Environmental Factors and Genetic Diversity as Drivers of Early Gonadal Maturation: A Gonadosomatic Index Based Investigation on Indian Shad, Tenualosa ilisha Population of Bangladesh

In recent years, attaining gonadal maturation in smaller Hilsa (Tenualosa ilisha) has become a burning issue for Hilsa fishery of Bangladesh. Causes of early maturation are not yet clearly understood. Along with environmental parameters, genetic differentiation within the population was hypothesized as the main driver, and therefore, assessing the correlation between gonadosomatic index (GSI) and environmental factors and analyzing genetic diversity were set as objectives of the present study. To address these complex issues, six diverse habitats across Bangladesh were chosen for Hilsa sample collection. For GSI, gonad was dissected from fresh fish and preserved in Bouin’s fluid for histological observation. Water quality parameters such as temperature, dissolved oxygen, pH, and salinity were also assessed. 35 fish from each habitat were used to extract and amplify DNA through the PCR technique, and genetic diversity was examined. Further, to draw a firm conclusion, the phylogenetic tree of the Hilsa population was developed by the unweighted pair-group method of arithmetic mean method based on the Cyt b gene of mitochondrial DNA. Results of GSI studies revealed that peak spawning months of T. ilisha were in October and February, where October showed the highest values in all six habitats. Histological examination showed different stages of gonadal development in different sizes and ages of Hilsa. Among all sampling sites, no statistical difference was observed for GSI value; however, smaller sized and aged Hilsa being ripped were evident in Gaglajur Haor and Kali River. Among the observed water quality parameters, temperature correlated with GSI strongly. Increased GSI was observed with temperature augmentation from downstream to upper stream, irrespective of body size and age. A perplex correlation between dissolved oxygen of observed habitats and GSI was executed. Other physico-chemical parameters viz. pH and salinity exhibited weak and moderate positive association with the GSI, respectively. Haplotype diversity of mitochondrial DNA divided the Hilsa population into three possible sub-populations, where the most distant group (Gaglajur Haor and Kali River) was subjected to early gonadal maturity. Results of this study make clear conclusions regarding the role of environmental and genetic factors on early gonadal maturations, pointing fingers at the curse of climate change and anthropogenic stressors for the migration of the Hilsa fishery of Bangladesh.

Influence of 17β-oestradiol (E2) and 17α-hydroxyprogesterone (OHP) on ovarian maturation in Penaeus semisulcatus De Haan, 1844 (Decapoda, Dendrobranchiata, Penaeidae) from the Mediterranean Sea off Alexandria, Egypt

This study searches to provide appropriate conditions to trigger spawning in Penaeus semisulcatus in aquaculture. The aim was to test the effect of 17β-oestradiol and 17α-hydroxyprogesterone on ovarian maturation and to study the correlation between the different stages of ovarian maturation and hormonal levels in the ovaries, hepatopancreas, and haemolymph. Female shrimps were collected from Ras El-Tin beach and the fishing port to Abu Qir Bay of the Mediterranean Sea off Alexandria, Egypt from June 2018-August 2019. Shrimps were individually weighed and subsequently killed by thermal shock at a maximum temperature of 45°C. Gonadosomatic and hepatosomatic indices were calculated. The correlation between the gonadosomatic and hepatosomatic indices indicates the presence of endogenous and exogenous vitellogenesis. The haemolymph, hepatopancreas, and ovaries were studied by stereo- and electron microscopy. From 15 females in each season the mentioned tissues were isolated for the extraction and quantification of steroid hormones during the different stages of gonadal maturation using radioimmunoassay. An ecdysteroid- and steroid-extraction protocol was applied. Aliquots were collected from the haemolymph to identify 17β-oestradiol and 17α-hydroxyprogesterone by mass spectrometry. Five stages of gonadal maturation were observed. The hepatopancreas was formed by a set of tubules lined with pseudostratified epithelium composed of five cell types, cell E (undifferentiated), cell F (fibrillar), cell B (vesicular), R (reabsorbing), and M (basal) cells. Steroidogenic tissues were small in size and numerous. The ultrastructure of the ovary and hepatopancreas were studied during oocyte development. There was a high concentration of 17β-oestradiol and 17α-hydroxyprogesterone in stages I and II of ovarian maturation, coinciding with the preparation phase for vitellogenesis. A progressive decrease of these steroid hormones occurred during the course of gonadal maturation, and reached its lowest concentration in the spawning period. The ecdysone concentration did not fluctuate in ripe females. Its level increases from stages I-IV of vitellogenesis onward.

Thermal Stress Affects Gonadal Maturation by Regulating GnRH, GnRH Receptor, APGWamide, and Serotonin Receptor Gene Expression in Male Pacific Abalone, Haliotis discus hannai During Breeding Season

Temperature has crucial effects on gonadal development and reproduction of abalone. To understand the impact of thermal stress on molecular and physiological processes triggering the regulation of reproduction, changes in the mRNA expression of neuroendocrine genes encoding two abalone gonadotropin-releasing hormone (Hdh-GnRH, Hdh-GnRH-like peptide), GnRH receptor (Hdh-GnRH-R), Hdh-APGWamide, serotonin receptor (5-HThdh), and a heat shock protein HSP70 were examined in ganglia and testis of male Pacific abalone (Haliotis discus hannai). Abalone were exposed to low water temperature (LWT) and high water temperature (HWT) in early and peak breeding seasons for 7 days. Then, gonadosomatic index (GSI) was calculated, relative gene expression was measured by qRT-PCR, and levels of testosterone in hemolymph were also measured by ELISA during the peak breeding season. GSI did not show any significant changes during the early breeding season. However, it was significantly decreased in LWT- or HWT-exposed abalone compared to the normal water temperature (NWT) group during the peak breeding season. In the early breeding season, changes of mRNA expression of all five genes were significant between LWT and HWT groups on day-7. In the peak breeding season, compared to the NWT group, the mRNA expressions of different genes were significantly decreased in different tissues both in LWT and HWT groups of abalone, such as Hdh-GnRH-like peptide in the cerebral ganglion (CG) and testis; Hdh-GnRH in the pleuropedal ganglion (PPG) and branchial ganglion (BG); Hdh-GnRH-R in the CG, PPG, and testis; and Hdh-APGWamide in the PPG and testis. Interestingly, the expression of 5-HThdh was significantly increased in the HWT group but decreased in the LWT group. Expression of HSP70 was significantly increased in the testis after exposure to HWT. Hemolymph levels of testosterone were significantly decreased in the HWT group compared to those in the NWT group. Altogether, these results denote that thermal stress has a repressive effect on gonadal maturation and reproduction by regulating the expression of Hdh-GnRH-like peptide, Hdh-GnRH, Hdh-GnRH-R, Hdh-APGWamide, 5-HThdh, and HSP70 genes and levels of hemolymph testosterone.

Investigations on Gonadosomatic Index and Gonad Histology of Barred Spiny eel Macrognathus pancalus Hamilton, 1822 from Upper Assam, India

Background: Macrognathus pancalus or barred spiny eel is a highly valued ornamental and food fish found in the Indian subcontinent. Due to the burgeoning population and their associated impacts, the population of the species is in rapid decline that necessitates time-bound intervention to conserve the species. The current investigation was undertaken to study the gonadosomatic index and gonad histology of Macrognathus pancalus collected from upper Assam, India.Methods: 500 samples of Macrognathus pancalus were collected from different water bodies such as wetlands, ponds, paddy fields, etc. from upper Assam, India during 2018-2019. Monthly samplings were carried out to evaluate the gonadosomatic index. The gonads were dissected out, measured and subsequently preserved for histological studies. Histological sections of preserved samples were prepared by employing accepted methodologies. The sections were then photographed using Leica DM 750 and gonadal staging was ascertained. Result: Gonadosomatic index studies revealed that it peaked in August in both males (3.45±0.18) and females (8.85±0.35), thereby indicating its spawning season. Minimum GSI values were observed during January in males (0.39±0.04) and in December in females (1.04±0.15) indicating the culmination of the breeding period and the advent of the preparatory phase. Macroscopic and microscopic examination unearthed five different phases of gonadal maturation. Gonadal staging through microscopic and macroscopic technique showed synchronicity with GSI values.

Multi-Parametric Portfolio to Assess the Fitness and Gonadal Maturation in Four Key Reproductive Phases of Brown Trout

Brown trout is an environmental freshwater sentinel species and is economically important for recreational fishing and aquaculture. Despite that, there is limited knowledge regarding morpho-physiological variations in adults throughout the reproductive cycle. Thus, this study aimed to analyze the fitness and gonadal maturation of cultured adult brown trout in four reproductive phases (spawning capable—December, regressing—March, regenerating—July, and developing—November). The systematic evaluation of males and females was based on biometric, biochemical, and hormonal parameters, along with a histomorphological grading of gonads and the immunophenotype location of key steroidogenic enzymes. The total weight and lengths reached the lowest levels in December. Gonad weights were higher in December and November, while the opposite pattern was found for liver weights. The lowest levels of cholesterol and total protein were also noted during those stages. The 11-ketotestosterone (11-KT) and testosterone (T) for males, and estradiol (E2) and T for females, mostly explained the hormonal variations. The immunohistochemistry of cytochrome P450c17 (CYP17-I), aromatase (CYP19), and 17β-hydroxysteroid dehydrogenase (17β-HSD) showed sex and site-specific patterns in the distinct reproductive phases. The sex- and season-specific changes generated discriminative multi-parameter profiles, serving as a tool for environmental and aquaculture surveys.