Estimates of body sizes at maturation and at sex change, and the spawning seasonality and sex ratio of the endemic Hawaiian grouper (Hyporthodus quernus, F. Epinephelidae)

A case study of the reproductive biology of the endemic Hawaiian grouper or hapu’upu’u (Hyporthodus quernus) is presented as a model for comprehensive future studies of economically important epinephelid groupers. Specimens were collected throughout multiple years (1978–81, 1992–93, and 2005–08) from most reefs and banks of the Northwestern Hawaiian Islands. The absence of small males, presence of atretic oocytes and brown bodies in testes of mature males, and both developed ovarian and testicular tissues in the gonads of five transitional fish provided evidence of protogynous hermaphroditism. No small mature males were collected, indicating that Hawaiian grouper are monandrous (all males are sex-changed females). Complementary microscopic criteria also were used to assign reproductive stage and estimate median body sizes (L50) at female sexual maturity and at adult sex change from female to male. The L50 at maturation and at sex change was 580 ±8 (95% confidence interval [CI]) mm total length (TL) and 895 ±20 mm TL, respectively. The adult sex ratio was strongly female biased (6:1). Spawning seasonality was described by using gonadosomatic indices. Females began ripening in the fall and remained ripe through April. A February–June main spawning period that followed peak ripening was deduced from the proportion of females whose ovaries contained hydrated oocytes, postovulatory follicles, or both. Testes weights were not affected by season; average testes weight was only about 0.2% of body weight—an order of magnitude smaller than that for ovaries that peaked at 1–3% of body weight. The species’ reproductive life history is discussed in relation to its management.

Sexual Reproduction and Early Development in the Estuarine Sea Anemone, Nematostella vectensis Stephenson, 1935

This dissertation: 1) determines the factor(s) responsible for spawning induction in NematosteJla vectensis; 2) isolates, describes, and documents the source of jelly from egg masses of N. vectensis; and 3) describes N. vectensis' early development. Namatostella vectensis were maintained on a 7-day mussel feeding/water change regime over 159 days. Within 36 hours of mussel feeding/water change. 69.1% of females and 78.5% of males spawned reliably. Through manipulation of feeding, water change, oxygen and nitrogenous waste concentrations, spawning induction was found to be triggered by the oxygen concentration associated with water change, and not by feeding. Ammonia, anemones' major waste product, inhibited this induction in a concentration-dependent manner. Female N. vectensis release eggs in a persistent jellied egg mass which is unique among the Actiniaria. The major component of this egg mass jelly was a positive periodic acid-Schiffs staining, 39.5-40.5 kD glycoprotein. Antibodies developed in rabbits against this glycoprotein bound to jelly of intact egg masses and to granules (~ 2.8 IJm in diameter) present in female anemone mesenteries and their associated filaments. Antibodies did not label male tissues. Nematostella vecfensis embryos underwent first karyokinesis -60 minutes following the addition of sperm to eggs. Second nuclear division took place, followed by first cleavage, 90-120 minutes later. Each of the 4 blastomeres that resulted from first cleavage contained a single nucleus. Arrangement of these blastomeres ranged from radial to pseudospiral. Embryonic development was both asynchronous and holoblastic. Following formation of the 4-cell stage, 71% of embryos proceeded to cleave again to form an 8-cell stage. In each of the remaining 29% of embryos, a fusion of from 2-4 blastomeres resulted in 4 possible patterns which had no affect on either cleavage interval timing or subsequent development. The fusion event was not due to ooplasmic segregation. Blastomeres isolated from 4-celled embryos were regulative and developed into normal planula larvae and juvenile anemones that were 1/4 the size of those that developed from intact 4-celled embryos. Embryos exhibiting the fusion phenomenon were examined at the fine structural level. The fusion phenomenon resulted in formation of a secondary syncytium and was not a mere compaction of blastomeres.

An allometric smoothing function to describe the relation between otolith and somatic growth over the lifespan of walleye pollock (Theragra chalcogramma)

We propose a new equation to describe the relation between otolith length (OL) and somatic length (fork length [FL]) of fish for the entire lifespan of the fish. The equation was developed by applying a mathematical smoothing method based on an allometric equation with a constant term for walleye pollock (Theragra chalcogramma) —a species that shows an extended longevity (>20 years). The most appropriate equation for defining the relation between OL and FL was a four-phase allometric smoothing function with three inflection points. The inflection points correspond to the timing of settlement of walleye pollock, changes in sexual maturity, and direction of otolith growth. Allometric smoothing functions describing the relation between short otolith radius and FL, long otolith radius and FL, and FL and body weight were also developed. The proposed allometric smoothing functions cover the entire lifespan of walleye pollock. We term these equations “allometric smoothing functions for otolith and somatic growth over the lifespan of walleye pollock.”

Sexual differentiation and gonad development in striped mullet (Mugil cephalus L.) from South Carolina estuaries*

This study examined the sexual differentiation and reproductive dynamics of striped mullet (Mugil cephalus L.) in the estuaries of South Carolina. A total of 16,464 specimens were captured during the study and histological examination of sex and maturity was performed on a subsample of 3670 fish. Striped mullet were sexually undifferentiated for the first 12 months, began differentiation at 13 months, and were 90% fully differentiated by 15 to 19 months of age and 225 mm total length (TL). The defining morphological characteristics for differentiating males was the elongation of the protogonial germ tissue in a corradiating pattern towards the center of the lobe, the development of primary and secondary ducts, and the lack of any recognizable ovarian wall structure. The defining female characteristics were the formation of protogonial germ tissue into spherical germ cell nests, separation of a tissue layer from the outer epithelial layer of the lobe-forming ovarian walls, a tissue bud growing from the suspensory tissue that helped form the ovary wall, and the proliferation of oogonia and oocytes. Sexual maturation in male striped mullet first occurred at 1 year and 248 mm TL and 100% maturity occurred at age 2 and 300 mm TL. Female striped mullet first matured at 2 years and 291 mm total length and 100% maturity occurred at 400 mm TL and age 4. Because of the open ocean spawning behavior of striped mullet, all stages of maturity were observed in males and females except for functionally mature females with hydrated oocytes. The spawning season for striped mullet recruiting to South Carolina estuaries lasts from October to April; the majority of spawning activity, however, occurs from November to January. Ovarian atresia was observed to have four distinct phases. This study presents morpholog ical analysis of reproductive ontogeny in relation to size and age in South Carolina striped mullet. Because of the length of the undifferentiated gonad stage in juvenile striped mullet, previous studies have proposed the possibility of protandric hermaphrodism in this species. The results of our study indicate that striped mullet are gonochoristic but capable of exhibiting nonfunctional hermaphroditic characteristics in differentiated mature gonads.

Relative pleopod length as an indicator of size at sexual maturity in slipper (Scyllarides squammosus) and spiny Hawaiian (Panulirus marginatus) lobsters

Body size at gonadal maturity is described for females of the slipper lobster (Scyllarides squammosus) (Scyllaridae) and the endemic Hawaiian spiny lobster (Panulirus marginatus) (Palinuridae) based on microscopic examination of histological preparations of ovaries. These data are used to validate several morphological metrics (relative exopodite length, ovigerous condition) of functional sexual maturity. Relative exopodite length (“pleopod length”) produced consistent estimates of size at maturity when evaluated with a newly derived statistical application for estimating size at the morphometric maturation point (MMP) for the population, identified as the midpoint of a sigmoid function spanning the estimated boundaries of overlap between the largest immature and smallest adult animals. Estimates of the MMP were related to matched (same-year) characterizations of sexual maturity based on ovigerous condition — a more conventional measure of functional maturity previously used to characterize maturity for the two lobster species. Both measures of functional maturity were similar for the respective species and were within 5% and 2% of one another for slipper and spiny lobster, respectively. The precision observed for two shipboard collection series of pleopod-length data indicated that the method is reliable and not dependent on specialized expertise. Precision of maturity estimates for S. squammosus with the pleopod-length metric was similar to that for P. marginatus with any of the other measures (including conventional evidence of ovigerous condition) and greatly exceeded the precision of estimates for S. squammosus based on ovigerous condition alone. The two measures of functional maturity averaged within 8% of the estimated size at gonadal maturity for the respective species. Appendage-to-body size proportions, such as the pleopod length metric, hold great promise, particularly for species of slipper lobsters like S. squammosus for which there exist no other reliable conventional morphological measures of sexual maturity. Morphometric proportions also should be included among the factors evaluated when assessing size at sexual maturity in spiny lobster stocks; previously, these proportions have been obtained routinely only for brachyuran crabs within the Crustacea.

Age, size, and sexual maturity of channeled whelk (Busycotypus canaliculatus) in Buzzards Bay, Massachusetts

With the southern New England lobster fishery in distress, lobster fishermen have focused more effort toward harvesting channeled whelk (Busycotypus canaliculatus). However, minimal research has been conducted on the life history and growth rates of channeled whelk. Melongenid whelks generally grow slowly and mature late in life, a characteristic that can make them vulnerable to overfishing as fishing pressure increases. We sampled channeled whelk from Buzzards Bay, Massachusetts, in August 2010 and in July 2011, studied their gonad development by histology, and aged them by examining opercula. Males had a slower growth rate and a lower maximum size than females. Male whelk reached 50% maturity (SM50) at 115.5 mm shell length (SL) and at the age of 6.9 years. Female whelk reached SM50 at 155.3 mm SL and at the age of 8.6 years. With a minimum size limit of 69.9 mm (2.75 in) in shell width, males entered the fishery at 7.5 years, a few months after SM50, but females entered the fishery at 6.3 years, approximately 2 years before SM50. Increased fishing pressure combined with slow growth rates and the inability to reproduce before being harvested can easily constrain the long-term viability of the channeled whelk fishery in Massachusetts.

Observation on the urinogenital papilla and sexual dimorphism in some Indian gobiids (Gobiidae: Teleostei)

A thorough comparative study on the urinogenital papilla and sexual dimorphism has been made for the first time in both the sexes of twelve Indian gobiids: Glassogobius giuris (Hamilton); Acentrogobius cyanomos (Bleeker); Eleotriodes muralis (Valenciennes); Parapocryptes serperaster (Richardson); Apocryptes bato (Hamilton); Scartclaos viridis (Hamilton); Boleophthalmus boddarti (Pallas), Periophthalmus schlosseri (Pallas); P. koelreuteri (Pallas); Taenioides anguillaris (Linnaeus); T. buchanani (Day); Odontamblyopus rubicundus (Hamilton). The urinogenital papilla, originating as a free muscular organ from the ventral surface of the body-wall and shortly behind anus, is present in both the sexes. It is an important organ of primary sex recognition in all species. In case of male the papilla is conical, broad at the base and in female it is either flattened, distally truncated or bluntly rounded. The presence of permanent colour mark over the specific region of the body surface is another secondary sexual character in a few species. Besides, colouration may also be a nuptial secondary sex character developed in some during peak breeding season. The enlargement and colouration of the organ is subject to seasonal variations parallel to the seasonal gonadal cycle. The histological architecture of the papilla shows a high degree of cellular specialization and an interrelationship to the urinary and genital ducts. The functional efficacy and significance of the papilla in the breeding biology of these fishes has been discussed.

Histology and hormonal effect of Ghrelin on ovary of Benni (Barbus sharpeyi) in order to study sexual maturity, zygote and larval generation

Benni (Barbus sharpeyi) is valuable fish that Khuzastan fisheries office propagated it artificially in Susangerd Fish Propagation Center every year. Pituitary gland is used for this aim but female fish lost their fertilization power after 2-3 years, so in present research, new hormone, that is called Ghrelin. The aims of this research are histology, hormonal, zygote and larval generation studies and comparing the results with each other. Ghrelin is a multifunctional peptidyl hormone which increases GTH-II in fish, amphibian, and birds and mammalian so its effect on Benni sexual maturation was studied. Human Ghrelin (hGRL) was obtained from ANASPEC, Canada, with 28 amino acids. In the present study, three levels of ghrelin including 0 (sham treatments), 0.10 (treatment 1) and 0.15 μg/g (treatment 2) body wt and one level of pituitary gland 4000 μg/g (pituitary treatment) with two replications were used. 56 specimens were injected intraperitonealy and their ghrelin level was evaluated immediately after injection and after 24 h. Control fish(n=16) were just injected by physiological saline. For hormonal studies sham and experimental fish(n=40) were anesthetized with MS-222 at a concentration of 250 mg l-1, and blood samples were collected and kept at 4ْC, then spun to collect serum. Serum samples were stores at -20ْC until the RIA for CTH-II. For histology studies immediately after injection a piece of ovary was collected from control fish (Sham zero) after being anesthetized. The sampled ovaries were fixed in Buin solution and embedded in paraffin, and stained to Sections of 5–6 μm using haematoxylin and eosin. The ovarian samples were performed with a compound microscope. Histology and micrometry studies had done. The mature oocytes had given from mature fish, then weighted and the working fecundity were counted. The mature oocytes fertilized, the eggs were incubated and the percentage of fertilization was calculated. After 72h the eggs hatched and the percentage of hatch was counted. The percentage of hindrance was calculated after 6 days. Hormonal results indicate that ghrelin and pituitary increase significantly the GTH-II level in comparison to sham. Macroscopic observations (before taking ovary) showed that ovaries with green colored have couple oval structure located in the abdominal cavity. Microscopic studies of dissected ovaries indicated simultaneous growth of 127 oocytes with 6 stages. The type of the ovary is asynchronous. The results indicated that both of the ghrelin treatment increased the percentage of mature follicles followed by decrease of immature follicles. There were significant differences (P<0.05) between the number of mature and immature follicles. Average diameter of follicle in both of the ghrelin treatment was significantly (P<0.05) declined in the stages of the vitellogenesis when the result compared to the other treatment. Just treatment 1 and pituitary treatment can give mature oocytes. The fecundity of pituitary treatment significantly increase in comparision to ghrelin treatment (P<0.05). In food-restricted fish where endogenous ghrelin levels are known to be increased, a chronic administration of ghrelin induces overt negative effect in releasing mature oocytes. The percentage of fertilization was significantly increase (P<0.05) in ghrelin t. in comparison to pituitary t. and the percentage of hatch was significantly increase (P<0.05) in pituitary t. in comparison to ghrelin t. There was no significant difference (P>0.05) in terms of percentage of hindrance between treatments. In conclusion, the present study demonstrated that ghrelin has positive effect on the level of GTH-II, oocyte maturation, ovarian vitellogenesis and the number of mature follicles of Barbus sharpeyi ovary. Increasing of the mature follicles number reduces their average diameter, indicating stimulating effect of ghrelin in sexual maturation of Barbus sharpeyi.The ghrelin and pituitary treatment have equal chance in the post-stage of spawning.

Study on biological characteristics, sexual maturation and reproduction of Liza abu in the water of Khozestan Province

As the most of the fish resources are known and exploited, protecting their generation is of the greatest importance. Aquaculture is one of the efficient procedures in protecting and reviving fish resources and knowing about the reproductive cycle and gonads development has an important role in approaching this aim. Liza abu belongs to the family Mugilidae that according to its resistance to the environmental condition and its fast growth , can be introduced as a fish with economical value. As there is no scientific data on the reproductive biology of this species , study on the reproductive biology and gonad development is considered as the aim of this research . For this purpose , 360 samples of this species were investigated during the period from February 2007 to January 2008 in Khozestan Province . After studing morphological and histological characteristics of gonad specimen , they were prepared through histological method. Samples were prepared through usual histological method and studied under light microscope. According to the results, the maturity stages of male and female Liza abu were separated to six different successive stages. In ovaries , these stages were as follow : In stage І, the oocytes were small , this stage was observed from July to October . In stage ІІ, considerable growth was observed in the oocytes . This stage was observed from October to January . In stage III, due to vitellogenesis, the maximum growth was observed and three layers of theca, granullosa and follicle cells were visible. This stage was observed during January and February . In stage IV, migration of germinal vesicle was observed and due to hydration of the oocytes , their diameter was increased. The ovaries were yellowish and in maximum size and ovules could be easily observed with naked-eye . This stage was observed in February and March . In stage V, spawning occured. This stage was observed in April . In stage VI, ovaries consisted of immature and atretic oocytes and also empty follicles. This stage was observed in May and June. In testes , these stages were as follow : In stage I , the testes were small in size and contained the spermatogonia which were the only cellular components.This stage was observed in August and September . In stage II (maturing virgin ) , the spermatogonia and the primary spermatocytes were visible. This stage was observed in October . In stage III (developing), intensive spermatogenesis was occured and the primary and the secondary spermatocytes were the most visible cells during this stage .This stage was observed from November to January. In stage IV(developed), cells of all stages of spermatogenesis could be seen but the secondary spermatocytes and spermatids were in large number. This stage was observed from January to March. In stage V , the testes were filled with sperms. This stage was observed in March and April .In stage VI, residual spermatozoa and the spermatogonia were visible in the testes. This stage was observed from May to August. According to cyclic changes in GSI, sexual maturation in breeding begins in January and spawning occurs in April. The ova diameter ranged from 30.75 μ in stage I to 472.19 μ in stage IV. In this study , the sex ratio was 1:2.7, and male and female percentage were 27.02% and 72.98% respectively. This means that females predominate males. In this study absolute fecundity was calculated and changing between 30805.44 to 431247.3 was observed and absolute fecundity was calculated 111275.3 in average.

The survey of sexual maturity in Mugil cephalus so measuring sex hormones and histologic studies

In this study the process of female gray mullet brooders was carried out by using histological study and masurment of sex steroids. Results of histological studies showed that oocyte of gray mullet brooders in Gomishan Rearing Center conditions of develop to the end of yolk globule stage. The results were observed with oocyte in chromatin nucleolar stage (first stage) with means of diameter of 20 p m, in August, perinucleolar stage (second stage) in September with mean diameter of 87 p m, yolk vesicle stage (third stage) in October with mean diameter 200 p m and yolk granules stage (forth stage) from October to November with average diameter of 180 — 650 p m. For the reason of stopping oocyte develop at the end of fourth stage, hormonal induction to final oocyte maturation and ovulation was used. For this purpose, carp pituitary , HCG and LRH-A2 with different combinations were used in two stages, second injection was used 24 hours after first injection. 15 females brooders were divided in 5 groups, different hormonal combinations were injected to four groups and to fifth group as control, only saline, was injected. The process of female brooder rippening in hormonal induction was studied via masurment of sex steroids including 17 a - hydroxy progestrone, estradio1-17)6 and testosterone. Blood samples were collected from caudal vein during first injection, 24, 30 and 48 hours after the first injection. At the same time, for distinguishing histological changes the sample has been attained from the gonads Sex stroid fluctuation patterns in different brooder groups that injected hormon were similar, however hormonal composition had similar effects. All brooder that their oocyte in the beginning of hormonal injection were At the end of fourth stage with oocyte diameter average of 600 p m received to final maturation and ovulation. The brooder that its oocytes were At the begining or mid-fourth stage did not show ovulation but hormonal induction caused oocyte develop at the beginning of fifth stage. Study of 17-hydroxy progestrone fluctuation showed that the maximum level of this steroid (0.347 ng/ml) measured 30 hours after the first injection and was significantly higher (p< 0.05) than those of control group. So, 17-hydroxy progestrone is probably precursor of maturation inducing steroid (MIS). However the maximum level of that observed was coincident with germinal vesicle breakdown, oil droplets coalescence and dissolution of yolk granuls The maximum levels of esteradiol— 17/0 and testosterone (3.778 and 16.801ng/ml,respectively) in spawned brooders,were observed 24 hours after the first injection. levels of those steroids were significantly higher (p<0.05) than control group. Maximum level of sex steroids in the brooders that did not spawn to the end of treatment was observed with more delay than those in spawned brooders. Therefor maximum level of 17a-hydroxy progestrone (0.264 ng/ml) in those brooders observed in fourth sampling time and the maximum levels of estradio1-17a and testosterone (2.944 and 18.993 ng/ml, respectivly)observed in third sampling time that was significantly higher (p<0.05) than those of control group. For the study of stress effect on brooders during the hormonal induction, level of cortisol was measured in every sampling time. level of cortisol had high fluctuation that showed handling level and stress effect on brooders. However maximum level of cortisol in majority of brooders was dominant in third sampling time that was coincident with final maturation.