Cell-Cultured Fish Meat via Scale-Up Expansion of Carassius auratus Skeletal Muscle Cells Using Edible Porous Microcarriers and Quality Evaluation.

Emerging technologies for cell-cultured fish meat as an environmentally friendly protein source for humans still have many obstacles, including large-scale production of high-quality cells, differentiation and bioassembly of cellular material, and improvement of the quality of meat products. Here, we used edible porous microcarriers as scaffolds to support scalable skeletal muscle cell expansion to prepare centimeter-scale cell-cultured fish (CCM) of Carassius auratus for the first time. The quality of CCM was assessed by analyzing the texture, nutrition, flavor, and safety. The results indicated that CCM demonstrated a softer texture than natural fish due to a high moisture content. CCM contained higher protein and lower fat contents, with no significant difference in energy from natural golden crucian carp meat (NGM). CCM had better digestible properties, and 17 volatile components were identified in CCM, ten cocontained compared to NGM. ELISA quantified penicillin, streptomycin, vitamin D, and insulin residues as risk factors in CCM. In conclusion, we utilized edible porous microcarriers to scale-up the expansion of Carassius auratus skeletal muscle cells and bioassembled high-quality CCM of Carassius auratus for the first time, which represents a state-of-the-art protocol applicable to different fish species and even to other economic animals and provides a theoretical basis for scaling up cell-cultured meat production.

Elements in cultured fishes in Chittagong, Bangladesh and risk assessment.

The concentrations of manganese, iron, copper, lead, nickel, cadmium, arsenic, copper and mercury were determined in 10 cultured fish species, which were collected from local markets of Chittagong, Bangladesh, in June 2021. Measurements were performed by atomic absorption spectrophotometry, after acid digestion of the samples. In some cases, the concentration of the investigated elements was more than the maximum limit set by the WHO. Although the concentration of toxic elements in fishes was relatively high in some species, no health risk has been identified in comparison to the estimated daily intake and the maximum limit. Calculated hazard indices were below 1, which indicates the investigated fish would not cause human health risks. Carcinogenic risk indices for Cr, As, and Cu in all species were considered to be significant.

Microplastics contamination in fish feeds: Characterization and potential exposure risk assessment for cultivated fish of Bangladesh.

Fish feed is becoming an increasingly vital source of nourishment for farmed fish, which are mainly coming from marine fish and agricultural sources. Anthropogenic particles, such as microplastics, are abundant in both marine fish and agricultural byproducts that are utilized to make fish feed. This study investigated whether fish feed could be a source of microplastic contamination, and revealed that a 20 weeks adult farmed tilapia fish might consume up to 268.45 ± 1.438 microplastic particles via fish feed where finisher type feeds were found to be mostly contributory in this number. The microplastics were initially observed with a stereomicroscope and FESEM-EDS. Polymeric composition of microplastics was determined to be polypropylene (PP), nylon-6 (NY-6), polyethylene terephthalate (PET), polystyrene (PS), polyvinyl alcohol (PVA), polyethylene (PE), high- and low-density polyethylene (HDPE, LDPE), ethylene vinyl acetate (EVA), polycarbonate (PC), poly vinyl acetate (PVAc), poly urethane (PU) and polyvinyl chloride (PVC) by FTIR. Results also revealed that the size of microplastic particles in all fish feed ranged from 14 µm to 4480 µm, with 550 ± 45.45 to 11,600 ± 56.1 microplastic particles/kg of fish feed. The FESEM-EDS data demonstrated to overlook the microplastic surface along with attachment of heavy metals onto that surface such as Pb, Ni, and Co in finisher type feed that could create additional health risks.

Occurrence and human health risk assessment of antibiotics in cultured fish from 19 provinces in China.

The occurrence of antibiotics and potential health risk of 300 cultured fish samples from 19 provinces in China were investigated. The levels of 28 antibiotics (15 fluoroquinolones, 4 tetracyclines, 8 macrolides and rifampin) in 8 fish species were measured through liquid chromatography electrospray tandem mass spectrometry. As a result, 10 antibiotics were detected with an overall detection frequency of 24.3%, and the individual detection frequency of antibiotics ranged from 0.33 to 16.7%. The extremely high concentrations (above 100 µg/kg) of doxycycline and erythromycin were found in the samples. Antibiotics with high detection frequency was noticed in largemouth bass (41.2%), followed by snakehead (34.4%) and bream (31.2%). Specifically, Heilongjiang, Xinjiang, Qinghai and Gansu presented high detection frequency values of more than 60%. Moreover, the highest mean concentration was observed in Shandong, and the concentration covered from 34.8 µg/kg to 410 µg/kg. Despite the high detection frequency and levels of antibiotics were found in samples, ingestion of cultured fish was not significantly related to human health risks in China, according to the calculated estimated daily intakes and hazard quotients. These results provided us the actual levels of antibiotics in cultured fish and human health risk assessment of consuming fishery products.

Element concentrations of cultured fish in the Black Sea: selenium-mercury balance and the risk assessments for consumer health.

Fish are a powerful model for risk-benefit analyses to explore the impact of elements on human health among all accessible species. The sea bream (Sparus aurata), sea bass (Dicentrarchus labrax), turbot (Scophthalmus maximus), and weights of > 1 kg large rainbow trout registered as "Turkish Salmon," (Oncorhynchus mykiss) are four economically important cultured species in the Black Sea In this research, it is aimed (1) to determine the value of the elements, (2) to determine total Hg and Se concentrations, Se/Hg molar ratios, and the HBVSe index, and (3) to calculate the consumer's possible health risk in the edible tissues of cultivated four economically important Black Sea fish species. Fish and diet samples from all species were obtained in 2020 from aquaculture locations in the Black Sea (Sinop, Samsun, and Trabzon cites of Turkey). At the end of the study, the elements in all edible tissues and all the parameters analyzed [Se/Hg, estimated daily intake (EDI), target hazard quotient (THQ), maximum allowable consumption rate (CRlim and CRmm)] in cultured fish tissues were below permissible values. Additionally, when compared with the data of the World Health Organization and The National Academy of Sciences, it determined that consuming cultured fish in the Black Sea adequately meets the daily elemental requirement.

Occurrence of quinolones in cultured fish from Shandong Province, China and their health risk assessment.

The residue levels of 6 quinolones in 160 cultured fish samples from Shandong Province, China were investigated using UPLC-MS/MS. The detection rate was 43.1% and enrofloxacin had the highest detection rate as well as the highest residue concentration. The violation rates were 2.50% for the sum of enrofloxacin and ciprofloxacin and 1.25% for ofloxacin. Among the 9 fish species, quinolone contamination problems should receive more attention in Carp, Grass carp, Crucian and Catfish. The health risk assessment showed that when calculated by the maximum concentration, the estimated daily intakes (EDIs) of Carp, Grass carp and Crucian for the high consumption group accounted for more than 10% of the acceptable daily intakes (ADIs), indicating that a large intake of these fish species might pose a potential health risk and health risk monitoring of quinolones in cultured fish should be continually performed.

Molecularly Imprinted Polymer for a Smart Dispersive Micro-Solid Phase Extraction Technique for Assessing Trace Level Aflatoxins in Cultured Fish.

Molecularly imprinted technology (MIT) consists of preparing materials exhibiting specific recognition cavities to selective mimic the target analytes. The prepared materials promote selective interactions with the targets and avoid interactions of concomitants from complex food, biological, clinical, and environmental matrices. This chapter provides information on a recent development of a vortex-assisted micro-solid phase extraction using a molecularly imprinted polymer (MIP) as an adsorbent for aflatoxins (AFs) determination in cultured fish. MIP particles were synthesized by precipitation polymerization using 5,7-dimethoxycoumarin as a dummy template, methacrylic acid as a functional monomer, divinylbenzene as a cross-linker, and 2,2-azobisisobutyronitrile as an initiator. Polymerization following the precipitation method guarantees homogeneous particle size distribution and the integrity of the imprinted cavities. The MIP microparticles were found to have 5 µm in diameter and a spherical shape. Important parameters such as sample extract pH, adsorption stirring speed and time, desorption stirring speed and time, elution solvent composition and volume, and polymer mass, were fully optimized. The pre-concentration method allows therefore the assessment of four major AFs (B1, B2, G1, and G2) present in cultured fish at very low levels, with pre-concentration factors from 15 to 50 depending of the volume of extract used for performing the dispersive micro-solid phase extraction (D-µ-SPE).

In vitro effects of bisphenol F on antioxidant system indicators in the isolated hepatocytes of rainbow trout (Oncorhyncus mykiss).

Bisphenol F (BPF) has been used frequently in the plastics industry and the production of daily consumer products as an alternative to bisphenol A (BPA). It was aimed herein to determine the cytotoxic effects of BPF on hepatocytes isolated from the liver of rainbow trout (Oncorhyncus mykiss) using lactate dehydrogenase (LDH) assay and antioxidant defence system indicators. The cultured hepatocytes were exposed to seven concentrations (0, 15.63, 31.25, 62.50, 125, 250, and 500 µM) of BPF for 24 h. According to the LDH assay, the percentage of cytotoxicity was increased dose dependently in the cells. The malondialdehyde content, which is indicative of lipid peroxidation, was increased significantly at BPF concentrations between 15.63 and 250 µM, whereas it remained unchanged with a concentration of 500 µM. The activities of superoxide dismutase were increased, while those of catalase were decreased with all of the BPF concentrations. Elevated levels of reduced glutathione content were determined with BPF concentrations between 15.63 and 250 µM, but decreased significantly with a concentration of 500 µM. Significant increases in the activities of the glutathione peroxidase were found in hepatocytes treated with BPF at concentrations of 31.25 to 500 µM. GST activity was only significantly increased with a BPF concentration of 250 µM. The results showed that the toxic mechanism of BPF was mainly based on cell membrane damage and oxidative stress, which have an influence on antioxidant defences. Therefore, BPF should be reconsidered as a safe alternative instead of BPA in the manufacturing of industrial or daily products.

New Species of Rhabdosynochus Mizelle and Blatz 1941 (Monogenoidea: Diplectanidae) From the Gills of the Cultured Centropomus viridis (Perciformes) in the Mexican Eastern Tropical Pacific.

PURPOSE: There are currently nine monogenoidean species of Rhabdosynochus infecting the gill lamellae of wild and cultured centropomid fishes from tropical and subtropical waters of the western Atlantic and eastern Pacific Oceans. The purpose of the present study was to describe the morphological distinctiveness of two new species of Rhabdosynochus found on the cultured Centropomus viridis collected from floating cages from the Mexican eastern Tropical Pacific in 2018. METHODS: monogenoideans were fixed with 4-5% formalin solution, observed and measured as temporary or permanent mounts stained with Gomori's trichrome, and mounted in Canada balsam. Other specimens were mounted on slides using a mixture of lactic acid (LA) and glycerin-ammonium picrate (GAP) and then remounted in Canada balsam to obtain measurements of the haptoral structures and copulatory complex. Illustrations were prepared with the aid of a drawing tube using a Leica microscope DM 2500 with Nomarski interference contrast. RESULTS: Rhabdosynochus viridisi n. sp. is mainly differentiated from all other congeneric species in the shape and size of their copulatory complexes, i.e., length 75-105 µm vs. 45-55 µm in R. alterinstitus, 26-44 µm in R. volucris, 19-22 µm in R. lituparvus, 21-37 µm in R. siliquaus, 48-75 µm in R. hargisi, 37-44 µm in R. hudsoni and 44-61 µm in R. guanduensis. Rhabdosynochus pacificus n. sp. differs from all other species of the genus in having an accessory piece (one subunit) distally twisted. CONCLUSIONS: Based on the morphometric differences of the two new species described above, the number of valid species of Rhabdosynochus has now increased to 11. These two new species of Rhabdosynochus represent the first described species of the genus on C. viridis.

Evaluation of the inhibition of mercury absorption by vegetable juices using a red sea bream intestine model.

The ability of vegetables to inhibit methylmercury absorption was verified, with the aim of lowering the mercury level in cultured fish. Vegetable juice was obtained from 17 varieties of commercial vegetables. A test solution containing 1 µg/g methylmercury, 10% vegetable juice, and 90% physiological saline (v/v) was introduced into the intestinal tract of red sea bream, and the mercury absorption rate was measured. A significant inhibitory effect was observed for green pepper, burdock, and red shiso, mainly in the fraction with a molecular weight >3 kDa. Frozen storage for one month did not affect the inhibitory effect of green pepper; however, the inhibitory effect of frozen burdock and red shiso were destroyed after one week and one month, respectively. During one month of storage in frozen conditions, the inhibitory effect of green pepper was observed in fractions larger than 100 kDa. Molecular weight distribution of the effective fraction varied among the vegetables.

Parasitological diagnosis in food fish produced in a fish farm, in the Zona da Mata, Minas Gerais, southeastern Brazil / Diagnóstico parasitológico em peixes para consumo produzidos em uma psicultura, na Zona da Mata, Minas Gerais, região sudeste do Brasil

Abstract This study identified the parasitic species in juvenile freshwater finfishes during the fattening stage, from a fish farm located in the Zona da Mata (MG), southeastern Brazil, and revealed both macro and microscopical lesions in fish gills. A total of 172 juvenile fishes of different species (Oreochromis niloticus, Ictalurus punctatus, Ctenopharyngodon idella, Cyprinus carpio, Astyanax bimaculatus and Brycon amazonicus) were transported to a laboratory in São Paulo city. The fish were sedated and then euthanized for parasitological analysis. All fish were infected by at least one parasite species. Ten different species of parasites were identified: Apiosoma sp., Epistylis sp., Ichthyobodo sp., trichodinids, Piscinoodinium pillulare, Ichthyophthirius multifiliis, Tetrahymena sp., monogeneans, Centrocestus formosanus metacercariae, and Dermocystidium sp. The best management practices and lack of sanitary control were also discussed.

Resumo Este trabalho identificou espécies parasitas em peixes de produção juvenis de água doce, durante a fase de engorda, oriundos de uma piscicultura da Zona da Mata (MG), na região sudeste do Brasil, além das lesões de brânquias, causadas tanto macro quanto microscopicamente. Um total de 172 peixes juvenis de diferentes espécies (Oreochromis niloticus, Ictalurus punctatus, Ctenopharyngodon idella, Cyprinus carpio, Astyanax bimaculatus e Brycon amazonicus) foram transportados para um laboratório na cidade de São Paulo. Os peixes foram anestesiados e eutanasiados para análise parasitológica. Todos os peixes estavam acometidos por pelo menos uma espécie de parasito. Dez diferentes espécies de parasitos foram identificadas: Apiosoma sp., Epistylis sp., Ichthyobodo sp., tricodinídeos, Piscinoodinium pillulare, Ichthyophthirius multifiliis, Tetrahymena sp., monogeneas, metacercárias de Centrocestus formosanus e Dermocystidium sp. As boas práticas de manejo e inadequado controle sanitário também foram discutidos.

Virulence-associated genes and antibiotic resistance patterns of Vibrio spp. isolated from cultured marine fishes in Malaysia.

BACKGROUND: Vibriosis is an important bacterial disease of cultured marine fishes worldwide. However, information on the virulence and antibiotic resistance of Vibrio spp. isolated from fish are scarce. This study investigates the distribution of virulence associated genes and antibiotic resistance patterns of Vibrio spp. isolated from cage-cultured marine fishes in Malaysia. RESULTS: A total of 63 Vibrio spp. isolated from 62 cultured marine fishes in various geographical regions in Peninsular Malaysia were analysed. Forty-two of the isolates (66.7%) were positive for all chiA, luxR and vhpA, the virulence genes produced by pathogenic V. harveyi. A total of 62 Vibrio isolates (98%) had tlh gene of V. parahaemolyticus, while flaC gene of V. anguillarum was detected in 43 of isolates (68%). Other virulence genes, including tdh, trh, hlyA and toxRvc were absent from any of the isolates. Multiple antibiotic resistance (MAR) was exhibited in all strains of Harveyi clade, particularly against ampicillin, penicillin, polypeptides, cephems and streptomycin. The MAR index ranged between 0.06 and 0.56, and 75% of the isolates have MAR index of higher than 0.20. Host species and geographical origin showed no correlation with the presence of virulence genes and the antibiotic resistance patterns of Vibrio spp. CONCLUSIONS: The study indicates that majority of Vibrio spp. isolated from cultured marine fishes possess virulence genes, but were not associated with human pathogen. However, the antibiotics resistance is a real concern and warrants ongoing surveillance. These findings represent an updated knowledge on the risk of Vibrio spp. to human health, and also provides valuable insight on alternative approaches to combat vibriosis in cultured fish.

TarSynFlow, a workflow for bacterial genome comparisons that revealed genes putatively involved in the probiotic character of Shewanella putrefaciens strain Pdp11.

Probiotic microorganisms are of great interest in clinical, livestock and aquaculture. Knowledge of the genomic basis of probiotic characteristics can be a useful tool to understand why some strains can be pathogenic while others are probiotic in the same species. An automatized workflow called TarSynFlow (Targeted Synteny Workflow) has been then developed to compare finished or draft bacterial genomes based on a set of proteins. When used to analyze the finished genome of the probiotic strain Pdp11 of Shewanella putrefaciens and genome drafts from seven known non-probiotic strains of the same species obtained in this work, 15 genes were found exclusive of Pdp11. Their presence was confirmed by PCR using Pdp11-specific primers. Functional inspection of the 15 genes allowed us to hypothesize that Pdp11 underwent genome rearrangements spurred by plasmids and mobile elements. As a result, Pdp11 presents specific proteins for gut colonization, bile salt resistance and gut pathogen adhesion inhibition, which can explain some probiotic features of Pdp11.

Pharmaceuticals and personal care products in water, sediments, aquatic organisms, and fish feeds in the Pearl River Delta: Occurrence, distribution, potential sources, and health risk assessment.

This study investigated the occurrence, distribution, and potential sources of 34 pharmaceuticals and personal care products (PPCPs) in water, sediments, aquatic organisms (fish and shellfish), and fish feeds from the mariculture areas of the Pearl River Delta (PRD). The health risk presented by this class of compounds was also assessed in relation to their intake via seafood consumption. Of the 34 PPCPs, a total of 9, 21, 14, and 28 PPCPs were detected in water, sediments, fish feeds, and aquatic organisms, respectively. Trimethoprim, norfloxacin, ofloxacin, and spectinomycin were detected in all matrices. The levels of PPCPs in water and sediment samples were relatively low. Spectinomycin, paracetamol, ciprofloxacin, norfloxacin, and ibuprofen were the most frequently detected PPCPs in feeds. Ibuprofen and ketoprofen were widely detected in aquatic organisms, with average concentrations of 562 and 267 ng/g wet weight, respectively. The residual levels of PPCPs in shellfish such as ME (mussel, Mytilus edulis) and OS (oyster, Ostrea gigas) were significantly higher (p < 0.05) than those in other species including CA (topmouth culter, Culter alburnus) and EO (orbfish, Ephippus orbis). Correlation analysis indicated that the medicated feeds were a potential source of PPCPs in the mariculture areas of the PRD, but other anthropogenic sources should not be ignored. Based on maximum residue limits and acceptable daily intake, the health risks presented to humans via seafood consumption are negligible. However, as multiple antibiotics were frequently detected in the mariculture environment, aquatic organisms, and feeds, the induction and dissemination of antimicrobial resistance associated with antibiotic usage in aquaculture would be of great concern. It is necessary to establish a centralized management system and control the use of veterinary drugs in mariculture to protect the aquaculture environment and ensure the safety of seafood.

Phthalate esters distribution in coastal mariculture of Hong Kong, China.

The aim of the study is to evaluate the impact of mariculture on phthalate esters speciation and distribution in sediments and cultured fish in the Hong Kong regions and near mainland China. Concentrations of ∑phthalate esters in mariculture surface sediments (0 to 5 cm) ranged from 0.20 to 54.3 mg/kg dw (mean 10.3 mg/kg dw), with the highest recorded at M2 (20.4 mg/kg dw). Concentrations of phthalate esters were not significantly (p > 0.05) enriched in surface and sediment cores at mariculture sites relative to the reference sediments, 1 to 2 km away in areas without mariculture activities. Among different congeners, only butyl benzyl phthalate (BBP) concentrations demonstrated a significant correlation (R2 = 0.40, p < 0.05) with TOC values of sediments. The median concentrations of di-2-ethylhexyl phthalate (DEHP) and di-n-butyl phthalate (DBP) in the sediments were 1.57 and 6.96 times higher than the environmental risk levels (ERL), which may pose environmental risks. Results of health risk assessments revealed that the cultured fish (snubnose pompano, orange-spotted grouper, and red snapper) were safe for consumption, in terms of phthalate esters. This is the first study to assess the differences of phthalate esters contamination between mariculture and natural coastal sediments.

Occurrence and risk assessment of fluoroquinolones and tetracyclines in cultured fish from a coastal region of northern China.

Occurrence and levels of 11 fluoroquinolones (FQs) and four tetracyclines (TC) in 14 cultured fish species from a coastal city in the northern China were investigated. Five FQs (ofloxacin, enoxacin, ciprofloxacin, enrofloxacin, and sarafloxacin) and oxytetracycline were detected. Lower detection frequencies of antibiotics were observed in the marine fish. The concentrations of ΣFQs ranged from not detectable (nd) to 130 ng/g wet weight (ww) (median, 7.2 ng/g ww), and the concentration range of ΣTCs was nd to 200 ng/g ww (median, nd ng/g ww). The Chinese snakehead contained the highest concentrations of ΣFQs (130 ng/g ww) and the small yellow croaker accumulated the highest concentrations of ΣTCs (200 ng/g ww), respectively. Although the calculated estimated daily intakes (EDI) suggested that the consumption of these cultured fish from this region was not associated with significant human health risks, this study provides useful information that will be helpful in the appropriate antibiotic use in aquaculture. To our knowledge, this can be the first report on the occurrence and levels of antibiotics in cage-cultured marine fish from the Bohai Rim region, China.

Human and veterinary pharmaceuticals in the marine environment including fish farms in Korea.

The occurrence trends and effects of 30 human and veterinary pharmaceuticals, including antibiotics, anthelmintics, anti-inflammatory drugs, and ß-blockers, in the marine environment, with a focus on seawater, sediment, cultured fish, and their feed collected from coastal and fish farm areas in the southern sea of Korea, were investigated. The concentrations of total pharmaceuticals in coastal area seawater (mean: 533ng/L) were higher than those in fish farm seawater (mean: 300ng/L), while the opposite trend (coastal area: 136ng/gdrywt

First records of Ambiphrya and Vorticella spp. (Protozoa, Ciliophora) in cultured Nile tilapia (Oreochromis niloticus) in the central region of Saudi Arabia.

The present study was carried out as part of an ongoing general survey seeking to uncover protozoan parasites infecting cultured tilapia in the central region of Saudi Arabia. In the sample of 400 specimens of tilapia (Oreochromis niloticus) 30 were infested with Ambiphrya ameiuri simultaneously with Vorticella sp. Morphometric criteria were used to describe and identify these species and this study presents the first records of these species among cultured fish in Saudi Arabia.

Presence of Contracaecum spp. in teleosts cultured and fished in Sardinia.

This study reports the results of the finding of Contracaecum spp. during a survey on endoparasites isolated from cultured and wild fish and also from some cephalopods caught in Sardinian waters. Contracaecum spp. is a nematode belonging to the Anisakidae, and is reported to cause zoonosis in humans. Nematodes were detected after visual inspection and enzymatic digestion and then identified by morphologic observation, which was confirmed by PCR. The results show that Contracaecum spp. were found in both fish caught from sea or lagoon, and in both cultured and wild fish: 33 of the parasitized samples were wild fish (24 caught in the sea and 9 in lagoons) and 11 were cultured ones. The prevalence of Contracaecum spp. was higher in Diplodus spp. (16.0%), Sparus aurata (15.8%) and Mullus spp. (14.6%). Larvae were also found by enzymatic digestion at muscular level in 5 species, with the highest prevalence in S. aurata (10.5%). The results of this study indicate that Contracaecum spp. was present in cultured fish such as S. aurata, Diplodus spp. and Dicentrarchus labrax. All cultured fish with parasites were collected from land-based semi-intensive tanks whose water came from an adjacent lagoon. Finally, the evidence that this parasite is found in both cultured and wild fish leads us to re-consider the zoonotic potential of Contracaecum spp., in particular when one bears in mind its dimensions at the L3 stage, when it is barely visible to the human eye.