Degeneration of postovulatory follicles in the Iberian sardine Sardina pilchardus: structural changes and factors affecting resorption

Inaccuracy in the aging of postovulatory follicles (POFs) and in estimating the effect of temperature on the resorption rate of POFs may introduce bias in the determination of the daily spawning age classes with the daily egg production method (DEPM). To explore the above two bias problems with f ield-collected European pilchard (Sardina pilchardus, known regionally as the Iberian sardine), a method was developed in which the time elapsed from spawning (POF age) was estimated from the size of POFs (i.e., from the cross-sectional area in histological sections). The potential effect of the preservative type and embedding material on POF size and the effect of ambient water temperature on POF resorption rate are taken into account with this method. A highly significant loglinear relationship was found between POF area and age; POF area shrank by approximately 50% per day. POFs were also shown to shrink faster at higher temperatures (approximately 3% per degree), but this temperature effect is unlikely to be an important source of bias in the assignment of females to daily spawning classes. The embedding material was also shown to influence the size of POFs, the latter being significantly larger in resin than in paraffin sections. In conclusion, the size of POFs provides an indirect, reliable estimation of the time elapsed from spawning and may thus be used to test both the validity of POF staging criteria for identifying daily classes of spawners and the effect of other factors (such as temperature and laboratory processing) in applications of the DEPM to S. pilchardus and other fish species.

The effect of temperature on the duration of spawning markers—migratory-nucleus and hydrated oocytes and postovulatory follicles—in the multiple-batch spawner Japanese flounder (Paralichthys olivaceus)

The duration of spawning markers (e.g. signs of previous or imminent spawnings) is essential information for estimating spawning frequency of fish. In this study, the effect of temperature on the duration of spawning markers (i.e., oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, as well as new postovulatory follicles) of an indeterminate multiple-batch spawner, Japanese f lounder (Paralichthys olivaceus), was evaluated. Cannulation was performed to remove samples of oocytes, eggs, and postovulatory follicles in individual females at 2–4 hour intervals over 27–48 hours. The duration of spawning markers was successfully evaluated in 14 trials ranging between 9.2° and 22.6°C for six females (total length 484–730 mm). The durations of spawning markers decreased exponentially with temperature and were seen to decrease by a factor of 0.16, 0.36, 0.30, and 0.31 as temperature increased by 10°C for oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, and new postovulatory follicles, respectively. Thus, temperature should be considered when estimating spawning frequency from these spawning markers, especially for those fish that do not spawn synchronously in the population.

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.

Reproductive biology of blue marlin (Makaira nigricans) in the western Pacific Ocean

The reproductive biology of blue marlin (Makaira nigricans) was assessed from 1001 fish (ranging from 121 to 275 cm in eye-to-fork length; EFL) caught by Taiwanese offshore longliners in the western Pacific Ocean from September 2000 to December 2001 and from 843 gonad samples from these fish, The overall sex ratio of the catch was approximately 1:1 dur ing the sampling period, but blue marlin are sexually dimorphic; females are larger than males. Reproductive activity (assessed by histology), a gonadosomatic index, and the distribution of oocyte diameters, indicated that spawning occurred predominantly from May to September. The estimated sizes-at-maturity (EFL50) were 179.76 ±1.01 cm (mean ±standard error) for females and 130 ±1 cm EFL for males. Blue marlin are multiple spawners and oocytes develop asynchronously. The proportion of mature females with ovaries containing postovulatory follicles (0.41) and hydrated oocytes (0.34) indicated that the blue marlin spawned once every 2–3 days on average. Batch fecundity (BF) for 26 females with the most advanced oocytes (≥1000 μm), but without postovulatory follicles, ranged from 2.11 to 13.50 million eggs (6.94 ± 0.54 million eggs). The relationships between batch fecundity (BF, in millions of eggs) and EFL and round weight (RW, kg) were BF = 3.29 × 10 –12 EFL5.31 (r2 = 0.70) and BF = 1.59 × 10–3 RW 1.73 (r2= 0.67), respectively. The parameters estimated in this study are key information for stock assessments of blue marlin in the western Pacific Ocean and will contribute to the conservation and sustainable yield of

Advances in methods for determining fecundity: application of the new methods to some marine fishes

Estimation of individual egg production (realized fecundity) is a key step either to understand the stock and recruit relationship or to carry out fisheries-independent assessment of spawning stock biomass using egg production methods. Many fish are highly fecund and their ovaries may weigh over a kilogram; therefore the work time can be consuming and require large quantities of toxic fixative. Recently it has been shown for Atlantic cod (Gadus morhua) that image analysis can automate fecundity determination using a power equation that links follicles per gram ovary to the mean vitellogenic follicular diameter (the autodiametric method). In this article we demonstrate the precision of the autodiametric method applied to a range of species with different spawning strategies during maturation and spawning. A new method using a solid displacement pipette to remove quantitative fecundity samples (25, 50, 100, and 200 milligram [mg]) is evaluated, as are the underlying assumptions to effectively fix and subsample the ovary. Finally, we demonstrate the interpretation of dispersed formaldehyde-fixed ovarian samples (whole mounts) to assess the presence of atretic and postovulatory follicles to replace labor intensive histology. These results can be used to estimate down regulation (production of atretic follicles) of fecundity during maturation.

Reproductive biology of female Rikuzen sole (Dexistes rikuzenius)*

The annual ovarian cycle, mode of maturation, age at maturity, and potential fecundity of female Rikuzen sole (Dexistes rikuzenius) from the North Pacific Ocean off the coast of Japan were studied by 1) histological examination of the gonads, 2) measurement and observation of the oocytes, and 3) by otolith aging. The results indicated that ovulation occurs from September to December and peaks between September and October. Vitellogenesis began again soon after the end of the current season. Maturity was divided into eight phases on the basis of oocyte developmental stages. Mature ovaries contained developing oocytes and postovulatory follicles but no recruiting oocytes, indicating that this species has group-synchronous ovaries and is a multiple spawner. Almost all females matured first at an age of 1+ year and spawned every year until at least age 8+ years. Potential fecundity increased exponentially with body length and the most fecund fish had 15 times as many oocytes as the least fecund fish. Potential fecundity and relative fecundity were both positively correlated with age from 1 to 6+ years, but were negatively correlated, probably because of senescence, in fish over 7 years. These results emphasize that the total productivity of a D. rikuzenius population depends not only on the biomass of females older than 1+ but also on the age structure of the population.

Movements and spawning of white marlin (Tetrapturus albidus) and blue marlin (Makaira nigricans) off Punta Cana, Dominican Republic

With a focus on white marlin (Tetrapturus albidus), a concurrent electronic tagging and larval sampling effort was conducted in the vicinity of Mona Passage (off southeast Hispaniola), Dominican Republic, during April and May 2003. Objectives were 1) to characterize the horizontal and vertical movement of adults captured from the area by using pop-up satellite archival tags (PSATs); and 2) by means of larval sampling, to investigate whether fish were reproducing. Trolling from a sportfishing vessel yielded eight adult white marlin and one blue marlin (Makaira nigricans); PSAT tags were deployed on all but one of these individuals. The exception was a female white marlin that was unsuitable for tagging because of injury; the reproductive state of its ovaries was examined histologically. Seven of the PSATs reported data summaries for water depth, temperature, and light levels measured every minute for periods ranging from 28 to 40 days. Displacement of marlin from the location of release to the point of tag pop-up ranged from 3l.6 to 267.7 nautical miles (nmi) and a mean displacement was 3.4 nmi per day for white marlin. White and blue marlin mean daily displacements appeared constrained compared to the results of other marlin PSAT tagging studies. White marlin ovarian sections contained postovulatory follicles and final maturation-stage oocytes, which indicated recent and imminent spawning. Neuston tows (n=23) yielded 18 istiophorid larvae: eight were white marlin, four were blue marlin, and six could not be identified to species. We speculate that the constrained movement patterns of adults may be linked to reproductive activity for both marlin species, and, if true, these movement patterns may have several implications for management. Protection of the potentially important white marlin spawning ground near Mona Passage seems warranted, at least until further studies can be conducted on the temporal and spatial extent of reproduction and associated adult movement.

The reproductive cycle of the thorny skate (Amblyraja radiata) in the western Gulf of Maine

The thorny skate (Amblyraja radiata) is a large species of skate that is endemic to the waters of the western north Atlantic in the Gulf of Maine. Because the biomass of thorny skates has recently declined below threshold levels mandated by the Sustainable Fisheries Act, commercial harvests from this region are prohibited. We have undertaken a comprehensive study to gain insight into the life history of this skate. The present study describes and characterizes the reproductive cycle of female and male thorny skates, based on monthly samples taken off the coast of New Hampshire, from May 2001 to May 2003. Gonadosomatic index (GSI), shell gland weight, follicle size, and egg case formation, were assessed for 48 female skates. In general, these reproductive parameters remained relatively constant throughout most of the year. However, transient but significant increases in shell gland weight and GSI were obser ved during certain months. Within the cohort of specimens sampled monthly throughout the year, a subset of females always had large preovulatory follicles present in their ovaries. With the exception of June and September specimens, egg cases undergoing various stages of development were observed in the uteri of specimens captured during all other months of the year. For males (n=48), histological stages III through VI (SIII−SVI) of spermatogenesis, GSI, and hepatosomatic index (HSI) were examined. Although there appeared to be monthly fluctuations in spermatogenesis, GSI, and HSI, no significant differences were found. The production and maintenance of mature spermatocysts (SVI) within the testes was observed throughout the year. These findings collectively indicate that the thorny skate is reproductively active year round.

Reproductive biology of carpenter seabream (Argyrozona argyrozona) (Pisces: Sparidae) in a marine protected area

The carpenter seabream (Argyrozona argyrozona) is an endemic South African sparid that comprises an important part of the handline fishery. A three-year study (1998−2000) into its reproductive biology within the Tsitsikamma National Park revealed that these fishes are serial spawning late gonochorists. The size at 50% maturity (L50) was estimated at 292 and 297 mm FL for both females and males, respectively. A likelihood ratio test revealed that there was no significant difference between male and female L50 (P>0.5). Both monthly gonadosomatic indices and macroscopically determined ovarian stages strongly indicate that A. argyrozona within the Tsitsikamma National Park spawn in the astral summer between November and April. The presence of postovulatory follicles (POFs) confirmed a six-month spawning season, and monthly proportions of early (0−6 hour old) POFs showed that spawning frequency was highest (once every 1−2 days) from December to March. Although spawning season was more highly correlated to photoperiod (r = 0.859) than temperature (r = −0.161), the daily proportion of spawning fish was strongly correlated (r= 0.93) to ambient temperature over the range 9−22oC. These results indicate that short-term upwelling events, a strong feature in the Tsitsikamma National Park during summer, may negatively affect carpenter fecundity. Both spawning frequency and duration (i.e., length of spawning season) increased with fish length. As a result of the allometric relationship between annual fecundity and fish mass a 3-kg fish was calculated to produce fivefold more eggs per kilogram of body weight than a fish of 1 kg. In addition to producing more eggs per unit of weight each year, larger fish also produce significantly larger eggs.

Reproductive dynamics of female spotted seatrout (Cynoscion nebulosus) in South Carolina

We describe reproductive dynamics of female spotted seatrout (Cynoscion nebulosus) in South Carolina (SC). Batch fecundity (BF), spawning frequency (SF), relative fecundity (RF), and annual fecundity (AF) for age classes 1−3 were estimated during the spawning seasons of 1998, 1999, and 2000. Based on histological evidence, spawning of spotted seatrout in SC was determined to take place from late April through early September. Size at first maturity was 248 mm total length (TL); 50% and 100% maturity occurred at 268 mm and 301 mm TL, respectively. Batch fecundity estimates from counts of oocytes in final maturation varied significantly among year classes. One-year-old spotted seatrout spawned an average of 145,452 oocytes per batch, whereas fish aged 2 and 3 had a mean BF of 291,123 and 529,976 oocytes, respectively. We determined monthly SF from the inverse of the proportion of ovaries with postovulatory follicles (POF) less than 24 hours old among mature and developing females. Overall, spotted seatrout spawned every 4.4 days, an average of 28 times during the season. A chronology of POF atresia for water temperature >25°C is presented. Length, weight (ovary-free), and age explained 67%, 65%, and 58% of the variability in BF, respectively. Neither RF (number of oocytes/g ovary-free weight) nor oocyte diameter varied significantly with age. However, RF was significantly greater and oocyte diameter was smaller at the end of the spawning season. Annual fecundity estimates were approximately 3.2, 9.5, and 17.6 million oocytes for each age class, respectively. Spotted seatrout ages 1−3 contributed an average of 29%, 39%, and 21% to the overall reproductive effort according to the relative abundance of each age class. Ages 4 and 5 contributed 7% and 4%, respectively, according to predicted AF values.

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.

Seasonal egg production of whitemouth croaker (Micropogonias furnieri) in the Río de la Plata estuary, Argentina-Uruguay

The reproductive biology of the whitemouth croaker (Micropogonias furnieri) inhabiting the estuarine waters of the Río de la Plata (Argentina-Uruguay) was studied by using histological analysis of the ovaries. Samples were collected during the spawning peak and the end of two breeding seasons (November 1995–Feb-ruary 1996 and November 1997–March 1998). Micropogonias furnieri is a multiple spawner with indeterminate annual fecundity. Spawning frequency, determined by using the percentage of females with postovulatory follicles, was about 31% in November 1995 and 25% in February 1996. At these frequencies, a female on average spawned a new batch of eggs every 3–4 days during the spawning season. Batch fecundity was fitted to a power function of length and a linear function of ovary-free female weight. The number of hydrated oocytes decreased at the end of the breeding season, coinciding with an increase of atresia. Annual egg production for a 40-cm-TL female was estimated to be between 3,300,000 and 7,300,000 eggs. In addition to the seasonal decrease in fecundity and spawning activity, a decline in egg size and weight toward the end of the breeding season was also observed.

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.

Age, growth, and reproduction of permit (Trachinotus falcatus) in Florida waters

We examined 536 permit (Trachinotus falcatus, 65–916 mm FL) collected from the waters of Florida Keys and from the Tampa Bay area on Florida’s Gulf coast to describe their growth and reproduction.Among permit that we sexed, females ranged from 266 to 916 mm in length (mean=617) and males ranged from 274 to 855 mm (mean=601). Ages of 297 permit ranging from 102 to 900 mm FL were estimated from thin-sectioned otoliths (sagittae). The large proportion of otoliths with an annulus on the margin and an otolith from an OTC-injected fish suggested that a single annulus was formed each year during late spring or early summer.Permit reach a maximum age of at least 23 years.Permit grew rapidly until an age of about five years, and then growth slowed considerably. Male and female von Bertalanffy growth models were not significantly different, and the sexes-combined growth model was FL=753.1(1–e –0.348(Age+0.585)). Gonad development was seasonal, and spawning occurred during late spring and summer over artificial and natural reefs at depths of 10–30 m. Ovaries that contained oocytes in the final stages of oocyte maturation or postovulatory follicles were found during May–July. We estimated that 50% of the females in the population had reached sexual maturity by 547 mm and an age of 3.1 years and that 50% of the males in the population had reached sexual maturity by 486 mm and an age of 2.3 years. Because Florida regulations restrict the maximum size of permit caught in recreational and commercial fisheries to 20-inch (508-mm), most fish harvested are sexually immature. With the current size selectivity of the fishery, the spawning stock biomass of permit could decrease quickly in response to moderate levels of fishing mortality; thus, the regulations in place in Florida to restrict harvest levels appear to be justified.

Reproductive biology, artificial propagation and larval rearing of two freshwater eels, Monopterus cuchia and Mastacembelus armatus

Studies on reproductive biology and artificial propagation including larval rearing of freshwater mud eel, Monopterus cuchia and spiny eel, Mastacembelus armatus were attempted. The gonadosomatic index (GSI) of mud eel ranged from 0.41 (August) to 5.52 (June) in males and 0.53 (August) to 7.61 (June) in females. In both cases the GSI showed a peak in June. Fecundity ranged from 228 (TL - 396 mm; W - 78g) to 5510 (TL - 865 mm; W - 630 g). In case of spiny eel, the GSI varied from 0.65 (August) to 8.30 (July) in males and 0.70 (August) to 10.46 (July) in females. GSI showed single peak in July. Fecundity ranged from 570 (TL - 240 mm; W - 30 g) to 10870 (TL - 601; W - 350g). Histology of the testes and ovaries of the eels were carried out to investigate the gonadal development stages during the reproductive months (August to November 2003). In case of male M. cuchia, the secondary primordial germ cells, primary spermatogonium, some spermatogonia A and clone of spermatogonium B in testis were observed in September. In October-males different sized lobules having spermatogonia, spermatocytes and spermatids were observed. In the ovary of M. cuchia, polygonal shaped oocytes were seen during September. The oogonia were reduced with dense and irregular shaped during October. Numerous pycnotic cells were visible during November. In male M. armatus numerous broken lobule walls were found in testes during September. In October, abundant primary germ cells, pycnotic nests of degenerating cells, spermatogonia and spermatids were observed. In females, ovaries had distinct yolk vesicles stage and yolk granules stages in August. In September, the follicular cells of the oogonia were ruptured, shrunk forming irregular shaped in October. Oogonia were also shrunk with thin, irregular shaped structure but broken parts of the ruptured follicular cells were scattered in case of M. armatus. Experimental attempts on artificial propagation indicated that both freshwater eels were difficult to breed using inducing agents like pituitary glands (PG) of 10, 20, 50, 100 and 150 mg per kg of body weight. Same doses were used for both sexes with equal sex-ratio. In both cases, brood fish died at higher doses of injection given at 100 and 150 mg PG/kg bodyweight. However, M. cuchia breed naturally in cisterns when provided with water hyacinths and tunnel in muddy bottom. M. cuchia fed with chopped cooked fish attained a mean weight of 18.75 ± 2.3 g and cent percent survival. While in case of M. armatus best growth by weight (12.0 ± 2.48 g) and cent percent survival were achieved using chopped raw fish. Car tyre was observed as best shelter for attaining the mean weight gain 22.53 ± 2.24 g and cent percent survival of M. cuchia. While PVC pipe was found to be the best shelter for M. armatus, where it attained the mean weight of 12.73 ± 1.88 g and cent percent survival.