Post-spawning Fish Research Articles

Development of a Simple Morphometric Model to Identify Sex in Chinook Salmon Returning to Spawn in the Yukon River

AbstractChinook Salmon Oncorhynchus tshawytscha support major fisheries in the Yukon River in Canada and the United States. Demographic data, including length, age, and sex composition of annual runs underpin management programs tasked with meeting escapement goals and providing for various fisheries within the drainage. Numerous sampling projects along the river collect these data annually, most without sacrificing fish. However, substantial error can occur when assigning sex based on observations of secondary sexual characteristics, particularly early in the spawning migration. We sought to develop a quantitative method of analyzing morphology of Chinook Salmon in the Yukon River that would achieve ≥95% accuracy of sex assignment with live fish. We collected snout lengths (SL) and mid‐eye‐to‐fork lengths (MEF) from 2,342 known‐sex fish from six regions of the drainage ranging from 40 to 2,000 km upstream from the sea, including a collection from postspawning fish. We fit three logistic regression models to the data using MEF, the ratio of SL to MEF, and regional group as covariates, including interactions and quadratic terms. Model selection was made using Akaike information criterion and cross‐validation of prediction performance. Accuracy rates of the final quantitative model ranged from 79% for a lower Yukon River group to 92% for the postspawning group. Though short of our accuracy goal, the quantitative method may provide an improvement over existing methods of sex assignments in some regions of the drainage. Lower assignment rates in the lower river may reflect the mixed‐stock composition of the fishery, where samples include fish with both short and long migrations ahead of them. However, we speculate that Chinook Salmon migrating to upper Yukon River spawning areas minimize the expression of secondary sexual characteristics, such as SL in males, and instead allocate their energy reserves to accomplishing the long migrations.

The Entrainment and Screening of Returning and Postspawning Adult Salmonids at Irrigation Canals of the Umatilla River, Oregon

Abstract Anadromous salmonids can be vulnerable to entrainment at diversion intake structures on streams, effectively trapping fish in irrigation canals and removing them from a population. Currently little is known about how the differences in timing and direction of movement among adult salmonids contribute to their risk of entrainment and how successful they are at escaping irrigation canals. Potential routes of escape include passing against water currents and through the headgate of an irrigation canal intake or by navigating through screen and bypass infrastructure primarily designed to return juvenile fish to a stream. In this study, passive integrated transponders (PIT tags) were used to track the movement of adult Chinook Salmon Oncorhynchus tshawytscha (n = 573), Coho Salmon Oncorhynchus kisutch (n = 39), and anadromous Rainbow Trout Oncorhynchus mykiss (steelhead, n = 853) as they entered areas of the Umatilla River basin (Oregon) with irrigation canals and as they attempted to escape irrigation canals after entrainment. Although adult steelhead and spring Chinook Salmon often encountered diversions at similar times, the vast majority of entrained adults were steelhead (94%). Between 2% and 8% of adult steelhead observed entering the area were entrained. The entrainment of steelhead was strongly associated with downstream movements and Umatilla River discharge below 40 m3/s. Many downstream-moving steelhead were postspawning fish (kelts). As a result, vulnerability of anadromous adults to entrainment differed by species due to the direction of their movements and how these movements coincide with canal operations and river flows. It is unlikely that the screened irrigation canals acted as an ecological sink; the majority of adult salmonids approached the screen and bypass infrastructure (≥88%) and later river detection confirmed that many had used that infrastructure to return to the river (≥47%). However, half of steelhead appeared to experience bypass delays at fish screens. Adult steelhead that approached the canal headgate after becoming trapped in the canal did not successfully return to the Umatilla River using this route. Unscreened irrigation canals elsewhere may disproportionally trap downstream-moving steelhead, like postspawning kelts, due to their propensity for entrainment and their difficulties escaping through the water intakes of irrigation canals. In streams with anadromous salmonids, fish screen and bypass infrastructure primarily designed to eliminate the permanent entrainment of juvenile fish can also prevent the removal of adult fish that may reproductively contribute to the population.

A follow-up study of selected biomarkers of health in cod Gadus morhua L. collected from the southern Baltic off the Polish coast.

Selected biomarkers of health were examined in 50 post-spawning cod Gadus morhua collected in November 2015 from the southern Baltic. The biomarkers included condition factor (CF), macroscopic lesions, histopathology of spleen, liver and gonads, and morphometry of follicular atresia and hepatic and splenic melanomacrophage cells (MMC). All fish appeared in good body condition. One fish had a dermal ulcer, and in seven, macroscopic nematodes were noted within body cavity. Microscopic lesions in the liver included biliary myxozoanosis, microsporidial and necrocentric granulomas, parasitic hepatitis, multifocal necrosis, foci of cellular alterations, spongiosis, peliosis and cytoplasmic fibrillar inclusions. The spleen and gonads had microsporidial and/or necrocentric granulomas. Some of the biomarkers showed differences as compared to spawning cod collected in May from the same location in 2012, most importantly values an order of magnitude lower for splenic MMC in post-spawning fish. In post-spawning fish, there were statistically significant correlations between MMC, CF, follicular atresia, parasitic hepatitis and microsporidiosis. This is the first comparison of biomarkers of health in post-spawning and spawning Baltic cod. Future studies need to examine the relationships of biomarkers to levels of pollutants in the environment and in tissues of cod.

Determination of the spawning history of Pacific saury Cololabis saira from rearing experiments: identification of post-spawning fish from histological observations of ovarian arterioles

Rearing experiments under controlled temperature conditions were carried out to establish a methodology for estimating the spawning history of the Pacific saury Cololabis saira. The experiments involved histological examination of ovarian arterioles at various maturational stages. We found that pre-spawning immature fish lacked Victoria blue (VB)-positive arterioles in the ovaries, as did maturing fish, while post-spawning females had VB-positive arterioles. The latter were also observed after 6 months when most spawning activity had ceased. The results indicate that VB-positive ovarian arterioles are a characteristic feature of actively spawning Pacific saury and that females with VB-positive arterioles can be classified as post-spawning fish. Females without VB-positive arterioles can be regarded as “practically unspawned” fish, and this group may include some fish that have spawned a number of times, in addition to completely unspawned fish. The methodology has many practical uses such as in the estimation of maturation rates for stock management.

Spermatogenesis and related plasma androgen and progestin level in wild male golden mahseer, Tor putitora (Hamilton, 1822), during the spawning season.

Testicular development and plasma levels of sex steroid [11-ketotestosterone (11-KT), testosterone (T) and 17,20β-dihydoxy-4-pregnen-3-one (17,20β-P)] were studied for the first time in wild golden mahseer, Tor putitora. Testicular development was investigated by macroscopic observation and histology of the gonads, whereas steroids were measured by enzyme-linked immunosorbent assay. Based on macroscopic observation and germ cell types present in gonad histology, the testes of T. putitora were divided into five developmental stages: immature [stage I; spermatogonia (SPG)], early spermatogenesis [stage II; SPG and spermatocytes (SPC)], late spermatogenesis [stage III; SPG, SPC, spermatids (SPD) and spermatozoa (SPZ)], spermiation (stage IV; SPZ) and post-spawning (stage V; SPG, SPD and SPZ). During the stage I of the testes, the lowest levels of plasma sex steroid and gonadosomatic index (I G) were recorded. The highest plasma level of T was 0.89±0.09ng/mL and 11-KT was 4.23±0.54ng/mL, which was during the stage III and IV, respectively. The peak in 11-KT was coincident with the peak in I G (1.65±0.12%). The lowest T and 11-KT levels were 0.25±0.02ng/mL and 0.47±0.09ng/mL, respectively, which was at stage I. Plasma levels of 17,20β-P increased significantly at stage III (1.04±0.06ng/mL) and stage IV testes (1.28±0.03ng/mL) and then declined in post-spawned fish. This indicates that 17,20β-P could also be a possible maturation-inducing steroid in this fish. The condition factor (K) significantly decreased during the testicular development and was lowest at spermiation stage (0.53±0.02%). The proportion of running male peaked concomitantly with the appearance of stage IV testes. Presence of germ cells of different developmental stages indicates that T. putitora male is a multiple spawner, and the information generated here is important for developing a captive breeding, culture and conservation programs for this endangered coldwater Himalayan fish species.

Post-mortem sporulation of Ceratomyxa shasta (Myxozoa) after death in adult Chinook salmon.

Ceratomyxa shasta (Myxozoa) is a common gastrointestinal pathogen of salmonid fishes in the Pacific Northwest of the United States. We have been investigating this parasite in adult Chinook salmon ( Oncorhynchus tshawytscha ) in the Willamette River, Oregon. In prior work, we observed differences in the pattern of development of C. shasta in adult salmon compared to juvenile salmon. Adult salmon consistently had large numbers of prespore stages in many of the fish that survived to spawn in the fall. However, myxospores were rarely observed, even though they were exposed and presumably infected for months before spawning. We evaluated the ability of C. shasta to sporulate following fish death because it is reported that myxosores are common in carcasses of Chinook salmon. We collected the intestine from 30 adult salmon immediately after artificial spawning and death (T0). A total of 23 fish were infected with C. shasta based on histology, but only a few myxospores were observed in 1 fish by histology. Intestines of these fish were examined at T0 and T7 (latter held at 17 C for 7 days) using quantified wet mount preparations. An increase in myxospore concentrations was seen in 39% of these fish, ranging between a 1.5- to a 14.5-fold increase. The most heavily infected fish exhibited a 4.6-fold increase from 27,841 to 129,352 myxospores/cm. This indicates, supported by various statistical analyses, that under certain conditions presporogonic forms are viable and continue to sporulate after death in adult salmon. Considering the life cycle of C. shasta and anadromous salmon, the parasite may have evolved 2, non-mutually exclusive developmental strategies. In young fish (parr and smolts), the parasite sporulates shortly after infection and is released into freshwater from either live or dead fish before their migration to seawater, where the alternate host is absent. The second strategy occurs in adult salmon, particularly spring Chinook salmon, which become infected upon their return to freshwater in the spring or early summer. For several months throughout the summer, only prespore stages are observed in most fish, even at the time of spawning. But once the fish dies, environmental conditions experienced by C. shasta change and viable presporogonic stages are induced to sporulate. As the post-spawned fish occur in the upper reaches of rivers, the myxospores would be released in a freshwater environment that would provide a reasonable opportunity for them to encounter their freshwater polychaete hosts, which reside downstream.

Physiological indices of seawater readiness in postspawning steelhead kelts

AbstractManagement goals to improve the recovery of steelhead (Oncorhynchus mykiss) stocks at risk of extinction include increasing the proportion of postspawning fish that survive and spawn again. To be successful, postspawning steelhead (kelts) migrating downstream to the ocean must prepare physiologically and physically for a seawater transition. We sampled blood, gill filaments, and evaluated the external condition of migrating kelts from an ESA‐listed population in the Snake/Columbia River system over two consecutive years to evaluate their physiological readiness for transition to seawater. We chose attributes often considered as measures of preparation for seawater in juveniles, including gill Na+,K+ ATPase activity, plasma electrolytes and hormones to consider factors related to external condition, size and sex. We found kelts in good external condition had plasma profiles similar to downstream‐migrating smolts. In addition, we found more than 80% of kelts ranked in good external condition had smolt‐like body silvering. We compared measures from migrating kelts with samples obtained from hatchery fish at the time of spawning to confirm that Na+, K+ ATPase activity in kelts was significantly elevated over spawning fish. We found significant differences in gill Na+, K+ ATPase activity in migrating kelts between the years of sampling, but little indication of influence of fish condition. We conclude that the postspawning steelhead sampled exhibited a suite of behaviours, condition and physiology characteristic of fish prepared for successful transition to a seawater environment.

Metabolic endocrine factors involved in spawning recovery and rematuration of iteroparous female rainbow trout (Oncorhynchus mykiss)

To determine how energy balance affects metabolic hormones hypothesized to play a role in the onset of a new reproductive cycle in iteroparous salmonids, food availability after spawning was restricted in female rainbow trout. These fish were compared with a control group that was fed a standard brood stock ration. Bodyweight, length, and muscle lipid content were determined, and blood was collected from fish at regular intervals; a subset of fish from each group was sacrificed at each sampling time for the collection of liver and ovary tissue, and to calculate hepatosomatic index (HSI) and gonadosomatic index (GSI). Plasma hormone levels were quantified by radioimmunoassay, and tissue gene expression levels were analyzed using q-RT-PCR. The experiment was conducted twice, using two-year-old and three-year-old post-spawned fish. Food-restriction arrested ovarian growth and development within 15–20weeks, as evidenced by lower GSI in restricted-ration fish. Food restriction also reduced Fulton’s condition factor, muscle lipid content, and specific growth rate from one month onward, and reduced HSI after 3months. In the liver, insulin-like growth factor (igf1 and igf2) gene expression was reduced in three-year-old food-restricted fish within 2months; however, no effect of ration on igf1 or igf2 expression was detected in two-year-old fish. In both years, IGF binding protein-1 (igfbp1) gene expression decreased over time in both treatment groups. Liver leptin (slepA1) gene expression was lower in two-year-old food-restricted fish at 4months. These results show that this feed restriction regime arrested reproductive development and affected factors associated with energy balance purported to play a role in initiating reproductive development within 2–4months after spawning.

Expression of vitellogenin receptor gene in the ovary of wild and captive Atlantic bluefin tuna (Thunnus thynnus)

The cDNA sequences of vitellogenin receptor proteins (VgR+ and VgR−), containing or lacking the O-linked sugar domain, were determined in Atlantic bluefin tuna (Thunnus thynnus L.). VgR− gene expression in the ovary was compared in captive-reared and wild Atlantic bluefin tuna during the reproductive cycle. Gonad samples from adult fish were sampled from 2008 to 2010 from stocks reared in captivity at different commercial fattening operations in the Mediterranean Sea and from wild individuals caught either by traditional tuna traps during their migration towards the spawning grounds in the Mediterranean Sea or by the long-line artisanal fishery. In addition, juvenile male and female Atlantic bluefin tuna were sampled from a farming facility, to obtain baseline information and pre-adulthood amounts of VgR−. The total length of VgR+ cDNA was 4006 nucleotides (nt) and that of VgR− was 3946nt. Relative amounts of VgR− were greater in juvenile females and in those adults having only previtellogenic oocytes (119±55 and 146±26 folds more than juvenile males, respectively). Amounts of VgR− were less in individuals with yolked oocytes (ripening stage, May–June) and increased after spawning in July (92±20 and 113±13 folds more than juvenile males in ripening and post-spawning fish, respectively). These data suggest that regulation of VgR− is not under oestrogen control. During the ripening period, greater VgR− gene expression was observed in wild fish than in fish reared in captivity, possibly because of (a) differences in water temperature exposure and/or energy storage, and/or (b) an inadequate diet in reared Atlantic bluefin tuna.

Migratory patterns of exotic brown trout Salmo trutta in south‐western Hokkaido, Japan, on the basis of otolith Sr:Ca ratios and acoustic telemetry

Acoustic telemetry and microchemical analysis of otolith strontium-calcium ratios were used to evaluate how exotic brown trout Salmo trutta have responded to Japanese riverine environments of south-western Hokkaido by observing their migratory patterns. The existence of anadromous S. trutta was also verified. Most S. trutta caught in rivers for otolith analysis were freshwater residents (95·6%), whereas those caught in the sea were mainly smolts (91·3%), which had just migrated from rivers during spring. Anadromous S. trutta (n = 6) were captured in rivers and in the sea, confirming the existence of mature pre- and post-spawning fish. According to telemetry results, both mature and immature S. trutta used the river in winter, and their estimated sea-run timings showed individual differences. Through the combination of these two methods, migratory patterns on various spatio-temporal scales were observed. This first documentation of the presence of both male and female anadromous S. trutta in the same region within Japan indicated the risk of further colonization of exotic S. trutta via oceanic migration.

Teleost maturation-inducing hormone, 17,20β-dihydroxypregn-4-en-3-one, peaks after spawning in Tinca tinca

During an eight month study of the reproductive cycle in two age groups, and in both sexes, of tench ( Tinca tinca L.), it was found that plasma concentrations of the presumptive ‘maturation inducing hormone (MIH)’ 17,20β-dihydroxypregn-4-en-3-one (17,20β-P) did not reach a peak during the spawning season, but as much as two months after spawning had ceased. The cessation of the spawning season was confirmed by histological examination of the gonads and by measurement of 11-ketotestosterone and 17β-estradiol in the plasma of males and females, respectively. Measurements were also made of the ‘alternative MIH’ 17,20β,21-trihydroxypregn-4-en-3-one in the older fish. However, this steroid did not show the same pattern as 17,20β-P. An assessment was made of the prevalence of primary spermatocytes in the testes of post-spawned fish – to test an alternative hypothesis that 17,20β-P might be involved in the stimulation of meiosis. However, there was no evidence for any increase in testis differentiation post-spawning. In fact the testes became increasingly undifferentiated as the autumn progressed. The role, if any, of this ‘unseasonal’ peak of 17,20β-P production remains to be determined.

Cytoskeletal ultrastructure and lipid composition of I-Z-I fraction in muscle from pre- and post-spawned female hake ( Merluccius hubbsi)

Myofibrils from pre- and post-spawned female hake were depleted of thin and thick filaments by KI-treatment and the cytoskeletal ultrastructure of the preparations was investigated by electron microscopy. Irrespective of the gonadal stage of the fish, transmission (TEM) and scanning (SEM) electron micrographs showed an extensive network of filaments connecting Z-structures, which appeared as two circular rings held together. Z-structures of KI-treated myofibrils from post-spawned hake were more compact than those from pre-spawned fish. This ultrastructural difference was absent when KI-treated myofibrils from hake in both gonadal conditions were prepared in the presence of a proteinase inhibitor cocktail (1 mM EDTA+1 mM PMSF+1 mM iodoacetic acid). The total lipid (TL) content of the I-Z-I fraction from fish in pre-spawned condition was about 3.7 mg/g I-Z-I and non-polar lipids (NPL) represent 86.5% of TL. TL and NPL contents of the I-Z-I fraction from post-spawned hake were about 50% and 78.2% lower than those obtained from this fraction in pre-spawning fish. No significant changes ( p>0.05) were observed in phospholipids (PL) and acidic phospholipids (AP). No significant differences ( p<0.01) were observed among the corresponding lipid fractions of I-Z-I purified from pre- and post-spawning fish in the presence of a protease inhibitor cocktail.

Proteolytic Activity of Muscle in Pre- and Post-Spawning Hake ( Merluccius hubbsi Marini) After Frozen Storage

The proteolytic activity of fillets from fresh fish and after frozen storage of hake muscle from pre- and post-spawned fish was investigated. The autolytic activity of fresh muscle extracts from pre-spawned muscle extracts was significantly higher than that of post-spawned, except for pH 3.4 and incubation temperature of 60 °C where no significant difference was found. In both gonadal conditions, the freezing of hake fillets affected the proteolytic activity of crude muscle extracts. After 50 d of frozen storage, the proteolytic activity values of crude muscle extracts remained higher than the fresh values, except for pre-spawned extracts incubated at 37 °C. At longer storage period (110 d), the proteolytic activity continued with similar or higher values compared with those observed for fresh muscle extracts. Densitometric analysis of crude muscle extracts indicated degradation of MHC in fresh and frozen storage at both gonadal conditions under incubation conditions of pH 6.2 and temperatures 37 and 60 °C, pH 8.5 and temperatures 37 and 60 °C, pH 3.4 and temperature 4 °C.

Aerobic scaling and resting metabolism in oviferous and post-spawning Barents Sea capelin Mallotus villosus villosus (Müller, 1776)

As part of the quantitative investigations into the bioenergetic relationships of the Barents Sea capelin, Mallotus villosus villosus (Müller, 1776), resting metabolic rates were examined in the oviferous and post-spawning fish in order to provide insights to aerobic scaling and the basal energetic costs associated with reproduction. Aerobic scaling of the different categories of sexually mature fish (body weight, W=13–54 g) could be expressed as: Q O 2 =0.106 W 1.049 (oviferous fish; N=11), Q O 2 =0.411 W 0.430 (post-spawning females; N=9), and Q O 2 =0.075 W 1.012 (post-spawning males; N=14), where Q O 2 is the oxygen consumption (ml O 2 h −1) per fish. The weight specific oxygen consumption of oviferous capelin was about 30% higher (∼125 ml O 2 kg −1 h −1) compared to those of the post-spawning fish (79–87 ml O 2 kg −1 h −1). The results are discussed in context with other empirical studies on the aerobic scaling and metabolic costs involved with the build-up of roe in the fish.

Response of individual shoaling Atlantic cod to ocean currents on the northeast Newfoundland Shelf

The movements of sonically-tagged Atlantic cod swimming within a large (80,000 mt) shoal of post-spawning fish on the northeast Newfoundland Shelf were related to the acoustically determined position of the shoal and event structure in the local ocean currents. For the first several days of the study, a period when currents were unfavorable for shoreward transport, the tagged fish maintained station. There was little net movement of individual fish and no change in the geographical location of the shoal. Then, apparently in response to a shoreward current event, the cod moved with the flow. The tagged cod behaved as members of the shoal, in that their net movement was similar to that of the shoal itself. These observations further support the hypothesis that cod on the northeast Newfoundland Shelf selectively use currents for transport during their shoreward feeding migration. The cue to initiate migration by post-spawning cod is apparently an organized onshore flow exceeding a threshold velocity and duration.

Influence of spawning on swimming performance and muscle contractile properties in the short‐horn sculpin

The total distance travelled during the first two kinematic stages of the escape response of short‐horn sculpin was significantly greater in post spawning fish (0·41 L) than in gravid fish (0·23 L). The maximum velocity of the snout during the C‐bend was significantly higher (5·6 L s−1) in postspawning fish than in gravid fish (3·8 L s−1). To investigate some of the mechanisms underlying changes in swimming performance, the contractile properties of fast muscle fibres were determined in fish of similar body length. The rate of tetanic force relaxation (time from last stimulus to 50% peak force) was 34% faster in gr avid than in postspawning fish. Maximum contraction velocity, determined by the slack‐test method, was significantly higher in gravid than in postspawning fish (6·8 v. 5·9 muscle lengths s−1). In contrast, both maximum isometric stress and power output (determined from the force–velocity relationship) were &gt;50% higher in fibres from postspawning than from gravid fish, even though myofibrillar protein and water contents were similar (120 mg g−1 wet mass and 86% of body mass, respectively). The results show that swimming performance and the contractile properties of fast muscle fibres vary with the reproductive cycle in short‐horn sculpin acclimated to the same photoperiodic and temperature regime.

夏季の中部北太平洋におけるサンマの成熟と日齢

The maturation process and age in days of female Pacific saury Cololabis saira (Brevoort) in the central North Pacific Ocean were examined based on histological observations and otolith daily growth increments. The maturation process of Pacific saury used in this study (243-342mm in knob length) was divided into 6 stages: early yolk vesicle stage, late yolk vesicle stage, yolk formation stage, maturation stage, atretic stage I, and atretic stage II. Atretic stage fish were considered to be post-spawning as many degenerated oocytes were observed. The large size group (more than 296mm), in which fish were more than 1.5 years old as estimated from daily growth increments in otoliths, contained spawning and post-spawning individuals, although all individuals from the medium size group (241-295mm), from 10 months to 1 year and 3 months old, were immature. Since post-spawning fish were seen to have recovered their initial condition factors, it is suggested that they survive beyond the first spawning season. Based on observations of asynchronously developing oocytes, it is considered that Pacific saury spawn several times during a single spawning season.

Manipulation of the seasonal reproductive cycle in winter flounder, Pleuronectes americanus, using a gonadotropic hormone releasing hormone

A gonadotropic hormone-releasing hormone analogue ([D-Ala6, Pro9-NHEt]LHRH, GnRH-A) was administered to the winter flounder, Pleuronectes americanus (Walbaum) at different stages of the reproductive cycle from September 1988 to July 1989. Both males and females responded to releasing hormone treatment throughout the year except for a brief summer period in post-spawned fish when the gonads were regressed. In females, GnRH-A markedly increased plasma estradiol-17β and testosterone levels and accelerated growth of the ovaries during the period of early gonadal recrudescence beginning in August. In prespawning females, GnRH-A treatment increased plasma testosterone levels in association with a stimulation of germinal vesicle migration and ultimately ovulation. During the fall, GnRH-A treatment of males increased plasma androgen levels (testosterone and 11-ketotestosterone) and stimulated growth of the testes. In February, a time when spermiation is initiated in only half of the males, the proportion of spermiating males was increased by treatment with releasing hormone, indicating that spermiation can be advanced in some males by ca. 2 mo.

Infection dynamics of Lemaeocera lusci on sand goby Pomatoschistus minutus in the Oosterschelde (The Netherlands)

Male and female sand gobies Pomatoschistus minutus (Pallas) from the Oosterschelde (The Netherlands) were examined for the crustacean mesoparasite Lernaeocera lusci (Bassett-Smith) from Apnl to July 1992. Prevalence increased steeply from 4 % in Apnl to 80% (females) and 57 % (males) in June. The peak abundance of recently attached infective stages (pennella larvae) on sand gobies was found in May. By June, about 70% of parasites were in the post-metamorphosis stage and possessed egg strings. Significant differences in abundance between males and females were found in June but not in May. This may be due to sex-specific differences in behaviour during spawning time. The majority of post-spawning fish probably d ~ e d in July. Fish of the new year class which were captured in July harboured no parasites.

In vitro characterization of gonadotropin-releasing hormone antagonists in goldfish, Carassius auratus.

The two native forms of GnRH, salmon GnRH and chicken GnRH-II, in the brain and pituitary of goldfish are both active in stimulating gonadotropin-II (GTH-II) and GH release. The objective of the present study was to characterize GnRH antagonists for their ability to inhibit sGnRH- and cGnRH-II-induced GTH-II and GH release in goldfish using a pituitary fragments perifusion system. Contrary to expectations, putative GnRH antagonists with D-Arg6 stimulated GTH-II and GH release in nearly all cases. [Ac-delta 3-Pro1,4FD-Phe2,D-Trp3,6]mammalian (m) GnRH inhibited sGnRH- and cGnRH-II-stimulated GTH-II release in a dose-dependent manner, with ED50 values of 242 +/- 48 and 169 +/- 17 nM, respectively. [Ac-delta 3-Pro1,4FD-Phe2,D-Trp3,6]mGnRH also inhibited GH release stimulated by sGnRH (ED50, 128 +/- 74 nM) and cGnRH-II (ED50, 157 +/- 67 nM). The degree of inhibition was higher in sexually regressed fish compared to postspawning fish. [D-p-Glu1,D-Phe2,D-Trp3,6]mGnRH suppressed both sGnRH- and cGnRH-II-induced GTH-II release with ED50 values of 326 +/- 96 and 249 +/- 74 nM, respectively. [Ac-delta 3-Pro1,4FD-Phe2,D-Trp3,6]sGnRH inhibited sGnRH and cGnRH-II stimulated GTH-II release, but stimulated GH release. On the other hand, [Ac-D(2)-Nal1,4Cl-D-Phe2,D-(3)Pal3,6,Arg5,D-A la10]mGnRH weakly stimulated GTH-II release, but strongly inhibited basal GH release. These results indicate that [Ac-delta 3-Pro1,4FD-Phe2,D-Trp3,6]mGnRH has clear antagonistic activity on sGnRH and cGnRH-II stimulation of GTH-II and GH release in vitro. The differential actions of a few GnRH analogs on GTH-II and GH release indicate that the properties of the GnRH receptors on GTH and GH cells may be different. The amino acid in position 6 plays an important role in determining the nature of intrinsic activity of GnRH peptides, and substitution of D-Arg6 normally produces agonistic analogs.