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.

Hypoxia alters the upper thermal limits and blood physiology in zebrafish, Danio rerio.

Hypoxic aquatic environments occur more frequently as a result of climate change, thereby exerting challenges on the physiological and metabolic functions of aquatic animals. In this study, a model fish, zebrafish (Danio rerio) was used to observe the climate-induced hypoxic effect on the upper thermal limit (critical thermal maximum; CTmax), hemoglobin, and blood glucose levels, and abnormalities of erythrocytes at cellular and nuclear level. The value of CTmax decreased significantly under hypoxia (39.10 ± 0.96 °C) compared to normoxia (43.70 ± 0.91 °C). At CTmax, hemoglobin levels were much lower (9.33 ± 0.60 g/dL) and blood glucose levels were significantly higher (194.20 ± 11.33 mg/L) under hypoxia than they were under normoxia and at the beginning of the experiment. Increased frequencies of abnormalities in the erythrocytes at both cellular (fusion, twin, elongated, spindle and tear drop shaped) and nuclear (micronucleus, karyopyknosis, binuclei, nuclear degeneration and notched nuclei) levels were also found under hypoxia compared to normoxia. These results suggest that hypoxic conditions significantly alter the temperature tolerance and subsequent physiology in zebrafish. Our findings will aid in the development of effective management techniques for aquatic environments with minimum oxygen availability.

Expression dynamics of the genes for the hypothalamo-pituitary-gonadal axis in tiger puffer (Takifugu rubripes) at different reproductive stages.

Tiger puffer, Takifugu rubripes, a commercially important long-distance migratory fish, return to specific spawning grounds for reproduction. To clarify reproductive neuroendocrine system of the tiger puffer, the changes in the expression levels of the genes encoding three gonadotropin-releasing hormones (GnRHs), gonadotropin-inhibitory hormone (GnIH), GnIH receptor (GnIH-R), kisspeptin and kisspeptin receptor in the brain and gonadotropin (GTH) subunits, growth hormone (GH) and prolactin (PRL) in the pituitary were examined in the tiger puffer captured in the wild at different reproductive stages, namely immature and mature fish of both sexes, and post-ovulatory females that were obtained by hormonal treatment. The amounts of three gnrh mRNAs, gnih, gnih-r, fshb and lhb were substantially increased in the mature fish compared to the immature fish, especially in the females, and these augmented expressions were drastically decreased in the post-ovulatory females. gh expression showed a slight increase in the mature males. In contrast, kiss2, kiss2r and prl did not show significant changes in the males but significantly decreased in the post-ovulatory females. The present results demonstrate the expression dynamics of the hypothalamo-pituitary-gonadal axis genes associated with the reproductive conditions and the possible involvement of the GnRH/GnIH/GTH system in the regulation of the sexual maturation and spawning in the wild tiger puffer.

Higher acclimation temperature affects growth of rohu (Labeorohita) through suppression of GH and IGFs genes expression actuating stress response.

Water temperature alone can affect the growth, metabolic rates and physiological responses of aquatic organisms. Our earlier study reported that higher temperature affects cellular and hemato-biochemical responses in rohu, Labeo rohita. In this backdrop, the present study assessed the effect of higher acclimation temperature on the regulatory mechanisms of growth and stress responses of juvenile L. rohita acclimatized in three temperature conditions (30 °C, 33 °C, and 36 °C) for a period of 30 days. The relative expression of genes for growth hormone (GH), insulin-like growth factors (IGF-1 and IGF-2) and heat shock proteins (hsp70 and hsp90) were measured by real-time quantitative PCR. The results revealed that the highest acclimation temperature (36 °C) significantly decreased the weight gain (WG) and specific growth rate (SGR), and increased the feed conversion ratio (FCR) compared to 30 °C (control), while increased WG, SGR and lowered FCR were observed in fish reared at the intermediate temperature (33 °C) compared to 30 °C. Similarly, the GH gene expression in the pituitary was significantly decreased and increased at 36 °C and 33 °C, respectively as compared to 30 °C. A significantly lower expression of IGF-1 and IGF-2, and higher expression of hsp70 and hsp90 were observed in the liver of fish at 36 °C. The results of the present study indicate that although slightly elevated temperature promotes the growth of juvenile L. rohita, the higher acclimation temperature may induce stress response and impair growth performance by suppressing GH/IGF system.

Hypoxia reduced upper thermal limits causing cellular and nuclear abnormalities of erythrocytes in Nile tilapia, Oreochromis niloticus.

Global warming is a threat across the world that leads to estimates of the upper thermal limits of ectothermic species. Increased water temperature up-regulates oxygen consumption and metabolic rates, and alters the physiological processes. In this study, we identified the critical thermal maxima (CTmax) and physiological responses under normoxia and hypoxia in Nile tilapia, Oreochromis niloticus. CTmax was 41.25 °C under hypoxia and 44.50 °C under normoxia. Compared to normoxia, lower values of hemoglobin (Hb) and red blood cells (RBCs) were observed at the CTmax under hypoxia. In contrast, higher values of white blood cells (WBCs) and blood glucose (Glu) levels were observed at the CTmax under hypoxia. Consequently, higher frequencies of micronucleus, cellular and nuclear abnormalities of erythrocytes were observed at the CTmax under hypoxia. These results suggest that high temperature tolerance and subsequent physiology are significantly affected by the oxygen supply in Nile tilapia. As climate vulnerability is intensifying day by day, this data will be helpful in successful management practice for the aquatic environment having low oxygen content.

Extreme warm acclimation temperature alters oxygen consumption, micronucleus formation in erythrocytes, and gill morphology of rohu (Labeo rohita) fingerlings.

Experiencing the seasonal variation and rapid global warming in the tropical climate is a common phenomenon which challenged the aquatic organisms to adapt the physiology and behavior. To investigate the effect of high-temperature acclimation, we selected Indian major carp, rohu (Labeo rohita), a commercially important freshwater aquaculture species. Oxygen consumptions, micronucleus formation in erythrocytes, and gill histopathology were observed in L. rohita fingerlings acclimated at three temperatures (30, 33, and 36 °C) for 30 days. Results showed that the highest acclimated temperature (36 °C) induced higher oxygen consumption and increased frequency of micronucleus formation in erythrocytes. Severity of different histological alterations (hyperplasia, epithelial necrosis, telangiectasis, epithelial lifting, and hypertrophy of chloride cells) in the gills was found to be increased in the highest acclimated temperature (36 °C). These findings indicate the temperature induced adaptive responses and climate vulnerability in a changing environment.

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.

Clupeids in the kaptai reservoir, a blessing or a curse: estimation of fisheries reference points.

Sustainable fisheries management depends on the degree of the present exploitation status of significant fish stocks. A recently developed fish stock assessment approach, CMSY, was used to estimate the fisheries reference points of data-limited Gudusia chapra and Corica soborna from the Kaptai reservoir using catch data, resilience, and exploitation records during the first and last year of the time series catch data. CMSY, along with a Bayesian state-space Schaefer production model (BSM), estimated maximum sustainable yield (MSY) as 2680 mt and 2810 mt, and 3280 mt and 3020 mt for the above stocks, respectively. The MSY range for both stocks was higher than the last catches meaning that both stocks are perfectly sustainable. The lower biomass B (4340 mt) for G. chapra estimated by CMSY and B MSY (4490 mt) indicates that the stock has started to be depleted. However, considering the precautionary fisheries management, the lower limit of MSY might be suggested to follow. Therefore, it could be suggested not to exceed the MSY limit (2680 mt) for the sustainability of G. chapra stock while it was 3020 mt for the C. soborna fishery. The intrinsic growth rate r was 0.862-1.19 yr-1 for G. chapra and 0.428-0.566 yr-1 for C. soborna, suggesting a high and medium increase of biomass in the existing population, respectively. A F/F MSY less than 1 and B/B MSY greater than 1 report both stocks at underfishing and underfished states. The study recommends enforcing strict and lawful actions regarding the net's mesh size to catch less small fish. Otherwise, negligence of this crucial management practice may bring severe threats to the sustainability of the whole reservoir resources and the reservoir ecosystem.

Macro and microanatomy of some organs of a juvenile male Ganges River dolphin (Platanista gangetica spp. gangetica).

Ganges River dolphins (Platanista gangetica spp. gangetica) are air-breathing, warm-blooded mammals endemic to the Ganges and Karnaphuli rivers of the Indian subcontinent. Nevertheless, very little basic histomorphological research has been conducted on this endangered species. Therefore, this study aimed to describe the morphological features of different organs of P. gangetica. Despite becoming aquatic animals, they showed similarities with terrestrial mammals, such as the pair of lungs and an apical bronchus in the respiratory system, which are pretty standard in ruminants and pigs. However, unlike the terrestrial animal, the tracheobronchial tree was stiffer due to circularly arranged anastomosing plates of the hyaline cartilaginous ring in the trachea, cartilaginous plates in the bronchiole, and thick alveolar septa. The digestive system showed a three-chambered mechanical and glandular stomach similar to the artiodactyles. However, the intestine showed smaller caecum like the monogastric mammal. The urogenital system showed lobulated kidneys, a urinary bladder, a fibroelastic penis with sigmoid flexure, and a long urethral process similar to some terrestrial ruminants. Considering the aquatic environment, all those modifications, unlike terrestrial mammals, are necessary for their adaptation. Thus, this research will broadly help our clinicians and conservationist to take further steps toward disease diagnosis and monitoring of marine health of this endangered species.

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.

Kisspeptin Exhibits Stimulatory Effects on Expression of the Genes for Kisspeptin Receptor, GnRH1 and GTH Subunits in a Gonadal Stage-Dependent Manner in the Grass Puffer, a Semilunar-Synchronized Spawner.

Kisspeptin has an important role in the regulation of reproduction by directly stimulating the secretion of gonadotropin-releasing hormone (GnRH) in mammals. In non-mammalian vertebrates, there are multiple kisspeptins (Kiss1 and Kiss2) and kisspeptin receptor types, and the two kisspeptins in teleosts have different effects depending on fish species and reproductive stages, serving reproductive and non-reproductive functions. In the grass puffer, Takifugu alboplumbeus, which has only a single pair of kiss2 and kissr2, both genes display seasonal, diurnal, and circadian oscillations in expression in association with the periodic changes in reproductive functions. To elucidate the role of kisspeptin in this species, homologous kisspeptin peptide (gpKiss2) was administered at different reproductive stages (immature, mature and regressed) and the expression levels of the genes that constitute hypothalamo-pituitary-gonadal axis were examined in male grass puffer. gpKiss2 significantly elevated the expression levels of kissr2 and gnrh1 in the brain and kissr2, fshb and lhb in the pituitary of the immature and mature fish. No noticeable effect was observed for kiss2, gnih, gnihr, gnrh2 and gnrh3 in the brain and gpa in the pituitary. In the regressed fish, gpKiss2 was ineffective in stimulating the expression of the gnrh1 and GTH subunit genes, while it stimulated and downregulated the kissr2 expression in the brain and pituitary, respectively. The present results indicate that Kiss2 has a stimulatory role in the expression of GnRH1/GTH subunit genes by upregulating the kissr2 expression in the brain and pituitary at both immature and mature stages, but this role is mostly ineffective at regressed stage in the grass puffer.

Alteration of Blood Parameters and Histoarchitecture of Liver and Kidney of Silver Barb after Chronic Exposure to Quinalphos.

Quinalphos (QP) is commonly used for pest control in the agricultural fields surrounding freshwater reservoirs. This study was conducted to evaluate the chronic toxicity of this pesticide on blood parameters and some organs of silver barb, Barbonymus gonionotus. Fish were exposed to two sublethal concentrations, 0.47 ppm and 0.94 ppm, of QP for a period of 28 days. All the blood parameters (red blood cell, hematocrit, and hemoglobin) and blood glucose except for white blood cells decreased with increasing concentration of toxicant and become significantly lower (p < 0.05) at higher concentration when compared with control. The derived hematological indices of mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were equally altered compared to control. Histoarchitectural changes of liver and kidney were observed after exposure to the QP. Hypertrophy of hepatocytes, mild to severe necrosis, ruptured central vein, and vacuolation were observed in the liver of treated groups. Highly degenerated kidney tubules and hematopoietic tissue, degeneration of renal corpuscle, vacuolization, and necrosis were evident in the kidney of treated groups. In conclusion, chronic exposure to QP at sublethal concentrations induced hematological and histological alterations in silver barb and offers a simple tool to evaluate toxicity derived alterations.