Astyanax Bimaculatus Research Articles

Integrative Taxonomy Suggests Resurrection of Species of the Astyanax bimaculatus Group (Characiformes, Characidae).

Using integrative tools can be effective for species identification, especially in complex groups like Astyanax. Astyanax bimaculatus group is composed of six valid species, including A. lacustris. "A. altiparanae", "A. asuncionensis", and "A. jacuhiensis" are considered as junior synonyms of A. lacustris. Seeking to test the operational taxonomic unit (OTU) status of the junior synonyms of A. lacustris ("A. altiparanae", "A. asuncionensis", and "A. jacuhiensis"), we used analyses through mitochondrial DNA (COI and Cytb), cytogenetic markers (classical and molecular), and morphometry ("truss network"). Analysis of mitochondrial DNA sequences separated A. lacustris from the other synonymized species. The cytogenetic and morphometric analyses did not corroborate the synonymization and suggest that besides A. lacustris, the OTUs A. altiparanae, A. asuncionensis, and A. jacuhiensis are valid species. The analysis of different characters proposed by the integrative taxonomy used on the same individuals could provide greater reliability and minimize the underestimation of biodiversity.

Gametogenesis and seminatural reproduction of the Amazon twospot astyanax Astyanax bimaculatus (Linnaeus, 1758) cultivated in an enriched environment

Environmental enrichment is used to provide well-being to the animals, such as fish, in captive conditions, mimicking their natural habitat. It may influence fish behavior, physiology, and survival. In terms of reproduction, however, the relationship between environment enrichment and successful reproduction in captivity is still poorly explored in fish species. Aiming to understand any possible benefits of structural enrichment on fish reproduction, 10-days-hatched larvae of the twospot astyanax Astyanax bimaculatus were raised for 18 weeks in tanks with different elements of structural environmental enrichment (PVC pipes, stones, and artificial plants). In the 5th month of life, those animals were hormonally induced to reproduce to assess gamete formation and offspring quality. Animals raised in a sterile-reared environment (non-enriched) showed earlier spawning than the enriched one, presenting significant quantities of Postovulatory follicle complexes (POCs) and cells in atresia in female ovaries, indicating possible reproductive dysfunction or stress, as well as a greater quantity of empty testicular lumen in males, indicating great release of sperm. On the contrary, animals cultivated in enriched environments showed gonads filled with semen in males and vitellogenic oocytes in females. Furthermore, offspring from the sterile-reared group presented significant rates of larval abnormality compared to the enriched group. In conclusion, the results of this study show that environmental enrichment can interfere with the reproduction of fish in captivity, mainly by preventing early maturation of gametes, which can result in low-quality offspring and, consequently, low production of fish species.

New parameters for in vitro development of cell lines of the species Astyanax bimaculatus (Linnaeus, 1758) and Geophagus proximus (Castelnau, 1855).

Intending to compare in vitro cell growth in different conditions, we established cell cultures using fin biopsies of two freshwater fishes, Astyanax bimaculatus and Geophagus proximus. Three different culture media (Leibovitz-L-15, Dulbecco's Modified Eagle Medium [DMEM], and 199) were employed, with or without the addition of AmnioMax, maintaining a standard temperature of 29°C. Based on the results obtained, we standardized a cell growth protocol in which medium 199 was less efficient for both species. Notably, G. proximus cells exhibited superior proliferation in DMEM and L-15 media, whereas A. bimaculatus cells demonstrated better parameters exclusively in the DMEM medium. Successful subculturing of cells with good proliferation index was observed, accompanied by preserved morphological characteristics. Therefore, the methodology outlined in this study represents an advancement in establishing fish cell cultures.

Comparative cytogenetics of microsatellite distribution in two tetra fishes Astyanax bimaculatus (Linnaeus, 1758) and Psalidodon scabripinnis (Jenyns, 1842).

The main cytogenetic studies of the Characidae family comprise the genera Astyanax and Psalidodon involving the use of repetitive DNA probes. However, for the microsatellite classes, studies are still scarce and the function of these sequences in the genome of these individuals is still not understood. Thus, we aimed to analyze and compare the distribution of microsatellite sequences in the species Astyanax bimaculatus and Psalidodon scabripinnis. We collected biopsies from the fins of A. bimaculatus and P. scabripinnis to perform cell culture, followed by chromosome extraction, and mapped the distribution of 14 microsatellites by FISH in both species. The diploid number observed for both species was 2n = 50, with an acrocentric B microchromosome in A. bimaculatus and a metacentric B chromosome in P.scabripinnis. Regarding FISH, 11 probes hybridized in the karyotype of A. bimaculatus mainly in centromeric regions, and 13 probes hybridized in P.scabripinnis, mainly in telomeric regions, in addition to a large accumulation of microsatellite hybridization on its B chromosome. Comparative FISH mapping of 14 microsatellite motifs revealed different patterns of distribution both in autosomes and supernumerary chromosomes of A.bimaculatus and P. scabripinnis, suggesting independent evolutionary processes in each of these species, representing excellent data on chromosome rearrangements and cytotaxonomy.

Reproductive toxicity induced by lead exposure: Effects on gametogenesis and sex steroid signaling in teleost fish

Lead (Pb) is an emerging contaminant widely distributed in aquatic environments, which has serious effects on human and animal health. In this study, we determined whether Pb exposure affects gametogenesis, sex steroids, estrogen (ERα and ERβ), and androgen (AR) receptors. Adult specimens of Astyanax bimaculatus were exposed in duplicate to 15, 50, and 100 μg/L of lead acetate, whereas the control group was not exposed. After 28 days of exposure, fish were euthanized and samples of the gonads, liver, and blood were collected for analysis. The results indicated a reduction in the gonadosomatic index as well as the diameters of the vitellogenic follicles and seminiferous tubules in the exposed groups. Morphometry of gametogenesis revealed inhibition of the secondary oocyte growth and a reduction in the number of spermatozoa in the 50 and 100 μg/L Pb-treated groups. In females, plasma 17β-estradiol (E2) increased following 15 and 50 μg/L Pb treatment, whereas males exhibited an increase in E2 and 11-ketotestosterone following treatment with 15 and 100 μg/L Pb, respectively. Vitellogenin was significantly reduced in females exposed to 100 μg/L Pb, but metallothionein levels were unchanged. ERα, ERβ, and AR were immunolocalized in the somatic and germ cells, with increased ovarian expression of ERα and Erβ in the 100 μg/L Pb-treated group, but no significant difference in AR among the groups. In males, only ERα increased in the 100 μg/L Pb-treated group. These results indicate that Pb exposure impairs gametogenesis, disrupts estrogen receptor signaling, and affects the expression of major reproductive biomarkers in A. bimaculatus.

Bioaccumulation and Depletion of the Antibiotic Sulfadiazine 14C in Lambari (Astyanax bimaculatus).

Antibiotics are present in the environment, primarily due to their release through wastewater treatment plants, agricultural practices, and improper disposal of unused medications. In the environment, these drugs can be bioaccumulated by organisms and transferred along the food chain. This is a problem when considering the consumption of fish meat. In the United States, legislation stipulates that the maximum residue limit for sulfadiazine (SDZ) should not exceed 100 μg kg-1. Lambari fishes have potential economic importance in aquaculture, as they are relatively easy to breed and can be raised in small-scale operations. Finally, studying the biology and ecology of lambari could provide valuable information about freshwater ecosystems and their inhabitants. The current work aimed to measure the bioaccumulation and depletion of the antibiotic SDZ 14C in lambari (Astyanax bimaculatus). For this purpose, the tests were divided into two stages; seven days of exposure and seven days of depletion, where one fish was randomly selected and sampled every day. In the exposure phase, the fish were fed the medicated feed three times a day at a concentration of 2.5 mg·g-1. The control fish were fed uncontaminated feed. For the depletion phase, the remaining lambari were transferred to clean tanks and fed uncontaminated feed three times a day. The fish samples were burned in the Oxidizer and the reading of radioactivity was performed in a liquid scintillation spectrometer. It is worth noting that on day 7 and day 14, the water in the aquariums was filtered through filter paper to collect the metabolic excrement. SDZ concentrations increased over the days and accumulation occurred in the fish, with day seven presenting the maximum accumulation value of 91.7 ng·g-1 due to feeding uptake. After the depletion phase on day 13, the value found was 0.83 ng·g-1. The bioconcentration factor calculated was 20 L·kg-1. After the bioaccumulation period, the concentrations of SDZ in the water and excreta were 4.5 µg·L-1 and 363.5 ng·g-1, respectively. In the depletion period, the concentrations in the water and excreta were 0.01 µg·L-1 and 5.96 ng·g-1, respectively. These results imply that there was little SDZ bioaccumulation in the fish, but that it was distributed in larger amounts in the water. This is due to the physicochemical properties of the molecule with the low Log P value. Regarding the maximum residue limit, the value was below the established value. This study contributes to understanding SDZ dynamics in an aquatic species native to Brazil.

Oogenesis and in vitro reproduction of the twospot astyanax Astyanax bimaculatus (Linnaeus, 1758) exposed to conspecific alarm substance

Stress situations can be essential to trigger reproduction in fish; however, it may also inhibit it. One of those situations involves the release of the conspecific alarm substance (CAS), a natural stressor, into the water by specific fish epidermal cells after a predator attack. Little is known about the effects of that substance on fish reproduction. This study aimed to evaluate the effects of CAS exposure on the oogenesis and reproduction of the twospot astyanax Astyanax bimaculatus before the hormonal induction for artificial reproduction. No macroscopic or cellular changes in the ovaries were observed for the females exposed to CAS, and the oocyte stages show all females in the same phase of maturation (Spawning Capable). Females exposed to CAS spawned 20 min before the females without exposure. On the other hand, they ovulated only once, whereas the females from the control group ovulated multiple times for approximately two hours after hormonal induction. Moreover, the precocious ovulation of the females submitted to CAS did not generate offspring, since all generated zygotes did not develop. In contrast, the control group females produced more than 11 thousand healthy larvae. Exposing the female fish to CAS during their reproductive management in captivity may reduce breeding success.

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Stress situations can be essential to trigger reproduction in fish; however, it may also inhibit it. One of those situations involves the release of the conspecific alarm substance (CAS), a natural stressor, into the water by specific fish epidermal cells after a predator attack. Little is known about the effects of that substance on fish reproduction. This study aimed to evaluate the effects of CAS exposure on the oogenesis and reproduction of the twospot astyanax Astyanax bimaculatus before the hormonal induction for artificial reproduction. No macroscopic or cellular changes in the ovaries were observed for the females exposed to CAS, and the oocyte stages show all females in the same phase of maturation (Spawning Capable). Females exposed to CAS spawned 20 min before the females without exposure. On the other hand, they ovulated only once, whereas the females from the control group ovulated multiple times for approximately two hours after hormonal induction. Moreover, the precocious ovulation of the females submitted to CAS did not generate offspring, since all generated zygotes did not develop. In contrast, the control group females produced more than 11 thousand healthy larvae. Exposing the female fish to CAS during their reproductive management in captivity may reduce breeding success.

A new species of Astyanax (Characiformes, Characidae) from the rio Apiacás, rio Teles Pires basin, with a discussion on its phylogenetic position

A new species of Astyanax Baird & Girard, 1854 is described from the rio Apiacás, a tributary of the rio Teles Pires, rio Tapajós basin, Mato Grosso state, Brazil. The new taxon can be distinguished from all congeners, except those belonging to the Astyanax bimaculatus species group and to the Astyanax orthodus species group, by the presence of a horizontally elongated to rounded humeral blotch. The new taxon can be readily distinguished from all species belonging to the A. bimaculatus species group and to the A. orthodus species group by presenting a distinct morphology in premaxillary and dentary teeth with conspicuous diastema (a teeth gap) between them. We also present a hypothesis about the phylogenetic relationships of the new taxon within Astyanax.

The effects of water temperature and hybridization on embryonic development and gametogenesis of two species of Amazonian tetra

The study of embryonic development in fish and how water temperature affects the life stage in this group is of pivotal importance to support works that aim at the conservation, management, and recovery of endangered or high economic valuable species. It allows a better evaluation of the species and the identification of morphophysiological changes related to the environmental factor in question. In addition, hybridization emerges as a biotechnological tool that may be applied as a technique for the creation of sterile animals that can be used as recipients for the germ cell transplantation approach, thus contributing to the conservation of different fish species. Based on this, we artificially reproduced two Amazonian fish species, Astyanax bimaculatus, and Moenkhausia oligolepis, and the resulting offspring of each species had their embryonic development followed at room temperature (25 °C) and at high temperature (30 °C). In addition, we performed interspecific crossbreeding between the species, and the offspring were also incubated and observed under the above conditions. The results showed that the high temperature accelerates the embryonic development of both, the offspring from A. bimaculatus and from hybridization. However, no larvae survived. Regarding M. oligolepis, the embryos did not develop, dying at 4:40 post-fertilization. For the hybridization, only the crossbreeding between females of A. bimaculatus and males of M. oligolepis resulted in normal larvae. They presented their own morphological characteristics, showing no total or partial characteristics from the parents, besides presenting atrophied and sterile gonads. Heat waters were proven to compromise the embryonic development of both Amazonian species. On the other side, the hybridization between those two Amazonian species has shown to be a tool with great potential for sterilization and the development of individuals suitable for use as surrogate breeders aiming for the ecological maintenance of various fish species.

Predation on pirarucu larvae by opportunistic fish species

ABSTRACT One of the constraints in pirarucu, Arapaima gigas farming is the reproduction in captivity, due to inconsistent number of spawning events per reproductive cycle, thus, unreliable availability of juvenile fish. This is a first-hand report on predation of pirarucu larvae by opportunistic fish species in a breeding pond. Fish samples were collected for stomach content analysis from a pirarucu-breeding pond with reported predation on the larvae. We obtained 61 fish of different species (Tetragonopterus sp., Astyanax bimaculatus, Moenkhausia intermedia, Hoplias malabaricus, and Apistogramma sp.). Tetragonopterus sp. stomachs had the highest frequency of occurrence of pirarucu larvae (83.3%), followed by A. bimaculatus (7.4%). The relationship between the feeding behaviour of these species and their predation upon pirarucu offspring is discussed.

Characterization of a new cytotype and ocurrence of a B microchromosome in two spot astyanax, Astyanax bimaculatus Linnaeus, 1758 (Characiformes: Characidae).

Although Astyanax bimaculatus is the most representative species of the genus in the Amazon region, there are no cytogenetic studies of A. bimaculatus species in Amazon region. Thus, we aimed to analyse the chromosome complements of specimens from this area using classic and molecular cytogenetic approaches. The results revealed the existence of a distinct cytotype and this is the first report of the occurrence of a B microchromosome in the species. Overall, these data indicate that the karyotypic evolution of this species is complex, involving the occurrence of chromosomal rearrangements.

Upper thermal tolerances and vulnerability to global warming in a Brazilian Caatinga fishAstyanax bimaculatus (Linnaeus, 1758) population

AbstractGlobal warming is predicted to increase both average temperatures and the frequency and duration of heat waves. Tropical ectotherms, particularly those living in warm environments are more prone to receive heat impacts. Then, it is crucial to identify those populations already exposed to high temperatures that may be at risk of decimation by ongoing warming. We assessed heating risk in a population of the fish species Astyanax bimaculatus, inhabiting hot pools in intermittent streams in the Brazilian Caatinga ecoregion. We experimentally examined the upper thermal tolerance (CTmax), and provided estimates of warming tolerance (WT), as the difference between CTmax and micro‐environmental peak temperatures (Tmax). Two CTmax estimates ranged between 37.6 and 40.6°C, whereas pond Tmax ranged between 34.6 and 41.3°C. This determines very narrow, even negative WTs, for these extreme heated A. bimaculatus populations, which ultimately may determine their local extinction in the coming decades.Abstract in Portuguese is available with online material.

Connections among Land Use, Water Quality, Biodiversity of Aquatic Invertebrates, and Fish Behavior in Amazon Rivers.

Rivers in the Amazon have among the greatest biodiversity in the world. The Xingu River, one of the tributaries of the Amazon River, has a length of 1640 km, draining 510,000 km2 in one of the most protected regions on the planet. The Middle Xingu region in Brazil has been highly impacted by mining and livestock farming, leading to habitat fragmentation due to altered water quality. Therefore, comparing two rivers (the preserved Xingu River and the impacted Fresco River) and their confluence, the aims of the present study were to (1) assess the land uses in the hydrographic basin; (2) determine the water quality by measurements of turbidity, total solids, and metals (Cd, Cu, Fe, Mn, Pb, Zn, and Hg); (3) compare the zooplankton biodiversity; and (4) to evaluate the avoidance behavior of fish (Astyanax bimaculatus) when exposed to waters from the Xingu and Fresco Rivers. Zooplankton were grouped and counted down to the family level. For the analysis of fish avoidance, a multi-compartment system was used. The forest class predominated at the study locations, accounting for 57.6%, 60.8%, and 63.9% of the total area at P1XR, P2FR, and P3XFR, respectively, although since 1985, at the same points, the forest had been reduced by 31.3%, 25.7%, and 27.9%. The Xingu River presented almost 300% more invertebrate families than the Fresco River, and the fish population preferred its waters (>50%). The inputs from the Fresco River impacted the water quality of the Xingu River, leading to reductions in local invertebrate biodiversity and potential habitats for fish in a typical case of habitat fragmentation due to anthropic factors.

Autochthonous and allochthonous dietary probiotics mitigate acute stress in Astyanax bimaculatus during transport

Autochthonous and allochthonous dietary probiotics mitigate acute stress in <i>Astyanax bimaculatus</i> during transport

Autochthonous and allochthonous lactic acid bacteria: action on the hematological and intestinal microbiota for two species of Astyanax genus.

The objective of this work was to evaluate the effect of autochthonous and allochthonous lactic acid bacteria (LAB) in two species of lambaris (Astyanax bimaculatus and Astyanax fasciatus), and to investigate the effects on intestinal microbiota and hematological changes. Two experiments were carried out, one for each lambari species, both assays were divided into three treatments: autochthonous LAB, allochthonous LAB and control. The 10% inoculum was included on diet in the LAB treatments and sterile medium for control. After 30 days for A. bimaculatus LAB indigenous changes all bacteria groups analyzed, while allochthonous LAB just decrease Staphylococcus spp. count. Though for A. fasciatus his autochthone LAB reduced the staphylococcal count. In hematology, for A. bimaculatus autochthonous LAB showed a higher number of thrombocytes, lymphocytes, monocytes and total leukocytes in the circulatory system than the control. Though for A. fasciatus his autochthone LAB showed a higher number of total lymphocytes and leukocytes than the control, while Lactobacillus sp. acting as an allochthone, it did not differ among treatments. In conclusion, both LAB (Lactobacillus sp. and L. lactis) promoted more beneficial changes in the microbiota and hematological profile when they act as an autochthone probiotic, demonstrating a probiotic-associated host.

High rDNA polymorphisms in Astyanax lacustris (Characiformes: Characidae): new insights about the cryptic diversity in A. bimaculatus species complex with emphasis on the Paraná River basin

Abstract This study aimed to identify species of Astyanax bimaculatus group from four Itaipu Reservoir tributaries (Paraná River Basin) by cytogenetics and molecular markers (COI) to investigate the possible occurrence of cryptic diversity in part of this basin. The four populations showed only one karyotype formula and simple AgNORs. FISH with 18S rDNA probe showed a high variation, and 5S rDNA probes evidenced simple sites in most of the specimens, although multiple sites are present in two specimens. The variations of 5S and 18S cistrons generated 13 cytotypes. The molecular data did not reveal cryptic diversity in the populations; however, its grouping with 82 sequences from other stretches of the Paraná River Basin originated three haplogroups (distances of 3.12% and 8.82%) and 33 haplotypes were identified. DNA Barcode suggests that cytogenetic variations represent a high polymorphism degree, and it identified the analyzed specimens as Astyanax lacustris, which confirms the morphological identification. Our data suggest that the cryptic diversity of this group in the tributaries of the Paraná River Basin is different than the proposed by the synonymizations of A. altiparanae and A. asuncionensis to A. lacustris. This study reinforces the importance of integrative cytogenetics and molecular methods for taxonomy.

HYDROCHEMICAL PARAMETERS AND BIOMETRIC DATA OF FISH SPECIES FROM THE BORORÉ, BILLINGS RESERVOIR, SÃO PAULO, BRAZIL

The artisanal fishing is an important activity in Billings Reservoir (São Paulo, Brazil) and there is a paucity of biometric data about the fish in this region. Thereby, it is important to provide data about ichthyofauna that can be used for future management actions. The aim of this study was to present some data about the water quality and biometric data on fish species collected at the Bororé site (Billings Reservoir). In general, the obtained hydrochemical data raised a concern about the water quality because some parameters were above the established limit for body water class 2. A total of 170 fish were captured, corresponding to seven species and one genus. Geophagus brasiliensis was the most representative species (36%), followed by Astyanax bimaculatus (25%) and A. eigenmanniorum (20%). The hepatosomatic index (HSI) data showed differences in energy displacement between the species; the highest HSI values were observed in Hoplias malabaricus (HSImale = 1.33 ± 0.20; HSIfemale = 1.17 ± 0.82) while the smallest HSI was observed in A. fasciatus (HSImale = 0.47 ± 0.14; HSIfemale = 0.68 ± 0.35).

Biomarker responses induced by bisphenol A on spermatogenesis in a Neotropical teleost fish are temperature-dependent

Bisphenol A (BPA) is an organic synthetic compound used in the plastic industry with endocrine disrupting activity. Although it is frequently found in surface waters, few studies have investigated its impact on fish gametogenesis, particularly when associated with natural stressors. In this regard, the present study evaluated BPA toxicity on spermatogenesis in the lambari Astyanax bimaculatus under controlled conditions and its interactive effects with water temperature. Adult specimens were exposed in duplicate to 40 μg/L and 400 μg/L BPA at 23 °C and 28 °C for 21 days; the control group did not receive BPA. Testicular samples were collected and analyzed using different cellular and molecular techniques. The results showed a significant reduction in the gonadosomatic index in the BPA-treated groups at both temperatures. A decrease in the testicular levels of 11-ketotestosterone was observed in the 400 μg/L BPA group at 23 °C, 17β-estradiol increased significantly in the treated groups at 28 °C, and vitellogenin showed no difference between the treatments. The morphometric analysis of spermatogenesis revealed a significant increase in the proportion of spermatogonia, spermatocytes, and Sertoli cells in the treated groups, with a higher proportion at 23 °C than at 28 °C. Otherwise, the proportion of spermatozoa was significantly lower in the BPA-treated groups, with a greater reduction at 23 °C. In addition, BPA also stimulated spermatogonial proliferation in the treated groups, but apoptosis was significantly increased in spermatids at 23 °C. Testis-ova, cell degeneration, and chromatin alterations in spermatids and Sertoli cells were observed in the germinal epithelium of the BPA-treated groups. The integrated biomarker response (IBR) index revealed that the analyzed endpoints are suitable for assessing estrogenic contamination. Taken together, our results indicate that the interactive effects of BPA and temperature contribute to the impairment of spermatogenesis in A. bimaculatus with more severe effects observed on sperm production at 23 °C than at 28 °C.

Can use of substrates affect water quality in aquatic organism culture?

The choice of suitable substrates has received special attention in aquaculture and fishkeeping ventures. The aim of this study was to evaluate the influence of different substrates on water quality and the development of lambari fish (Astyanax bimaculatus) and freshwater shrimp (Macrobrachium potiuna). Two experiments were performed sequentially, one with fish and the other with shrimp. In both experiments, three different substrates were evaluated (shell gravel, river gravel, and dolomite), and the control was without any substrate. The water parameters evaluated were as follows: dissolved oxygen, saturation, oxide-reduction potential, temperature, pH, alkalinity, conductivity, ammonia, nitrite, nitrate, and phosphorus. It was observed that both dolomite and shell gravel interfered with the parameters that are related to the acid-base balance of the water and the availability of calcium and magnesium. The productive performance of lambari fish was not affected considering the different substrates evaluated; however, the productive performance of freshwater shrimp was lower in the absence of substrates in relation to the other treatments.