Immunological and molecular diagnostic techniques in fish health: present and future prospectus.

Fish health management is critical to aquaculture and fisheries as it directly affects sustainability and productivity. Fish disease diagnosis has taken a massive stride because of advances in immunological and molecular diagnostic tools which provide a sensitive, quick, and accurate means of identifying diseases. This review presents an overview of the main molecular and immunological diagnostic methods for determining the health of fish. The immunological techniques help to diagnose different fish diseases by detecting specific antigens and antibodies. The application of immunological techniques to vaccine development is also examined in this review. The genetic identification of pathogens is made possible by molecular diagnostic techniques that enable the precise identification of bacterial, viral, and parasitic organisms in addition to evaluating host reactions and genetic variation associated with resistance to disease. The combination of molecular and immunological methods has resulted in the creation of novel techniques for thorough evaluation of fish health. These developments improve treatment measures, pathogen identification and provide new information about the variables affecting fish health, such as genetic predispositions and environmental stresses. In the framework of sustainable fish farming and fisheries management, this paper focuses on the importance of these diagnostic techniques that play a crucial role in protecting fish populations and the aquatic habitats. This review also examines the present and potential future directions in immunological and molecular diagnostic techniques in fish health.

Effects of metformin on wild fathead minnows (Pimephales promelas) using in-lake mesocosms in a boreal lake ecosystem.

Due to its widespread use for the treatment of Type-2 diabetes, metformin is routinely detected in surface waters globally. Laboratory studies have shown that environmentally relevant concentrations of metformin can adversely affect the health of adult fish, with effects observed more frequently in males. However, the potential risk to wild fish populations has yet to be fully elucidated and remains a topic of debate. To explore whether environmentally relevant metformin exposure poses a risk to wild fish populations, the present study exposed wild fathead minnows (Pimephales promelas) to 5 or 50 µg/L metformin via 2 m diameter in-lake mesocosms deployed in a natural boreal lake in Northern Ontario at the International Institute for Sustainable Development - Experimental Lakes Area (IISD-ELA). Environmental monitoring was performed at regular intervals for 8-weeks, with fish length, weight (body, liver and gonad), condition factor, gonadosomatic index, liver-somatic index, body composition (water and biomolecules) and hematocrit levels evaluated at test termination. Metabolic endpoints were also evaluated using liver, brain and muscle tissue, and gonads were evaluated histologically. Results indicate that current environmental exposure scenarios may be sufficient to adversely impact the health of wild fish populations. Adult male fish exposed to metformin had significantly reduced whole body weight and condition factor and several male fish from the high-dose metformin had oocytes in their testes. Metformin-exposed fish had altered moisture and lipid (decrease) content in their tissues. Further, brain (increase) and liver (decrease) glycogen were altered in fish exposed to high-dose metformin. To our knowledge, this study constitutes the first effort to understand metformin's effects on a wild small-bodied fish population under environmentally relevant field exposure conditions.

Novel protective aspects of dietary polyphenols against pesticidal toxicity and its prospective application in rice-fish mode: A Review.

The rice fish system represents an innovative and sustainable approach to integrated farming, combining rice cultivation with fish rearing in the same ecosystem. However, one of the major challenges in this system is the pesticidal pollution resulting from various sources, which poses risks to fish health and overall ecosystem balance. In recent years, dietary polyphenols have emerged as promising bioactive compounds with potential chemo-preventive and therapeutic properties. These polyphenols, derived from various plant sources, have shown great potential in reducing the toxicity of pesticides and improving the health of fish within the rice fish system. This review aims to explore the novel aspects of using dietary polyphenols to mitigate pesticidal toxicity and enhance fish health in the rice fish system. It provides comprehensive insights into the mechanisms of action of dietary polyphenols and their beneficial effects on fish health, including antioxidant, anti-inflammatory, and detoxification properties. Furthermore, the review discusses the potential application methods of dietary polyphenols, such as direct supplementation in fish diets or through incorporation into the rice fields. By understanding the interplay between dietary polyphenols and pesticides in the rice fish system, researchers can develop innovative and sustainable strategies to promote fish health, minimize pesticide impacts, and ensure the long-term viability of this integrated farming approach. The information presented in this review will be valuable for scientists, aqua-culturists, and policymakers aiming to implement eco-friendly and health-enhancing practices in the rice fish system.

Metal bioaccumulation and histological alterations in Oreochromis andersonii at Itezhi-tezhi Dam downstream of a mining area in the Central Province of Zambia.

The Itezhi-tezhi Dam on the Kafue River in Zambia is a major capture fishery. However, the upstream reaches of the Kafue River receive effluents from copper mines. It was unclear whether fish health in the dam is adversely affected due to the mining effluents. We investigated the health status of fish in Itezhi-tezh Dam using a histology-based fish health assessment protocol with Oreochromis andersonii as a bioindicator. Fish were sampled in the Itezhi-tezh Dam and at a reference site further upstream on the Kafue River before it enters the mining region. Metal bioaccumulation, biometric indices and histological alterations in the gills, gonads, hearts, kidneys and livers were assessed. The findings revealed significantly higher copper and selenium sediment concentrations (p = 0.02843 and p = 0.02107 respectively), bioaccumulation of copper and selenium, and increased histological alterations in the gills, kidneys and livers of fish in the Itezhi-tezhi Dam.

Acipenserid herpesvirus 2 genome and partial validation of a qPCR for its detection in white sturgeon Acipenser transmontanus.

White sturgeon Acipenser transmontanus is the primary species used for caviar and sturgeon meat production in the USA. An important pathogen of white sturgeon is acipenserid herpesvirus 2 (AciHV-2). In this study, 4 archived isolates from temporally discrete natural outbreaks spanning the past 30 yr were sequenced via Illumina and Oxford Nanopore Technologies platforms. Assemblies of approximately 134 kb were obtained for each isolate, and the putative ATPase subunit of the terminase gene was selected as a potential quantitative PCR (qPCR) target based on sequence conservation among AciHV-2 isolates and low sequence homology with other important viral pathogens. The qPCR was repeatable and reproducible, with a linear dynamic range covering 5 orders of magnitude, an efficiency of approximately 96%, an R2 of 0.9872, and an analytical sensitivity of 103 copies per reaction after 35 cycles. There was no cross-reaction with other known viruses or closely related sturgeon species, and no inhibition by sturgeon DNA. Clinical accuracy was assessed from white sturgeon juveniles exposed to AciHV-2 by immersion. Viral culture (gold standard) and qPCR were in complete agreement for both cell culture negative and cell culture positive samples, indicating that this assay has 100% relative accuracy compared to cell culture during an active outbreak. The availability of a whole-genome sequence for AciHV-2 and a highly specific and sensitive qPCR assay for detection of AciHV-2 in white sturgeon lays a foundation for further studies on host-pathogen interactions while providing a specific and rapid test for AciHV-2 in captive and wild populations.

Poison in the water: Arsenic's silent assault on fish health.

Arsenic occurs across the world in freshwater and marine environments, menacing the survival of aquatic organisms. Organic and inorganic forms of this substance can be found, in which the inorganic form is more hazardous than the organic form. Most aquatic bodies contain inorganic arsenic species, but organic species are believed to be the dominant form of arsenic in the majority of fish. Natural and anthropogenic both are the sources of water contamination with arsenic. Its bioaccumulation and transfer from one trophic level to another in the aquatic food chain make arsenic a vital environmental issue. Continuous exposure to low concentrations of arsenic in aquatic organisms including fish leads to its bioaccumulation, which may affect organisms of higher trophic levels including large fishes or humans. Humans can be exposed to arsenic through the consumption of fish contaminated with arsenic. Hence, the present review facilitates our understanding about sources of arsenic, its bioaccumulation, food chain transfer, and its effect on the fish health. Also, "Poison in the Water: Arsenic's Silent Assault on Fish Health" serves as a wake-up call to recognize the pressing need to address arsenic contamination in water bodies. By understanding its devastating impact on fish health, we can strive to implement sustainable practices and policies that safeguard our precious aquatic environments and ensure the well-being of both wildlife and human communities that depend on them.

Mucosal and systemic physiological changes underscore the welfare risks of environmental hydrogen sulphide in post-smolt Atlantic salmon (Salmo salar).

Atlantic salmon (Salmo salar) might encounter toxic hydrogen sulphide (H2S) gas during aquaculture production. Exposure to this gas can be acute or chronic, with heightened levels often linked to significant mortality rates. Despite its recognised toxicity, our understanding of the physiological implications of H2S on salmon remains limited. This report details the mucosal and systemic physiological consequences in post-smolt salmon reared in brackish water at 12 ppt after prolonged exposure to elevated H2S levels over 4 weeks. The fish were subjected to two concentrations of H2S: 1 µg/L (low group) and 5 µg/L (high group). An unexposed group at 0 µg/L served as the control. Both groups exposed to H2S exhibited incremental mortality, with cumulative mortality rates of 4.7 % and 16 % for the low and high groups, respectively. Production performance, including weight and condition factors, were reduced in the H2S-exposed groups, particularly in the high group. Mucosal response of the olfactory organ revealed higher tissue damage scores in the H2S-exposed groups, albeit only at week 4. The high group displayed pronounced features such as increased mucus cell density and oedema-like vacuoles. Transcriptome analysis of the olfactory organ unveiled that the effects of H2S were more prominent at week 4, with the high group experiencing a greater magnitude of change than the low group. Genes associated with the extracellular matrix were predominantly downregulated, while the upregulated genes primarily pertained to immune response. H2S-induced alterations in the metabolome were more substantial in plasma than skin mucus. Furthermore, the number of differentially affected circulating metabolites was higher in the low group compared to the high group. Five core pathways were significantly impacted by H2S regardless of concentration, including the phenylalanine, tyrosine, and tryptophan biosynthesis. The plasma levels of phenylalanine and tyrosine were reduced following exposure to H2S. While there was a discernible distinction in the skin mucus metabolomes among the three treatment groups, only one metabolite - 4-hydroxyproline - was significantly impacted by H2S. Furthermore, this metabolite was significantly reduced in the plasma and skin mucus of H2S-exposed fish. This study underscores that prolonged exposure to H2S, even at concentrations previously deemed sub-lethal, has discernible physiological implications that manifest across various organisational levels. Given these findings, prolonged exposure to H2S poses a welfare risk, and thus, its presence must be maintained at low levels (<1 µg/L) in salmon land-based rearing systems.

Connecting thiamine availability to the microbial community composition in Chinook salmon spawning habitats of the Sacramento River basin.

Thiamine deficiency complex (TDC) is a major emerging threat to global populations of culturally and economically important populations of salmonids. Salmonid eggs and embryos can assimilate exogenous thiamine, and evidence suggests that microbial communities in benthic environments can produce substantial amounts of thiamine. We therefore hypothesize that natural dissolved pools of thiamine exist in the surface water and hyporheic zones of riverine habitats where salmonids with TDC migrate, spawn, and begin their lives. To examine the relationship between dissolved thiamine-related compounds (dTRCs) and their microbial source, we determined the concentrations of these metabolites and the compositions of microbial communities in surface and hyporheic waters of the Sacramento River, California and its tributaries. Here we determine that all dTRCs are present in femto-picomolar concentrations in a range of critically important salmon spawning habitats. We observed that thiamine concentrations in the Sacramento River system are orders of magnitude lower than those of marine waters, indicating substantial differences in thiamine cycling between these two environments. Our data suggest that the hyporheic zone is likely the source of thiamine to the overlying surface water. Temporal variations in dTRC concentrations were observed where the highest concentrations existed when Chinook salmon were actively spawning. Significant correlations were seen between the richness of microbial taxa and dTRC concentrations, particularly in the hyporheic zone, which would influence the conditions where embryonic salmon incubate. Together, these results indicate a connection between microbial communities in freshwater habitats and the availability of thiamine to spawning TDC-impacted California Central Valley Chinook salmon.IMPORTANCEPacific salmon are keystone species with considerable economic importance and immeasurable cultural significance to Pacific Northwest indigenous peoples. Thiamine deficiency complex has recently been diagnosed as an emerging threat to the health and stability of multiple populations of salmonids ranging from California to Alaska. Microbial biosynthesis is the major source of thiamine in marine and aquatic environments. Despite this importance, the concentrations of thiamine and the identities of the microbial communities that cycle it are largely unknown. Here we investigate microbial communities and their relationship to thiamine in Chinook salmon spawning habitats in California's Sacramento River system to gain an understanding of how thiamine availability impacts salmonids suffering from thiamine deficiency complex.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues / Respostas fisiológicas de tambaqui (Colossoma macropomum) alimentado com dietas suplementadas com silagem de resíduos de pescado e vegetais

The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

A procura por insumos que substituam produtos de alto custo, como óleo e farinha de peixe, na fabricação de rações para uso na aquicultura é crescente, sendo necessário garantir a qualidade nutricional das dietas. A silagem produzida a partir de resíduos de pescado e de vegetais apresenta-se como uma alternativa de baixo custo e eficiente fonte proteica. O objetivo do presente estudo foi avaliar as respostas fisiológicas e bioquímicas de alevinos de tambaqui alimentados com quatro níveis de inclusão de silagem em ração comercial com 28% de proteína bruta, em dois períodos: 45 e 90 dias. Cada tratamento foi realizado em três repetições, com 10 tambaquis em cada caixa experimental de 100 L. Ao término de cada período estabelecido, amostras sanguíneas foram coletadas de cinco animais de cada repetição para determinação das variáveis hematológicas e bioquímicas. Peso, comprimento total e índices hepatossomático e lipossomático de espécimes alimentados com silagem não mostraram alterações significativas em ambos os períodos de avaliação, bem como os valores de hematócrito. A concentração de hemoglobina de tambaqui após 45 dias de alimentação aumentou quando foi fornecida dieta com inclusão de 20% de silagem. A contagem de eritrócitos, volume corpuscular médio e hemoglobina corpuscular média não apresentaram alterações entre os tratamentos, em ambos os períodos. As proteínas totais aumentaram significativamente no plasma de tambaquis que receberam dietas com inclusão de 5 e 10% de silagem, avaliados após 45 dias de alimentação. Evidenciou-se que os grupos com inclusão de silagem na dieta não apresentaram alterações significativas nos parâmetros avaliados, assim não sendo prejudiciais à higidez do tambaqui. Portanto, o uso da silagem como suplemento alimentar durante o cultivo do tambaqui é uma alternativa sustentável para produtores, por promover a redução do descarte de resíduos de pescado e da agricultura.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues

Abstract The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

Resumo A procura por insumos que substituam produtos de alto custo, como óleo e farinha de peixe, na fabricação de rações para uso na aquicultura é crescente, sendo necessário garantir a qualidade nutricional das dietas. A silagem produzida a partir de resíduos de pescado e de vegetais apresenta-se como uma alternativa de baixo custo e eficiente fonte proteica. O objetivo do presente estudo foi avaliar as respostas fisiológicas e bioquímicas de alevinos de tambaqui alimentados com quatro níveis de inclusão de silagem em ração comercial com 28% de proteína bruta, em dois períodos: 45 e 90 dias. Cada tratamento foi realizado em três repetições, com 10 tambaquis em cada caixa experimental de 100 L. Ao término de cada período estabelecido, amostras sanguíneas foram coletadas de cinco animais de cada repetição para determinação das variáveis hematológicas e bioquímicas. Peso, comprimento total e índices hepatossomático e lipossomático de espécimes alimentados com silagem não mostraram alterações significativas em ambos os períodos de avaliação, bem como os valores de hematócrito. A concentração de hemoglobina de tambaqui após 45 dias de alimentação aumentou quando foi fornecida dieta com inclusão de 20% de silagem. A contagem de eritrócitos, volume corpuscular médio e hemoglobina corpuscular média não apresentaram alterações entre os tratamentos, em ambos os períodos. As proteínas totais aumentaram significativamente no plasma de tambaquis que receberam dietas com inclusão de 5 e 10% de silagem, avaliados após 45 dias de alimentação. Evidenciou-se que os grupos com inclusão de silagem na dieta não apresentaram alterações significativas nos parâmetros avaliados, assim não sendo prejudiciais à higidez do tambaqui. Portanto, o uso da silagem como suplemento alimentar durante o cultivo do tambaqui é uma alternativa sustentável para produtores, por promover a redução do descarte de resíduos de pescado e da agricultura.

Cytotoxicity and immunological impact of Trypanosoma sp. infection on blood parameters of wild African catfish, Clarias gariepinus.

Fish trypanosomiasis is a common blood parasitic disease transmitted by aquatic invertebrates, such as leeches. This study aims to shed light on the cytotoxicity of Trypanosoma sp. on erythrocytes and its impacts on the innate immune response (serum lysozyme activity, nitric oxide production, phagocytic activity, serum total protein, and globulin) in wild African catfish, Clarias gariepinus. One hundred catfish were examined using blood smears stained with Giemsa and confirmed with PCR. The prevalence of infection was found to be 10% by microscope detection and 15% by PCR. The morphological identification of Trypanosoma as Trypanosoma mukasai was determined. Additionally, this study included previously undescribed features of Trypanosoma, such as the width of the anterior and posterior body, the length of the posterior pale region, and the number of folds. Various alterations in erythrocytes were observed, totaling 54.57%. Nuclear abnormalities, including fragmented nuclei, eccentric nuclei, and micronuclei, were also reported. Infected fish showed a reduction in serum total protein and globulin levels, while nitric oxide production, lysozyme activity, and phagocytic activity exhibited a significant increase compared to non-infected fish. We believe that our findings will contribute valuable data to the morphological and molecular identification of Trypanosoma sp. in African catfish, as well as their cytotoxic impact.

Accelerated eutrophication alters fish and aquatic health: a quantitative assessment by using integrative multimarker, hydrochemical, and GIS modelling method in an urban lake.

The ramifications of anthropogenic activities on the environment and the welfare of aquatic life in lakes worldwide are becoming increasingly alarming. There is a lack of research in the Indian Himalayas on fish biomarker responses to stressful aquatic conditions and the use of environmetric modelling in GIS. Our research evaluates the environmental health of urban lakes in multiple basins using multi-biomarker endpoints (13 features) in Schizothorax niger and hydrochemical characterization (9 features) of water. The study covers 31 grids, each at a distance of 1 km2. This study demonstrated a statistically significant (P = 0.001) increase in white blood cells (WBC), mean cell size (MCH), helminth infection, and health assessment index score (HAIS) score in fish from a highly eutrophic cluster or basin compared to a reference cluster, which is indicative of environmental stress in fish. Based on hydrochemical similarities, the lake water datasets were divided into three categories using hierarchical cluster analysis (HCA). In the PCA analysis, the first three principal components were responsible for 78.1% of the data's variance. The first principal component (PC1) accounted for 57.4% of the variance and had a strong positive loading from ammonia, total phosphate, pH, nitrates, and total alkalinity for water quality parameters. Additionally, PC1 had a favourable loading from WBC, helminth infection (%), and the health assessment index score (HAIS) for biological endpoints. These findings are in alignment with the results of the multivariate analysis. The trophic state index (TSI) showed a significant (P < 0.05) increase in Cluster 1, which includes the peripheral areas of Hazratbal and Gagribal side (> 70), compared to the reference cluster. The multiple regression model indicates that ammonia, phosphate, and nitrate significantly impact the general health of fish (R2 > 0.7). A novel methodology for monitoring water quality fluctuations across different basins and clusters is presented in this study. By integrating fish health biomarkers and GIS technology, we have developed a comprehensive approach to evaluate the overall well-being of aquatic habitat. This technique may prove beneficial in the management of urban lentic water bodies in the Kashmir Himalayas and other comparable water systems around the globe, while also supporting sustainable practices.

Captivating Colors, Crucial Roles: Astaxanthin's Antioxidant Impact on Fish Oxidative Stress and Reproductive Performance.

Fish, constantly exposed to environmental stressors due to their aquatic habitat and high metabolic rates, are susceptible to oxidative stress. This review examines the interplay between oxidative stress and fish reproduction, emphasizing the potent antioxidant properties of astaxanthin. Our primary objective is to highlight astaxanthin's role in mitigating oxidative stress during critical reproductive stages, leading to improved gamete quality, ovary development, and hormone levels. We also explore its practical applications in aquaculture, including enhanced pigmentation and overall fish health. We conducted a comprehensive literature review, analyzing studies on astaxanthin's antioxidant properties and its impact on fish reproduction. Astaxanthin, a carotenoid pigment, effectively combats reactive oxygen species, inhibiting lipid peroxidation and maintaining membrane integrity. It significantly enhances reproductive success in fish and improves overall fish health in aquaculture settings. This review reveals astaxanthin's multifaceted benefits in fish health and reproduction, offering economic advantages in aquaculture. Future research should delve into species-specific responses, optimal dosages, and the long-term effects of astaxanthin supplementation to inform sustainable aquaculture strategies.

The impacts of synthetic and cellulose-based fibres and their associated dyes on fish hosts and parasite health.

Plastic pollution is now a ubiquitous feature of freshwater systems and the majority of this is fibrous. Here, we test the effects of plastic and cellulose-based fibres (polyester, cotton, and bamboo from commercial clothing) on fish host-parasite interactions using a freshwater fish host-parasite model system (guppy Poecilia reticulata-Gyrodactylus turnbulli). For uninfected fish, polyester exposure was associated with significantly higher mortality rates compared with the other two fibre types. For infected fish, whilst polyester and cotton exposure were not associated with any significant changes to parasite burdens, fish exposed to bamboo fibres had significantly reduced maximum parasite burdens compared with fish not exposed to any fibres, indicating that the bamboo fibres and/or associated dyes conferred some degree of resistance or tolerance. Whilst unable to determine the exact nature of the chemical dyes, when testing off-host parasite survival on exposure to the fibre dyes, cotton and particularly polyester dyes were associated with higher parasite mortality compared to bamboo. Overall, we add to the growing body of evidence which shows that polyester microplastic fibres and their associated dyes can be detrimental for both fish and parasite survival, and we highlight the need for increased transparency from textile industries on the chemical identity of fabric dyes.

Wearable Bioimpedance-Based Deep Learning Techniques for Live Fish Health Assessment under Waterless and Low-Temperature Conditions.

(1) Background: At present, physiological stress detection technology is a critical means for precisely evaluating the comprehensive health status of live fish. However, the commonly used biochemical tests are invasive and time-consuming and cannot simultaneously monitor and dynamically evaluate multiple stress levels in fish and accurately classify their health levels. The purpose of this study is to deploy wearable bioelectrical impedance analysis (WBIA) sensors on fish skin to construct a deep learning-based stress dynamic evaluation model for precisely estimating their accurate health status. (2) Methods: The correlation of fish (turbot) muscle nutrients and their stress indicators are calculated using grey relation analysis (GRA) for allocating the weight of the stress factors. Next, WBIA features are sieved using the maximum information coefficient (MIC) in stress trend evaluation modeling, which is closely related to the key stress factors. Afterward, a convolutional neural network (CNN) is utilized to obtain the features of the WBIA signals. Then, the long short-term memory (LSTM) method learns the stress trends with residual rectification using bidirectional gated recurrent units (BiGRUs). Furthermore, the Z-shaped fuzzy function can accurately classify the fish health status by the total evaluated stress values. (3) Results: The proposed CNN-LSTM-BiGRU-based stress evaluation model shows superior accuracy compared to the other machine learning models (CNN-LSTM, CNN-GRU, LSTM, GRU, SVR, and BP) based on the MAPE, MAE, and RMSE. Moreover, the fish health classification under waterless and low-temperature conditions is thoroughly verified. High accuracy is proven by the classification validation criterion (accuracy, F1 score, precision, and recall). (4) Conclusions: the proposed health evaluation technology can precisely monitor and track the health status of live fish and provides an effective technical reference for the field of live fish vital sign detection.

Optimisation of trace mineral supplementation in diets for Atlantic salmon smolt with reference to holistic fish performance in terms of growth, health, welfare, and potential environmental impacts.

The aquafeed ingredient inventory is ever changing, from marine to plant based, and recently evolving to incorporate increasing amounts of low trophic, side stream and circular economy based raw materials, each one contributing with variable amounts and qualities of macro- and micronutrients. Meeting the micronutrient requirement of farmed fish for healthy and efficient growth under normal and challenging conditions is of paramount importance. In this study we run a trial based on a 2 × 4 factorial design with three replications for each dietary treatment, where Atlantic salmon smolt were fed one of 8 experimental diets supplemented with either organic or inorganic mineral premixes (copper, iron, manganese, selenium, and zinc) at four dietary inclusion levels. We saw a trend for higher growth rate in the organic mineral groups irrespective of the dietary mineral levels. Mineral digestibility was negatively correlated with increasing mineral supplementation levels for all tested minerals but Se which increased with the increasing supplementation in the inorganic and up to the 2nd inclusion level in the organic mineral groups. Increasing mineral supplementation affected retention efficiency of Zn, Mn, Cu and Fe while mineral source affected only the retention of Se which was higher in the organic mineral groups. Moreover, fish obtained higher EPA and DHA in their body and increased slaughter yield in the organic as compared to the inorganic mineral groups and corroborated that trace mineral inclusion levels play a key role on salmon fillet's technical quality. More effects from different origin and dietary inclusion levels of trace minerals were seen on fillet yield, fillet technical and nutritional quality, bone strength, skin morphology, organ mineralization and midgut transcriptome.

Improving the diagnosis of Streptococcus iniae using a novel probe-based qPCR assay combined with an enrichment step.

Streptococcus iniae is a bacterial pathogen that causes streptococcosis, leading to significant losses in fish aquaculture globally. This study reported a newly developed probe-based quantitative polymerase chain reaction (qPCR) method for the detection of S. iniae. The primers and probes were designed to target the lactate oxidase gene. The optimized method demonstrated a detection limit of 20 copies per reaction and was specific to S. iniae, as evidenced by no cross-reactivity when assayed against genetic materials extracted from 23 known aquatic animal pathogens, and fish samples infected with Streptococcus agalactiae or Streptococcus dysgalactiae. To validate the newly developed qPCR protocol with field samples, fish specimens were systematically investigated following the Food and Agriculture Organization of the United Nations & Network of Aquaculture Centres in Asia-Pacific three diagnostic levels approach, which integrated basic and advanced techniques for disease diagnosis, including observation of gross signs (level I), bacterial isolation (level II), qPCR and 16S rDNA sequencing (level III). The result showed that 7/7 affected farms (three Asian seabass farms and four tilapia farms) experiencing clinical signs of streptococcosis were diagnosed positive for S. iniae. qPCR assays using DNA extracted directly from fish tissue detected S. iniae in 11 out of 36 fish samples (30.6%), while 24 out of 36 samples (66.7%) tested positive after an enrichment step, including apparently healthy fish from affected farms. Bacterial isolation of S. iniae was only successful in a proportion of clinically diseased fish but not in healthy-looking fish from the same farm. Overall, the newly developed qPCR protocol combined with enrichment would be a useful tool for the diagnosis and surveillance of S. iniae infections in fish populations, thereby aiding in the disease control and prevention.

Selection and Characterization of Bacteriocinogenic Lactic Acid Bacteria from the Intestine of Gilthead Seabream (Sparus aurata) and Whiting Fish (Merlangius merlangus): Promising Strains for Aquaculture Probiotic and Food Bio-Preservation.

This study sought to evaluate the probiotic properties and the food preservation ability of lactic acid bacteria isolates collected from the intestines of wild marine fishes (gilthead seabream (Sparus aurata) (n = 60) and whiting fish (Merlangius merlangus) (n = 40)) from the Mediterranean sea in the area of Mostaganem city, Algeria. Forty-two isolates were identified as: Enterococcus durans (n = 19), Enterococcus faecium (n = 15), Enterococcus faecalis (n = 4), Lactococcus lactis subp. lactis (n = 3), and Lactobacillus plantarum (n = 1). All isolates showed inhibition to at least one indicator strain, especially against Listeria monocytogenes, Staphylococcus aureus, Paenibacillus larvae, Vibrio alginolyticus, Enterococcus faecalis, Bacillus cereus, and Bacillus subtilis. In all collected isolates, PCR analysis of enterocin-encoding genes showed the following genes: entP (n = 21), ent1071A/B (n = 11), entB (n = 8), entL50A/B (n = 7), entAS48 (n = 5), and entX (n = 1). Interestingly, 15 isolates harbored more than one ent gene. Antimicrobial susceptibility, phenotypic virulence, and genes encoding virulence factors were investigated by PCR. Resistance to tetracycline (n = 8: tetL + tetK), erythromycin (n = 7: 5 ermA, 2 msrA, and 1 mef(A/E)), ciprofloxacin (n = 1), gentamicin (n = 1: aac(6')-aph(2″)), and linezolid (n = 1) were observed. Three isolates were gelatinase producers and eight were α-hemolytic. Three E. durans and one E. faecium harbored the hyl gene. Eight isolates showing safety properties (susceptible to clinically relevant antibiotics, free of genes encoding virulence factors) were tested to select probiotic candidates. They showed high tolerance to low pH and bile salt, hydrophobicity power, and co-culture ability. The eight isolates showed important phenotypic and genotypic traits enabling them to be promising probiotic candidates or food bio-conservers and starter cultures.

Catch of the Day: New Serum Amyloid A (SAA) Antibody Is a Valuable Tool to Study Fish Health in Salmonids.

Serum amyloid A (SAA) proteins belong to a family of acute-phase reactants, playing an integral role in defending the organism from pathological damage. Despite a wealth of data on the regulation of SAA transcripts in teleosts, there is only limited information on these proteins' abundance in fish. The aim of this study is to characterise SAA protein levels in salmonids using a newly developed antibody specific to salmonid SAA. The salmonid SAA antibody detected SAA and accurately discriminated between stimulated and control specimens from rainbow trout macrophage cell line (RTS-11) in vitro, as well as rainbow trout challenged with Aeromonas salmonicida- or flagellin-stimulated Atlantic salmon in vivo. The presence of SAA protein was analysed in RTS-11 cell line supernatants, liver, and spleen samples using ELISA, immunoblotting, and immunohistochemistry. This study is the first to characterise SAA protein levels in salmonids in vivo and in vitro. The newly developed salmonid SAA antibody was able to discriminate between stimulated and unstimulated specimens, showing that it can be used to study the acute-phase response in salmonids with the potential to be further developed into assays to monitor and evaluate health in wild and farmed fish.

Health of wild fish exposed to contaminants of emerging concern in freshwater ecosystems utilized by a Minnesota Tribal community.

Fish serve as indicators of exposure to contaminants of emerging concern (CECs)-chemicals such as pharmaceuticals, hormones, and personal care products-which are often designed to impact vertebrates. To investigate fish health and CECs in situ, we evaluated the health of wild fish exposed to CECs in waterbodies across northeastern Minnesota with varying anthropogenic pressures and CEC exposures: waterbodies with no human development along their shorelines, those with development, and those directly receiving treated wastewater effluent. Then, we compared three approaches to evaluate the health of fish exposed to CECs in their natural environment: a refined fish health assessment index, a histopathological index, and high-throughput (ToxCast) in vitro assays. Lastly, we mapped adverse outcome pathways (AOPs) associated with identified ToxCast assays to determine potential impacts across levels of biological organization within the aquatic system. These approaches were applied to subsistence fish collected from the Grand Portage Indian Reservation and 1854 Ceded Territory in 2017 and 2019. Overall, 24 CECs were detected in fish tissues, with all but one of the sites having at least one detection. The combined implementation of these tools revealed that subsistence fish exposed to CECs had histological and macroscopic tissue and organ abnormalities, although a direct causal link could not be established. The health of fish in undeveloped sites was as poor, or sometimes poorer, than fish in developed and wastewater effluent-impacted sites based on gross and histologic tissue lesions. Adverse outcome pathways revealed potential hazardous pathways of individual CECs to fish. A better understanding of how the health of wild fish harvested for consumption is affected by CECs may help prioritize risk management research efforts and can ultimately be used to guide fishery management and public health decisions. Integr Environ Assess Manag 2023;00:1-18. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).