Endocrine disruptive action of diclofenac and caffeine on Astyanax altiparanae males (Teleostei: Characiformes: Characidae)

Diclofenac (DCF) and caffeine (CAF) are persistent pharmaceuticals that occur in mixtures in the aquatic ecosystems causing effects in the reproductive physiology of aquatic organisms. This study evaluated the physiological reproductive responses of Astyanax altiparanae males exposed to nominal concentrations of DCF (3.08 mg L− 1) and CAF (9.59 mg L− 1) separately and combined, for 96 h. The steroids profile, estrogenic biomarker vitellogenin (vtgA), testes and liver morphology, and also mortality of males were assessed. DCF and CAF degradation was 5% of the initial concentration for 24 h. The LC50 of the DCF and CAF were 30.8 mg L− 1 and 95.9 mg L− 1, respectively. Males exposed to DCF and CAF exhibited a reduction of 17β-Estradiol (E2) concentration compared to control (CTL). Similarly, testosterone (T) was also reduced in the DCF treatment, but this response was not observed in 11-Ketotestosterone (11-KT). Males exposed to DCF + CAF combined did not exhibit differences in T, E2 and 11-KT steroids. The vtgA gene expression and the sperm concentration did not change among the treatments. Moreover, acute exposure revealed a hypertrophy of hepatocytes cells in the DCF and DCF + CAF treatments. In conclusion, DCF and CAF, isolated, exhibit an endocrine disruptive activity in A. altiparanae male, an opposite response observed with the mixture of both compounds that abolishes the endocrine disruptive effects. DCF seems to be more toxic for this species, altering also hepatocytes morphology.Fil: Godoi, Filipe G.A.. Universidade de Sao Paulo; BrasilFil: Muñoz Peñuela, Marcela. Universidade de Sao Paulo; BrasilFil: Olio Gomes, Aline D.. Universidade de Sao Paulo; BrasilFil: Tolussi, Carlos E.. Universidade Anhembi-Morumbi; BrasilFil: Brambila Souza, Gabriela. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Branco, Giovana S.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Lo Nostro, Fabiana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Moreira, Renata. Universidade de Sao Paulo; Brasi

Embryonic development and larval stages of Steindachneridion parahybae (Siluriformes: Pimelodidae): implications for the conservation and rearing of this endangered Neotropical species

Steindachneridion parahybae is a freshwater catfish endemic to the Paraíba do Sul River and is classified as an endangered Neotropical species. An increasing number of conservation biologists are incorporating morphological and physiological research data to help conservation managers in rescue these endangered species. This study investigated the embryonic and larval development of S. parahybae in captivity, with emphasis in major events during the ontogeny of S. parahybae. Broodstocks were artificially induced to reproduce, and the extrusion occurred 200-255 degree-hours after hormonal induction at 24°C. Larval ontogeny was evaluated every 10 minutes under microscopic/stereomicroscopic using fresh eggs samples. The main embryogenic development stages were identified: zygote, cleavage, including the morula, blastula, gastrula phase, organogenesis, and hatching. The extruded oocytes showed an average diameter of 1.10 ± 0.10 mm, and after fertilization and hydration of eggs, the average diameter of eggs increased to about 1.90 ± 0.60 mm, characterized by a large perivitelline space that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). Cell division started about 2 minutes after fertilization (AF), resulting in 2, 4, 8 (4 x 2 arrangement of cells), 16 (4 x 4), 32 (4 x 8) and 64 (2 x 4 x 8) cells. Furthermore, the blastula and gastrula stages followed after these cells divisions. The closed blastopore occurred at 11 h 20 min AF; following the development, the organogenetic stages were identified and subdivided respectively in: early segmentation phase and late segmentation phase. In the early segmentation phase, there was the establishment of the embryonic axis, and it was possible to distinguish between the cephalic and caudal regions; somites, and the optic vesicles developed about 20 h AF. Total hatching occurred at 54 h AF, and the larvae average length was 4.30 ± 0.70 mm. Gradual yolk sac reduction was observed during the first two days of larval development. The first feeding occurred at the end of the second day. During the larval phase, cannibalism, heterogeneous larval growth and photophobia were also observed. This information will be important in improving the artificial reproduction protocols of S. parahybae in controlled breeding programs.<br>Steindachneridion parahybae é um bagre de água doce, endêmico do rio Paraíba do Sul e é classificado como espécie neotropical ameaçada. Um número crescente de biólogos conservacionistas estão incorporando dados de pesquisas morfológicas e fisiológicas para ajudar os gestores de conservação no resgate destas espécies ameaçadas de extinção. Este estudo investigou o desenvolvimento embrionário e larval de S. parahybae em cativeiro, com ênfase nos principais eventos durante a ontogenia de S. parahybae. Reprodutores foram artificialmente induzidos à reprodução e a extrusão ocorreu com 200-255 horas-graus após a indução hormonal a 24°C. A ontogenia larval foi avaliada a cada 10 minutos sob microscópio/ estereomicroscópio, utilizando-se amostras de ovos recém coletados. Os principais estágios de desenvolvimento embrionário foram identificados: zigoto, clivagem, incluindo as fases, mórula, blástula e gástrula, organogênese e eclosão. Os ovócitos extrusados apresentaram uma média de diâmetro de 1,10 ± 0,10 mm e depois da fertilização e hidratação dos ovos, a média de diâmetro dos ovos aumentou para 1,90 ± 0,60 mm, caracterizado pelo grande espaço vitelínico que persistiu até o desenvolvimento do embrião, duplo córion e os polos (animal e vegetal). A divisão celular iniciou-se aproximadamente dois minutos após a fertilização (AF), resultando em 2, 4, 8 (4 x 2 arranjo das células), 16 (4 x 4), 32 (4 x 8) e 64 (2 x 4 x 8) células. Adicionalmente, seguiram as fases de blástula e gástrula depois das divisões celulares. O fechamento do blastóporo ocorreu às 11 h 20 min AF; seguindo o desenvolvimento, os estágios de organogênese foram identificados e subdivididos, respectivamente em: fase de divisão inicial e fase de divisão avançada. Na fase de divisão inicial, depois do estabelecimento do eixo do embrião, foi possível distinguir as regiões cefálica e caudal; os somitos e as vesículas ópticas se desenvolveram com aproximadamente 20 h AF. A eclosão total ocorreu com cerca de 54 h AF e o comprimento médio foi de 4,30 ± 0,70 mm. A redução gradual do saco vitelínico foi observada durante os primeiros dois dias de desenvolvimento larval. A primeira alimentação ocorreu no final do segundo dia. Durante a fase larval, canibalismo, crescimento larval heterogêneo e fotofobia também foram observados. Estas informações serão importantes para aperfeiçoar o protocolo de reprodução artificial em S. parahybae em programas de reprodução controlada

Embryonic development and larval stages of Steindachneridion parahybae (Siluriformes: Pimelodidae): implications for the conservation and rearing of this endangered Neotropical species

Steindachneridion parahybae is a freshwater catfish endemic to the Paraíba do Sul River and is classified as an endangered Neotropical species. An increasing number of conservation biologists are incorporating morphological and physiological research data to help conservation managers in rescue these endangered species. This study investigated the embryonic and larval development of S. parahybae in captivity, with emphasis in major events during the ontogeny of S. parahybae. Broodstocks were artificially induced to reproduce, and the extrusion occurred 200-255 degree-hours after hormonal induction at 24°C. Larval ontogeny was evaluated every 10 minutes under microscopic/stereomicroscopic using fresh eggs samples. The main embryogenic development stages were identified: zygote, cleavage, including the morula, blastula, gastrula phase, organogenesis, and hatching. The extruded oocytes showed an average diameter of 1.10 ± 0.10 mm, and after fertilization and hydration of eggs, the average diameter of eggs increased to about 1.90 ± 0.60 mm, characterized by a large perivitelline space that persisted up to embryo development, the double chorion, and the poles (animal and vegetative). Cell division started about 2 minutes after fertilization (AF), resulting in 2, 4, 8 (4 x 2 arrangement of cells), 16 (4 x 4), 32 (4 x 8) and 64 (2 x 4 x 8) cells. Furthermore, the blastula and gastrula stages followed after these cells divisions. The closed blastopore occurred at 11 h 20 min AF; following the development, the organogenetic stages were identified and subdivided respectively in: early segmentation phase and late segmentation phase. In the early segmentation phase, there was the establishment of the embryonic axis, and it was possible to distinguish between the cephalic and caudal regions; somites, and the optic vesicles developed about 20 h AF. Total hatching occurred at 54 h AF, and the larvae average length was 4.30 ± 0.70 mm. Gradual yolk sac reduction was observed during the first two days of larval development. The first feeding occurred at the end of the second day. During the larval phase, cannibalism, heterogeneous larval growth and photophobia were also observed. This information will be important in improving the artificial reproduction protocols of S. parahybae in controlled breeding programs

Mobilization of energetic substrates in the endangered catfish Steindachneridion parahybae (Siluriformes: Pimelodidae): changes in annual reproductive cycle in captivity

ABSTRACT This study aimed at analyzing the energetic substrate (ES) in the main storage tissues of Steindachneridion parahybae, throughout the reproductive cycle in captivity. Differently from wild, in captivity, feeding is not interrupted during the reproductive period, the females do not spawn spontaneously, and they are sedentary. Adult females were sampled monthly and based on their histology and gonadosomatic index (GSI), ovaries were classified into: previtellogenic (PRV), vitellogenic (VTG), and regression (REG) stages. Ovaries at the VTG stage showed higher protein and lipids levels than at the PRV stage with a positive correlation between these substrates and the GSI. Muscle was the main source of proteins transferred to the ovaries, according to the negative correlation between these organs. Lipids remained unchanged in the liver, which is an important supplier in vitellogenesis, a pattern that probably occurs due to the continuous feeding. Muscular glycogen levels were higher at the VTG and REG than at the PRV stages. Plasma triglycerides were also higher during REG, while glucose levels were more elevated during the VTG stage. These results suggest that with constant food supply, the pattern of deposition of ES in S. parahybae is different from that described for other wild potamodromous species

Environmental pollution affects molecular and biochemical responses during gonadal maturation of Astyanax fasciatus (Teleostei: Characiformes: Characidae)

Endocrine disrupting compounds (EDCs) have the potential to alter fish reproduction at various levels of organization. The aim of this study was to assess the impact of a natural environment with heavily anthropogenic influence on the physiological processes involved in reproduction in the freshwater fish lambari (Astyanax fasciatus) using different biomarkers. Adult males and females were collected in different seasons from two distinct sites in the same watershed: Ponte Nova Reservoir (PN) considered a pristine or small anthropogenic influence reference point; and Billings Reservoir (Bil), subjected to a large anthropogenic impact. Biological indices, such as hepatosomatic index and gonadosomatic index (GSI), gonadal histomorphology, fecundity, and biomarkers such as plasma levels of estradiol (E2) as well as hepatic gene expression of its alfa nuclear receptor (ERα), were analyzed. Hepatic vitellogenin (VTG) gene expression was evaluated in both sexes, as an indicator of xenoestrogen exposure. Females collected at PN presented a typical annual variation reflected in GSI, whereas for those sampled at Bil the index did not change through the seasons. The higher concentration of E2 in males collected at Bil during spring/2013, together with the detection of VTG gene expression, suggest the presence of EDCs in the water. These EDCs may have also influenced fecundity of females from Bil, which was higher during winter and spring/2013. Gene expression of ERα and ovarian morphology did not differ between fish from both sites. Water conditions from Bil reservoir impacted by anthropic activity clearly interfered mainly with biomarkers of biological effect such as plasma E2 levels and absolute and relative fecundity, but also altered biomarkers of exposure as VTG gene expression. These facts support the notion that waterborne EDCs are capable of causing estrogenic activity in A. fasciatus.Fil: Tolussi, Carlos E.. Universidade de Sao Paulo; BrasilFil: Olio Gomes, Aline Dal. Universidade de Sao Paulo; BrasilFil: Kumar, Anupama. CSIRO Land and Water; AustraliaFil: Ribeiro, Cristiele S.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Lo Nostro, Fabiana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Ecotoxicología Acuática; ArgentinaFil: Bain, Peter A.. CSIRO Land and Water; AustraliaFil: Souza, Gabriela B. de. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Da Cuña, Rodrigo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Ecotoxicología Acuática; ArgentinaFil: Honji, Renato Massaaki. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Moreira, Renata. Universidade de Sao Paulo; Brasi