Freshwater Fish Research Articles

Prevalence and antimicrobial susceptibility characterization of Escherichia coli isolated from the intestines of freshwater fish from the Mindu dam in Morogoro, Tanzania

Background: The Mindu dam, an open system, serves as a microbial reservoir, housing uropathogenic microorganisms like Escherichia coli. Morogoro municipality relies on the dam for water and fish. This study aimed to assess the antimicrobial susceptibility of Escherichia coli isolated from fish and water in Mindu dam. Methods: In May 2019, a cross-sectional study was conducted in Morogoro municipality. Selected antimicrobials, including ampicillin, tetracycline, cefoxitin, erythromycin, trimethoprim, gentamicin, enrofloxacin, and nalidixic acid, were tested using the disc diffusion method. E. coli ATCC 25922 served as a positive control for quality assurance during bacterial isolation. The potential source of antimicrobial contamination was identified through the multiple antibiotic resistance index. Results: Out of 148 samples, E. coli was found in 24, none in water. E. coli showed high resistance (≥50.0%) to erythromycin (62.5%), nalidixic acid (79.2%), and ampicillin (75%). Additionally, 58.0% of isolates displayed multi-drug resistance across antimicrobial classes, with a multiple antibiotic resistance index ranging from 0.00 to 0.75. Conclusion: Antimicrobial-resistant E. coli in the food chain may heighten the spread of complex urinary tract infections in the municipality. It underscores the necessity for robust municipal surveillance of antimicrobial resistance and effective antimicrobial stewardship for enhanced infection prevention and control.

A Comparative Evaluation of eDNA Metabarcoding Primers in Fish Community Monitoring in the East Lake

East Lake in Wuhan, China, harbors a high number of freshwater fish species of great conservation value, concurrently serving as vital resources for local livelihoods. However, the ecosystem is threatened by an array of anthropogenic activities, thus requiring consistent monitoring of the local fish community to enable more efficacious conservation management. In place of conventional surveying methods, we undertook the first analysis of the fish distribution within East Lake via metabarcoding of environmental DNA (eDNA). The accuracy and efficacy of eDNA metabarcoding rely heavily upon selecting an appropriate primer set for PCR amplification. Given the varying environmental conditions and taxonomic diversity across distinct study systems, it remains a challenge to propose an optimal genetic marker for universal use. Thus, it becomes necessary to select PCR primers suitable for the composition of fish in the East Lake. Here, we evaluated the performance of two primer sets, Mifish-U and Metafish, designed to amplify 12S rRNA barcoding genes in fishes. Our results detected a total of 116 taxonomic units and 51 fish species, with beta diversity analysis indicating significant differences in community structure diversity between the six sampling locations encompassing East Lake. While it was difficult to accurately compare the species-level discriminatory power and amplification bias of the two primers, Mifish outperformed Metafish in terms of taxonomic specificity for fish taxa and reproducibility. These findings will assist with primer selection for eDNA-based fish monitoring and biodiversity conservation in the East Lake and other freshwater ecosystems.

A multidisciplinary study on Clinostomum infections in Nile tilapia: micro-morphology, oxidative stress, immunology, and histopathology

Yellow grub disease, caused by Clinostomum metacercaria, is an endemic zoonotic infection in freshwater fish, responsible for Halzoun syndrome transmitted through the consumption of raw infected fish. This study aimed to conduct a multidisciplinary investigation integrating detailed morphology, oxidative stress, immunology, and histopathology alteration to advance our understanding of Clinostomum infection. In this annual study, 400 Nile tilapia (Oreochromis niloticus) were collected from the Nile River at Al Bahr Al Aazam, Giza Governorate to assess Clinostomum infection prevalence. Of the examined fish, 160 individuals (40.0%) harboured larval Clinostomum infections. Clinostomum metacercariae were observed in various anatomical locations, with 135 fish (33.8%) in buccal cavities, 21 fish (5.25%) in gill chambers, and 4 fish (1.0%) on the skin. Infection intensity ranged from 2 to 12 cysts per fish, averaging 5 cysts, notably with skin infections characterized by a single cyst in each fish. Macroscopic encysted metacercariae were collected from buccal cavities, gills, and skin. Micro-morphology revealed distinct features in C. complanatum, including an elliptical oral sucker with collar-like rings and large sensory papilla-like structures, contrasting with the absence of these features in C. phalacrocoracis. Oxidative stress, assessed through malondialdehyde (MDA) and nitric oxide levels, revealed an elevation in MDA to 35.13 ± 6 nmol/g and nitric oxide to 25.80 ± 3.12 µmol/g in infected fish. In infected fish, MHC-I gene expression increased approximately 13-fold, MHC-II peaked at 19-fold, and IL-1β significantly upregulated by 17-fold, compared to control levels. Histopathology detailed associated lesions, such as cyst encapsulation and eosinophilic infiltration. Clinstomiasis and its impacts on fish hosts offer crucial insights to control this emerging fish-borne zoonotic disease, threatening wildlife, aquaculture, and human health.

Taxonomic revision of the Pirate Perches, Aphredoderus, (Percopsiformes: Aphredoderidae) with descriptions of two new species.

Pirate Perches, Aphredoderus, are a widespread lowland freshwater fish native to the Eastern half of the United States. Aphredoderus was thought to contain a single species divided into an Eastern and Western subspecies on either side of the Appalachian Mountains with a widespread intergrade zone through much of the Eastern Gulf of Mexico and Southern Atlantic drainages. We use morphology and genetic data from specimens spanning the entire range of the genus to determine species limits within Aphredoderus. We find evidence of five species, four of which exhibit widespread sympatry in the Southeastern United States. We elevate A. sayanus sayanus and A. sayanus gibbosus to species, redescribe A. mesotrema, supplement previous descriptions, and describe two new species, A. retrodorsalis, and A. ornatus.

Henneguya (Cnidaria: Myxobolidae) species infecting Oligosarcus jenynsii (Characiformes: Characidae) in a Neotropical shallow lake from Argentina: morphological and molecular characterisation.

Two previously undescribed myxozoan species, Henneguya sardellae sp. n. and H. margaritae sp. n., found infecting connective tissues of the Neotropical characid fish Oligosarcus jenynsii (Günther) from Argentina are morphologically and molecularly characterised. Mature spores of H. sardellae sp. n. are ellipsoid, with two, straight and visibly fused caudal appendages cleaved at its blunt terminal end; measuring 33.5 ± 1.2 (30.9-35.5) μm in total length, spore body 17.5 ± 0.6 (16.3-18.6) µm, 7.8 ± 0.4 (7.0-8.8) µm wide and 6.9 ± 0.2 (6.6-7.2) µm thick, with two elongated, unequally-sized polar capsules situated at anterior end, and 11-13 turns of polar tubules. Mature spores of H. margaritae sp. n. are pyriform, with two caudal appendages visible fused together and much longer than spore body, with unequal endings; measuring 35.9 ± 2.8 (29.2-40.7) µm in total length, spore body 11.5 ± 0.9 (9.2-13.0) µm long, 5.8 ± 0.4 (5.1-6.7) µm wide and 5.5 ± 0.2 (5.1-5.8) µm thick, with two polar capsules similar in size, pyriform polar capsules containing polar tubules with 4-5 coils. Both species showed a membraneous sheath surrounding the spore body and caudal appendages; in H. sardellae sp. n. this feature can deploy laterally. Phylogenetic analyses based on SSU rDNA sequences showed that H. sardellae sp. n. and H. margaritae sp. n. clustered with other myxobolids parasitising Characiformes in Brazil, Cichliformes in Mexico and Cyprinodontiformes in Mexico and the United States. The description of these two new species of Henneguya as the first described species of the genus that parasitise freshwater fish in Argentina highlights the importance of further research on the diversity and distribution of myxozoans in this region.

Conservation and Population Status of Freshwater Fishes from Kelekar River, South Sumatra, Indonesia

This study assessed the conservation and population status of the fish species collected from the Kelekar river. This study was conducted between May 2022 and June 2023. A total of 1.267 fish belonging to 49 species and 19 families were recorded. Cyprinidae was found to be the richest family (18 species) followed by Osphronemidae (5 species), Channidae (4 species), Siluridae (3 species), Bagridae, Claridae, Pangasidae, Pristolepidae (2 species), and others (1 species). The Shannon-Weiner diversity index was 3.547. The Simpson’s dominance index was 0.035, and the Evenness index was 0.921. The biodiversity index of the Kelekar river was in the high category. Base on the latest IUCN red list, one species as data deficient, forty-three species as least concern, two species as near threatened, two species as vulnerable, and one species as critically endangered. The IUCN based population status of fish species present in the Kelekar River was unknown (35%), stable (47%), and decreasing (18%).

Comparative analyses of gut microbiota reveal ammonia detoxification and nitrogen assimilation in Cyprinus carpio var. specularis.

The complex niche of fish gut is often characterized by the associated microorganisms that have implications in fish gut-health nexus. Although efforts to distinguish the microbial communities have highlighted their disparate structure along the gut length, remarkably little information is available about their distinct structural and functional profiles in different gut compartments in different fish species. Here, we performed comparative taxonomic and predictive functional analyses of the foregut and hindgut microbiota in an omnivorous freshwater fish species, Cyprinus carpio var. specularis, commonly known as mirror carp. Our analyses showed that the hindgut microbiota could be distinguished from foregut based on the abundance of ammonia-oxidizing, denitrifying, and nitrogen-fixing commensals of families such as Rhodospirillaceae, Oxalobacteraceae, Nitrosomonadaceae, and Nitrospiraceae. Functionally, unique metabolic pathways such as degradation of lignin, 2-nitrobenzoate, vanillin, vanillate, and toluene predicted within hindgut also hinted at the ability of hindgut microbiota for assimilation of nitrogen and detoxification of ammonia. The study highlights a major role of hindgut microbiota in assimilating nitrogen, which remains to be one of the limiting nutrients within the gut of mirror carp.

Assessment of microplastic contamination in commercially available fishes

Plastics have widespread applications for human use, but their disposal poses a significant threat to living organisms and these plastics end up in the marine environment. They will be fragmented into small pieces as a result of ultraviolet exposure, climatic changes, and temperature changes; Microplastics (MPs) are plastics that are less than 5 mm in size. The level of MP (Microplastic) pollution in commercially harvested fish from different habitant in Vellore, India is currently unknown. Therefore, this study aimed to determine the presence and characteristics of ingested or inhaled MPs in marine and freshwater fishes highly consumed by the local population. Fish gills and gastrointestinal tracts were aseptically dissected and digested (30% hydrogen peroxide), then filtered and examined under a microscope for the presence of MPs. Further analysis was performed on the samples using Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR) and Scanning Electron Microscopy (SEM) with Energy Dispersive X-Ray (EDAX). Of the samples analysed, a total of 875 MPs were recovered from 32 fishes, with 478 from marine fishes and 397 from freshwater fishes. The most common colours of the MPs were blue and black, while stereo microscopy analysis revealed that the majority of MPs were fibers (91%), followed by fragments (8%) and a small number of films. The ATR-FTIR analysis identified polyvinyl alcohol (39.76%), polyethylene (16.51%), methylcellulose (12.84%) and styrene (9.07%), as the predominant types of MPs in the fish samples. This study highlights the significant impact of MP pollution on marine ecosystems. The research provides insight into the nature and extent of MPs in fish from both marine and freshwater habitats, with an aim for policies and interventions aimed to reduce plastic pollution in the locality.

EDNA Metabarcoding: Gaps of Publicly Available Reference Databases of Freshwater Vertebrates in Sri Lanka

Environmental DNA (eDNA) has become a rapidly emerging technology during the last decade. Although eDNA applications are used worldwide, none of the eDNA metabarcoding studies have been conducted solely in Sri Lanka yet. Detection rate of species through DNA metabarcoding technique varies with the availability of gene sequences in reference databases. Therefore, the main objective of this study was to assess the data gaps of local species in two selected reference databases (the NCBI database and the BOLD database). Local freshwater vertebrates were considered for this analysis by dividing them into three major categories as freshwater fish (97 species), freshwater amphibians (40 species) and freshwater reptiles (7 species). The availability of COI gene sequence and CytB gene sequence of each species were recorded. In the NCBI database, reference gene sequences were available only for 17.53% (17 species) of freshwater fish, 7.50% (3 species) of freshwater amphibians and 14.29% (1 species) of freshwater reptiles. In the BOLD database, reference gene sequences were available for 52.58% (51 species) of freshwater fish, 17.50% (7 species) of freshwater amphibians and 42.86% (3 species) of freshwater reptiles. Endemic species were analyzed separately. This revealed the availability of reference gene sequences for 4.92% (3 species) of endemic freshwater fish in the NCBI database. However, none of the reference gene sequence was found for endemic freshwater amphibians and reptiles. In the BOLD database, reference gene sequences were available for 36.07% (22 species) of endemic freshwater fish and 33.33% (1 species) of endemic freshwater reptiles but none for endemic freshwater amphibians. According to the results, large gaps of publicly available reference CO1 gene sequences were observed in both NCBI and BOLD databases. Not only the availability of reference gene sequences, but also the submitted countries of those sequences were also analyzed as a part of this study. Asia has contributed the most according to the country-vice analysis. Among them, many are in the South Asian region. Therefore, this study highlights the need of expanding reference databases for greater local representation as well as the importance of forging collaborations with the developed countries in the same region in order to increase international cooperation and funding to carry on further studies on neglected taxa and under‐represented taxa in Sri Lanka. 
 Keywords: eDNA (environmental DNA), Metabarcoding, Reference databases, Reference gene sequences

Global freshwater fish invasion linked to the presence of closely related species

In the Anthropocene, non-native freshwater fish introductions and translocations have occurred extensively worldwide. However, their global distribution patterns and the factors influencing their establishment remain poorly understood. We analyze a comprehensive database of 14953 freshwater fish species across 3119 river basins and identify global hotspots for exotic and translocated non-native fishes. We show that both types of non-native fishes are more likely to occur when closely related to native fishes. This finding is consistent across measures of phylogenetic relatedness, biogeographical realms, and highly invaded countries, even after accounting for the influence of native diversity. This contradicts Darwin’s naturalization hypothesis, suggesting that the presence of close relatives more often signifies suitable habitats than intensified competition, predicting the establishment of non-native fish species. Our study provides a comprehensive assessment of global non-native freshwater fish patterns and their phylogenetic correlates, laying the groundwork for understanding and predicting future fish invasions in freshwater ecosystems.

Broad-scale perspective on body size-trophic position patterns of freshwater fishes at the intraspecific and community levels from individual-based data in China

As predators are generally larger than their prey, positive body size and trophic position relationships are typically assumed, and these relationships have helped in estimating and predicting the effects of global environmental change on the trophic dynamics of aquatic ecosystems. However, current efforts to confirm the generality of body size-trophic position relationships have focused mainly on interspecific patterns using species-aggregated and averaged data, and little effort has been devoted to assessing how trophic position scales with body size at the intraspecific and community levels as well as exploring its ecological drivers, particularly for fishes in freshwater ecosystems. To fill this gap, we present a broad-scale study of body size-trophic position relationships in freshwater ecosystems at both the intraspecific and community levels using an individual-based body size and isotopic signature dataset that includes 2564 samples of 65 fish species in China. Our results indicate that body size-trophic position patterns at both the intraspecific and community levels can be positive, negative or insignificant. Non-significant patterns predominated at both levels, with slightly more than 30 % showing positive relationships, which highlights that the positive body size-trophic position pattern is not universally prevalent and that body size should be used with caution as a proxy for the trophic position of fishes at both the intraspecific and community levels in freshwater ecosystems. No factor was related to body size-trophic position patterns at the community level, while habitat type, elevation, temperature and fishing pressure were identified as the key determinants of body size-trophic position patterns at the intraspecific level, implying that fishing and climate warming can shift the food-web size structure of freshwater ecosystems by affecting population trophic dynamics.

Assessment of Heavy Metal Accumulation in Freshwater Fishes in Irrigation Tanks, Anuradhapura


 Sri Lanka is well known for its dense freshwater irrigation tanks, especially in dry zones, which play an important role in sustaining both the environment and human livelihoods by providing essential resources such as drinking water, irrigation for agriculture, and nutrition through fish proteins. However, heavy metal contamination poses a significant threat to the sustainability of these valuable freshwater irrigation tanks due to their persistence and non-biodegradability. These heavy metals tend to accumulate in living organisms and biomagnified and have harmful effects on high tropic level organisms like humans who rely on aquatic fish species via food chains. This study was carried out to assess the heavy metal accumulation in muscle and liver parts of freshwater fishes, Tilapia (Oreochromis sp.), and striped snakehead; Lula (Channa striata) in Kammalakkulama, Hammillewa, Kirindegama, and Madawalagama tanks in Anuradhapura district, Sri Lanka. The Inductively coupled plasma mass spectrometry (ICP-MS) was used to analyse the randomly collected fish samples for total Cadmium (Cd), Arsenic (As), Lead (Pb), and Mercury (Hg) in the tanks. The results indicated a significant difference (p<0.05) in the detected concentrations of heavy metal levels in tested samples among the four tanks. The highest concentrations of Cd (2.098 mg/kg) and As (0.366 mg/kg) were recorded in the liver of Tilapia in the Kammalakkulama tank. The highest Pb (5.509 mg/kg) and Hg (2.015 mg/kg) concentration was recorded in the Tilapia muscle in the Hammillewa tank and the Lula muscle in the Madawalagama tank respectively exceeding the WHO/FAO standards. The accumulation levels of these heavy metals were higher in the fish liver compared to their muscle in most of the samples. Arsenic (As) showed a significant positive linear relationship (R2=0.531) between their dry body weight and accumulated concentration in the Tilapia muscle. The canonical discriminant analysis showed a significant differentiation among the four tanks concerning heavy metal levels in fish tissues.
 
 Keywords: Channa striata, Heavy metals, Irrigation tanks, ICP-MS, Oreochromis sp.

Digest: Habitat and competition drive diversification in Gondwanan fish.

Why do some clades diversify more than others? And what are the roles of abiotic and biotic factors in driving such unequal diversification? To explore this, Burns et al. (2024) investigate variation in body morphology in a group of freshwater fish, Characiformes, across South America and Africa. They find that habitat features such as slope, elevation, and flow rates have played a major role in the diversification of morphological variation in the Characiformes. The Neotropical lineages residing in significantly more diverse slope and elevation habitats were found to display more body shape diversity than African lineages. The authors also report that competition might have limited the morphological variation in African Characiformes compared to the Neotropical lineages. This study highlights the importance of exploring both abiotic and biotic factors when examining diversification.

Biomarker selection depends on gene function and organ: the case of the cytochrome P450 family genes in freshwater fish exposed to chronic pollution.

Pollution and its effects have been of major concern in recent decades. Many strategies and markers have been developed to assess their effects on biota. Cytochrome P450 (CYP) genes have received significant attention in this context because of their relationship with detoxification and activation of exogenous compounds. While their expression has been identified as a pollution exposure biomarker, in most cases, it has been tested only after acute exposures and for CYP genes associated with exogenous compounds. To elucidate CYP gene expression patterns under chronic pollution exposure, we have used the silverside Basilichthys microlepidotus as a model, which inhabits the Maipo River Basin, a freshwater system with different pollution levels. We performed next-generation RNA sequencing of liver and gill tissues from polluted and non-polluted populations. We found most CYP genes were not dysregulated by pollution, and the seven genes that were present and differentially expressed in liver and gill were mainly downregulated. Three CYP genes associated with exogenous compounds showed differential expression in the gill, while four CYP genes associated with endogenous compounds showed differential expression in the liver. The findings presented here highlight the importance of CYP genes, his family, tissues and his interaction in the context of pollution biomarkers use.

Influence of host phylogeny and water physicochemistry on microbial assemblages of the fish skin microbiome.

The skin of fish contains a diverse microbiota that has symbiotic functions with the host, facilitating pathogen exclusion, immune system priming, and nutrient degradation. The composition of fish skin microbiomes varies across species and in response to a variety of stressors, however, there has been no systematic analysis across these studies to evaluate how these factors shape fish skin microbiomes. Here, we examined 1922 fish skin microbiomes from 36 studies that included 98 species and nine rearing conditions to investigate associations between fish skin microbiome, fish species, and water physiochemical factors. Proteobacteria, particularly the class Gammaproteobacteria, were present in all marine and freshwater fish skin microbiomes. Acinetobacter, Aeromonas, Ralstonia, Sphingomonas and Flavobacterium were the most abundant genera within freshwater fish skin microbiomes, and Alteromonas, Photobacterium, Pseudoalteromonas, Psychrobacter and Vibrio were the most abundant in saltwater fish. Our results show that different culturing (rearing) environments have a small but significant effect on the skin bacterial community compositions. Water temperature, pH, dissolved oxygen concentration, and salinity significantly correlated with differences in beta-diversity but not necessarily alpha-diversity. To improve study comparability on fish skin microbiomes, we provide recommendations for approaches to the analyses of sequencing data and improve study reproducibility.

Effects of lipid extraction on stable isotope ratios of carbon and nitrogen in muscles of freshwater fish

ABSTRACT The extraction of lipids by the Folch method from the muscles of all the fish studied led to statistically significant differences in the values of δ 15N. At the same time, lipid extraction led to a statistically significant increase in δ 13C in pike and roach, and to a statistically insignificant decrease in δ 13C in perch and bream. Thus, lipid extraction cannot serve as a universal method of sample preparation for the analysis of the isotopic composition of carbon (13C/12C) and nitrogen (15N/14N) in fish muscles. The differences between the δ 13C values in the samples before and after lipid extraction were statistically investigated by different models. It is shown that mathematical correction method models can be used, but the results are depending on the fish types.

A Comparative Study of Freshwater Fish Burgers Made from Three Amazonian Species: Omega 3 Fortification and Sodium Reduction.

This study aimed to formulate burgers made from three Amazonian fish species: pacu (Pyaractus brachypomus), boquichico (Prochilodus nigricans), and bujurqui (Chaetobranchus flavescens), focusing on sodium reduction and fortification with fish oil microparticles (FOM) rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The proximal composition, sodium and calcium content, instrumental texture profile, fatty acid profile, sensory profile, and overall liking were evaluated. Differences in proximal composition and fatty acid profiles between the fillets were reflected in the burgers. Fortification with FOM increased EPA and DHA in the burgers; thus, they can be considered "high in omega-3 fatty acids" and reduced the n-6/n-3 ratio below 4. There were sensory attributes that could be related to lipid oxidation but reduced overall liking for less than 10% of consumers. Nevertheless, certain sensory attributes (grilled, characteristic, aromatic, tasty, tender, and juicy) had a positive impact on the overall liking of more than 20% of consumers, yielding adequate scores (between 5.60 and 5.71) on the 9-point hedonic scale. The production process must be optimized by knowing the fish fillet quality in depth, improving the FOM and burgers' oxidative stability, and achieving an adequate sensory and hedonic profile by employing consumers' vocabulary to characterize new products.

Formation of Male Sex Tilapia Larvae (Oreochromis niloticus) Using Natural Methyltestosterone Hormone

Aims: Obtaining a productive tilapia, by changing the sex of the tilapia larvae into male monosex. This was achieved by determining the effect of the duration of administrating the natural methyltestosterone (MT) hormone on the male sex of tilapia using the Immersion method.
 Methodology: The study was conducted at the Northern Region Marine and Fisheries Service Branch, Wanayasa Freshwater Fish Seed Development Center, Purwakarta Regency, West Java Province, Indonesia. The experimental design used a completely randomized design (CRD) with 5 treatments and 3 replications. The study used the Immersion method where untreated tilapia and the four hormones tilapia. The treatments are as follows:
 A = untreated tilapia
 B = tilapia treated in 17a-MT (5h-100mg/L)
 C = tilapia treated in MT (5h-350mg/L)
 D = tilapia treated in MT (10h-350mg/L)
 E = tilapia treated in MT (15h-350mg/L)
 Results: Tilapia larvae used were aged 6 days after hatching and reared for 40 days, then the male and SR sex were observed. The results of this study showed that the duration of immersion of tilapia larvae using natural MT significantly affected the male tilapia which resulted in the highest male percentage at 15 h of immersion of 80.2%, then followed by 10-h immersion of 75.6% with the same SR value of about 90%.
 Conclusion: Administration of natural MT using cow testis meal by immersion method at different durations significantly affects male sexual development in 6-day-old tilapia larvae. Based on the obtained data analyses, the percentage of male tilapia in the 40-day experimental study shows the most effective duration in treatment E (15 h) with a measure of 350 mg/l, which is 80.2%. Further studies regarding the relationship between sex change and the growth of sex-changed fish as an effect of immersion duration.

Apparent migration and natal homing of a small minnow in a large ancient lake

Migratory freshwater fish species are particularly susceptible to population decline because they rely on multiple habitats and open passages to complete their life cycle. Understanding the migratory patterns of cyprinids is thus essential for their management and conservation. Gnathopogon caerelescens, a small cyprinid species endemic to Lake Biwa (Japan), is one of the most important targets of inland fisheries. However, their abundance has dramatically decreased. By using an extensive alizarin complexone-mark-and-recapture program for of G. caerelescens juveniles, together with the collaboration of commercial fishermen, this cyprinid fish was demonstrated as: (1) migrating from a nursery lagoon toward the main lake and moving along the near-shore from spring to autumn; (2) migrating from the near-shore shallow habitats to off-shore deep habitats from autumn to winter; and (3) migrating to a nursery lagoon from off-shore deep habitats from winter to spring. These results highlighted the importance of protecting individual spawning/nursery habitats, and maintaining connectivity between each spawning/nursery habitat and the lake system.

Molecular identification, phylogenetic analysis and histopathological study of pathogenic free-living amoebae isolated from discus fish (Symphysodon aequifasciatus) in Iran: 2020–2022

Free-living amoebae (FLA) are capable of inhabiting diverse reservoirs independently, without relying on a host organism, hence their designation as “free-living”. The majority of amoebae that infect freshwater or marine fish are amphizoic, or free-living forms that may colonize fish under particular circumstances. Symphysodon aequifasciatus, commonly referred to as the discus, is widely recognized as a popular ornamental fish species. The primary objective of the present study was to determine the presence of pathogenic free-living amoebae (FLA) in samples of discus fish. Fish exhibiting clinical signs, sourced from various fish farms, were transferred to the ornamental fish clinic. The skin, gills, and intestinal mucosa of the fish were collected and subjected to culturing on plates containing a 1% non-nutrient agar medium. The detection of FLA was conducted through morphological, histopathological and molecular methods. The construction of the phylogenetic tree for Acanthamoeba genotypes was achieved using the maximum likelihood approach. The molecular sequence analysis revealed that all cultures that tested positive for FLA were T4 genotype of Acanthamoeba and Acanthamoeba sp. The examination of gill samples using histopathological methods demonstrated the presence of lamellar epithelial hyperplasia, significant fusion of secondary lamellae, and infiltration of inflammatory cells. A multitude of cysts, varying in shape from circular to elliptical, were observed within the gills. The occurrence of interlamellar vesicles and amoeboid organisms could be observed within the epithelial tissue of the gills. In the current study, presence of the Acanthamoeba T4 genotype on the skin and gills of discus fish exhibiting signs of illness in freshwater ornamental fish farms was identified. This observation suggests the potential of a transmission of amoebic infection from ornamental fish to humans, thereby highlighting the need for further investigation into this infection among ornamental fish maintained as pets, as well as individuals who interact with them and their environment.