Lunar Age-Dependent Oscillations in Expression of the Genes for Kisspeptin, GnIH, and Their Receptors in the Grass Puffer during the Spawning Season.

Grass puffer is a semilunar-synchronized spawner: spawning occurs on beaches only for several days of spring tide around new moon (lunar age 0) and full moon (lunar age 15) every 2 weeks from spring to early summer. To investigate the role of kisspeptin and gonadotropin-inhibitory hormone (GnIH) in the semilunar-synchronized spawning, lunar age-dependent expression of the genes encoding kisspeptin (kiss2), kisspeptin receptor (kissr2), GnIH (gnih), GnIH receptor (gnihr), gonadotropin-releasing hormone 1 (GnRH1) (gnrh1), and three gonadotropin (GTH) subunits (gpa, fshb, lhb) was examined in the male grass puffer, which was kept in an aquarium under natural light condition in a lunar month during the spawning period. In the brain, both kiss2 and kissr2 showed lunar variations with a peak at lunar age 10, while both gnih and gnihr showed semilunar variations with two peaks at lunar age 0 and 20. On the other hand, gnrh1 showed semilunar variation with two peaks at lunar age 0 and 15. In the pituitary, kiss2, kissr2, gnih, and gnihr showed similar variations to those shown in the brain. The fshb and lhb mRNA levels showed semilunar variations with two peaks at lunar age 0 and 15. The present study shows lunar and semilunar oscillations of kiss2/kissr2 and gnih/gnihr expressions, respectively, with their peaks around spring tide in the brain and pituitary along with the semilunar expressions of gnrh1 and the pituitary GTH subunit genes. These results suggest that the lunar age-dependent expressions of the kisspeptin, GnIH, and their receptor genes may be primarily important in the control of the precisely timed semilunar spawning of the grass puffer.

High temperature acclimation alters upper thermal limits and growth performance of Indian major carp, rohu, Labeo rohita (Hamilton, 1822).

Increase in water temperature due to anthropogenic and climatic changes is expected to affect physiological functions of fish. In this study, we determined high temperature tolerance (CTmax) of a common aquacultured Indian major carp, rohu, Labeo rohita fingerlings (15.96 ± 0.72 g BW, 11.56 ± 0.42 cm TL) followed by acclimatization at three temperatures (30, 33, 36 °C). To determine the CTmax, we analyzed the major hemato-biochemical indices - hemoglobin (Hb), red blood cell (RBC), white blood cell (WBC), blood glucose levels, and erythrocytic nuclear abnormalities (ENAs) and erythrocytic cellular abnormalities (ECAs) of peripheral erythrocytes in the fish sampled at the start and end point at each acclimated temperature. Significantly decreased CTmax of the fish was found at 36 °C compared to 30 °C and 33 °C. The fish in the highest (36 °C) temperature were found with significantly lower Hb and RBC content and significantly higher WBC and blood glucose levels than that of the fishes in the lowest (30 °C) temperature both at the start and end points. The highest frequencies of ENAs and ECAs were found in the highest (36 °C) temperature group compared to the lowest (30 °C) temperature group at both the points. We also evaluated growth performance of the rohu fingerlings reared in the three temperatures for 60 days. The growth parameters - final weight gain, percent weight gain and specific growth rate were the highest at 33 °C and the lowest at 36 °C. The present study revealed that the highest temperature (36 °C) tested here may be hazardous to rohu and the temperature should be kept below 36 °C in the aquaculture setting to avoid physiological damage and growth and production loss to the fish.

Determinants of arsenic methylation efficiency and urinary arsenic level in pregnant women in Bangladesh.

BACKGROUND: Prenatal inorganic arsenic (iAs) exposure is associated with pregnancy outcomes. Maternal capabilities of arsenic biotransformation and elimination may influence the susceptibility of arsenic toxicity. Therefore, we examined the determinants of arsenic metabolism of pregnant women in Bangladesh who are exposed to high levels of arsenic. METHODS: In a prospective birth cohort, we followed 1613 pregnant women in Bangladesh and collected urine samples at two prenatal visits: one at 4-16 weeks, and the second at 21-37 weeks of pregnancy. We measured major arsenic species in urine, including iAs (iAs%) and methylated forms. The proportions of each species over the sum of all arsenic species were used as biomarkers of arsenic methylation efficiency. We examined the difference in arsenic methylation using a paired t-test between first and second visits. Using linear regression, we examined determinants of arsenic metabolism, including age, BMI at enrollment, education, financial provider income, arsenic exposure level, and dietary folate and protein intake, adjusted for daily energy intake. RESULTS: Comparing visit 2 to visit 1, iAs% decreased 1.1% (p <  0.01), and creatinine-adjusted urinary arsenic level (U-As) increased 21% (95% CI: 15, 26%; p <  0.01). Drinking water arsenic concentration was positively associated with iAs% at both visits. When restricted to participants with higher adjusted urinary arsenic levels (adjusted U-As > 50 µg/g-creatinine) gestational age at measurement was strongly associated with DMA% (ß = 0.38, p <  0.01) only at visit 1. Additionally, DMA% was negatively associated with daily protein intake (ß = - 0.02, p <  0.01) at visit 1, adjusting for total energy intake and other covariates. CONCLUSIONS: Our findings indicate that arsenic metabolism and adjusted U-As level increase during pregnancy. We have identified determinants of arsenic methylation efficiency at visit 1.

Effects of high and low temperature on expression of GnIH, GnIH receptor, GH and PRL genes in the male grass puffer during breeding season.

Gonadotropin-inhibitory hormone (GnIH) is a multifunctional hypophysiotropic neurohormone and has a stimulatory role in the control of reproduction in the grass puffer. To clarify the neuroendocrine mechanisms underlying the effect of changes in water temperature on reproduction in fish, we previously revealed that, in parallel to gonadal regression, both low and high temperature significantly decreased the expressions of the genes encoding kisspeptin (kiss2), kisspeptin receptor (kiss2r), gonadotropin-releasing hormone 1 (gnrh1) in the brain and gonadotropin (GTH) subunits (fshb and lhb) in the pituitary of sexually mature male grass puffer. In this study, we examined the changes in expression of gnih and GnIH receptor gene (gnihr) in the brain and pituitary along with the genes for growth hormone (gh) and prolactin (prl) in the pituitary of male grass puffer exposed to low temperature (14 °C), normal temperature (21 °C, as initial control) and high temperature (28 °C) conditions for 7 days. The levels of gnih and gnihr mRNAs were significantly decreased in both low and high temperature conditions compared to normal temperature in the brain and pituitary. Similarly, the gh mRNA levels were significantly decreased in both low and high temperature conditions. The prl mRNAs showed no significant changes at high temperature, whereas drastically decreased at low temperature possibly by dysfunctional cold stress. Taken together, the present results suggest that, in addition to the inhibitory effect of temperature changes on the Kiss2/GnRH1/GTH system, the suppression of GnIH/GH system may also be involved in the termination of reproduction by high temperature at the end of breeding season.

Anomalous Temperature Interdicts the Reproductive Activity in Fish: Neuroendocrine Mechanisms of Reproductive Function in Response to Water Temperature.

Fish are poikilotherm and small changes in water temperature can greatly affect physiological processes including reproduction, which is regulated by complex neuroendocrine mechanisms that respond to climatic events. This review provides evidence that anomalous high and low temperature may directly affect reproduction in fish by suppressing the expression of genes in the reproductive neuroendocrine system. The grass puffer, Takifugu alboplumbeus, is an excellent animal model for studying the thermal regulation of reproduction, for they exhibit periodic spawning activities, which are synchronized with seasonal, lunar and daily cycles. In the grass puffer, the expression of the genes encoding gonadotropin-releasing hormone (GnRH) 1, kisspeptin, gonadotropin-inhibitory hormone (GnIH) and their receptors were markedly suppressed in the diencephalon of fish exposed to high temperature (28°C) when compared to normal temperature (21°C), followed by the decrease in the pituitary mRNA levels for follicle-stimulating hormone (FSH), luteinizing hormone (LH) and growth hormone (GH). On the other hand, the exposure to low temperature (14°C) also inhibited the expression of gnrh1, kiss2, gnih and their receptor genes in the brain and fshb, lhb, gh and prl in the pituitary. Taken together, it is plausible that anomalous high and low temperature may be a proximate driver of termination of reproduction by suppressing the activity of the reproductive GnRH/kisspeptin/GnIH system, possibly through direct action of temperature signals at transcription level.

Gut probiotic bacteria of Barbonymus gonionotus improve growth, hematological parameters and reproductive performances of the host.

This study aimed to isolate and identify probiotic bacteria from the gut of Barbonymus gonionotus and evaluate their effects on growth, hematological parameters, and breeding performances of the host. Five probiotic bacteria viz. Enterococcus xiangfangensis (GFB-1), Pseudomonas stutzeri (GFB-2), Bacillus subtilis (GFB-3), Citrobacter freundii (GFB-4), and P. aeruginosa (GFB-5) were isolated and identified using 16S rRNA gene sequencing. Application of a consortium of probiotic strains (1-3 × 1.35 × 109 CFU kg-1) or individual strain such as GFB-1 (1.62 × 109 CFU kg-1), GFB-2 (1.43 × 109 CFU kg-1), GFB-3 (1.06 × 109 CFU kg-1), GFB-4 (1.5 × 109 CFU kg-1) or GFB-5 (1.43 × 109 CFU kg-1feed) through feed significantly improved growth, histological and hematological parameters and reproductive performances of B. gonionotus compared to untreated control. Moreover, the application of these probiotics significantly increased gut lactic acid bacteria and activities of digestive enzymes but did not show any antibiotic resistance nor any cytotoxicity in vitro. The highest beneficial effects on treated fishes were recorded by the application of GFB-1, GFB-2, GFB-3, and a consortium of these bacteria (T2). This is the first report of the improvement of growth and health of B. gonionotus fishes by its gut bacteria.