Effects of stress on the immune system of fish

The effects of stress on the immune system of various fish species including dab Limanda limanda, flounder Platichthys flesus, sea bass Dicentrarchus labrax and gobies Zosterisessor ophiocephalus, were investigated from laboratory and field experiments, using various assays to measure immunocompetence, correlated with histological and ultrastructural observations. Modulation of the immune system was demonstrated at tissue, cellular and biochemical levels following exposure to various stressors. The spleen somatic index was depressed in dab stressed in the laboratory and gobies collected from polluted sites in the Venice Lagoon. Differential blood cell counts consistently showed an increase in phagocytes and decrease in thrombocytes in fish exposed to various stressors. Phagocytic activity from spleen and kidney adherent cells was stimulated in dab stressed by transportation but depressed in fish exposed to chemical pollutants. Respiratory burst activity in phagocytic cells was also stimulated in stressed dab but depressed in sea bass exposed to cadmium. The results are discussed in relation to current concepts on stress in fish and the regulation of the immune system.

Age, growth, and spawning season of red bream (Beryx decadactylus) off the southeastern United States

Red bream (Beryx decadactylus) is a commercially important deep-sea benthopelagic fish with a circumglobal distribution on insular and continental slopes and seamounts. In the United States, small numbers are caught incidentally in the wreckfish (Polyprion americanus) fishery which operates off the southeastern coast, but no biological information exists for the management of the U.S. red bream population. For this study, otoliths (n=163) and gonads (n=161) were collected from commercially caught red bream between 2003 and 2008 to determine life history parameters. Specimens ranged in size from 410 to 630 mm fork length and were all determined to be mature by histological examination of the gonads. Females in spawning condition were observed from June through September, and reproductively active males were found year-round. Sectioned otoliths were difficult to interpret, but maximum age estimates were much higher than the 15 years previously reported for this species from the eastern North Atlantic based on whole-otolith analysis. Estimated ages ranged from 8 to 69 years, and a minimum lifespan of 49 years was validated by using bomb radiocarbon dating. Natural mortality was estimated at 0.06/yr. This study shows that red bream are longer lived and more vulnerable to overfishing than previously assumed and should be managed carefully to prevent overexploitation.

Annual fecundity, batch fecundity, and oocyte atresia of Atka mackerel (Pleurogrammus monopterygius) in Alaskan waters

Annual potential fecundity, batch fecundity, and oocyte atresia were estimated for Atka mackerel (Pleurogrammus monopterygius) collected in Alaskan waters during 1993−94. Atka mackerel were assumed to be determinate spawners on the basis of decreasing fecundity after batch spawning events. Histological examination of the ovaries indicated that oocytes in the vitellogenic stage and higher had been spawned in the current spawning season. For an average female of 40 cm, potential annual fecundity was estimated to be 41,994 eggs, average batch size (i.e., batch fecundity) was estimated to be 6689 eggs, and there were 6.13 batches per spawning season. Atresia was estimated by examining postspawning specimens and was found to be substantial. The average amount of atresia for a 40-cm fish was estimated to be 11,329 eggs, resulting in an estimated realized fecundity of only 30,664 eggs and 4.64 batches of eggs per spawning season.

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.

Reproductive biology, spawning season, and growth of female rex sole (Glyptocephalus zachirus) in the Gulf of Alaska

Rex sole (Glyptocephalus zachirus) have a wide distribution throughout the North Pacific, ranging from central Baja California to the western Bering Sea. Although rex sole are an important species in the commercial trawl fisheries off the U.S. West Coast, knowledge of their reproductive biology is limited to one study off the Oregon coast where ovaries were analyzed with gross anatomical methods. This study was initiated to determine reproductive and growth parameters specific to rex sole in the Gulf of Alaska (GOA) stock. Female rex sole (n=594) ranging in total length from 166 to 552 mm were collected opportunistically around Kodiak Island, Alaska, from February 2000 to October 2001. All ovaries were analyzed by using standard histological criteria to determine the maturity stage. Year-round sampling of rex sole ovaries confirmed that rex sole are batch spawners and have a protracted spawning season in the GOA that lasts at least eight months, from October to May; the duration of the spawning season and the months of spawning activity are different from those previously estimated. Female rex sole in the GOA had an estimated length at 50% maturity (ML50) of 352 mm, which is greater than the previously estimated ML50 at southern latitudes. The maximum age of collected female rex sole was 29 years, and the estimated age at 50% maturity (MA50) in the GOA was 5.1 years. The von Bertalanffy growth model for rex sole in the GOA was significantly different from the previously estimated model for rex sole off the Oregon coast. This study indicated that there are higher growth rates for rex sole in the GOA than off the Oregon coast and that there are differences in length at maturity and similarity in age at maturity between the two regions.

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.

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.

Pawpaw seed as fertility control agent on male Nile tilapia

To find out if pawpaw (Carica papaya) seeds can induce sterility in male Nile tilapia (Oreochromis niloticus) and to determine if sterility so induced is reversible or otherwise, mature male tilapia of mean weight 40 g were treated for 30 days with a low dose (4.9 g/kg/day) and a high dose (9.8 g/kg/day) of ground pawpaw seeds incorporated into their feed. Fish of similar sizes in the control experiment were fed with feed that did not contain pawpaw seed. The pawpaw seeds induced permanent sterility in the fish that received the high dose, while sterility in the low dose treatment was reversible. Fish in the control experiment spawned two weeks into the experiment and again in the fifth week. Fish in the low dose treatment spawned three weeks after the treatment had been discontinued. Histological sections of the testes showed that pawpaw seeds produced swollen nuclei in the low dose treatment and disintegrated cells in the high dose treatment. The study showed that pawpaw seeds, which are easy to obtain, can be incorporated into fish feeds and used by farmers to control prolific breeding of Nile tilapia.

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.

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.

Inshore spawning of cobia (Rachycentron canadum) in South Carolina

We documented inshore spawning of the recreationally important cobia (Rachycentron canadum) in Port Royal Sound (PRS) and St. Helena Sound (SHS), South Carolina, during the period from April to June in both 2007 and 2008. Histological analysis of ovaries confirmed the presence of actively spawning females inshore, and gonadosomatic index (GSI) values from females collected inshore (mean=7.8) were higher than the values from females caught offshore (mean=5.6); both of these mean values indicate that spawning occurred locally. Additionally, we conducted an ichthyoplankton survey in 2008 and found cobia eggs and larvae as far as 10 and 15 km inshore from the mouths of SHS and PRS, respectively. A study of egg development that we conducted in 2007 and 2008 using hatchery-reared cobia eggs provided descriptions of embryological development of cobia. Comparison of visual and quantitative characteristics of the field-collected eggs with those of the hatchery-reared eggs allowed positive identification of eggs collected in plankton samples. The ages of field-collected eggs and presence of females with hydrated oocytes in PRS and SHS observed in our ichthyoplankton survey and histological analysis indicated that wild cobia spawn in the afternoon and early evening. The inshore migration of cobia from April to June, the presence of actively spawning females, significantly higher GSI values, and the collection of eggs inside PRS and SHS all confirm that these estuaries provide spawning habitat for cobia. Because of the potential for heavy exploitation by recreational anglers as cobia move inshore to spawn in South Carolina, current management strategies may require review.

Staging ovaries of Haddock (Melanogrammus aeglefinus): implications for maturity indices and field sampling practices

We build on recent efforts to standardize maturation staging methods through the development of a field-proof macroscopic ovarian maturity index for Haddock (Melanogrammus aeglefinus) for studies on diel spawning periodicity. A comparison of field and histological observations helped us to improve the field index and methods, and provided useful insight into the reproductive biology of Haddock and other boreal determinate fecundity species. We found reasonable agreement between field and histological methods, except for the regressing and regenerating stages (however, differentiation of these 2 stages is the least important distinction for determination of maturity or reproductive dynamics). The staging of developing ovaries was problematic for both methods partly because of asynchronous oocyte hydration during the early stage of oocyte maturation. Although staging on the basis of histology in a laboratory is generally more accurate than macroscopic staging methods in the field, we found that field observations can uncover errors in laboratory staging that result from bias in sampling unrepresentative portions of ovaries. For 2 specimens, immature ovaries observed during histological examination were incorrectly assigned as regenerating during macroscopic staging. This type of error can lead to miscalculation of length at maturity and of spawning stock biomass, metrics that are used to characterize the state of a fish population. The revised field index includes 3 new macroscopic stages that represent final oocyte maturation in a batch of oocytes and were found to be reliable for staging spawning readiness in the field. The index was found to be suitable for studies of diel spawning periodicity and conforms to recent standardization guidelines.

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.

Translocation as a strategy to rehabilitate the queen conch (Strombus gigas) population in the Florida Keys

Queen conch (Strombus gigas) stocks in the Florida Keys once supported commercial and recreational fisheries, but overharvesting has decimated this once abundant snail. Despite a ban on harvesting this species since 1985, the local conch population has not recovered. In addition, previous work has reported that conch located in nearshore Keys waters are incapable of spawning because of poor gonadal condition, although reproduction does occur offshore. Queen conch in other areas undergo ontogenetic migrations from shallow, nearshore sites to offshore habitats, but conch in the Florida Keys are prevented from doing so by Hawk Channel. The present study was initiated to determine the potential of translocating nonspawning nearshore conch to offshore sites in order to augment the spawning stock. We translocated adult conch from two nearshore sites to two offshore sites. Histological examinations at the initiation of this study confirmed that nearshore conch were incapable of reproduction, whereas offshore conch had normal gonads and thus were able to reproduce. The gonads of nearshore females were in worse condition than those of nearshore males. However, the gonadal condition of the translocated nearshore conch improved, and these animals began spawning after three months offshore. This finding suggests that some component of the nearshore environment (e.g., pollutants, temperature extremes, poor food or habitat quality) disrupts reproduction in conch, but that removal of nearshore animals to suitable offshore habitat can restore reproductive viability. These results indicate that translocations are preferable to releasing hatchery-reared juveniles because they are more cost-effective, result in a more rapid increase in reproductive output, and maintain the genetic integrity of the wild stock. Therefore, translocating nearshore conch to offshore spawning aggregations may be the key to expediting the recovery of queen conch stocks in the Florida Keys.

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.