Neotropical Fish Species Research Articles

Transferability of heterologous microsatellite primers in Brycon gouldingi

ABSTRACT: Brycon gouldingi is a species of neotropical fish of socioeconomic and environmental importance in the Tocantins-Araguaia Basin. Genetic studies on this species are still limited, making it difficult to evaluate the population structure and genetic diversity in natural and captive stocks. Here, we aimed to evaluate the transferability of heterologous microsatellite primers in B. gouldingi. A total of 30 primers for eight species were evaluated: Brycon hilarii, Brycon opalinus, Brycon cephalus, Brycon orbignyanus, Prochilodus lineatus, Prochilodus argenteus, Piaractus mesopotamicus, and Colossoma macropomum. The primers that showed the best amplification patterns were applied to 20 specimens of B. gouldingi, and their genetic parameters were assessed. Among the 30 primers, seven showed satisfactory transferability, six of which belonged to the genus Brycon: Bh13 (B. hilarii), BoM5, BoM13 (B. opalinus), Borg9, Borg13, and Borg59 (B. orbignyanus), and one belonged to P. argenteus (Par80). The primers for the other species tested showed non-specificity or monomorphism; and were therefore, excluded from the analyses. The number of alleles ranged between two (Borg13 and Borg59) and three (Bh13, BoM5, BoM13, Borg9 and Par80), with sizes varying between 103 bp (BoM5) and 430 bp (Borg9). Four primers showed evidence of null alleles (BoM13, Borg9, Borg13, and Par80), which could probably be attributed to the respective Hardy-Weinberg deviation. Thus, seven primers were validated for cross-amplification in B. gouldingi, which may be used in future studies involving this species.

Histological evaluation of vital organs of the livebearer Jenynsia multidentata (Jenyns, 1842) exposed to glyphosate: A comparative analysis of Roundup® formulations

Roundup formulations are herbicides whose active principle is glyphosate. However, these formulations are potentially more toxic to non-target organisms than pure glyphosate. This study aimed to evaluate and compare the toxic potential of the Roundup formulations through histological alterations in fish. Thus, males and females of the neotropical fish species Jenynsia multidentata (Jenyns, 1842) were exposed for 24 or 96 h to the Roundup Original® (RO), Roundup Transorb® (RT) or Roundup WG® (RWG) formulations, at a fixed concentration of 0.5 mg/L of glyphosate. This concentration is close to the maximum glyphosate limits found in the environment and is non-lethal to J. multidentata. The three formulations caused histological damage to the liver, gills and brain of J. multidentata, which increased over the exposure time. Differences in the histological alterations between females and males were observed in the liver and brain. Females were more tolerant to RO and RT than RWG. Males did not exhibit these differences in sensitiveness with formulations. The RWG caused more damage in the liver and gills and RT in the brain. Overall, there were differences in the toxicity of RO, RT and RWG and the toxic effect was presented through histological damage, reinforcing the usefulness of histological biomarkers for Roundup® toxicity. The comparison of the toxic potential of glyphosate-based herbicides is important because it could give support to the governmental organizations to set protective rules for the water ecosystems and human health, as well as to reduce the use of highly toxic formulations in agriculture.

The effect of fasting on nutritional status, organs size and isotopic composition in a Neotropical fish species (Jenynsia multidentata)

Digestive flexibility is considered an adaptive response that allows individuals to minimise maintenance costs during periods of food scarcity. Here, we evaluated changes in body condition and digestive (and energy storage) organs size during short-term (7 days) fasting in Jenynsia multidentata. Also, we analysed changes in the isotopic composition and C:N ratio (a proxy of lipids content) in the intestine, liver and muscle. Fasted animals were compared against two feeding groups: field animals and animals fed ad libitum in the laboratory. We found that intestinal length was larger in feeding and field animals than in fasting fish, while intestine and liver masses were heavier in feeding animals, followed by field animals and then by fasting individuals. Liver and intestine δ15N was higher in fasting fish than in field animals, but δ13C did not differ between both groups. Finally, body condition, C:N ratios and perivisceral fat mass, were greater in feeding and field individuals than in fasting animals. This way, our results indicate that (i) changes in digestive and energy storage organs size during fasting in fish are ubiquitous and noticeable, and (ii) nutritional status of animals should be taken in account for a proper interpretation of stable isotope results, particularly for splanchnic tissues (such as intestine and liver).

The Response of Neotropical Fish Species (Brazil) on the Water Pollution: Metal Bioaccumulation and Genotoxicity.

The streams and rivers of the Upper Paraná River Basin have been seriously affected by impacts of high population density around the basin area. Fishes are widely used as models to assess the health of aquatic ecosystems, being considered as bioindicators of environmental pollution. In this context, our objective was to assess the potential genotoxic and mutageniceffects of the polluted water in three native fish species (Astyanax lacustris, Hypostomus ancistroides, and Rhamdia quelen) from Tarumã Microbasin, Upper Paraná River, Brazil. We also investigated the concentration of metals in water and in fish muscle to verify bioavailability and bioaccumulation of metals. For both less impacted sites (LI) and impacted sites (IMP) of the microbasin, the concentrations of metals were above the maximum limit allowed by Brazilian legislation (Resolution CONAMA 357/2005), except for Pb, total Cr, and Cu at LI sites and total Cr at IMP sites. A. lacustris showed a higher frequency of micronuclei (MN) at IMP sites compared with LI sites (p < 0.0001). We found no significant differences in MN frequency between site classes for H. ancistroides and R. quelen (p > 0.05). There were no significant differences between site classes regarding to nuclear abnormalities in erythrocytes frequencies (p > 0.05). A. lacustris from IMP sites had higher concentrations of Pb, Cu, Fe, Zn, and Ni in muscle tissue (p < 0.05), whereas H. ancistroides from IMP sites had higher concentration of Cr, Cu, and Ni (p < 0.0001) and R. quelen showed higher concentration of Cd, Fe, and Ni at these sites (p < 0.0001). So, the chosen biomarkers are able to identify the environmental risk of the water pollution.

Behavior and histopathology as biomarkers for evaluation of the effects of paracetamol and propranolol in the neotropical fish species Phalloceros harpagos.

Pharmaceutical drugs in the aquatic environment can induce adverse effects on nontarget organisms. This study aimed to assess the short-term effects of sublethal concentrations of both paracetamol and propranolol on the fish Phalloceros harpagos, specifically light/dark preference, swimming patterns, skin pigmentation, histopathology, and liver glycogen levels. Fish were acutely exposed to sublethal concentrations of both paracetamol (0.008, 0.08, 0.8, 8, 80mgL-1) and propranolol (0.0001, 0.001, 0.01, 0.1, 1mgL-1) under controlled conditions. For scototaxis, a significant preference for the dark compartment was observed for the group exposed to the highest concentration of paracetamol (80mgL-1). Propranolol exposure significantly altered the swimming pattern, especially in fish exposed to the 0.001mgL-1 concentration. Pigmentation was reduced in propranolol-exposed fish (0.1, 1mgL-1). The lowest concentration of propranolol (0.0001mgL-1) induced a decrease of histochemical reaction for hepatic glycogen. These data demonstrate that pharmaceuticals can induce sublethal effects in nontarget organisms, even at low concentrations, compromising specific functions of the individual with ecological relevance, such as energy balance and behavior.

Dams cause genetic homogenization in populations of fish that present homing behavior: Evidence from a demogenetic individual-based model

Few studies have assessed the impact of dams on the distribution of the genetic diversity of Neotropical fish species, and some of them suggest that the genetic structure of fish metapopulations is not affected by damming. However, the effects of dams on the genetic structure of populations may hinge on the biological characteristics of the species, such as homing behavior. Dams prevent some individuals from migrating from their feeding area to their natal spawning area, constraining them to reproduce in the tributaries that are closest to the barrier, and which are already being used as breeding grounds by other subpopulations. In these places, subpopulations may interbreed, thus disrupting the overall genetic structure of the metapopulation. To test this hypothesis, we developed a demogenetic Individual-Based Model that simulates the impact of permeable and impermeable dams on the genetic structure of homing populations, in both the long-term and the short-term. Our results support that genetic homogenization occurs among the subpopulations in the short-term, especially near the dam. The impact of a dam on the genetic structure is also detectable in the long-term (100 generations after the addition of the dam) when the dam is permeable (symmetrical or asymmetrical). However, when the barrier is not permeable, the genetic differentiation among the subpopulations is reestablished in the long-term. Our results suggest that conservation programs should be developed to avoid/mitigate genetic homogenization of populations of fish that display homing behavior in the short-term, and to reestablish the pre-dam genetic structure of these populations in the long-term.

The effect of water temperature on muscle cellularity and gill tissue of larval and juvenile Lophiosilurus alexandri, a Neotropical freshwater fish

The effect of temperature on muscle development and gill tissue were investigated in larval and juvenile of Lophiosilurus alexandri, a carnivorous Neotropical fish species. Larvae and juveniles were reared at temperatures of 23, 26, 29 and 32 °C for 15 and 35 days, respectively. The larvae were fed Artemia nauplii, while the juveniles received a formulated diet. In both stages, high temperatures resulted in increased fiber diameter of white muscle. Moreover, a pronounced effect of hypertrophy was observed in later stages of development. No alterations to minimum fiber diameter, and similar mosaic standards of muscle growth dynamics, were observed in larvae kept at 23–32 °C, suggesting that hyperplasia was not effected during early stages of development. Gills were analyzed by Degree of Tissue Change (DTC), which is based on the severity of lesions. Significant morphological changes were observed in gills when juveniles were kept at 32 °C, which was considered moderate tissue damage. Temperature modulates cellularity in L. alexandri larvae and juveniles, with pronounced alterations of hypertrophy of white muscle fibers in later stages of development when exposed to elevated temperatures. Higher temperatures may also induce muscle growth dynamics with increased recruitment of new white fiber, increased fiber diameter in the early stages and induced gill lesions.

A cost‐and‐time effective procedure to develop SNP markers for multiple species: A support for community genetics

Abstract Multispecies population genetics is an emerging field that provides insight relevant to conservation biology and community ecology. However, to date, this approach is limited to species with available genetic resources. The use of thousands of single‐nucleotide polymorphism (SNP) markers developed from recent genotyping‐by‐sequencing (GBS) technologies is a roadmap for the study of nonmodel species, but remains cost prohibitive when several, distantly related species are involved. We aimed to overcome this issue using a single HiSeq3000 run of restriction‐site associated DNA sequencing (RAD‐Seq) to retrieve SNP markers for 40 diverse species including plants, invertebrates, fish, and mammals. We developed a Python‐based pipeline to isolate c. 100–500 high‐quality SNP markers for each species that could be genotyped through classical PCR amplification methods. To assess the quality of these markers, we validated our approach on c. 160 of the characterized SNPs for each of 18 Neotropical fish species from the river Maroni (French Guiana, South America), using the MassARRAY iPLEX platform from Agena Bioscience (San Diego, CA, USA). A run of the pipeline applying stringent filtering parameters enabled the successful design of between 130 and 3,492 SNP markers for 30 of the 40 study species. Relaxing pipeline parameters allows for an increase in the number of detected SNPs. Across the 18 species from French Guiana, an average of 85% of markers were successfully amplified, polymorphic, and scored in ≥90% of individuals (c. 200 individuals per species). The great majority (&gt;98%) of these markers were at Hardy–Weinberg equilibrium in each sampling site from the river Maroni. This SNP discovery was performed at the cost of c. $US110 for each of the 40 species. Genotyping was performed at the cost of c. $US6000 for each of the 18 fish species with an average of 200 individuals per species. This strategy was found cost‐and‐time efficient to develop hundreds of SNP markers for a large range of nonmodel species, which can be used to investigate ecological and evolutionary questions that do not require whole‐genome coverage.

Biology, ecology and biogeography of the South American silver croaker, an important Neotropical fish species in South America

The South American silver croaker is a popular fish that has recently received substantial attention from scientists, mainly due to its importance as source of animal protein and as a key fisheries species. However, little is known about the conditions that explain its historical and current spatial distribution, both in its native habitat and where it is a successful invasive species. The aim of the present study was to explore the ecological information available for this species, to then critically examine ecological theories related to the conditions underpinning its success. To this end, an exhaustive literature search was conducted with the immediate aim of investigating whether the success of South American silver croaker was driven by species-climate or species–human interactions. The non-native populations were found to occupy climate niche spaces different from those observed in their native ranges. In addition, it was clear that humans played a role in facilitating the large-scale dispersion of silver croaker, and assisted as agents of impact driving the observed current and, probably, the future spatial distribution, which we can predict from our data and from the pattern of propagule pressure. Overall, the current biogeography of this species illustrates how the construction of dams, along with the introduction and stocking of non-native species, overfishing and other human activities can alter fish populations and assemblages. Such processes can reduce native species, increase the abundance and distribution of invasive species, as well as cause changes in life-history traits and genetic variability, all with long-term socioeconomic consequences.

Spatial distribution and habitat use by early fish stages in a dammed river basin, Southern Brazil

Fish diversity loss is threatened by the construction of dams as they prevent the regular natural dispersal among populations. Thus, conservation of key riverine habitats for fish reproduction may be essential for the recruitment of new native species of fish. The present study aimed to identify key habitats for fish spawning and early development in the Paranapanema River basin, as well as to determine the taxonomic composition, reproductive and life-history strategy, and to report spatial distribution of eggs, larvae and juveniles. The importance of lagoons, tributaries, and sub-tributaries was evaluated in the Paranapanema River basin between October 2012 and March 2013. Eggs and larvae samples were collected at dawn and dusk with conical plankton nets (0.5 mm mesh size), whereas juveniles were captured during the day with seine and sieve (0.5 cm mesh size). A total of 547 eggs, 904 larvae and 1 228 juveniles were captured. We observed that 2 larvae and 288 juveniles of non-migratory species, parental care, and equilibrium life-history strategy, predominated in lagoons and tributaries. On the other hand, 13 larvae and 60 juveniles of short migratory distance, no parental care, and periodic life-history strategy predominated in sub-tributaries. The highest densities of eggs were recorded in tributaries and sub-tributaries (Tukey’s test, P = 0.001 and P = 0.03, respectively), and the highest densities of larvae were recorded for lagoons and tributaries (P = 0.005 and P = 0.0001, respectively). Captures of eggs and larvae were higher at night; while the highest catches per unit effort of juveniles were recorded for tributaries and sub-tributaries. Fish species that adopt different life-history strategies can use diverse types of habitats during the early stages. Lagoons, tributaries and sub-tributaries of the Paranapanema River play different roles in the reproductive success of fish fauna in a heavily modified basin. The preservation of spawning and nursery areas trapped between reservoirs is necessary for Neotropical fish species recruitment and survival. Rev. Biol. Trop. 66(2): 605-621. Epub 2018 June 01.

Assessment of biochemical alterations in the neotropical fish species Phalloceros harpagos after acute and chronic exposure to the drugs paracetamol and propranolol.

Over time, many pollutants of anthropogenic origin have caused the contamination of aquatic ecosystems. Among several characteristics, these compounds can reach the trophic chain, causing deleterious interactions with the biota. Pharmaceutical substances can be included in this scenario as emerging contaminants that reach the aquatic environment because of direct human and veterinary usage, and release by industrial effluents, as well as through domestic dumping of surplus drugs. The effects of these compounds on exposed organisms have been studied since the 1990s, but ecotoxicological data for such chemicals are still scarce especially concerning aquatic organisms from tropical regions. Paracetamol and propranolol were selected for this study since they are frequently found in surface waters. Paracetamol is a drug used as analgesic and antipyretic, while propranolol, a β-blocker, is used in the treatment of hypertension. The objective of this study was to assess the toxic effects of these substances on the neotropical freshwater fish Phalloceros harpagos after acute (96h) and chronic (28days) exposures. In order to understand the effects of these drugs on P. harpagos, biochemical markers were selected, including the enzymes involved in oxidative stress, xenobiotic metabolism, and neurotransmission (catalase, glutathione-S-transferase, and cholinesterase activities, respectively). After acute exposure, no significant alterations were observed for catalase activity, suggesting the absence of oxidative stress. On the contrary, significant alterations in glutathione-S-transferases activity were described for the higher concentrations of both pharmaceuticals after acute exposure. In addition, acute exposure to paracetamol caused a significant increase of cholinesterase activity. None of the tested pharmaceuticals caused significant changes in catalase or cholinesterase activities after chronic exposure. Glutathione S-transferases activity was significantly increased for propranolol following chronic exposure, indicating the potential involvement of phase II detoxification pathway.

Cytogenetics characterization of Crenuchus spilurus (Günther, 1863): a remarkable low diploid value within family Crenuchidae (Characiformes)

Conventional and molecular cytogenetic analyses were performed in specimens of the Neotropical Crenuchus spilurus freshwater fish species from a single location (Caete River, Brazil). All specimens presented diploid values of 2n = 38 chromosomes (12 m + 4sm + 2st + 20a), the lowest reported for family Crenuchidae up to now. A single pair of nucleolar organizing regions (NORs) was detected in the subtelocentric chromosome pair no. 9 by silver-staining and fluorescence in situ hybridization (FISH) with 18S rDNA sequence-specific probe. Two pairs of 5S rRNA gene clusters were found either interstitial or terminally located in the long arms of the acrocentric chromosome pairs nos. 10 and 13. Heterochromatic regions were clearly observed in the short arms of the NOR-bearing chromosome pair and weakly-positive to the pericentromeric regions of most acrocentric chromosomes. Additionally, no sex chromosomes were identified in the surveyed specimens. Crenuchidae have signals of several mechanisms involved in karyotype diversification within this family: differential location of heterochromatin-rich regions, multiplication, and translocation of rDNA clusters, presence/absence of sex chromosomes, macrostructural changes in morphology and number of chromosomes. This variety of karyotype patterns reveals the importance of widening cytogenetic studies to more taxa for better know the chromosomal evolution occurred in this group.

Co-exposure to titanium dioxide nanoparticles (NpTiO2) and lead at environmentally relevant concentrations in the Neotropical fish species Hoplias intermedius.

Growing production and utilization of titanium dioxide nanoparticles (NpTiO2) invariably lead to their accumulation in oceans, rivers and other water bodies, thus increasing the risk to the welfare of this ecosystem. The progressive launch of these nanoparticles in the environment has been accompanied by concern in understanding the dynamics and the toxic effect of these xenobiotic in different ecosystems, either on their own or in tandem with different contaminants (such as organic compounds and heavy metals), possibly altering their toxicity. Nevertheless, it remains unknown if these combined effects may induce damage in freshwater organisms. Therefore, this study aimed to analyze the consequences caused by NpTiO2, after a waterborne exposure of 96 h to a Neotropical fish species Hoplias intermedius, as well as after a co-exposure with lead, whose effects for fish have already been well described in the literature. The characterization of NpTiO2 stock suspension was carried out in order to provide additional information and revealed a stable colloidal suspension. As a result, NpTiO2 showed some genotoxic effects which were observed by comet assay in gill, kidney and brain cells. Also, the activity of brain acetylcholinesterase (AChE) has not changed, but the activity of muscle AChE decreased in the group exposed only to PbII. Regarding the hepatic antioxidant system, catalase (CAT) did not show any change in its activity, whereas that of superoxide dismutase (SOD) intensified in the groups submitted only to PbII and NpTiO2 alone. As for lipid peroxidation, there was a decrease in the group exposed to the NpTiO2 alone and to the co-exposed group (NpTiO2+PbII). As far as metallothionein is concerned, its concentration rose for the co-exposed group (NpTiO2+PbII) and for the group exposed to PbII alone. Overall, we may conclude that NpTiO2 alone caused DNA damage to vital tissues. Also, some impairment related to the antioxidant mechanism was described but it is probably not related to the DNA damage observed, suggesting that the genotoxic effect observed may be due to a different mechanism instead of ROS production.

Chromosomal Evolution in Lower Vertebrates: Sex Chromosomes in Neotropical Fishes.

Fishes exhibit the greatest diversity of species among vertebrates, offering a number of relevant models for genetic and evolutionary studies. The investigation of sex chromosome differentiation is a very active and striking research area of fish cytogenetics, as fishes represent one of the most vital model groups. Neotropical fish species show an amazing variety of sex chromosome systems, where different stages of differentiation can be found, ranging from homomorphic to highly differentiated sex chromosomes. Here, we draw attention on the impact of recent developments in molecular cytogenetic analyses that helped to elucidate many unknown questions about fish sex chromosome evolution, using excellent characiform models occurring in the Neotropical region, namely the Erythrinidae family and the Triportheus genus. While in Erythrinidae distinct XY and/or multiple XY-derived sex chromosome systems have independently evolved at least four different times, representatives of Triportheus show an opposite scenario, i.e., highly conserved ZZ/ZW system with a monophyletic origin. In both cases, recent molecular approaches, such as mapping of repetitive DNA classes, comparative genomic hybridization (CGH), and whole chromosome painting (WCP), allowed us to unmask several new features linked to the molecular composition and differentiation processes of sex chromosomes in fishes.

Impact of waterborne and trophic mercury exposures on cardiac function of two ecologically distinct Neotropical freshwater fish Brycon amazonicus and Hoplias malabaricus

Metal pollutants have been considered one of the main factors underlying the depletion of biodiversity in natural populations unbalancing aquatic environments. The aim of this study was to evaluate the effects of exposure to inorganic Hg on myocardial contractility and the electrocardiogram (ECG) of two ecologically distinct Neotropical fish species, namely: matrinxã (Brycon amazonicus) and trahira (Hoplias malabaricus). Matrinxãs were exposed to a sublethal concentration of 0.1mgL−1 of Hg in water for 96h. Trahiras were exposed to dietary Hg doses (0.45mg of Hg, each 4days, for 30days) using juvenile B. amazonicus as the prey vehicle. Hg exposures decreased myocardial isometric twitch force development, harmed contraction/relaxation dynamics and cardiac pumping capacity (CPC), and reduced the relative contribution of the calcium stored in the sarcoplasmic reticulum (SR) to excitation contraction (EC) coupling in both fish species. Analysis of the ECG revealed that Hg impaired electrical conduction across the heart, inducing first degree atrioventricular block and lengthening the plateau phase of action potential duration. In trahira trophic doses of Hg induced a marked bradycardia, increasing the duration of the ventricular action potential and delaying atrial and ventricular depolarization. These findings indicate that both acute and long-term Hg exposure, by different routes is cardiotoxic to matrinxã and trahira. Hg potently impaired intracellular calcium kinetics in the cardiomyocytes, myocardium contractility, and electrical conduction across the heart, all of which can be implicated in decreased cardiac output and putative heart failure.

Morphology and Histology of the Pirarucu (Arapaima gigas) Digestive Tract

The present study describes the morphology and histology of the digestive tract of juvenile pirarucu, which is a Neotropical fish species with great potential for aquaculture in the Amazon region. Three size classes of pirarucu juveniles from commercial fish farms were sampled for histological examination of the oesophagus, stomach, pyloric caeca, intestine and liver. The morphological and histological features of the pirarucu digestive tract are similar to other carnivorous teleosts and enable the species to ingest, store and digest large food items. Acidic and neutral mucins are secreted by the mucosa of the oesophagus, stomach, pyloric caeca and intestine to protect the mucosal epithelium and to lubricate and facilitate food passage along the digestive tract. Complex transverse folds are predominant in the intestinal mucosa and most likely contribute to optimization of the digestion and nutrient absorption processes, thereby offsetting the relatively short length of the intestine. The exocrine pancreas appears diffuse in the liver and mesentery around the pyloric region of the stomach, pyloric caeca and initial part of the intestine. The digestive tract morphology and thickness become increasingly developed with the size of pirarucu.

Dietary lyophilized colostrum alters distribution of goblet cells and the intestinal epithelium of Piaractus mesopotamicus

The intestinal epithelium is responsible for the first line of defense, directly influenced by bioactive, dietary molecules. The lyophilized bovine colostrum (LBC) is a nutraceutical and a surrogate protein source for farmed fish. This study aimed at evaluating distribution of goblet cells and morphometric changes in the intestinal epithelium of juvenile, neotropic Characin pacu Piaractus mesopotamicus fed diets containing 0%, 10% or 20% LBC for either 30 or 60days. Muscle layer thickness (MLT); partial volume of the absorptive mucosa (Vv); number of goblet cells containing acidic and neutral mucins and the acidic subtypes – sialomucin and sulphomucin, were studied in three intestinal segments (S1, S2, S3 and rectum). Fish fed 20% dietary LBC had enlarged MLT at 60days in S2 segment, and increased number of goblet cells containing neutral and acidic sulphomucin in S1, as well as number of goblet cells containing sulphomucin and sialomucin and total number of goblet cells in S2 at 60days. Juveniles fed 10% LBC diets had reduced number of goblet cells containing neutral mucin, acidic mucin and total number of goblet cells at 60days. The number of goblet cells containing sialomucin was higher in juveniles fed 20% LBC diets than those fed 0% and 10% LBC diets at 60days. Dietary LBC influenced the development of intestinal muscle layer and the volume of absorptive mucosa of juvenile pacu, improving capacity of nutrient and macromolecules absorption in S2 segment. The presence of unidentified substances from the bovine colostrum determined a reaction in intestinal epithelium with subsequent increased production of mucin. Statement of RelevanceAlternatives for fish nutrition are placed annually in the world market with the expectation of getting a quick performance at minimum cost. As an alternative animal food, lyophilized bovine colostrum may be innovative when added as a source of protein and bioactive peptides in fish food, such as neotropical fish species. The lyophilization process preserves the quality and unique features of colostrum, like antioxidant, immunological and antimicrobial proprieties. Additionally, the present work contributes to a better understanding of basic histology of neotropical fish species, that still demands for basic studies about enteric physiology. The presence and distribution of sialomucins, sulphomucins and neutral mucins in the intestinal epithelium of an omnivorous fish may contribute to the evaluation of nutritional alternatives that enable greater production efficiency at the national level. In this context, the present manuscript is complementary to others already published papers, which includes the study of the feasibility of using bovine colostrum as an alternative protein source for fish, as well as the consequences of the presence of biological active molecules, such as IGF-1, on the enteric histology and morphology.

Lethal dose and clinical signs of Aeromonas hydrophila in Arapaima gigas (Arapaimidae), the giant fish from Amazon

Aeromonas hydrophila is causing substantial economic losses in world aquaculture. This study determined the tolerance limit (LD50-96h) of A. hydrophila in Arapaima gigas, and also investigated the clinical signs after intradermal inoculation. Arapaima gigas fingerlings were inoculated intraperitoneally with 0 (control), 1.0×105, 1.0×106, 1.0×107, 1.0×109 and 1.0×1010CFU/mL of A. hydrophila for the determination of LD50-96h, which was 1.8×108CFU/mL. In another trial with intradermal inoculation of 1.8×108CFU/mL A. hydrophila, there was a 91.6% of mortality between 8 and 23h, and several clinical signs were found. As follows: depigmentation in the tegument, lesions in the tail and fins, loss of balance, reduction of respiratory movements, hemorrhagic foci, necrotic hemorrhages in the kidney, liver and swim bladder, splenomegaly, ascites in the abdominal cavity and hyperemia, enlargement of the gall bladder, among other clinical signs observed. The results showed that A. gigas has a relative tolerance to A. hydrophila when compared to other Neotropical fish species.

Habitat heterogeneity on feeding habit of two sympatric and congeneric characidae fishes in two tropical reservoirs

Abstract Food flexibility and omnivory are important features pronounced in Neotropical freshwater fish species, particularly for Astyanax species. Traditionally most fish diet resources are known to be originated in the aquatic environment, however recent studies have pointed to the importance of allochthonous sources. Besides, the colonization of macrophytes, common at several tropical reservoirs, may enhance insectivory in fish diet expanding or concentrating the area of available resources for feeding. Here we employed stomach contents analysis of two sympatric Astyanax species to access the importance of habitat differentiations as spatial complexity in two tropical reservoirs with different environmental features. The NMDS analysis indicated separation in the diet of these species between reservoirs (Stress= 8.28%). Additionally, analysis of variance revealed a significative difference in the volume of food itens ingested between the reservoirs (Anova one-way F(1, 132)=4.4446; p= 0.037). This points out the importance of environmental conditions on the composition of the diet of fishes. This study highlighted the insectivorous feeding habit of Astyanax species and revealed different feeding strategies between sympatric fishes despite high niche overlap in both environments. Habitat heterogeneity increasing food resources availability plays an important role in the diet strategy of these Astyanax species and on their constant maintenance in the two different reservoirs.

Application of the physical habitat simulation for fish species to assess environmental flows in an Atlantic Forest Stream in South-eastern Brazil

ABSTRACT The physical habitat simulation sub-routine of the Instream Flow Incremental Methodology (IFIM) uses hydraulic modeling and suitability indices of target fish species to predict how differences in-stream flows affect the microhabitat occupation by fish species. This habitat modelling approach was adopted to assess the ecological effects of running flows on three neotropical fish species of different orders (Bryconamericus ornaticeps , Ancistrus multispinis and Geophagus brasiliensis ).The study encompassed two reaches of an Atlantic Forest stream in Southeastern Brazil where topographic and hydraulic (depth, velocity and type of substrate) characteristics were measured to implement one-dimensional hydraulic simulation. Sub aquatic observation of fish was performed to collect data on microhabitat use and these data were used to develop habitat suitability curves that were used in the habitat simulation to obtain the habitat suitability index (HSI) and weighted usable area (WUA) versus flow curves. Upon these curves minimum and optimum environmental flows for the target fish species were proposed. Bryconamericus ornaticeps and A. multispinis selected microhabitats around 0.6 m depth, whereas G. brasiliensis showed a wider suitable range (0.35-0.9 m). All the three species were mainly observed in microhabitat with low flow velocity (0.1 m/s). Bryconamericus ornaticeps selected more frequently coarse substrate (e.g. boulders) but it appeared also over sandy substrate, whereas A. multispinis and G. brasiliensis selected preferably boulders. The range of 0.65-0.85 m3/s was found as the optimum to meet the needs of the three fish species. Our results agree with the necessary objective information to perform grounded management actions in the frame of a management program aiming at ecosystem conservation. Thereby it can be considered a successful pilot study in environmental flow assessment in an Atlantic Forest stream of Brazil.