Exposure to salinity induces oxidative damage and changes in the expression of genes related to appetite regulation in Nile tilapia (Oreochromis niloticus).

Variations in water salinity and other extrinsic factors have been shown to induce changes in feeding rhythms and growth in fish. However, it is unknown whether appetite-related hormones mediate these changes in Nile tilapia (Oreochromis niloticus), an important species for aquaculture in several countries. This study aimed to evaluate the expression of genes responsible for appetite regulation and genes related to metabolic and physiological changes in tilapia exposed to different salinities. Moreover, the study proposed to sequence and to characterize the cart, cck, and pyy genes, and to quantify their expression in the brain and intestine of the fish by quantitative polymerase chain reaction (qPCR). The animals were exposed to three salinities: 0, 6, and 12 parts per thousand (ppt) of salt for 21 days. Furthermore, lipid peroxidation, reactive oxygen species, DNA damage, and membrane fluidity in blood cells were quantified by flow cytometry. The results indicated an increased expression of cart, pyy, and cck and a decreased expression of npy in the brain, and the same with cck and npy in the intestine of fish treated with 12 ppt. This modulation and other adaptive responses may have contributed to the decrease in weight gain, specific growth rate, and final weight. In addition, we showed oxidative damage in blood cells resulting from increasing salinity. These results provide essential data on O. niloticus when exposed to high salinities that have never been described before and generate knowledge necessary for developing biotechnologies that may help improve the production of economically important farmed fish.

Chronic cold exposure modulates genes related to feeding and immune system in Nile tilapia (Oreochromis niloticus).

Nile tilapia is the fourth most produced species in the global aquiculture panorama. This species requires water temperatures higher than 16 °C to grow and survive, and so, little is known about the effects of low temperatures on genes related to food intake and inflammatory responses. This study brought insights about the modulation of genes in different tissues of Nile tilapia chronically exposed to low temperatures. Thus, sixty animals were divided in two experimental groups: a control group in which the animals remained at the optimum temperature of 24 °C; and an exposed to cold group, in which a decrease in the water temperature was applied until reaching 15 °C. These conditions were maintained for 28 days. Blood samples were collected for flow cytometry analysis, while brain, spleen, liver, and kidney tissues were collected for total RNA extraction, followed by quantitative PCR (RT-qPCR). For genes related to feeding process pathway, it was observed an upregulation in pyy and a downregulation of npy and cart gene expression. Also, pro-inflammatory cytokine genes were modulated in the spleen, kidney and liver with a higher expression of il-1b and tnfα and a reduction in the il-8 and nf-κß gene expressions in the group exposed to 15 °C. The fish exposed to cold presented higher serum cortisol levels than the ones from control group. The blood cell analysis showed a lower level of membrane fluidity and a higher DNA fragmentation and cell disruption in the group exposed to cold. These findings suggest an important effect of a stressful situation in the tilapia organism due to cold exposure. This study brings insights on tilapia wellbeing under low temperature stress. It can be a first step to understanding the appropriate way to cope with cold impacts on aquaculture.

GH Overexpression Alters Spermatic Cells MicroRNAome Profile in Transgenic Zebrafish.

Overexpression of growth hormone (GH) in gh-transgenic zebrafish of a highly studied lineage F0104 has earlier been reported to cause increased muscle growth. In addition to this, GH affects a broad range of cellular processes in transgenic fish, such as morphology, physiology, and behavior. Reports show changes such as decreased sperm quality and reduced reproductive performance in transgenic males. It is hypothesized that microRNAs are directly involved in the regulation of fertility potential during spermatogenesis. The primary aim of our study was to verify whether gh overexpression disturbs the sperm miRNA profile and influences the sperm quality in transgenic zebrafish. We report a significant increase in body weight of gh-transgenic males along with associated reduced sperm motility and other kinetic parameters in comparison to the non-transgenic group. MicroRNA transcriptome sequencing of gh-transgenic zebrafish sperms revealed expressions of 186 miRNAs, among which six miRNA were up-regulated (miR-146b, miR-200a-5p, miR-146a, miR-726, miR-184, and miR-738) and sixteen were down-regulated (miR-19d-3p, miR-126a-5p, miR-126b-5p, miR-22a-5p, miR-16c-5p, miR-20a-5p, miR-126b-3p, miR-107a-3p, miR-93, miR-2189, miR-202-5p, miR-221-3p, miR-125a, miR-125b-5p, miR-126a-3p, and miR-30c-5p) in comparison to non-transgenic zebrafish. Some of the dysregulated miRNAs were previously reported to be related to abnormalities in sperm quality and reduced reproduction ability in other species. In this study, an average of 134 differentially expressed miRNAs-targeted genes were predicted using the in silico approach. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that the genes of affected pathways were primarily related to spermatogenesis, sperm motility, and cell apoptosis. Our results suggested that excess GH caused a detrimental effect on sperm microRNAome, consequently reducing the sperm quality and reproductive potential of zebrafish males.

Transfection of exogenous DNA complexed to cationic dendrimer induces alterations of bovine sperm microRNAome.

MicroRNAs have been hypothesized to be involved in the regulation of male fertility potential. The primary aim of our study was to demonstrate the effects of transfection with dendrimer nanostructure on the parameters of bovine sperm quality and to investigate whether the microRNA profile could be disturbed after cationic dendrimer-mediated exogenous DNA transfection of bovine spermatozoa. The binding of exogenous DNA was significantly increased when dendrimer-based transfection was implemented. However, cationic dendrimer transfection induced detrimental changes in the kinetics and sperm quality parameters, such as membrane integrity, acrosome reaction, and mitochondrial membrane potential, when compared to the control group. Sperm microRNA sequencing revealed 218 known and 106 novel microRNAs in the sperm samples, among which nine were dysregulated after transfection (one was upregulated and eight were downregulated), in comparison to the non-transfected sperm. All the dysregulated microRNAs were related to sperm quality and embryonic development. These results suggest that the transfection process using the dendrimer nanostructure has an impact on the quality and microRNA profile of bovine sperm.

Estudo de ovários fetais equinos: uma abordagem histológica / Study of fetal equine ovary: an histological approach

O estudo do desenvolvimento embriológico de órgãos reprodutivos representa uma ferramenta importante para a compreensão das particularidades da espécie equina, assim como ampliar os conhecimentos sobre o desenvolvimento de folículos antrais que podem ser utilizados para produção in vitro de embriões. O presente trabalho teve como objetivo descrever as alterações morfológicas de ovários fetais equinos ocorrentes durante o desenvolvimento gestacional. Foram utilizados fetos equinos provenientes de abatedouro. Imediatamente após o abate foi realizada medição da distância cefalococcígea para o cálculo da idade gestacional em dias (dias de gestação - DG) e dissecação dos fetos para a retirada e fixação dos ovários. Foram realizadas avaliações macroscópicas e histológicas pelas técnicas de Hematoxilina-eosina e PAS. Foram utilizados 19 fetos com idade de 50-269 dias de gestação, distribuídos em 9 grupos de DG. Na avaliação macroscópica foi observada diferença entre o volume dos ovários de acordo com a idade gestacional, sendo observado maior volume ovariano entre os dias 210 a 269 de desenvolvimento gestacional. Na avaliação histológica foi observado epitélio de revestimento cúbico e distinção entre as camadas cortical e medular nos ovários fetais a partir de 50-89 DG. As principais características identificadas foram à distinção morfológica entre as camadas cortical e medular nos ovários fetais equinos: os cordões ovígeros surgiram no intervalo de 150-179 DG; o córtex iniciada como uma camada delgada e por volta do 210º dia de gestação apresentou espessamento pelo aumento da quantidade de tecido conjuntivo rico em vasos sanguíneos, o qual se manteve até os estágios finais da gestação; a região medular adquiriu uniformidade estrutural por volta do 150º dia, mostrando células similares em tamanho e vasos sanguíneos de calibre maior. A partir desta etapa, as células medulares diminuíram em tamanho e feixes de tecido conjuntivo denso tornaram esta zona parcialmente dividida.(AU)

The study of embryological development of reproductive organs is an important tool for understanding the particularities of the equine species. Also it is important for increasing knowledge about the development of antral follicles that can be used for in vitro production of embryos. This study aimed to describe the morphological changes of fetal equine ovaries occurring during gestational development. It was used equine fetuses from a slaughterhouse. After slaughter, it was performed the measurement of cefalococcígea distance to calculate the gestational age (days of pregnancy - DP). Afterwards, the ovaries were grossly evaluated, removed and fixed. Then, they were histologically assessed by hematoxylin-eosin and PAS techniques. It was used 19 fetuses aged 50 to 269 days of gestation, distributed in 9 groups divided according to DP. In the grossly evaluation, it was observed differences between the volume of the ovaries according to gestational age, being observed bigger/larger ovarian volume between 210 days to 269 DP. During the histological evaluation, it was observed a simple cuboidal epithelium and a distinction between the cortical and medullary layers in fetal ovaries from 50-89 DP. The main characteristics identified were: the morphological distinction between the cortical and medullary layers in equine fetal ovaries; the ovigerous cords emerged in the interval between 150 and 179 DP; the cortex initiated as a thin layer with an increased amount of connective tissue rich in blood vessels by the 210 DP, structural uniformity acquired in the medular region by the 150 DP, showing similar cells in size and blood vessels of larger caliber. From this stage, the cells in this region had decreased in size and dense connective tissue bundles, which made this zone partially divided.(AU)

Histopathological Analysis of UVB and IR Interaction in Rat Skin.

To determine the chronic skin effects caused by the interaction of infrared and ultraviolet B radiations, male Rattus norvegicus (Wistar) (2 months old) were exposed for 15 days to infrared radiation (600-1500 nm, with a peak at 1000 nm, n = 12) for 30 min (1080 J cm(-2) ) (IRo); to ultraviolet B radiation (peak emission at 313 nm, n = 9) for 90 min (55.08 J cm(-2) ) (UVB); to infrared radiation followed after 90 min by ultraviolet B (n = 6) (IRUVB) and to ultraviolet B followed after 90 min by infrared radiation (n = 9) (UVBIR). Skin samples were collected and histopathological analysis showed the presence of acanthosis, parakeratotic and orthokeratotic hyperkeratosis, intraepidermal pustules, keratin pearls, detachment of epidermis, collagen necrosis, inflammatory infiltrate, vasodilation, basal cell vacuolization and superficial dermis degeneration both in UVB and UVBIR treatments. IRUVB animals showed the same characteristics as above except for parakeratotic hyperkeratosis, keratin pearls and superficial dermis degeneration. To conclude, infrared radiation exposure after ultraviolet B irradiation increases skin damage without protecting the tissue, while infrared radiation exposure before ultraviolet B irradiation showed a protective effect against ultraviolet skin damage.

Age, growth, and reproductive aspects of the dusky grouper Mycteroperca marginata (Actinopterygii: Epinephelidae) in a man-made rocky habitat in southern Brazil

The dusky grouper, Mycteroperca marginata, is an important commercial marine fish that is currently considered an endangered species worldwide and listed as overexploited in Brazil. Although its reproductive biology has been studied elsewhere, no information is available for populations in its southernmost distribution limit on the Brazilian coast. The present work investigates age structure, growth and reproduction of individuals inhabiting a pair of rocky jetties, about 4.5 km long each, located in the mouth of Patos Lagoon in the municipality of Rio Grande, in the southwestern Atlantic Ocean. A total of 130 individuals obtained between February 2007 and May 2009 were analyzed, with total length ranging from 260 to 800 mm. The growth coefficient found in the present study (K: 0.069) was lower than values reported for dusky grouper populations from the Mediterranean Sea (0.087) and southeast Africa (0.09). This difference could be related to higher average water temperatures in these regions compared to the current study site or due to poorer habitat quality in terms of the shelters available for dusky grouper individuals to establish their territories. Microscopic ovarian analyses of individuals during three years of sampling revealed only the occurrence of immature and resting females. The absence of ripe or post-spawning individuals, associated with low GSI values (<0.2) over the studied months, indicates that the dusky grouper did not use this man-made rocky substrate as a spawning site. The current L50 estimate of 451.3 mm indicates that most individuals captured in this area are immature. Such fishing pressure on immature specimens, associated with its slow growth rates and complex reproductive characteristics, threatens sustainability of the current dusky grouper fishery at the study site.

A garoupa-verdadeira, Mycteroperca marginata, é uma espécie de importância comercial que atualmente é considerada como ameaçada mundialmente e sobre-explotada no Brasil. Embora sua biologia reprodutiva tenha sido estudada em outras regiões, nenhuma informação está disponível sobre populações que habitam o extremo sul da sua distribuição na costa brasileira. O presente trabalho investiga a estrutura de idade, crescimento e reprodução de indivíduos que habitam dois molhes de pedra, com cerca de 4,5 km de extensão cada, localizados na desembocadura da laguna dos Patos no município de Rio Grande, no Atlântico Sudoeste. Foram analisados 130 indivíduos obtidos entre fevereiro de 2007 e maio de 2009, com comprimento total entre 260 e 800 mm. O coeficiente de crescimento no presente estudo (K: 0,069) foi inferior aos valores reportados para oMediterrâneo (0,087) e sudeste da África (0,09). Essa diferença pode estar relacionada a temperaturas médias maiores nessas regiões comparadas ao presente local de estudo ou devido a menor qualidade de habitat em termo de abrigos para os indivíduos de garoupa-verdadeira estabelecerem seus territórios. Análises microscópicas dos ovários dos indivíduos durante esses três anos de amostragens revelaram somente a ocorrência de indivíduos imaturos e fêmeas em repouso. A ausência de indivíduos ovados ou em pós-desova, associada com baixos valores do IGS (<0,2), ao longo de todos os meses de coleta, indica que a garoupa-verdadeira não utiliza esse substrato rochoso construído pelo homem como um local de desova. A estimativa de um L50 de 451,3 mm indica que a maioria dos indivíduos capturados nessa região é imatura. Pressão de pesca sobre indivíduos imaturos, associada com a sua lenta taxa de crescimento e complexo modo reprodutivo, ameaça a sustentabilidade da atual pesca da garoupa-verdadeira na área de estudo.

Diet and food consumption of the pearl cichlid Geophagus brasiliensis (Teleostei: Cichlidae): relationships with gender and sexual maturity

Despite the importance of the pearl cichlid Geophagus brasiliensis for the aquarium fish trade worldwide and its wide distribution, many aspects of its biology, such as the relationships between its feeding ecology and reproductive behavior, are not fully understood in natural conditions on its native habitat. In this paper, we investigated its diet focusing on how differences in diet and food consumption are related to differences in gender and sexual maturity. The digestive tract of each individual was dissected and had its content analyzed, whereas each gonad was microscopically analyzed to determine gender (male/female) and sexual maturity (immature/mature). A total of 28 females and 31 males were analyzed. Mature individuals were more common than immature specimens both for males (64.50 percent) and females (64.30 percent). The analysis of 52 individuals with non-empty digestive tracts revealed a diet comprised of 27 items. According to the Index of Alimentary importance ( percentIAi), the most important food items in the diet were Gastropoda (37.30 percent), fragments of vascular plants (15.16 percent), detritus (10.14 percent), Amphipoda (9.24 percent), and fish scales (6.29 percent). Mature males had more empty stomachs (65.00 percent) when compared to immature males (27.27 percent) and immature (55.56 percent) and mature females (40.00 percent). Also, mature females seemed to have more food consumption (greater mean values of total volume) in their digestive tracts than mature males. Some hypotheses are proposed in order to distinguish if this gender-based difference in food consumption in mature individuals of the pearl cichlid could be associated with the development of primary and secondary sexual characteristics or with asymmetrical time invested in parental care activities.

Apesar da sua ampla distribuição e importância para o comércio aquariofilista, muitos aspectos da biologia do cará Geophagus brasiliensis no seu habitat natural, como a relação entre sua ecologia alimentar e comportamento reprodutivo, ainda não são bem conhecidos. Nesse trabalho foi investigada sua dieta, enfocando como as diferenças na dieta e no consumo alimentar estão relacionadas ao sexo (macho/fêmea) e a maturidade (imaturo/maturo). O trato digestório de cada indivíduo foi dissecado e teve seu conteúdo analisado, enquanto cada gônada foi analisada microscopicamente para determinar o sexo e a maturidade. Um total de 28 fêmeas e 31 machos foram analisados. Indivíduos maturos foram mais comuns do que espécimes imaturos, tanto para machos (64,50 por cento) quanto para fêmeas (64,30 por cento). A análise de 52 indivíduos com alimento no trato revelou uma dieta composta por 27 itens. De acordo com o Índice de Importância Alimentar ( por centoIAi), os itens alimentares mais importante na dieta foram Gastropoda (37,30 por cento), fragmentos de plantas vasculares (15,16 por cento), detrito (10,14 por cento), Amphipoda (9,24 por cento) e escamas de peixe (6,29 por cento). Machos maturos tiveram mais estômagos vazios (65,00 por cento) quando comparados com machos imaturos (27,27 por cento) e fêmeas imaturas (55,56 por cento) e maturas (40,00 por cento). Além disso, fêmeas maturas apresentaram maior consumo de alimentos (valor médio do volume total de alimentos) do que machos maturos. Algumas hipóteses são propostas na tentativa de explicar se essa diferença no consumo de alimento entre machos e fêmeas maturos está relacionada ao desenvolvimento de caracteres sexuais primários e secundários, ou com a assimetria na quantidade de tempo empregado em atividades relacionadas ao cuidado parental.