. Coastal lagoons are highly vulnerable to microplastic and microbiological pollution, yet limited understanding of the risks complicates management. Here, we present the main bacterial groups, including potentially pathogenic species, identified on microplastics in waters, sediments, and commercial fish from Ciénaga Grande de Santa Marta (CGSM), the largest coastal lagoon in Colombia. DNA metabarcoding allowed identifying 1760 bacterial genera on microplastics, with Aeromonas and Acinetobacter as the most frequent and present in all three matrices. The greatest bacterial richness and diversity were recorded on microplastics from sediments, followed by waters and fish. Biochemical analyses yielded 19 species of potentially pathogenic culturable bacteria on microplastics. Aeromonas caviae was the most frequent and, along with Pantoea sp., was found on microplastics in all three matrices. Enterobacter roggenkampii and Pseudomonas fluorescens were also found on microplastics from waters and fish. We propose management strategies for an Early Warning System against microbiological and microplastic pollution risks in coastal lagoons, illustrated by CGSM. This includes forming inter-institutional alliances for research and monitoring, accompanied by strengthening governance and health infrastructures.

The Abrolhos Nominally Herbivorous Coral Reef Fish Acanthurus chirurgus, Kyphosus sp., Scarus trispinosus, and Sparisoma axillare Have Similarities in Feeding But Species-Specific Microbiomes.

Coral reefs rely heavily on reef fish for their health, yet overfishing has resulted in their decline, leading to an increase in fast-growing algae and changes in reef ecosystems, a phenomenon described as the phase-shift. A clearer understanding of the intricate interplay between herbivorous, their food, and their gut microbiomes could enhance reef health. This study examines the gut microbiome and isotopic markers (δ13C and δ15N) of four key nominally herbivorous reef fish species (Acanthurus chirurgus, Kyphosus sp., Scarus trispinosus, and Sparisoma axillare) in the Southwestern Atlantic's Abrolhos Reef systems. Approximately 16.8 million 16S rRNA sequences were produced for the four fish species, with an average of 317,047 ± 57,007 per species. Bacteria such as Proteobacteria, Firmicutes, and Cyanobacteria were prevalent in their microbiomes. These fish show unique microbiomes that result from co-diversification, diet, and restricted movement. Coral-associated bacteria (Endozoicomonas, Rhizobia, and Ruegeria) were found in abundance in the gut contents of the parrotfish species Sc. trispinosus and Sp. axillare. These parrotfishes could aid coral health by disseminating such beneficial bacteria across the reef. Meanwhile, Kyphosus sp. predominantly had Pirellulaceae and Rhodobacteraceae. Four fish species had a diet composed of turf components (filamentous Cyanobacteria) and brown algae (Dictyopteris). They also had similar isotopic niches, suggesting they shared food sources. A significant difference was observed between the isotopic signature of fish muscular gut tissue and gut contents, pointing to the role that host genetics and gut microbes play in differentiating fish tissues.

Bioactive compound from marine seagrass Streptomyces argenteolus TMA13: combatting fish pathogens with time-kill kinetics and live-dead cell imaging.

Actinobacteria, pervasive in aquatic and terrestrial environments, exhibit a filamentous morphology, possess DNA with a specific G + C content and production of numerous secondary metabolites. This study, focused on actinobacteria isolated from marine seagrass, investigating their antibacterial activity against fish pathogens. Among 28 isolates, Streptomyces argenteolus TMA13 displayed the maximum zone of inhibition against fish pathogens-Aeromonas hydrophila (10 mm), Aeromonas caviae (22 mm), Edwardsiella tarda (17 mm), Vibrio harveyi (22 mm) and Vibrio anguillarum (12 mm) using the agar plug method. Optimization of this potent strain involved with various factors, including pH, temperature, carbon source and salt condition to enhance both yield production and antibacterial efficacy. In anti-biofilm assay shows the maximum percentage of inhibition while increasing concentration of TMA13 extract. Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) assays with TMA13 crude extract demonstrated potent activity against fish pathogens at remarkably low concentrations. Time-kill kinetics assay showcased growth curve variations over different time intervals for all fish pathogens treated with a 100 µg/ml concentration of crude extract, indicating a decline in cells viability and progression into the death phase. Additionally, fluorescence microscopic visualization of bacterial cells exposed to the extracts emitting green and red fluorescence, enabling live-dead cell differentiation was also studied. Further characterization of the crude extract through GC-MS and FT-IR analyses performed and identified secondary metabolites with functional groups exhibiting significant antibacterial activity. This study elucidates the capacity of Streptomyces argenteolus TMA13 to enhance the production of antibiotic compounds effective against fish pathogens.

Histological biomarkers and microbiological parameters of an estuarine fish from the Brazilian Amazon coast as potential indicators of risk to human health.

This study aimed to isolate and identify pathogenic bacteria in the intestinal tract, skin, and muscles of Sciades herzbergii; detect histopathological changes in the gill and liver; and use these biomarkers for the assessment of potential risks to human health. Fish were sampled during the rainy and dry seasons at two points in São Marcos Bay, Maranhão, Brazil: Ilha dos Caranguejos (IC) and Porto Grande (PG). Isolation and quantification were carried out using COLItest®. Colonies were subjected to identification and phenotypic investigation of antimicrobial resistance using Vitek®. Gill and liver samples were subjected to routine histological examination. The results indicated the presence of Klebsiella pneumoniae and Escherichia coli, the latter of which showed phenotypic resistance to norfloxacin and gentamicin. Fish caught at PG exhibited more extensive gill and liver damage than fish caught at IC. The findings suggest that histological changes in target organs of S. herzbergii may be influenced by infection with pathogenic bacteria.

Selenized non-Saccharomyces yeasts and their potential use in fish feed.

Selenium (Se) is a vital trace element, essential for growth and other biological functions in fish. Its significance lies in its role as a fundamental component of selenoproteins, which are crucial for optimal functioning of the organism. The inclusion of Se in the diets of farmed animals, including fish, has proved invaluable in mitigating the challenges arising from elemental deficiencies experienced in captivity conditions due to limitations in the content of fishmeal. Supplementing diets with Se enhances physiological responses, particularly mitigates the effects of the continuous presence of environmental stress factors. Organic Se has been shown to have higher absorption rates and a greater impact on bioavailability and overall health than inorganic forms. A characteristic feature of yeasts is their rapid proliferation and growth, marked by efficient mineral assimilation. Most of the selenized yeasts currently available in the market, and used predominantly in animal production and aquaculture, are based on Saccharomyces cerevisiae, which contains selenomethionine (Se-Met). The object of this review is to highlight the importance of selenized yeasts. In addition, it presents metabolic and productive aspects of other yeast genera that are important potential sources of organic selenium. Some yeast strains discussed produce metabolites of interest such as lipids, pigments, and amino acids, which could have applications in aquaculture and further enrich their usefulness.

Mercury concentration in larvae of Eustrongylides sp (Nematoda: Dioctophymatoidea) from fish of the Brazilian Amazon / Concentración de mercurio en larvas de Eustrongylides sp (Nematoda: Dioctophymatoidea) en peces de la Amazonía brasileña

Abstract Introduction: Chemical pollution represents a great concern to aquatic organisms, especially fish. Metals enter the aquatic environment from a variety of sources, including natural biogeochemical cycles and anthropogenic sources such as industrial and residential effluents, mining and atmospheric sources. Objective: To describe the Eustrongylides sp. larvae and the interaction with their fish hosts as indicators of mercury (Hg) contamination in the Brazilian Amazon, and the distribution of Hg in the internal organs of fish species Hoplias malabaricus and Pygocentrus nattereri collected in oxbow lakes on the Tapajós River, in the municipality of Santarém, in the state of Pará. Methods: Total Hg was analyzed using the Direct Hg Analyzer - DMA-80. Concentrations of Hg in Eustrongylides sp. were compared with those found in the tissues/organs of the hosts H. malabaricus and P. nattereri. Hg concentrations in the host/parasite system were statistically compared using Principal Component Analysis. The bioconcentration factor (BCF) was calculated to assess the bioaccumulation capacity of metals in Eustrongylides sp. larvae, comparing the concentration of Hg in the parasite with that accumulated in the musculature of infected hosts. Results: Hg concentrations in all tissues/organs analyzed were higher in the parasitic species Eustrongylides sp. larvae when compared with those found in tissues/organs of H. malabaricus and P. nattereri. There was an inversely proportional relationship, showing that when Eustrongylides sp. larvae are present, the concentration in the parasite is higher than in the musculature of host fish H. malabaricus and P. nattereri. The BCF of Hg was found by comparing Eustrongylides sp. larvae/H. malabaricus muscle and was observed during a flood (BCF Hg = 15 364). Conclusions: The results confirm the greater bioaccumulative capacity of Eustrongylides sp. compared to its host. The data indicated the viability of using Eustrongylides sp. larvae in biomonitoring programs. It is worth mentioning that fish samples for Hg analysis must be free of parasites since their presence can alter the results.

Resumen Introducción: La contaminación química del hábitat acuático representa un gran peligro para organismos acuáticos, especialmente para peces. Los metales ingresan al ambiente acuático desde una variedad de fuentes, incluidos los ciclos biogeoquímicos naturales y fuentes antropogénicas, como efluentes industriales y residenciales, minería y fuentes atmosféricas. Objetivo: Describir las especies de Eustrongylides sp. y la interacción con sus peces hospederos como indicadores de contaminación por mercurio en la Amazonía brasileña, y la distribución en los órganos internos de las especies de peces Hoplias malabaricus y Pygocentrus nattereri recolectadas en cochas del Río Tapajós, en el municipio de Santarém, del estado de Pará. Métodos: El Hg total se analizó utilizando el Direct Hg Analyzer - DMA-80. Las concentraciones de Eustrongylides sp. se compararon con las encontrados en los tejidos/órganos de los hospederos H. malabaricus y P. nattereri. Las concentraciones en el sistema hospedero/parásito se compararon estadísticamente utilizando el análisis de componentes principales. Se calculó el factor de bioconcentración (BCF) para evaluar la capacidad de bioacumulación de metales en larvas de Eustrongylides sp., comparando la concentración en el parásito con la acumulada en la musculatura de los hospederos infectados. Resultados: Las concentraciones de Hg en todos los tejidos/órganos analizados fueron mayores en las larvas de la especie parasitaria Eustrongylides sp. en comparación con las encontradas en los tejidos/órganos de H. malabaricus y P. nattereri. Hubo una relación inversamente proporcional, mostrando que cuando las larvas de Eustrongylides sp. están presentes, la concentración en el parásito es mayor que en la musculatura de los peces hospederos H. malabaricus y P. nattereri. El BCF de Hg se encontró comparando Eustrongylides sp. larvas/ músculo H. malabaricus y se observó durante una inundación (BCF Hg = 15 364). Conclusiones: Los resultados confirman la mayor capacidad bioacumulativa de Eustrongylides sp. en comparación con su hospedero. Los datos indicaron la viabilidad de utilizar larvas de Eustrongylides sp. en programas de biomonitoreo. Cabe mencionar que las muestras de pescado para análisis de Hg deben estar libres de parásitos ya que su presencia puede alterar los resultados.

Resistance of Aeromonas hydrophila isolates to antimicrobials and sanitizers / Resistência de isolados de Aeromonas hydrophila a antimicrobianos e sanitizantes

In recent decades, Aeromonas hydrophila has emerged as a foodborne bacterial pathogen of public health concern, especially as it exhibits resistance to the major chemical sanitizers commonly used in the food industry. Meanwhile, this pathogen may be spread from diseased fish flesh through the contamination of equipment contact surfaces during food processing, thus posing a food safety risk. Thise determined the susceptibility profiles of retail fish-borne A. hydrophila isolates to 24 common antibiotics and five major sanitizers used in the food industry. The polymerase chain reaction technique was used to confirm all A. hydrophila isolates to the species level, and the agar diffusion method was applied to determine the antimicrobial susceptibility profiles. All isolates were confirmed to be A. hydrophila species. This bacterium was observed to have resistance to multiple antibiotics, with the highest resistance index being for those of the beta-lactam class. Additionally, the isolates showed high resistance to four of the five chemical sanitizers tested, with the highest resistance rate being toward sodium hypochlorite. The results suggested that A. hydrophila isolates with multiple resistance to the antimicrobials and main sanitizers used in the food industry can be found in retail fish sold in the Cuiabá region of Mato Grosso, Brazil.

Aeromonas hydrophila emergiu nas últimas décadas como um patógeno humano relevante. O fato desse patógeno alimentar emergente apresentar resistência antimicrobiana é um desafio considerável para as Agências de Saúde devido à resistência desta espécie aos principais sanitizantes químicos comumente utilizados na indústria alimentícia em que representa um risco à segurança dos alimentos, uma vez que pode contribuir para a disseminação desta bactéria na carne de peixes e levar à contaminação da superfície de contato dos equipamentos durante o processamento dos alimentos. Este estudo teve como objetivo determinar os padrões de suscetibilidade de isolados de A. hydrophila de peixes comercializados no varejo a 24 antibióticos comuns e cinco principais sanitizantes utilizados nas indústrias alimentícias. Neste estudo, todos os isolados de A. hydrophila foram confirmados em nível de espécie pela técnica de Reação em Cadeia da Polimerase (PCR). A técnica de difusão em ágar foi utilizada para determinar o perfil de suscetibilidade antimicrobiana frente aos 24 antibióticos testados e para avaliar a suscetibilidade aos principais sanitizantes utilizados na indústria alimentícia. A partir dos resultados, todos os isolados foram confirmados como sendo da espécie A. hydrophila pela técnica de PCR molecular. Observou-se A. hydrophila com perfil de resistência a múltiplos antibióticos, em que os da classe dos Beta-Lactâmicos foram os antimicrobianos com maior índice de resistência. Além disso, a suscetibilidade aos sanitizantes apresentou alta resistência em quatro dos cinco sanitizantes testados, sendo o hipoclorito de sódio foi o sanitizante químico com maior índice de resistência entre os isolados deste estudo. Os resultados sugerem que isolados de A. hydrophila com perfil de resistência a antimicrobianos e aos principais sanitizantes utilizados na indústria alimentícia podem ser encontrados em peixes comercializados no varejo da região de Cuiabá/Mato Grosso.

Doenças de peixes ornamentais no contexto brasileiro / Diseases of ornamental fish in the Brazilian context

O comércio de peixes ornamentais é um setor diversificado da aquicultura, cuja indústria contém alta diversidade de espécies. No Brasil, há uma diversidade de pelo menos 374 espécies de peixes de água doce e 223 espécies de peixes marinhos anunciados para venda em plataformas sociais (Borges et al., 2021). O mercado de peixes ornamentais no Brasil é organizado da seguinte maneira (Figura 1): • Setor produtivo: Consiste em fazendas aquícolas ou centros de captura de peixes silvestres. As regiões Sudeste e Sul são as maiores responsáveis pela produção aquícola em cativeiro de peixes ornamentais no Brasil. A região Norte é a principal responsável pela coleta de peixes silvestres e pela exportação. Os problemas relacionados com os surtos de doenças são um dos maiores gastos dos produtores de peixes ornamentais tanto no Brasil quanto na Flórida, EUA (Boldt et al., 2022). No Brasil, alguns produtores relatam que apenas um terço dos juvenis de algumas espécies chegam à fase de comercialização (Viadanna, nota pessoal). Já os peixes silvestres, além de alta mortalidade devido ao transporte, quando capturados e sem devido manejo, vão inserir novas doenças ao sistema produtivo por meio do agrupamento dos espécimes, fazendo com que a dispersão de agentes infecciosos seja facilitada. Ressalta-se que é comum o uso da mesma água para várias espécies, tornando possível a transmissão desses agentes infecciosos, que podem causar doença leve em uma espécie e grave em outra.(AU)

Principais aspectos sanitários na produção de peixes ornamentais / Main sanitary aspects in production of ornamental fish

Sabe-se que, para lidar com peixes, é necessário se atentar não apenas aos aspectos nutricionais e genéticos, mas sobretudo às questões ambientais, como controle da qualidade de água, e sanitários, como cuidados relacionados aos agentes externos e patogênicos. Qualquer desequilíbrio dentro de todas essas variáveis pode levar ao aparecimento de enfermidades. Dessa forma, como proceder para 6. Principais aspectos sanitários na produção de peixes ornamentais pixabay.com evitar a instalação de doenças em uma produção de peixes ornamentais? A resposta é a prevenção. Infelizmente, na maioria dos casos, a prevenção só vai ser buscada pelos produtores após a instalação de algum problema sanitário na propriedade. Portanto, cabe ao profissional especialista contratado não apenas solucionar o problema, mas também identificar pontos críticos que possam minimizar ou inviabilizar uma nova ocorrência do problema na propriedade.(AU)

Collective behavior and virulence arsenal of the fish pathogen Piscirickettsia salmonis in the biofilm realm.

Piscirickettsiosis is a fish disease caused by the Gram-negative bacterium Piscirickettsia salmonis. This disease has a high socio-economic impact on the Chilean salmonid aquaculture industry. The bacterium has a cryptic character in the environment and their main reservoirs are yet unknown. Bacterial biofilms represent a ubiquitous mechanism of cell persistence in diverse natural environments and a risk factor for the pathogenesis of several infectious diseases, but their microbiological significance for waterborne veterinary diseases, including piscirickettsiosis, have seldom been evaluated. This study analyzed the in vitro biofilm behavior of P. salmonis LF-89T (genogroup LF-89) and CA5 (genogroup EM-90) using a multi-method approach and elucidated the potential arsenal of virulence of the P. salmonis LF-89T type strain in its biofilm state. P. salmonis exhibited a quick kinetics of biofilm formation that followed a multi-step and highly strain-dependent process. There were no major differences in enzymatic profiles or significant differences in cytotoxicity (as tested on the Chinook salmon embryo cell line) between biofilm-derived bacteria and planktonic equivalents. The potential arsenal of virulence of P. salmonis LF-89T in biofilms, as determined by whole-transcriptome sequencing and differential gene expression analysis, consisted of genes involved in cell adhesion, polysaccharide biosynthesis, transcriptional regulation, and gene mobility, among others. Importantly, the global gene expression profiles of P. salmonis LF-89T were not enriched with virulence-related genes upregulated in biofilm development stages at 24 and 48 h. An enrichment in virulence-related genes exclusively expressed in biofilms was also undetected. These results indicate that early and mature biofilm development stages of P. salmonis LF-89T were transcriptionally no more virulent than their planktonic counterparts, which was supported by cytotoxic trials, which, in turn, revealed that both modes of growth induced important and very similar levels of cytotoxicity on the salmon cell line. Our results suggest that the aforementioned biofilm development stages do not represent hot spots of virulence compared with planktonic counterparts. This study provides the first transcriptomic catalogue to select specific genes that could be useful to prevent or control the (in vitro and/or in vivo) adherence and/or biofilm formation by P. salmonis and gain further insights into piscirickettsiosis pathogenesis.

Quality assessment of amazonian fish from fish farming stored on ice / Avaliação da qualidade de pescados amazônicos provenientes de piscicultura armazenados em gelo

The cultivation of natives Amazonian fish such as tambaqui (Colossoma macropomum), pirapitinga (Piaractus brachypomus), tambatinga (female, Colossoma macropomum X Piaractus brachypomus, male) and Pintando da Amazônia (female, Pseudoplatystoma sp. X Leiarius marmoratus, male) temmoratus, increased in recent years, making studies related to the validity of these foods after processing necessary. Therefore, the aimed of the study was to evaluate the sensory, microbiological (Staphylococcus coagulase positive count and presence of Salmonella spp.) and chemical (pH and Total Volatile Bases (TVB) quality of fresh gutted fish stored on ice. Nine specimens of each species were used, all cultivated in an excavated tanks. The fasting time ranged from 16 to 48 hours, depending on the species, the fish was slaughtered by means of cold shock. The fish was gutted and stored in the fish fridge. Sensory analysis was performed by a trained team available. For other analyses, it was sent to a laboratory specializing in food analysis. The sensory analysis of fresh eviscerated tambaqui was characteristic of the species until the 15th day of storage. Fresh eviscerated pirapitinga presented until the 16th day. The fresh eviscerated tambatinga hybrid was kept until the 14th day of storage. And for the Pintado da Amazônia, the organoleptic characteristics were presentable until the 12th day of storage. The microbiological and physical-chemical analyzes were in accordance with Brazilian legislation in all specimens of the analyzed species. It was observed that the longer the fasting period, there was a numerical increase in the pH of the fish meat.(AU)

O cultivo de peixes nativos da Amazônia como tambaqui (Colossoma macropomum), pirapitinga (Piaractus brachypomus), tambatinga (fêmea, Colossoma macropomum X Piaractus brachypomus, macho) e o pintado da Amazônia (fêmea, Pseudoplatystoma sp. X Leiarius marmoratus, macho) tem aumentado nos últimos anos, fazendo com que estudos relacionados a validade desses alimentos após o processamento sejam necessários. Por isso, o objetivo do estudo foi avaliar os parâmetros de qualidade senso-rial, microbiológica (contagem de Staphylococcus coagulase positiva e presença de Salmonella spp.) e química (pH e Bases Voláteis Totais (BVT)) deses pescados eviscerados frescos armazenados em gelo. Foram utilizados 9 exemplares de cada espécie, todos de origem de cultivo em viveiro escavado. O tempo de jejum variou de 16 a 48 horas, dependendo da espécie, o abate do pescado foi por meio de choque térmico. O pescado foi eviscerado e armazenado no frigorifico de pescado. Analise sensorial foi realizada por uma equipe treinada disponível. Para as demais análises foi enviado para um laboratório especializado em análises de alimentos. A análise sensorial do tambaqui eviscerado fresco estava característica da espécie até o 15°dia de estocagem. A pirapitinga evisce-rado fresco apresentou até o 16°dia. O hibrido tambatinga eviscerado fresco foi até o 14°dia de estocagem. E para o pintado da Amazônia as características organolépticas estavam apresentáveis até o dia 12° dia de armazenamento. As analises microbiológicas e físico químicas estavam de acordo com a legislação brasileira em todos os exemplares das espécies analisadas. Foi observado que quanto maior o tempo de jejum, houve um aumento numérico no pH da carne dos pescados.(AU)

Multiplex PCR assay for correct identification of the fish pathogenic species of Edwardsiella genus reveals the presence of E. anguillarum in South America in strains previously characterized as E. tarda.

AIMS: Develop a species-specific multiplex PCR to correctly identify Edwardsiella species in routine diagnostic for fish bacterial diseases. METHODS AND RESULTS: The genomes of 62 Edwardsiella spp. isolates available from the National Center for Biotechnology Information (NCBI) database were subjected to taxonomic and pan-genomic analyses to identify unique regions that could be exploited by species-specific PCR. The designed primers were tested against isolated Edwardsiella spp. strains, revealing errors in commercial biochemical tests for bacterial classification regarding Edwardsiella species. CONCLUSION: Some of the genomes of Edwardsiella spp. in the NCBI platform were incorrectly classified, which can lead to errors in some research. A functional mPCR was developed to differentiate between phenotypically and genetically ambiguous Edwardsiella, with which, we detected the presence of Edwardsiella anguillarum affecting fish in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that the misclassification of Edwardsiella spp in Brazil concealed the presence of E. anguillarum in South America. Also, this review of the taxonomic classification of the Edwardsiella genus is a contribution to the field to help researchers with their sequencing and identification of genomes, showing some misclassifications in online databases that must be corrected, as well as developing an easy assay to characterize Edwardsiella species in an end-point mPCR.

Piscine mycobacteriosis in the ornamental fish trade in Trinidad and Tobago.

The freshwater ornamental fish trade represents a major contributor to the livelihoods of many producers in Trinidad and Tobago, with stocks destined for local, regional and international markets. A review of clinical cases presented to the Aquatic Animal Health Unit at the University of the West Indies, School of Veterinary Medicine for the period September 2010 to December 2012 suggested that piscine mycobacteriosis may be widespread throughout the local ornamental fish industry. Thus, to determine the prevalence of mycobacteriosis in ornamental fish sold in pet stores, a total of 122 specimens were sourced from 24 retail suppliers across Trinidad. Fish were killed and internal organs were examined for lesions suggestive of granulomas. All wet-mount slides were acid-fast stained, regardless of the presence or absence of observed granuloma-like lesions. Histological analysis was performed on one randomly selected whole specimen from each facility. Mycobacterium sp. was identified using real-time PCR detecting the 16S rRNA gene in tissue samples. Associations between parasitism, facility biosecurity and presence of positive animals were determined. The prevalence of Mycobacterium sp. infection was 61 ± 7% (74/122), with positive specimens being acquired from 54.2% (13/24) of facilities examined. Further, 100% of facilities did not employ optimum biosecurity measures.

Different response of Acipenser gueldenstaedtii CRP/SAP and SAA to bacterial challenge and chronic thermal stress sheds light on the innate immune system of sturgeons.

Sturgeons are chondrostean fish critically endangered due to anthropogenic loss and degradation of natural habitat and overfishing for meat and caviar production. Consequently, sturgeon aquaculture has extensively developed lately, being Russian sturgeon (Acipenser gueldenstaedtii) the second most important species reared for caviar production. However, Russian sturgeon aquaculture in subtropical countries, such as Uruguay, confronts difficulties because fish have to endure excessive summertime warm temperatures, which weaken their innate defences facilitating opportunistic infections. To address this problem, we look for identifying putative acute phase proteins (APPs), which might be robust serum biomarkers of both infection and chronic thermal stress, applied to monitoring Russian sturgeon health status in farms. We focused on the C-Reactive Protein/Serum Amyloid P (CRP/SAP) pentraxin since the pentraxin family includes well-known APPs, better characterised in mammals than fish. We identified A.gueldenstaedtii CRP/SAP (AgCRP/SAP), as a member of the universal CRP/SAP pentraxin sub-family, and studied AgCRP/SAP involvement in sturgeon response to bacterial challenge and chronic thermal stress, in comparison with A. gueldenstaedtii Serum Amyloid A (AgSAA), a previously described positive APP. Results showed that AgCRP/SAP is a constitutive serum component that remained constant upon Aeromonas hydrophila challenge and chronic thermal stress. Contrastingly, serum AgSAA was subjected to regulation by bacterial and thermal stress challenges, showing a 50-fold increase and 3-fold decline in serum levels, respectively. Overall, results highlight the potential value of AgSAA, but not of AgCRP/SAP, as a biomarker of bacterial infection and the need to continue searching for robust chronic thermal stress biomarkers in sturgeons.

Avaliação das condições higiênico-sanitárias na comercialização de carne e pescado no mercado municipal da cidade de Pio XII, estado do Maranhão, Brasil / Evaluation of the hygienic-sanitary conditions in the commercialization of meat and fish in the municipal market in the city of Pio XII, state of Maranhão, Brazil

Avaliaram-se as condições higiênico-sanitárias na comercialização de carne e pescado no mercado municipal de Pio XII - MA, através da aplicação de uma lista de verificação adaptada de acordo com a RDC nº 216/2004 da ANVISA (Agência Nacional de Vigilância Sanitária), dotada de um total de 33 itens distribuídos em 8 categorias distintas, em 17 boxes de comercialização, no período de agosto a setembro de 2021. Realizou-se a aplicação de um questionário para verificar o perfil socioeconômico dos vendedores do mercado de Pio XII - MA, com aprovação no Comitê de Ética em Pesquisa (CONEP), da Universidade Estadual do Maranhão - UEMA com o número de CAAE 25016719.8.0000.5554. Foram verificados que 13% dos boxes do mercado estavam em péssimas condições higiênico-sanitárias e o restante foi classificado como ruim (20 a 49%), sendo que o percentual de conformidades de nenhum deles ultrapassou 27,0%, não sendo nenhum estabelecimento classificado como regular (50 a 69%), bom (70 a 90%) ou excelente (91 a 100%). Em relação aos resultados do perfil socioeconômico dos comerciantes, 100% eram do sexo masculino, com 17,6% apresentavam-se na faixa etária de 30 a 39 anos, 41,2% tinham idade de 40-49 anos e 41,2%, com mais de 50 anos. Quanto à escolaridade, predominaram os níveis não alfabetizados e ensino fundamental com percentual de 40,2% para cada um. A renda familiar da maioria (58,8%) se concentrou no intervalo de 1 a 3 salários mínimos. Pode-se concluir que a atividade de comercialização de carne e pescado no Mercado Municipal de Pio XII - MA vem sendo realizada de forma inadequada, e a falta de conhecimento dos comerciantes sobre as Boas Práticas de Fabricação e a realização de treinamento adequado são fatores contribuem de forma significativa para o agravo dessa situação, assim como a origem da carne comercializada, que é de abate clandestino.

The hygienic-sanitary conditions in the marketing of meat and fish in the municipal market of Pio XII - MA were evaluated through the application of a checklist adapted according to the RDC No. 216/2004 of ANVISA (National Health Surveillance Agency), with a total of 33 items distributed into 8 distinct categories, in 17 commercialization boxes, in the period from August to September 2021. A questionnaire was applied to verify the socioeconomic profile of the vendors in the market of Pio XII - MA, with approval in the Research Ethics Committee (CONEP), of the Maranhão State University - UEMA with the number CAAE 25016719.8.0000.5554. It was verified that 13% of the market stalls were in very bad hygienic-sanitary conditions and the rest were classified as bad (20 to 49%), and the percentage of compliance of none of them exceeded 27.0%, with no establishment classified as regular (50 to 69%), good (70 to 90%) or excellent (91 to 100%). Regarding the results of the socioeconomic profile of the merchants, 100% were male, with 17.6% in the 30 to 39 age bracket, 41.2% were 40-49 years old, and 41.2% were over 50 years old. As for education, non-literate and elementary school levels predominated, with a percentage of 40.2% for each. The family income of the majority (58.8%) was concentrated in the range of 1 to 3 minimum wages. It can be concluded that the activity of meat and fish commercialization in the Municipal Market of Pio XII - MA has been performed in an inadequate way, and the lack of knowledge of traders about the Good Manufacturing Practices and the performance of proper training are factors that contribute significantly to the worsening of this situation, as well as the origin of the meat commercialized, which is from clandestine slaughter.

Antimicrobial resistance profile of Aeromonas spp. isolated from asymptomatic Colossoma macropomum cultured in the Amazonas State, Brazil / Perfil de resistência antimicrobiana de Aeromonas spp. isoladas de Colossoma macropomum assintomáticos no Estado do Amazonas, Brasil

Bacterial diseases are important factors that limit productivity in aquaculture. To reduce negative economic impacts, fish farmers use antimicrobials, often indiscriminately, and this action has led to bacterial resistance to drugs. The objectives of this study were to isolate and identify the main putative pathogenic bacterial species in tambaqui (Colossoma macropomum), establish the profile of resistance to antimicrobials by the methods of disc diffusion, and determine the minimum inhibitory concentration (MIC) values. Two hundred and ninety asymptomatic fish were collected between March and November 2015 from ten fish farms in the Amazonas state (Brazil). Of the total strains recovered from tambaqui, seven were identified as Aeromonas spp. by sequencing the 16S rRNA gene. These seven isolates showed resistance to ampicillin, 28% to erythromycin, and 28% to sulfonamide. Additionally, the seven isolates showed a MIC higher than the range evaluated for amoxicillin, penicillin, novobiocin, tylosin tartrate, and clindamycin, and 85% showed resistance to erythromycin. The results of this study indicate the need to increase the awareness of fish farmers and, most importantly, the government, about the lack of drug regulations for use in aquaculture, and good management practices, so the indiscriminate prophylactic and systemic use of antimicrobials be inhibited.

As doenças bacterianas são fatores importantes que limitam a produtividade na aquicultura. Para reduzir os impactos econômicos negativos, os piscicultores utilizam antimicrobianos, muitas vezes de forma indiscriminada, e essa ação tem levado à resistência bacteriana aos medicamentos. Os objetivos deste estudo foram isolar e identificar as principais bactérias com potencial putativo para o tambaqui (Colossoma macropomum), e estabelecer o perfil de resistência a antimicrobianos pelos métodos de difusão em disco e valores de concentração inibitória mínima (CIM). Duzentos e noventa peixes assintomáticos foram coletados entre março e novembro de 2015, em dez pisciculturas do estado do Amazonas (Brasil). Do total de cepas recuperadas de tambaqui, sete foram identificadas como Aeromonas spp. pelo sequenciamento do gene 16S rRNA. Esses sete isolados apresentaram resistência à ampicilina, 28% à eritromicina e 28% à sulfonamida. Além disso, os sete isolados apresentaram CIM superior à faixa avaliada para amoxicilina, penicilina, novobiocina, tartarato de tilosina e clindamicina, e 85% apresentaram resistência à eritromicina. Os resultados deste estudo indicam a necessidade de aumentar a conscientização dos piscicultores e, principalmente, do poder público, a falta de regulamentação de medicamentos para uso na aquicultura e sobre as boas práticas de manejo, para que o uso profilático e sistêmico de antimicrobianos de forma indiscriminada seja inibido.

Bioaccumulation of organochlorine pesticides, Procamallanus sp. (Baylis, 1923) infections, and microbial colonization in African Snakehead fish sampled from Lekki lagoon, Lagos, Nigeria / Bioacumulação de pesticidas organoclorados, infecções por Procamallanus sp. (Baylis, 1923) e colonização microbiana em peixes africanos Snakehead coletados na lagoa Lekki, Lagos, Nigéria

The exponential rise in the Nigerian population has necessitated the use of agrochemicals for enhanced agricultural yields to meet the ever-rising demand for food. However, agrochemicals such as organochlorine pesticides (OCPs) have caused several devastating health and ecological challenges. The study was therefore aimed at assessing the bioaccumulation of OCPs and the associated parasitological and microbial susceptibility in P. obscura to determine the possible ecological impacts of the chemical. A total of 106 specimens of Parachanna obscura fish species were sampled between July and November 2019 from Lekki Lagoon in Lagos, Nigeria. Four culture media, namely nutrient agar (NA), MacConkay agar (MCA), eosin methylene blue (EMB), and sabouraud dextrose agar (SDA) were employed in microbial culture. These microbes were subjected to ceftazidime, ceftriaxone, cefuroxime, gentamicin, ofloxacin, augmentin, nitrofurantoin, ciprofloxacin, and erythromycin to test for resistance, susceptibility and intermediate statuses before and after curing. OCPs were tested in the water, sediment, and tissues of P. obscura using gas chromatography flame ionization detector (GC-FID). P. obscura sampled in the lagoon had poor growth exponent which was characterized by negative allometry (slenderness) in the sampled fish. Although the incidence of parasitic infection in the fish was not alarming, the situation might be aggravated if the prevalent anthropogenic activities persist, resulting in immunosuppression. Regulation of anthropogenic activities in the catchment area is recommended to forestall the prognosis of health and environmental hazards associated with the agricultural, industrial, pharmaceutical, and municipal activities around the lagoon. Bacteria that conferred the most resistance to the majority of the antibiotics were Staphylococcus sp., Micrococcus sp. Escherichia coli and Klebsiella sp., testing positive to plasmid profile. They conferred high resistance to the antibiotics before plasmid curing but became highly susceptible post- plasmid curing. This implies that the gene for resistance in the bacteria isolates was plasmid-mediated, that is, they were obtained from the environment. In the event of an outbreak of waterborne diseases such as cholera, typhoid, dysentery, and diarrhea, there may be non-response to treatment among the infected inhabitants. The incidence of antibiotic resistance in bacteria colonies recorded in this study is of great public health concern, given the possibility of the antibiotic-resistant bacteria strains being passed to humans through fish consumption, resulting in increased multi-drug resistance in humans. Regulation of anthropogenic activities around the lagoon is recommended to forestall prognosis of health and environmental hazards associated with OCPs from agricultural, industrial, pharmaceutical, and municipal sources.(AU)

O aumento exponencial da população nigeriana exigiu o uso de agroquímicos para aumentar a produção agrícola e, assim, atender à crescente demanda por alimentos. No entanto, agroquímicos como pesticidas organoclorados (OCPs) causaram vários problemas de saúde e ecológicos. Portanto, o estudo teve como objetivo avaliar a bioacumulação de OCPs e a suscetibilidade parasitológica e microbiana associada em Parachanna obscura, a fim de determinar os possíveis impactos ecológicos desse produto químico. Foi amostrado um total de 106 espécimes de P. obscura entre julho e novembro de 2019 da lagoa Lekki, em Lagos, Nigéria. Quatro meios de cultura, como o ágar nutritivo (NA), o ágar MacConkay (MCA), o ágar eosina azul de metileno (EMB) e o ágar sabouraud dextrose (SDA), foram empregados na cultura microbiana. Esses micróbios foram submetidos a ceftazidima, ceftriaxona, cefuroxima, gentamicina, ofloxacina, augmentin, nitrofurantoína, ciprofloxacina e eritromicina para testar resistência, suscetibilidade e status intermediário antes e depois da cura. Os OCPs foram testados na água, sedimentos e tecidos de P. obscura usando um detector de ionização de chama por cromatografia em fase gasosa (GC-FID). Os peixes amostrados de P. obscura da lagoa apresentaram um expoente de crescimento ruim, caracterizado por alometria negativa (esbelteza). Embora a incidência de infecção parasitária nos peixes não tenha sido alarmante, a situação pode ser agravada se as atividades antropogênicas prevalecentes persistirem, resultando em imunossupressão. Recomenda-se a regulamentação de atividades antropogênicas na área de captação para prevenir o prognóstico de riscos à saúde e ecológicos associados a atividades agrícolas, industriais, farmacêuticas e municipais ao redor da lagoa. As bactérias que conferiram maior resistência à maioria dos antibióticos foram Staphylococcus sp., Micrococcus sp., Escherichia coli e Klebsiella sp., com teste positivo para o perfil plasmidial. Elas conferiram alta resistência aos antibióticos antes da cura do plasmídeo, mas se tornaram altamente suscetíveis após a cura dele. Isso implica que o gene de resistência nos isolados de bactérias foi mediado por plasmídeo, ou seja, foi obtido do ambiente. No caso de surtos de doenças transmitidas pela água, como cólera, febre tifoide, disenteria e diarreia, pode haver não resposta ao tratamento entre os habitantes infectados. A incidência de resistência a antibióticos nas colônias de bactérias registradas neste estudo é de grande preocupação para a saúde pública, dada a possibilidade de que as cepas de bactérias resistentes a antibióticos sejam transmitidas aos seres humanos por meio do consumo de peixes, resultando em maior resistência a múltiplas drogas em seres humanos. Recomenda-se a regulamentação de atividades antropogênicas ao redor da lagoa para impedir o prognóstico de riscos à saúde e ecológicos associados aos OCPs de fontes agrícolas, industriais, farmacêuticas e municipais.(AU)

Granulomatous bacterial diseases in fish: An overview of the host's immune response.

Bacterial diseases represent the main impediment to the development of fish aquaculture. Granulomatous diseases caused by bacteria lead to fish culture losses by high mortality rates and slow growth. Bacteria belonging to genera Streptococcus spp., Mycobacterium sp., Nocardia sp., Francisella sp., and Staphylococcus sp. have been implicated in the development of granulomatous processes. The granuloma formation and the fish's immune response continue to be the subject of scientific research. In fish, the first defense line is constituted by non-specific humoral factors through growth-inhibiting substances such as transferrin and antiproteases, or lytic effectors as lysozyme and antimicrobial peptides, and linking with non-specific phagocyte responses. If the first line is breached, fish produce antibody constituents for a specific humoral defense inhibiting bacterial adherence, as well as the mobilization of non-phagocytic host cells and counteracting toxins from bacteria. However, bacteria causing granulomatous diseases can be persistent microorganisms, difficult to eliminate that can cause chronic diseases, even using some immune system components to survive. Understanding the infectious process leading to granulomatosis and how the host's immune system responds against granulomatous diseases is crucial to know more about fish immunology and develop strategies to overcome granulomatous diseases.