Repeated evolution of eye loss in Mexican cavefish: Evidence of similar developmental mechanisms in independently evolved populations.

Evolution in similar environments often leads to convergence of behavioral and anatomical traits. A classic example of convergent trait evolution is the reduced traits that characterize many cave animals: reduction or loss of pigmentation and eyes. While these traits have evolved many times, relatively little is known about whether these traits repeatedly evolve through the same or different molecular and developmental mechanisms. The small freshwater fish, Astyanax mexicanus, provides an opportunity to investigate the repeated evolution of cave traits. A. mexicanus exists as two forms, a sighted, surface-dwelling form and at least 29 populations of a blind, cave-dwelling form that initially develops eyes that subsequently degenerate. We compared eye morphology and the expression of eye regulatory genes in developing surface fish and two independently evolved cavefish populations, Pachón and Molino. We found that many of the previously described molecular and morphological alterations that occur during eye development in Pachón cavefish are also found in Molino cavefish. However, for many of these traits, the Molino cavefish have a less severe phenotype than Pachón cavefish. Further, cave-cave hybrid fish have larger eyes and lenses during early development compared with fish from either parental population, suggesting that some different changes underlie eye loss in these two populations. Together, these data support the hypothesis that these two cavefish populations evolved eye loss independently, yet through some of the same developmental and molecular mechanisms.

Parallel evolution of regressive and constructive craniofacial traits across distinct populations of Astyanax mexicanus cavefish.

Life in complete darkness has driven the evolution of a suite of troglobitic features in the blind Mexican cavefish Astyanax mexicanus, such as eye and pigmentation loss. While regressive evolution is a hallmark of obligate cave-dwelling organisms, constructive (or augmented) traits commonly arise as well. The cavefish cranium has undergone extensive changes compared with closely-related surface fish. These alterations are rooted in both cranial bones and surrounding sensory tissues such as enhancements in the gustatory and lateral line systems. Cavefish also harbor numerous cranial bone asymmetries: fluctuating asymmetry of individual bones and directional asymmetry in a dorsal bend of the skull. This asymmetry is mirrored by the asymmetrical patterning of mechanosensory neuromasts. We explored the relationship between facial bones and neuromasts using in vivo fluorescent colabeling and microcomputed tomography. We found an increase in neuromast density within dermal bone boundaries across three distinct populations of cavefish compared to surface-dwelling fish. We also show that eye loss disrupts early neuromast patterning, which in turn impacts the development of dermal bones. While cavefish exhibit alterations in cranial bone and neuromast patterning, each population varied in the severity. This variation may reflect observed differences in behavior across populations. For instance, a bend in the dorsal region of the skull may expose neuromasts to water flow on the opposite side of the face, enhancing sensory input and spatial mapping in the dark.

Toxicity of Cyanopeptides from Two Microcystis Strains on Larval Development of Astyanax altiparanae.

Absorption and accumulation of bioavailable cyanobacterial metabolites (including cyanotoxins) are likely in fish after senescence and the rupturing of cells during bloom episodes. We determined the toxicity of cyanopeptides identified from two strains of Microcystis (M. panniformis MIRS-04 and M. aeruginosa NPDC-01) in a freshwater tropical fish, Astyanax altiparanae (yellowtail tetra, lambari). Aqueous extracts of both Microcystis strains were prepared in order to simulate realistic fish exposure to these substances in a freshwater environment. Both strains were selected because previous assays evidenced the presence of microcystins (MCs) in MIRS-04 and lack of cyanotoxins in NPDC-01. Identification of cyanobacterial secondary metabolites was performed by LC-HR-QTOF-MS and quantification of the MC-LR was carried out by LC-QqQ-MS/MS. MIRS-04 produces the MCs MC-LR, MC-LY and MC-HilR as well as micropeptins B, 973, 959 and k139. NPCD-01 biosynthetizes microginins FR1, FR2/FR4 and SD-755, but does not produce MCs. Larval fish survival and changes in morphology were assessed for 96 h exposure to aqueous extracts of both strains at environmentally relevant concentrations from 0.1 to 0.5 mg (dry weight)/mL, corresponding to 0.15 to 0.74 µg/mL of MC-LR (considering dried amounts of MIRS-04 for comparison). Fish mortality increased with concentration and time of exposure for both strains of Microcystis. The frequencies of morphological abnormalities increased with concentration in both strains, and included abdominal and pericardial oedema, and spinal curvature. Results demonstrate that toxicity was not solely caused by MCs, other classes of cyanobacterial secondary metabolites contributed to the observed toxicity.

Spatial and temporal biomarkers responses of Astyanax jacuhiensis (Cope, 1894)(Characiformes: Characidae) from the middle rio Uruguai, Brazil

Due to intense agricultural activity in the rio Uruguai (South Brazil), there is the potential for aquatic contamination by agrochemicals. In this region, there are many reservoirs to meet the water demand for rice fields, forming lentic environments. In line with this information, the aim of this study was to show a comparative analysis of some biomarkers, such as lipid peroxidation (TBARS), gluthatione S-transferase (GST), non-protein thiols (NPSH), amino acids (AA) and piscine micronucleus tests (MNE) in Astyanax jacuhiensis from lentic and lotic environments in the middle rio Uruguai region, comparing warm and cold seasons. Eight pesticides were found in water samples, with propoxur having the highest concentration found in both environments and seasons. Fish from the warm season showed higher levels of biochemical biomarkers, and fish from the cold season showed higher levels of MNE and AA. TBARS and AA presented higher levels in fish from the river, while GST, NPSH, MNE and AA presented higher levels in fish from dams. These environments have different characteristics in terms of redox potential, aeration, sedimentation, trophic structure, agrochemicals input and others, which may affect the physiological and biochemical responses of fish in against adverse situations.

Devido à intensa atividade agrícola no rio Uruguai (Sul do Brasil), há potencial para contaminação aquática por agrotóxicos. Há muitos reservatórios para atender a demanda de água de campos de arroz, formando ambientes lênticos. De acordo com estas informações, o objetivo do presente estudo foi mostrar uma análise comparativa de alguns biomarcadores como a peroxidação lipídica (TBARS), glutationa S-transferase (GST), tióis não-protéicos (NPSH), aminoácidos (AA) e teste písceo de micronúcleos (MNE) em Astyanaxjacuhiensis amostrados em ambientes lóticos e lênticos da região do médio rio Uruguai, comparando estações quentes e frias. Oito pesticidas foram encontrados em amostras de água, sendo propoxur a maior concentração encontrada em ambos os ambientes e estações. Peixes da estação quente apresentaram maiores níveis de biomarcadores bioquímicos e peixes da estação fria apresentaram maiores níveis de MNE e AA. TBARS e AA apresentaram maiores níveis nos peixes de rio, enquanto GST, NPSH, MNE e AA apresentaram níveis mais elevados em peixes da represa. Estes ambientes têm características diferentes, com potencial redox, aeração, sedimentação, estrutura trófica, a entrada de agroquímicos e outros que podem afetar as respostas fisiológicas e bioquímicas de peixe contra situação adversa.

Dermal lip protuberances associated with aquatic surface respiration in juveniles of the piscivorous characid Salminus brasiliensis (Actinopterygii: Characidae)

Some South American freshwater fishes can improve their capability of aquatic surface respiration (ASR) by developing dermal lip protuberances in the lower jaw. This adaptation was thought to be limited to omnivorous or herbivorous fishes. The present work provides the first evidence that juveniles of a piscivorous characid, Salminus brasiliensis, develop dermal lip protuberances during periods of hypoxia in floodplain ponds of the Salado River, in Argentina. The protuberance of S. brasiliensis involves dermal portions of both jaws exhibiting lateral lobes on the sides of the mouth arranged in the vertical plane. Water dissolved oxygen concentrations less than or equal to 1.05 mgl-1 were associated with a remarkable increase in lip protuberance. The lateral lobes of the protuberance in this species may limit the access of water to the anterior portion of the mouth which is positioned nearer to the oxygenated surface water during ASR. Finally, ASR, complemented with the development of dermal lip protuberances, can be considered a valuable strategy to survive in hypoxic environments, even for carnivorous fishes with elevated oxygen requirements.(AU)

Alguns peixes de água doce da América do Sul podem melhorar o desempenho da respiração aquática superficial (RAS) desenvolvendo protuberâncias dérmicas labiais na mandíbula. Considerava-se que essa adaptação era limitada aos peixes onívoros ou herbívoros. Este trabalho apresenta os primeiros indícios de que juvenis de um caracídeo piscívoro, Salminus brasiliensis, desenvolvem protuberâncias dérmicas labiais durante períodos de hipoxia em lagoas de várzea do rio Salado, Argentina. As protuberâncias de S. brasiliensis envolvem porções dérmicas das duas mandíbulas e exibem lóbulos laterais ao lado da boca dispostas no plano vertical. Concentrações de oxigênio dissolvido na água, inferiores ou iguais a 1,05 mgl-1, foram associadas a um aumento notável no desenvolvimento das protuberâncias labiais. Nesta espécie os lóbulos laterais da protuberância podem limitar o acesso de água à porção anterior da boca, que está posicionada mais perto da superfície da água oxigenada durante a RAS. Finalmente, a RAS, complementada com o desenvolvimento de protuberâncias dérmicas labiais, pode ser considerada uma valiosa estratégia para a sobrevivência em ambientes hipóxicos, mesmo para peixes carnívoros com elevada demanda de oxigênio.(AU)