An atlas of reproductive development in rockfishes, genus Sebastes

The genus Sebastes consists of over 100 fish species, all of which are viviparous and long-lived. Previous studies have presented schemes on the reproductive biology of a single targeted species of the genus Sebastes, but all appear to possess a similar reproductive biology as evidenced by this and other studies. This atlas stages major events during spermatogenesis, oogenesis, and embryogenesis, including atresia, in six species of Sebastes (S. alutus, S. elongatus, S. helvomaculatus, S. polyspinis, S. proriger, and S. zacentrus). Our study suggests that the male reproductive cycle of Sebastes is characterized by 11 phases of testicular development, with 10 stages of sperm development and 1 stage of spermatozoa atresia. Ovarian development was divided into 12 phases, with 10 stages of oocyte development, 1 stage of embryonic development, and 1 stage of oocyte atresia. Embryonic development up to parturition was divided into 33 stages following the research of Yamada and Kusakari (1991). Reproductive development of all six species examined followed the developmental classifications listed above which may apply to all species of Sebastes regardless of the number of broods produced annually. Multiple brooders vary in that not all ova are fertilized and progress to embryos; a proportion of ova are arrested at the pre-vitellogenic stage. Reproductive stage examples shown in this atlas use S. elongates for spermatic development, S. proriger for oocyte development, and S. alutus for embryological development, because opportunistic sampling only permitted complete analysis of each respective developmental phase for those species. The results of this study and the proposed reproductive phases complement the recommended scheme submitted by Brown-Peterson et al. (2011), who call for a standardization of terminology for describing reproductive development of fishes.

Oogenesis and spawn formation in the invasive lionfish, Pterois miles and Pterois volitans

The Indo-Pacific lionfish, Pterois miles and P. volitans, have recently invaded the U.S. east coast and the Caribbean and pose a significant threat to native reef fish communities. Few studies have documented reproduction in pteroines from the Indo-Pacific. This study provides a description of oogenesis and spawn formation in P. miles and P. volitans collected from offshore waters of North Carolina, U.S.A and the Bahamas. Using histological and laboratory observations, we found no differences in reproductive biology between P. miles and P. volitans. These lionfish spawn buoyant eggs that are encased in a hollow mass of mucus produced by specialized secretory cells of the ovarian wall complex. Oocytes develop on highly vascularized peduncles with all oocyte stages present in the ovary of spawning females and the most mature oocytes placed terminally, near the ovarian lumen. Given these ovarian characteristics, these lionfish are asynchronous, indeterminate batch spawners and are thus capable of sustained reproduction throughout the year when conditions are suitable. This mode of reproduction could have contributed to the recent and rapid establishment of these lionfish in the northwestern Atlantic and Caribbean.

Use of skeletochronological analysis to estimate the age of leatherback sea turtles Dermochelys coriacea in the western North Atlantic

Although growth rate and age data are essential for leatherback management, estimates of these demographic parameters remain speculative due to the cryptic life history of this endangered species. Skeletochronological analysis of scleral ossicles obtained from 8 captive, known-age and 33 wild leatherbacks originating from the western North Atlantic was conducted to characterize the ossicles and the growth marks within them. Ages were accurately estimated for the known-age turtles, and their growth mark attributes were used to calibrate growth mark counts for the ossicles from wild specimens. Due to growth mark compaction and resorption, the number of marks visible at ossicle section tips was consistently and significantly greater than the number visible along the lateral edges, demonstrating that growth mark counts should be performed at the tips so that age is not underestimated. A correction factor protocol that incorporated the trajectory of early growth increments was used to estimate the number of missing marks in those ossicles exhibiting resorption, which was then added to the number of observed marks to obtain an age estimate for each turtle. A generalized smoothing spline model, von Bertalanffy growth curve, and size-at-age function were used to obtain estimates of age at maturity for leatherbacks in the western North Atlantic. Results of these analyses suggest that median age at maturation for leatherbacks in this part of the world may range from 24.5 to 29 yr. These age estimates are much greater than those proposed in previous studies and have significant implications for population management and recovery.

Testicular development in migrant and spawning bluefin tuna (Thunnus thynnus (L.)) from the eastern Atlantic and Mediterranean

Testis histological structure was studied in bluefin tuna (Thunnus thynnus) from the eastern Atlantic and Mediterranean during the reproductive season (from late April to early June). Testicular maturation was investigated by comparing samples from bluefin tuna caught on their eastward reproductive migration off Barbate (Strait of Gibraltar area) with samples of bluefin tuna fished in spawning grounds around the Balearic Islands. Histological evaluations of cross sections showed that the testis consists of two structurally different regions, an outer proliferative region where germ cells develop synchronously in cysts, and a central region made up of a well-developed system of ducts that convey the spermatozoa produced in the proliferative region to the main sperm duct. Ultrastructural features of the different stages of the male germ cell line are very similar to those described in other teleost species. The bluefin tuna testis is of the unrestricted spermatogonial testicular type, where primary spermatogonia are present all along the germinative portion of the lobules. All stages of spermatogenesis were present in the gonad tissue of migrant and spawning bluefin tuna, although spermatids were more abundant in spawning fish. The testis size was found to increase by a factor of four (on average) during migration to the Mediterranean spawning grounds, whereas the fat bodies (mesenteric lipid stores associated with the gonads) became reduced to half their weight, and the liver mass did not change significantly with sexual maturation. Linear regression analysis of the pooled data of migrant and spawning bluefin tuna revealed a significant negative correlation between the gonad index (IG) and the fat tissue index (IF), and a weaker positive correlation between the gonad index (IG) and the liver index (IL). Our analyses indicate that the liver does not play a significant role in the storage of lipids and that mesenteric lipid reserves constitute an important energy source for gametogenesis in bluefin tuna.

Factors influencing the timing and frequency of spawning and fecundity of the goldlined seabream (Rhabdosargus sarba) (Sparidae) in the lower reaches of an estuary

We have studied the reproductive biology of the goldlined seabream (Rhabdosargus sarba) in the lower Swan River Estuary in Western Australia, focusing particularly on elucidating the factors influencing the duration, timing, and frequency of spawning and on determining potential annual fecundity. Our results demonstrate that 1) Rhabdosargus sarba has indeterminate fecundity, 2) oocyte hydration commences soon after dusk (ca. 18:30 h) and is complete by ca. 01:30−04:30 h and 3) fish with ovaries containing migratory nucleus oocytes, hydrated oocytes, or postovulatory follicles were caught between July and November. However, in July and August, their prevalence was low, whereas that of fish with ovaries containing substantial numbers of atretic yolk granule oocytes was high. Thus, spawning activity did not start to peak until September (early spring), when salinities were rising markedly from their winter minima. The prevalence of spawning was positively correlated with tidal height and was greatest on days when the tide changed from flood to ebb at ca. 06:00 h, i.e., just after spawning had ceased. Because our estimate of the average daily prevalence of spawning by females during the spawning season (July to November) was 36.5%, individual females were estimated to spawn, on average, at intervals of about 2.7 days and thus about 45 times during that period. Therefore, because female R. sarba with total lengths of 180, 220, and 260 mm were estimated to have batch fecundities of about 4500, 7700, and 12,400 eggs, respectively, they had potential annual fecundities of about 204,300, 346,100 and 557,500 eggs, respectively. Because spawning occurs just prior to strong ebb tides, the eggs of R. sarba are likely to be transported out of the estuary into coastal waters where salinities remain at ca. 35‰. Such downstream transport would account for the fact that, although R. sarba exhibits substantial spawning activity in the lower Swan River Estuary, few of its early juveniles are recruited into the nearshore shallow waters of this estuary.

Radiometric validation of age, growth, and longevity for the blackgill rockfish (Sebastes melanostomus)

As nearshore fish populations decline, many commercial fishermen have shifted fishing effort to deeper continental slope habitats to target fishes for which biological information is limited. One such fishery that developed in the northeastern Pacific Ocean in the early 1980s was for the blackgill rockfish (Sebastes melanostomus), a deep-dwelling (300−800 m) species that congregates over rocky pinnacles, mainly from southern California to southern Oregon. Growth zone-derived age estimates from otolith thin sections were compared to ages obtained from the radioactive disequilibria of 210Pb, in relation to its parent, 226Ra, in otolith cores of blackgill rockfish. Age estimates were validated up to 41 years, and a strong pattern of agreement supported a longevity exceeding 90 years. Age and length data fitted to the von Bertalanffy growth function indicated that blackgill rockfish are slow-growing (k= 0.040 females, 0.068 males) and that females grow slower than males, but reach a greater length. Age at 50% maturity, derived from previously published length-at-maturity estimates, was 17 years for males and 21 years for females. The results of this study agree with general life history traits already recognized for many Sebastes species, such as long life, slow growth, and late age at maturation. These traits may undermine the sustainability of blackgill rockfish populations when heavy fishing pressure, such as that which occurred in the 1980s, is applied.

Reproduction in the protogynous black grouper Mexico (Mycteroperca bonaci (Poey)) from the southern Gulf of Mexico

An analysis was made of sexual pattern, spawning season, sizes at sexual maturation, and sex change in black grouper (Mycteroperca bonaci) from the southern Gulf of Mexico. Samples were taken between 1996 and 2000, from industrial and small-craft commercial fi sheries, in offshore and inshore waters of the continental shelf of the Yucatan Peninsula (Campeche Bank), including the shallow waters of National Marine Park Alacranes Reef. For all collected specimens (n=1229), sex and maturation condition were determined by histological analysis of the gonads. The offshore sample consisted of 75.1% females, 24.3% males, and 0.6% transitional-stage fish. All individuals collected from inshore waters were females. Gonadal structure and population structure characteristics for Campeche Bank black grouper were consistent with the characteristics of monandric protogynous hermaphrodism for a serranid fish. Sexually active males and females were observed year-round, although ripening females, with stage-III, -IV, and -V vitellogenic oocytes in the ovaries, dominated in samples taken between December and March. In addition, peak occurrence of ripe-running females with hyaline oocytes or postovulatory follicles (or both) in the ovaries was recorded in January and February. A few precocious females began spawning in October and November, and others were still in spawning condition in May and June. Fifty percent maturity of females was attained at 72.1 cm fork length (FL). Median size at sexual inversion was 103.3 cm FL, and 50% of the females measuring 111.4 cm FL had transformed into males. The southern Gulf of Mexico grouper fishery was considered deteriorated and lacked a well-defined management strategy. Results of the present study provide helpful information on black grouper reproduction in this area and could help Mexican authorities choose appropriate management strategies for this fishery, such as minimum size limit, closed fishing season, and protection of spawning aggregations.

Fecundity and spawning season of striped mullet (Mugil cephalus L.) in South Carolina estuaries

Fecundity in striped mullet (Mugil cephalus) from South Carolina correlated highly with length and weight, but not with age. Oocyte counts ranged from 4.47 × 105 to 2.52 × 106 in 1998 for fish ranging in size from 331 mm to 600 mm total length, 2.13 × 105to 3.89 × 106in 1999 for fish ranging in size from 332 mm to 588 mm total length, and 3.89 × 105 to 3.01 × 106 in 2000 for fish ranging in size from 325 mm to 592 mm total length. The striped mullet in this study had a high degree of variability in the size-at-age relation-ship; this variability was indicative of varied growth rates and compounded the errors in estimating fecundity at age. The stronger relationship of fecundity to fish size allowed a much better predictive model for potential fecundity in striped mullet. By comparing fecundity with other measures of reproductive activity, such as the gonadosomatic index, histological examination, and the measurement of mean oocyte diameters, we determined that none of these methods by themselves were adequate to determine the extent of reproductive development. Histological examinations and oocyte diameter measurements revealed that fecundity counts could be made once developing oocytes reached 0.400 μm or larger. Striped mullet are isochronal spawners; therefore fecundity estimates for this species are easier to determine because oocytes develop at approximately the same rate upon reaching 400 μm. This uniform development made oocytes that were to be spawned easier to count. When fecundity counts were used in conjunction with histological examination, oocyte diameter measurements, and gonadosomatic index, a more complete measure of reproductive potential and the timing of the spawning season was possible. In addition, it was determined that striped mullet that recruit into South Carolina estuaries spawn from October through April.

Length and age at maturity of female petrale sole (Eopsetta jordani) determined from samples collected prior to spawning aggregation

The problem of bias in female petrale sole age and length-at-maturity relationships caused by sampling from spawning aggregations was investigated. Samples were collected prior to aggregation, and histological methods were used to determine maturity status. Mature and immature fish were classified by inspecting oocytes for the presence of yolk in September, when substantial divergence in yolked and unyolked oocyte diameters had been observed. Comparison of macroscopic and microscopic assessment of maturity showed that maturity status cannot be determined accurately by using macroscopic inspection during the summer. Female petrale sole from the central Oregon coast were 50% mature at 33 cm and 5 years of age. Comparison of data from our study with data used in recent petrale sole stock assessments showed that both sampling bias and the use of samples from sea-sons when status cannot be accurately determined have likely caused errors in fitted maturity relationships.

Reproductive engineering in crustacean aquaculture

Crustacean aquaculture industry in India suffers greatly from lack of technological developments. A major constraint in this enterprise is the limitation of seed stock availability. A critical appraisal is made of the techniques used in the manipulation of reproductive processes in order to augment year-round production of seeds. A new possibility of induced ovarian maturation in crustaceans is by administering steroid hormones of vertebrate source. Environmental factors are known to govern the gametogenic cycle of marine crustaceans. Cryopreservation of male gametes and artificial insemination by way of spermatophore transfer could solve some of the problems of mating under laboratory conditions.

Length-weight relationships and relative conditions of two sciaenids Otolithes cuvieri Trewavas and Johnius elongatus Mohan from the Karachi coast

Estimates of length-weight relationship in Otolithes cuvieri justify separate equations for males (log W =-5.0100+3.1365 log L) and females (log W =-5.2000+3.1006 log L). Relative condition factor "Kn" was found to be 0.877-1.946 in males and 0.879-1.328 in females. High "Kn" values during March to September at 180-220 mm TL in either sexes are indicative of the maturation of gonads. Separate equations for length-weight relationship are also justified for males (log W = -5.1126 + 3.0690 log L) and females (log W = -5.6400 + 3.3070 log L) of Johnius elongatus . "Kn" values were found to be 0.924-1.894 for males and 0.894-1.087 for females. High "Kn" values during January-May and August-September at 130 mm TL onward are indicative of gonadal maturation.

Some aspects of association and development of Lytocestus indicus Moghe in catfish Clarias batrachus (Lin.)

Some aspects of association and development of the caryophyllid cestode Lytocestus indicus Moghe, 1925 infecting the catfish Clarias batrachus (Linn.) from the Kailla Beel of Mymensingh, Bangladesh were studied. About 33.14% of Clarias batrachus were infected with a mean intensity of 3.75, mean density 1.25. The infection was not found throughout the year. Two seasonal occurrence of this cestode were observed, one in April-May and the other in August-September. However, maturation period of the worm coincided with the maturation of the host. The worm was found attached to the wall of the intestine of the host. At the site of attachment tissue layers were compressed due to mechanical injuries. Prevalence and mean intensity of infection increased with length groups. No variation in infection was significantly observed in different sexes of the host examined.

Binding of radio-iodinated gonadotropin to the ovary of exotic carp (Cyprinus carpio Lin.)

Heterologous murrel gonadotropic hormone (m-GtH) binds to common carp oocyte plasma membrane and enhances steroid secretion. With increasing concentration of radio-labelled hormone the receptor binding is also found to increase linearly up to a certain concentration and then decrease. The [¹²⁵I] murrel GtH binding characteristics to a preparation of common carp ovarian plasma membrane shows saturability with high affinity. Scat chard plot analysis gave dissociation constant (Kd) of 0.81 X 10(super -9) M and maximum binding capacity (MBC) of 22.05 f mole/mg protein.

Domestication of an endangered fish species Nandus nandus (Ham.). Pt. 1. Laboratory rearing of young fish up to sexual maturity

An experiment was conducted for rearing of Meni, Nandus nandus in laboratory condition for seven months with the objective to select appropriate feed for the species and to develop a rearing technique of the species up to the stage of sexual maturation. Different trials were conducted using artificial feed (35.5% protein), dead fresh kachki (Carica soborna), dead fresh prawn (Macrobrachium lamarrei) and live prawn (Macrobrachium lamarrei). The provision of bottom sediment did not significantly influence the growth of fish. Between dead fresh kachki and dead fresh prawn, the fish preferred dead fresh prawn. The fish was found to be reluctant to take dead fresh kachki and prawn as food unless they became very hungry. The fish was found actively feeding on live prawn. The FCR of the prawn as food for N. nandus was found to be 2.5. From the study, it was observed that in laboratory rearing N. nandus preferred live prawn as food than artificial feed, dead fresh kachki and dead fresh prawn. The fish fed on live prawn became sexually matured (eggs or white milt extruded by gentle pressure on the abdomen of the fish) in the laboratory at the end of the experiment.

Artificial insemination in penaeid shrimps

Artificial Insemination (AI) is a tool for genetic manipulation in the shrimp stocks. It is seen as one of the means for propagating shrimp culture to new areas by controlled reproduction. Attempts at artificial insemination in the dominant closed-thelycum penaeid shrimps species of the area viz. Metapenaeus affinis and Metapenaeus brevicornis were induced in wild adult stocks collected off Mumbai coast. Female specimens were subjected to unilateral eyestalk ablation by pinching so as to induce moulting and maturation. AI was performed two days after moulting on these females when the cuticle was still soft and flexible. Moulting also ensured rejection of initial spermatophores, if present. Response of males to electrical stimulation for spermatophore expulsion was spontaneous. Use of tissue glue for spermatophore retention was found to be unnecessary. Latency period ranged between10-16 days, while spawning occurred within 10-12 days of spermatophore transfer. Three partial spawning were recorded viz., two in Metapenaeus affinis and one in Metapenaeus brevicornis with an average spawning and hatching rates of 30% and 72.3% respectively. Average survival from first nauplius (N1) to one-day old post-larva (PLI) was a meager 3.43%. Use of AI in genetic manipulation of shrimp stocks for aquacultural purposes is indicated.