Skeletal deformities and meristic trait variations are common in the intertidal fish Bathygobius cocosensis (Perciformes-Gobiidae).

Extreme environmental conditions, such as temperature, can lead to meristic trait variation and skeletal deformities, which may have major impacts on individual fitness. As intertidal ecosystems experience rapid temperature and physicochemical fluctuations, intertidal fish living and reproducing in these environmental conditions may have phenotypes influenced by such variable environments. The impact of intertidal variability on fish development, however, has not been previously investigated. Skeletal deformities and meristic traits were assessed for Bathygobius cocosensis, a common intertidal fish living across the Indo-Pacific region, using a clearing and staining method on 72 individuals. Over 87% of individuals presented meristic variation and over 70% exhibited at least one type of skeletal deformity, mostly recorded in the caudal fin area. The unexpected prevalence of skeletal deformities among this intertidal fish population suggests that such deformities may be suitable markers to evaluate an individual's stress exposure during development and the subsequent fitness effects.

Deformities in reared cobia, Rachycentron canadum L. and grouper, Epinephelus marginatus, in São Paulo state coast, Brazil: case report / Deformidades em cobias, Rachycentron canadum L, e garoupas, Epinephelus marginatus, cultivadas na costa do estado de São Paulo, Brasil: relato de caso

Skeletal deformities are frequent problems in cultured fish populations due mainly to unfavorable abiotic conditions, inappropriate nutrition, and genetic factors. These may cause erratic swimming, decrease of conversion rate, growth, and market value. The aim of this case report was to present the occurrence of deformities in juvenile cobia and groupers from an offshore marine farm in Ilhabela, São Paulo, Brazil. Nine cobias and seven juvenile groupers, that presented deformities, were euthanized and fixed in 10% formaldehyde. They were tagged, processed, and stained with "Alcian Blue" (AB) and "Alizarin Red-S" (ARS) for visualization of cartilage and bone tissue, respectively. After evisceration, radiographic examination was performed. The deformities observed in cobia were atrophy of the operculum, disorganization of the gill lamellae, deformity of the lower jaw, kyphosis, and scoliosis. Groupers presented lower jaw deformity and saddleback syndrome. The farm studied in this case report had poor management standards, fed commercial feed for freshwater herbivorous fish and cooked sardines, high inbreeding and absence of adequately trained professionals to manage these species. The sum of these factors must have directly influenced the appearance of these deformities. It is necessary to control this situation to minimize the incidence of malformations that have economic impacts to farmers.(AU)

As deformidades esqueléticas são problemas frequentes em populações de peixes cultivados devido, principalmente, a condições abióticas desfavoráveis, nutrição inapropriada e fatores genéticos. Estas podem causar natação errática, diminuição do índice de conversão, do crescimento e do valor no mercado. O objetivo deste relato de caso foi apresentar a ocorrência de deformidades em cobias e garoupas juvenis provenientes de uma fazenda marinha em sistema offshore, em Ilhabela, São Paulo, Brasil. Nove cobias e sete garoupas juvenis, que apresentavam deformidades, foram sujeitos à eutanásia e fixados em formol a 10%. Os espécimes foram registrados, processados e corados com "Alcian Blue" (AB) e "Alizarin Red-S" (ARS) para visualização de tecido cartilaginoso e ósseo, respectivamente. Os peixes foram eviscerados para o exame radiográfico. As deformidades observadas em cobia foram: atrofia do opérculo, desorganização das lamelas branquiais, deformidade do maxilar inferior, cifose e escoliose. As garoupas apresentaram deformidade do maxilar inferior e síndrome de Saddleback. Os peixes deste relato de caso estiveram sob manejo deficiente, alimentados com ração comercial para peixes herbívoros de água doce e sardinhas cozinhadas, alta consanguinidade e ausência de profissionais adequadamente treinados para o manejo dessas espécies. A soma desses fatores influenciou diretamente na aparição dessas deformidades, precisando ser controlados para minimizar a incidência de malformações que tenham impacto econômico para os produtores.(AU)

Effects of manganese on fat snook Centropomus parallelus (Carangaria: Centropomidae) exposed to different temperatures

This study evaluates the effects of exposure to manganese (Mn2+) for 96 hours at two different temperatures (24 and 27°C) on juveniles of Centropomus parallelus through the activities of glutathione S-transferase (GST) and catalase (CAT), micronuclei test (MN) and comet assay. The GST activity did not show any significant difference between the groups exposed to Mn2+ and the respective control groups; in contrast, a major increase in the CAT activity was observed at 27°C in the group exposed to Mn2+ compared to the control group. The genotoxic analyses showed that in all animals exposed to Mn2+, the number of red cells with micronuclei increased significantly compared to the respective control groups. There was also a significant increase in the incidence of DNA damage in the groups exposed to Mn2+. At a temperature of 24ºC, animals exposed to Mn2+ had more DNA damage than those at 27°C. It is likely that the increase in temperature can also induce oxidative stress. Thus, we conclude that manganese is toxic to the fat snook juveniles, causing genotoxic damage, and when associated with an increase in temperature, manganese can also provoke an increase in oxidative stress.(AU)

Este estudo avaliou os efeitos da exposição ao manganês (Mn2+), após 96 horas, a duas temperaturas (24 e 27°C) em juvenis de Centropomus parallelus por meio de análises bioquímicas (atividade das enzimas glutationa S-transferase (GST) e catalase (CAT)) e genotóxicas (teste do micronúcleo e ensaio cometa). A atividade da GST não mostrou diferença significativa entre os grupos expostos ao Mn2+ e os seus respectivos grupos controle, enquanto que um aumento significativo na atividade da CAT foi observado a 27°C no grupo exposto ao Mn2+, quando comparado ao grupo controle. As análises genotóxicas mostraram que os animais expostos ao Mn2+ tiveram aumento significativo na quantidade de células com micronúcleo em relação aos seus grupos de controles. Houve também aumento significativo na incidência de danos ao DNA nos grupos expostos a esse contaminante. Na temperatura de 24°C, os animais expostos ao Mn2+ tiveram maior quantidade de danos no DNA em relação a 27°C. É provável que o aumento da temperatura também possa induzir o estresse oxidativo. Assim, concluímos que o manganês é tóxico para os juvenis de robalo, causando dano genotóxico, e quando associado a um aumento da temperatura, também pode provocar um aumento no estresse oxidativo.(AU)

Severe malformations of eelpout (Zoarces viviparus) fry are induced by maternal estrogenic exposure during early embryogenesis.

Pregnant eelpout were exposed via the water to known endocrine disrupting compounds (EDCs) to clarify if EDCs could be causing the increased eelpout fry malformation frequencies observed in coastal areas receiving high anthropogenic input. The presence of a teratogenic window for estrogen-induced malformations was also investigated by starting the exposure at different times during eelpout pregnancy. Both 17α-ethinylestradiol (EE2) (17.8 ng/L) and pyrene (0.5 µg/L) significantly increased fry malformation frequency whereas 4-t-octylphenol (4-t-OP) up to 14.3 µg/L did not. Vitellogenin was significantly induced by EE2 (5.7 and 17.8 ng/L) but not by 4-t-OP and pyrene. A critical period for estrogen-induced fry malformations was identified and closed between 14 and 22 days post fertilization (dpf). Exposure to 17ß-estradiol (E2) between 0 and 14 dpf caused severe malformations and severity increased the closer exposure start was to fertilization, whereas malformations were absent by exposure starting later than 14 dpf. Data on ovarian fluid volume and larval length supported the suggested teratogenic window. Larval mortality also increased when exposure started right after fertilization.

Effect of salinity on survival, growth and biochemical parameters in juvenile Lebranch mullet Mugil liza (Perciformes: Mugilidae)

Teleost fish growth may be improved under isosmotic condition. Growth and metabolic performance of juvenile Mugil liza (isosmotic point: 12‰) were evaluated after 40 days in different salinities (0, 6, 12 and 24‰). Tests were performed in quadruplicate (30 fish/tank; 0.48 ± 0.1 g body weight; 3.27 ± 0.1 cm total length) under controlled water temperature (28.2 ± 0.1ºC) and oxygen content (>90% saturation). Fish were fed on artificial diet (50% crude protein) four times a day until apparent satiation. Results showed that salinity influenced juvenile mullet growth. Fish reared at salinity 24‰ grew better than those maintained in freshwater (salinity 0‰). Gill Na+,K+-ATPase activity and whole body oxygen consumption showed an U-shape-type response over the range of salinities tested, with the lower values being observed at the intermediate salinities. Although no significant difference was observed in liver glycogen content at different salinities, it tended to augment with increasing salinity. These findings indicate that energy demand for osmorregulation in juvenile M. liza can be minimized under isosmotic condition. However, the amount of energy spared is not enough to improve fish growth. Results also suggest that M. liza is able to alternate between different energy-rich substrates during acclimation to environmental salinity.

O crescimento de peixes teleósteos pode ser melhorado em condição isosmótica. O crescimento e o desempenho metabólico de juvenis da tainha Mugil liza (ponto isosmótico: salinidade de 12‰) foram avaliados após 40 dias de cultivo em diferentes salinidades (0, 6, 12 e 24‰). Os testes foram realizados em 4 réplicas (30 peixes/tanque; 0,48 ± 0,1 g de peso corporal; 3,27 ± 0,1 cm de comprimento total) em condições controladas de temperatura (28,2 ± 0,1ºC) e conteúdo de oxigênio (>90% saturação). Os peixes foram alimentados quatro vezes ao dia com dieta artificial (50% de proteína bruta) até a saciedade aparente. Os resultados mostraram que a salinidade influenciou o crescimento dos juvenis da tainha. Os peixes cultivados na salinidade 24‰ cresceram melhor que aqueles mantidos na água doce (salinidade 0‰). A atividade da Na+,K+-ATPase branquial e o consumo corporal de oxigênio mostraram uma resposta do tipo em forma de U, na faixa de salinidade testada, com os menores valores sendo observados nas salinidades intermediárias. Apesar de não ter sido observada diferença significativa no conteúdo de glicogênio entre os peixes mantidos nas diferentes salinidades, este parâmetro tendeu a aumentar com o incremento da salinidade. Estes achados indicam que a demanda energética para osmorregulação em juvenis de M. liza podem ser minimizados em condição isosmótica. Entretanto, a quantidade de energia poupada não é suficiente para melhorar o crescimento. Os resultados também sugerem que M. liza é capaz de alternar entre diferentes substratos ricos em energia durante a aclimatação à salinidade da água.

Morphological deformities in the osseous structure in spotted sorubim Pseudoplatystoma coruscans (Agassiz & Spix, 1829) with vitamin C deficiency.

Vitamin C is essential for fish diets because many species cannot syntethize it. This vitamin is needed for bone and cartilage formation. Moreover, it acts as antioxidant and improve the immunological system. The present work investigated the effects of vitamin C diet supplementation to spotted sorubim (Pseudoplatystoma coruscans) fingerlings by frequency of bone and cartilage deformation. Ascorbyl poliphosphate (AP) was used as source of vitamin C in the diets for spotted sorubim fingerlings during three months. Six diets were formulated: one diet control (0 mg/kg of vitamin C) and 500, 1,000, 1,500, 2,000 and 2,500 mg AP/kg diets. Fishes fed without vitamin C supplementation presented bone deformation in head and jaws, and fin fragilities. Thus, 500 mg AP/kg diet was enough to prevent deformation and the lack of vitamin C supplementation worsening the development of fingerlings.

Hypoxia impairs embryo development and survival in black bream (Acanthopagrus butcheri).

Coastal environments are threatened by the increasing frequency, extent and severity of hypoxic events. Hypoxia affects vast areas around the world and often causes fish kills, reduced abundance, altered distribution, low benthic biomass and declines in fisheries. In Australia, many fisheries are based on sparid fishes and in the southern states black bream (Acanthopagrus butcheri) is important to both the recreational and commercial sectors. This species completes its entire life cycle in estuaries and annual recruitment is highly variable and very likely influenced by environmental conditions during the spawning season. In a laboratory-based experiment, fertilised black bream eggs (embryos) were exposed to five different levels of dissolved oxygen (DO). The DO levels were maintained in small test wells using nitrogen gas in a novel chamber design. Embryo development was assessed over a 2-day period and hatched larvae were observed until Day 2 post-hatch. Significant differences (p<0.05) were observed in embryonic development and survival as a function of DO level. In severely hypoxic conditions (30% saturation) survival to 1 day was reduced and no hatching occurred. In moderately hypoxic conditions (45-55%S), both precocious and delayed hatching was observed and hatch rates were reduced, whilst the number of hatched larvae with deformities increased, resulting in reduced larval lengths. No larvae survived to Day 2 post-hatch when held in hypoxic conditions (<55%S). This study demonstrates the detrimental effect that severe hypoxia can have on the early development of black bream which could result in reduced recruitment and lowered abundance. Other species that share similar early life histories may also be at risk.

Developmental disorders in larvae of eelpout (Zoarces viviparus) from German and Swedish Baltic coastal waters.

Pregnant eelpout were collected for several years at German and Swedish Baltic coastal stations with different impact of pollutants and the prevalence of fry abnormalities compared between sites. Mortality of apparently normal developed larvae was observed with high variability in both German and Swedish sites. Malformation of larvae was a prominent disorder at all German stations with a prevalence ranging from about 50% up to a maximum of 90% in almost all analysed samples. At the Swedish stations malformation rates were significantly lower (range: 0-6%). Growth retardation of fry was clearly more prominent in eelpout from German coastal waters as well. Taken together the field data suggests that impaired larval development was associated with the state of environmental pollution at the sampling locations.

Perinotochordal connective sheet of gilthead sea bream larvae (Sparus aurata, L.) affected by axial malformations: an histochemical and immunocytochemical study.

BACKGROUND: Spinal malformations in adult teleosts occur under natural conditions and, more frequently, in culture exploitations. Skeletal deformities are linked with dysfunctions in collagen metabolism. We studied axial deviations appearing in early larval stages of cultured sea bream (Sparus aurata, L.). METHODS: To evaluate connective tissue components of normal and lordotic fish we used histochemistry (alcian blue, picrosirius-polarization, clorhydric orcein, fuchsin resorcin), immunohistochemistry (anti-collagen I, II, III, and IV), and specific enzymatic digestions. The results were evaluated by semiquantitative methods. RESULTS: Lordosis appeared before a vertebral column was developed, thus affecting the only skeletal structure present in the animal body, the notochord. At this stage the animal depends on the vitelline sac and an inflated swim-bladder is missing. The region of the curvature showed strong alterations in the arrangement of the muscle bundles and irregularities in notochord and perinotochordal collagen sheet. Histochemical and immunocytochemical analysis of the perinotochordal sheet revealed the presence of type II collagen, non-sulfated glycosaminoglycans, and elastic fibers in normal and lordotic specimens. Low collagen-proteoglycan interactions occurred in lordotic animals. CONCLUSIONS: Lordosis in Sparus aurata originated during embryonic development and was characterized by disorganized connective tissue and muscle bundles. No major differences in connective tissue constituents were seen with respect to normal specimens.