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

Background: 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. Methods: 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. Results and Discussion: 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. Conclusion: 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.

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.

First determination and analysis of the complete mitochondrial genome of X-ray tetra Pristella maxillaris (Ulrey, 1894) (Actinopteri, Characidae).

The complete mitochondrial genome of the X-ray tetra (Pristella maxillaris, Ulrey, 1894) was determined by using next-generation sequencing technology, and its mitochondrial genome characteristics were analyzed. The sequence total length was 16,753 bp, and the A + T content was 57.44%. The position and composition of the 37 genes were consistent with those of other Characidae species in this family. There are 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and one control region. Except for ND2 and COX1, which use ATT and GTG as start codons, respectively, all other protein-coding genes use ATG as the start codon. COX1 uses AGG as the stop codon; ATP6 and COX3 use incomplete TA as the stop codon; COX2 and ND4 use incomplete T as the stop codon; ATP8 uses TAG as the stop codon, and the other seven protein-coding genes use TAA as the stop codon. Based on the concatenated nucleotide sequences of 13 protein-coding genes from 18 Characidae species, phylogenetic analysis revealed that P. maxillaris belongs to the family Characidae and is most closely related to Hyphessobrycon amandae. Determining the mitogenomes of P. maxillaris improves our understanding of the phylogeny and evolution of Characidae.

Mitogenome of Knodus borki (Cypriniformes: Characidae): genomic characterization and phylogenetic analysis.

BACKGROUND: The taxonomic status of Knodu in the family Characidae is not yet clear. This study aimed to address this by sequencing and annotating Knodu borki Zarske, 2008. MATERIALS AND RESULTS: K. borki Zarske, 2008 was sequenced using a Hiseq platform and the complete mitogenome was assembled in SPAdes v3.15.2 and SOAPdenovo2 v.2.01. The mitogenome of K. borki from Guangzhou, the first sequenced species of the genus Knodu, is 16,837 bp in length and contains 13 protein-coding genes (PCGs), two ribosomal (r) RNAs, 22 transfer (t) RNAs, and one D-loop. Among these 37 genes, 28 are encoded by the heavy strand, while nine are encoded by the light strand. Twenty-one of the tRNAs can form typical cloverleaf secondary structures, except tRNA-Ser1, which lacks dihydrouridine arms. All PCGs have the same start codon (ATG), with the exception of COI (GTG). Four PCGs (ND1, ATP8, ND4L, and ND5) have TAA as the stop codon, ND6 has TAG as the stop codon, COI has AGG as the stop codon, and the remaining seven genes have incomplete stop codons of TA-/T-(ND2, COII, COIII, ND3, ND4, and Cyt b as T-, ATP6 as TA-). Phylogenetic analysis showed that K. borki belongs to the family Characidae. CONCLUSIONS: Our findings demonstrate that K. borki belongs to the family Characidae, due to consistency with the morphological identification. This study provides molecular information for further research on the phylogeny of the genus Knodus and for analyses of the taxonomic status of Characidae.

Mitochondrial genomes of five Hyphessobrycon tetras and their phylogenetic implications.

To date, the taxonomic status and phylogenetic affinities within Hyphessobrycon, even among other genera in Characidae, remain unclear. Here, we determined five new mitochondrial genomes (mitogenomes) of Hyphessobrycon species (H. elachys, H. flammeus, H. pulchripinnis, H. roseus, and H. sweglesi). The mitogenomes were all classical circular structures, with lengths ranging from 16,008 to 17,224 bp. The type of constitutive genes and direction of the coding strand that appeared in the mitogenomes were identical to those of other species in Characidae. The highest value of the Ka/Ks ratio within 13 protein-coding genes (PCGs) was found in ND2 with 0.83, suggesting that they were subject to purifying selection in the Hyphessobrycon genus. Comparison of the control region sequences among seven Hyphessobrycon fish revealed that repeat units differ in length and copy number across different species, which led to sharp differences in mitogenome sizes. Phylogenetic trees based on the 13 PCGs did not support taxonomic relationships, as the Hyphessobrycon fish mixed with those from other genera. These data were combined to explore higher level relationships within Characidae and could aid in the understanding of the evolution of this group.

Effects of intraspecific chemical cues on the behaviour of the bloodfin tetra Aphyocharax anisitsi (Ostariophysi: Characidae).

Chemical communication can induce a multitude of behaviours when detected by fish olfactory systems, from parental care, predation and alarm signalling, to foraging, schooling, reproduction, and migration. Chemical cues provide information that visual traits cannot and fish can respond to chemical cues without any additional sensory cue. In this way, pheromones play an essential role in the fitness of fishes. Given that Aphyocharax anisitsi inhabits environments characterized by cloudy and highly vegetated waters, it is interesting to evaluate the olfactory contribution in their communication. Here, we investigated the relevance of chemical cues in the types of behaviours triggered in A anisitsi by two experimental contexts: 1) non-social and olfactory context (conspecific-chemical cues), and 2) social context (conspecific female or male presence). Non-social context experiments suggest that males of A. anisitsi respond to both male and female-chemical cues even in the absence of other sensory inputs. The high olfactory sensitivity of characids in general and of A. anisitsi, in particular, could facilitate vital functions, such as foraging and conspecific recognition in habitats that impose severe restrictions on the visual system.

Carbofuran affects behavior and metabolism of the Atlantic Forest lambari Deuterodon iguape, a native species from Brazil.

One of the major causes of aquatic biodiversity loss is the contamination of the environment by pesticides. Even though there is a considerable amount of studies on the subject, there are still few that deal with the effects of carbofuran on native species in Brazil. Although carbofuran is widely used in Brazil, its action on native organisms, such as the Atlantic Forest lambari Deuterodon iguape, has not yet been studied. This work aimed to evaluate the effects of exposure to carbofuran on the fish D. iguape, considering the behavior and specific oxygen consumption as end points. Opercular movements, dorsal fin movements, and swimming speed were analyzed as behavioral parameters. To assess specific oxygen consumption, fish were subjected to concentrations of 0.0, 0.05, 0.1, 0.25, and 0.5 mg/L, for 24 h. For behavior analysis, fish remained exposed to carbofuran at concentrations of 0.0, 0.01, 0.05, 0.1, and 0.5 mg/L, in periods of 0, 2, 24, and 48 h. The behavior was studied through filming, analyzed with the free software, Tracker 4.92 (Open Source Physics). The results demonstrate an increase in opercular movements (18% ± 2.65) and a decrease in dorsal fin movements (- 21.2% ± 2.97), as well as in swimming speed (- 58.3% ± 1.83) of the experimental groups compared to the control group. There was an increase in oxygen consumption of 58.4% in fish exposed to the highest concentration of carbofuran. Thus, it is concluded that carbofuran altered D. iguape's behavior and oxygen consumption. The species was sensitive to carbofuran concentrations and can be used as a bioindicator.

Mitochondrial Genome Structures and Phylogenetic Analyses of Two Tropical Characidae Fishes.

The Characidae family contains the largest number of tropical fish species. Morphological similarities make species identification difficult within this family. Here, the complete mitogenomes of two Characidae fish were determined and comparatively analyzed with those of nine other Characidae fish species. The two newly sequenced complete mitogenomes are circular DNA molecules with sizes of 16,701 bp (Hyphessobrycon amandae; MT484069) and 16,710 bp (Hemigrammus erythrozonus; MT484070); both have a highly conserved structure typical of Characidae, with the start codon ATN (ATG/ATT) and stop codon TAR (TAA/TAG) or an incomplete T--/TA-. Most protein-coding genes of the 11 Characidae mitogenomes showed significant codon usage bias, and the protein-coding gene cox1 was found to be a comparatively slow-evolving gene. Phylogenetic analyses via the maximum likelihood and Bayesian inference methods confirmed that H. amandae and H. erythrozonus belong to the family Characidae. In all Characidae species studied, one genus was well supported; whereas other two genera showed marked differentiation. These findings provide a phylogenetic basis for improved classification of the family Characidae. Determining the mitogenomes of H. erythrozonus and H. amandae improves our understanding of the phylogeny and evolution of fish species.

New insights into patterns and rates of tooth replacement in serrasalmid and characid fishes, with implications for the subsistence fishery of Peru's remote ribereños villages.

We explored patterns, rates and unexpected socio-ecological consequences of tooth replacement in serrasalmids and characids of the Peruvian Amazon using microcomputed tomography. Of 24 specimens collected in February 2019, representing a mix of red-bellied piranha Pygocentrus nattereri, redeye piranha Serrasalmus rhombeus, silver dollar fish Ctenobrycon hauxwellianus and mojara Astyanax abramis, six individuals possessed edentulous jaw quadrants. On average, 22.9% of fish collected per day from these species featured incomplete dentition, a value three to five times higher than anticipated based on replacement rates estimated from captive fish, differences that may be driven by ontogeny, seasonality or environmental quality.

Ecological impacts of pesticides on Astyanax jacuhiensis (Characiformes: Characidae) from the Uruguay river, Brazil.

Brazilian freshwater ecosystems are continuously exposed to pesticides and domestic sewage. The Uruguay River was chosen for this study because of its international importance, as it flows through Brazil, Argentina, and Uruguay. It receives contaminants such as pesticides and domestic residues. Thus, the aim of this study to assess the accumulation of pesticides in muscle of the fish Astyanax jacuhiensis, its biochemical responses, and the presence of pesticides in water. In total, seven pesticides were registered in water from both river sites. Eight pesticides were detected in fish muscle. The biochemical responses showed that brain lipid peroxidation (LPO) and protein carbonyl (PC) in A. jacuhiensis were higher in the summer. Muscle showed the highest LPO levels in the spring and the highest PC in the summer. Liver LPO and PC levels were higher in the spring and summer. In the gills, the PC was higher in the spring and the LPO in the spring and winter. In the brain and in the gills, glutathione-S-transferase activity was high in the summer and autumn. Catalase activity was lower during the winter and spring. Non-protein thiol (NPSH) levels were lower in the brain in the winter and spring. Muscle tissue showed lower NPSH in the winter (site 1). Liver NPSH showed increased levels in liver in the spring and winter (site 2). The biochemical results clearly is related to pesticides and/or to the presence of other contaminants in the water such as metals or domestic sewage. The accumulation of pesticides in fish muscle added evidence that pesticides have been used in the area surrounding the Uruguay River. In conclusion, the biomarkers assayed in the present study could be used in future investigations considering other sampling sites along Uruguay River.

Riverscape properties contribute to the origin and structure of a hybrid zone in a Neotropical freshwater fish.

Understanding the structure of hybrid zones provides valuable insights about species boundaries and speciation, such as the evolution of barriers to gene flow and the strength of selection. In river networks, studying evolutionary processes in hybrid zones can be especially challenging, given the influence of past and current river properties along with biological species-specific traits. Here, we suggest that a natural hybrid zone between two divergent lineages of the sexually dimorphic Neotropical fish Nematocharax venustus was probably established by secondary contact as a result of a river capture event between the Contas and Pardo river basins. This putative river capture is supported by hydrogeological evidence of elbows of capture, wind gaps and geological faults. The morphological (colour pattern) and genetic (mtDNA and RADseq) variation reveal a clinal transition between parental lineages along the main river, with predominance of F2 hybrids at the centre of the hybrid zone, absence of early generation backcrosses and different levels of hybridization in the tributaries. We highlight that different sources of information are crucial for understanding how the riverscape spatial history influences the connectivity between and within river systems and, consequently, the dynamics of gene flow between freshwater lineages/species.

Mitochondrial genomes of four American characins and phylogenetic relationships within the family Characidae (Teleostei: Characiformes).

Mitochondrial genome sequencing has become widely used in numerous fields, including systematics, phylogeny, and evolutionary genomics. To elucidate phylogenetic relationships among members of the family Characidae, we sequenced the mitogenomes of four species within this family, namely, Aphyocharax rathbuni, Hyphessobrycon herbertaxelrodi, Hyphessobrycon megalopterus, and Prionobrama filigera. The mitogenomes were found to be 16,678-16,841 bp and encode 37 typical mitochondrial genes (13 protein-coding, 2 ribosomal RNA, and 22 transfer RNA genes). Gene arrangements in the studied species are consistent with those in the inferred ancestral fish. Most protein-coding genes in these mitogenomes have typical ATN start codons and TAR or an incomplete stop codon T-. Phylogenetic relationships based on Bayesian inference and maximum-likelihood methods indicated that A. rathbuni, H. herbertaxelrodi, H. megalopterus, and P. filigera belong to the Characidae family. Of the 15 Characidae species studied, three pairs were of the same genus, but the results for only one pair were well supported. This phylogenetic classification is inconsistent with those described in previous morphological and taxonomic studies on this family. Thus, systematic classification of the Characidae requires further examination. Our findings yield new mitogenomic data that will provide a basis for future phylogenetic and taxonomic studies.

Anatomy and ontogenetic changes of the facial and gular musculature of the tetra Astyanax brucutu: A remarkable case of adaptation to durophagy.

Astyanax brucutu is a peculiar species of Neotropical tetra endemic from the Chapada Diamantina, a large plateau in northeastern Brazil. Individuals of this species undergo a dramatic ontogenetic shift in their diet that is accompanied by equally remarkable changes in their feeding apparatus. Whereas juveniles of A. brucutu feed mostly on algae, adults feed almost exclusively on an endemic species of hydrobiid snail and other associated living organisms that inhabit their dead shells (including infaunal invertebrates and algae). Skeletal adaptations associated with this change in diet were previously reported, but until now, the changes in the musculature remained mostly unknown. The present paper describes the facial and gular muscles, as well as the buccal ligaments of A. brucutu in different life stages, and identifies the major ontogenetic changes in these systems associated with the diet shift in the species. Such changes primarily involve expansions of specific portions of the adductor mandibulae and associated tendons and ligaments that likely represent adaptations to increase the biting power necessary to crush copious amounts of shells ingested by larger individuals of A. brucutu. Those adaptations are absent in specimens of any size of Astyanax cf. fasciatus, a sympatric congener lacking durophagous feeding habits. Anatomical comparisons and landmark-based principal components analysis (PCA) suggest that most specializations to durophagy in A. brucutu arose by peramorphosis. We also found that several of the muscular specializations of adults of A. brucutu are paralleled in species of Creagrutus and Piabina, two other characid genera distantly related to Astyanax, but that also feed on hard food items.

Immune responses induced by inactivated vaccine against Aeromonas hydrophila in pacu, Piaractus mesopotamicus.

Aeromonas hydrophila is responsible for outbreaks of a severe infectious disease in fish farms around the world and is one of the major causes of economic losses to the neotropical fish farmers. This study assessed the induction of immune responses and protection against A. hydrophila in pacu, Piaractus mesopotamicus, vaccinated through intraperitoneal and immersion route with inactivated virulent strain. Fish were randomly distributed in three vaccinated groups: intraperitoneal (i.p.) route; immersion; and immersion + booster; and control group (unvaccinated). All vaccination protocols used the concentration of 1.7 × 108 CFU mL-1 of inactivated A. hydrophila., and an oil adjuvant was used for vaccine prepararion for i.p. route vaccination. Blood and skin mucus from 9 fishes per treatment were collected at 14, 28, 42 and 84 days post-vaccination (DPV) for determination of lysozyme concentration in skin mucus, as well as antibodies anti-A. hydrophila in blood serum and skin mucus. Fish were challenged at 84 DPV with homologous and virulent strain of A. hydrophila for evaluation of resistance against bacterial infection. The results demonstrated that vaccination with inactivated A. hydrophila suspension by i.p. or immersion resulted in significant increase of skin mucus lysozyme and specific antibody levels in serum and skin mucus, at 28 and 42 DPV, and this increase in innate and adaptive immunity remained significant in pacu vaccinated through i.p. route up to 84 DPV. Although no significant differences were observed in the survival study, pacu vaccinated through i.p. route presented 31,33% of relative percentage survival (RPS) in LD50-96h when compared unvaccinated fish challenged at 84 DPV. The results observed in this study indicate that vaccination programs with inactivated A. hydrophila, including booster doses by i.p. or immersion routes, could result in more effective protection in pacu against this bacteriosis, by increasing innate and adaptive mucosal and systemic immune responses.

Determination of the ideal volume of activating solution and the optimal ratio of spermatozoa per oocyte for Prochilodus brevis fertilization.

Prochilodus brevis is a rheophilic species with a threatened natural population that promotes studies aimed at optimizing reproduction in captivity. The correct quantity of inseminating dose and activating solution volume significantly improves fertilization rates, thereby increasing productivity in captivity. The objective of this study was to determine the proportion of sperm per oocyte and the ideal volume of activating solution to be used in the assisted fertilization of P. brevis. Gametes were collected and fertilization performed in two steps. In step 1, the ideal proportion of spermatozoa was determined based on the fertilization rate:oocyte by testing six doses of semen: D1 = 30 × 103, D2 = 150 × 103, D3 = 300 × 103, D4 = 3 × 106, D5 = 5 × 106, and D6 = 10 × 106. In step 2, the fertilization and hatching rates were evaluated using different volumes of activating solution (V1 - 25 ml, V2 - 50 ml, V3 - 75 ml,V4 - 100 ml, V5 - 125 ml, and V6 - 150 ml). A linear regression equation was estimated from steps 1 and 2. The Student-Newman-Keuls test was used to compare the means. In step 1, the percentage of fertilization increased linearly, reaching a plateau of 51.69%. In step 2, the best fertilization rates were obtained with an estimated ideal volume of 75.64 ml per 2 ml of oocytes. Therefore, the proportion of 928,410.29 sperm:oocyte, associated with the volume of 75.64 ml of water per 2 ml of oocytes, provided the maximum reproductive performance for P. brevis.

Habitat and community structure modulate fish interactions in a neotropical clearwater river

Species interactions can modulate the diversity and enhance the stability of biological communities in aquatic ecosystems. Despite previous efforts to describe fish interactions in tropical rivers, the role of habitat characteristics, community structure, and trophic traits over these interactions is still poorly understood. To investigate among-habitat variation in substratum feeding pressure and agonistic interactions between fishes, we used remote underwater videos in three habitats of a clearwater river in the Central Western, Brazil. We also performed visual surveys to estimate the abundance and biomass of fishes and proposed a trophic classification to understand how these variables can affect fish interactions. Community structure was the main factor affecting the variation in the interactions among the habitats. Biomass was the main variable determining which habitat a fish will feed on, while species abundance determined with how many other species it will interact in the agonistic interaction networks for each habitat. Specific habitats are not only occupied, but also used in distinct ways by the fish community. Overall, our results demonstrate the importance of the heterogeneity of habitats in tropical rivers for the interactions performed by the fishes and how the intensity of these interactions is affected by community structure.(AU)

Interações realizadas por peixes podem modular a diversidade e assegurar a estabilidade de comunidades em rios tropicais. Apesar dessa importância, poucos estudos relacionam as interações ecológicas com as características do habitat, estrutura da comunidade e atributos das espécies de peixes. Por meio de filmagens remotas subaquáticas nós verificamos como a pressão alimentar dos peixes sobre a comunidade bentônica e as interações agonísticas entre peixes são influenciadas por essas características do habitat e da comunidade em um rio tropical de água clara na região Centro-Oeste do Brasil. Também realizamos censos visuais para estimar a abundância e a biomassa dos peixes e propusemos uma classificação funcional para entender como essas variáveis podem afetar as interações dos peixes. A estrutura da comunidade foi o principal fator que afetou a variação nas interações entre os habitats. A biomassa dos peixes determinou em qual hábitat um peixe se alimentará, enquanto a abundância das espécies determinou com quantas outras espécies elas interagem nas redes de interações agonísticas de cada habitat. Habitats específicos não são apenas ocupados, mas também utilizados de maneiras distintas pela comunidade de peixes. Nossos resultados demonstram a importância da heterogeneidade de habitats para as interações realizadas pelos peixes em rios tropicais e como a intensidade dessas interações é afetada pela estrutura da comunidade.(AU)

Fishes community composition and patterns of species distribution in Neotropical streams / Composição da comunidade de peixes e padrões de distribuição de espécies em riachos Neotropiacais

Abstract: The ichthyofauna of streams in the Neotropical region is not yet fully known. This study aims to investigate the ichthyofauna composition of six streams of the Ijuí River sub-basin, Rio Grande do Sul State, inserted in the Uruguay River basin, as to contribute to the knowledge of fishes species richness and distribution in the south of Brazil. Sampling was carried out between July 2015 and May 2016, bimonthly, using the technique of electric fishing to collect the fishes. Spatial variations (per sampled stream) in the ichthyofauna composition were tested with a permutational multivariate analysis of variance. In total, we collected 5,029 individuals from 55 species, 13 families and five orders. From these species, 17 are endemic to the Uruguay River basin. Five species alone represented approximately 70% of the ichthyofauna abundance sampled. Our hypothesis that the fish community composition is not homogeneous along the streams sampled was confirmed and we observed that species complexity increases from the upstream closest area to the downstream according to the river continuum concept.

Resumo: A ictiofauna de riachos na região Neotropical ainda não é totalmente conhecida. Este estudo objetiva investigar a composição da ictiofauna de seis riachos da sub-bacia do rio Ijuí, estado do Rio Grande do Sul, inseridos na bacia do rio Uruguai, de forma a contribuir no conhecimento da riqueza e distribuição de espécies de peixes no sul do Brasil. A amostragem foi realizada entre julho de 2015 e maio de 2016, bimestralmente, utilizando a técnica da pesca elétrica. Variações espaciais (por riacho amostrado) na composição da ictiofauna foram testadas com uma análise de variância multivariada permutacional. No total, coletamos 5029 indivíduos de 55 espécies, 13 famílias e cinco ordens. Destas espécies, 17 são endêmicas da bacia do rio Uruguai. Cinco espécies sozinhas representaram aproximadamente 70% da abundância da ictiofauna amostrada. Nossa hipótese de que a composição da comunidade de peixes não é homogênea ao longo dos riachos amostrados foi confirmada e observamos que a complexidade das espécies aumenta de áreas à montante em direção à jusante, conforme o conceito do rio contínuo.

Stream fish from recently deforested basins in the Meridional Amazon, Mato Grosso, Brazil / Peixes de riachos de bacias recentemente desmatadas na Amazônia Meridional, Mato Grosso, Brasil

Abstract: The replacement of tropical forests to production systems is one of the leading causes of riverine ecosystem alterations. However, current assemblages' composition may also result from the time since these transformations have begun. Therefore, the knowledge of diversified historical scenarios can facilitate the accomplishment of actions that involve the aquatic environments recovery. In this study, an inventory of stream fish was carried out in basins whose deforestation was intensified in the last 20 years, to compose a baseline for ecological and taxonomic studies. The habitat, physical and chemical variables, and the fish assemblages from 60 streams in the northwest region of the state of Mato Grosso, in the Aripuanã and Juruena river basins, were sampled with standardized procedures. For a total of 130 species, a numerical predominance of small-sized Characidae and great rarity were registered, with 50 species represented by less than ten individuals and 19 singletons. Approximately 15% of the sampled taxa were identified only at the generic level, and for several taxa, more detailed taxonomic and molecular studies are required in order to achieve satisfactory identifications. None threatened species were so far reported. On the other hand, two specimens of non-native species were sampled. Although habitat quality is higher in forested streams, no differences in the species richness were registered when compared to the pasture with riparian forest streams or to more deforested streams. However, abundance was greater in these last two streams groups as a result of small-sized characins dominance.

Resumo: A substituição de florestas tropicais por sistemas de produção representa uma das principais fontes de alteração nos ecossistemas de riachos. Contudo, a composição atual das assembleias também depende do tempo decorrente desde o início dessas transformações e, desta forma, o conhecimento de cenários históricos variados pode facilitar a realização de ações que envolvam a recuperação de ambientes aquáticos. Neste estudo, foi realizado o inventário dos peixes de riachos em bacias cujo desmatamento foi intensificado nos últimos 20 anos, para compor uma linha de base que possa ser usada em estudos ecológicos e taxonômicos. Foram amostradas as variáveis do hábitat, físicas e químicas e os peixes de 60 riachos da região noroeste do estado de Mato Grosso, nas bacias dos rios Aripuanã e Juruena. No total, 130 espécies foram registradas, com predominância numérica de pequenos caracídeos e grande número de espécies raras, sendo 50 espécies representadas por menos que dez indivíduos e 19 por apenas um indivíduo. Aproximadamente 15% dos táxons amostrados foram identificados somente no nível genérico e vários precisam de estudos taxonômicos e moleculares mais detalhados para alcançar identificações satisfatórias. As espécies não-nativas foram representadas por dois exemplares e nenhuma espécie sabidamente ameaçada foi registrada. Embora a qualidade do hábitat seja superior nos riachos florestados, não houve diferenças na riqueza das assembleias quando comparada aos riachos de microbacias de pastagem, porém com faixa ripária florestada, ou com maior desmatamento. Contudo, a abundância foi maior nesses dois grupos de riachos, como resultado da dominância de caracídeos de pequeno porte.

Histopathological alterations in gills of a fish (Astyanax bifasciatus) in neotropical streams: negative effects of riparian forest reduction and presence of pesticides.

The reduction of riparian vegetation around aquatic environments causes several physicochemical alterations and favors the entry of pesticides via surface runoff. Such changes have negative effects on aquatic organisms. In this study, we evaluated histopathological alterations in gills of Astyanax bifasciatus to test the hypothesis that more severe histopathological alterations occur in gills of fish from streams with higher agricultural impact from the surrounding area. The specimens were collected by electrofishing in seven streams of the lower Iguaçu basin between August 2015 and February 2016. The gills were processed according to routine histological methods and examined by light microscopy. The histopathological alterations, mainly stage II (lamellar aneurysm and total fusion of lamellae), were observed in fish collected in streams with higher agricultural activity. In these streams, the histopathological index indicated slight to moderate organ lesions. In contrast, in streams with more vegetation cover, fish collected presented stage I histopathological alterations (lamellar edema and lamellar hyperplasia), and the HI indicated normal functioning of the gills. In addition, chloride and acid mucous cells were more abundant in the gills of fish collected in rural streams. Our findings demonstrate that more severe histopathological alterations were registered in fish collected from streams with intense agricultural activity in the surrounding area. Therefore, it highlights that vegetation cover around the streams is a positive force for the conservation and health of aquatic organisms.

Comparison and optimization for DNA extraction of archived fish specimens.

The DNA extracted from museum alcohol-fixed specimens can be a valuable source of information for solving taxonomic, phylogenetic, ecological and conservational questions. However, this type of DNA, also called ancient DNA, is routinely obtained in small portions and highly fragmented. We have tested two different extraction kits in museum type-specimens of the fish family Characidae. Aiming to increase the DNA yield, we made modifications on a Qiagen manufacturer protocol, in the elution step. Also, to overcome the issue of DNA fragmentation, we applied our efforts in Sanger sequencing, to find a highly variable and, in result, informative COI fragment. Based on our results, there is no correlation between amount of the DNA extracted and the age of the sample. The Sanger sequencing generated sequences which are useful in solving taxonomic puzzles. Here are presented the customization and guidelines that allowed us to recover DNA from the archived fish specimens. •DNA extraction from archived fish specimens is more effective when using silica columns.•Change of the elution times from minutes in room temperature to 24 h in freezer greatly improved the DNA yielded.•Short but highly variable sequences replace the need to sequence the entire gene to identify a species.