Activin and its receptors in the goldfish

Activin (_AA, _AB and _BB) is a dimeric protein that belongs to the transforming growth factor- (TGF-) superfamily of growth factors and is involved in the regulation of many physiological and developmental processes. Recently, we have demonstrated that porcine activin stimulated goldfish gonadotropin-II (GTH-II) and growth hormone (GH) secretion from dispersed pituitary cells in static culture and pituitary fragments in perifusion. The action of activin in the goldfish is unique in that it has an acute stimulatory effect on the secretion of GTH-II and GH, whereas in mammals activin usually exhibits long-term stimulatory actions on FSH secretion. The action mechanism of activin is different from that of gonadotropin-releasing hormone (GnRH). Using domain-specific antibodies against mammalian activin subunits, we subsequently demonstrated the existence of immunoreactive activin subunits (_A and _B) in the goldfish ovary, testis, pituitary and brain, suggesting endocrine, paracrine and autocrine roles for activin in the regulation of goldfish reproduction. Both activin _A and _B subunits have been cloned from goldfish genome by polymerase chain reaction (PCR). Using the PCR fragments as probes, we have cloned a full length cDNA coding for activin _B subunit from the goldfish ovary. Both activin _A and _B subunits show high homology to those of other vertebrates with the _B subunit much more conserved (93 and 98% identity with human and zebrafish _B subunit, respectively). The identity of the cloned _B subunit was further confirmed by expression in the Chinese hamster ovary (CHO) cells and detection of the specific activity of activin in the culture medium. The messenger RNA of activin _B subunit is expressed in a variety of goldfish tissues including ovary, testis, brain, pituitary, kidney and liver, suggesting a wide range of physiological roles for activin in the goldfish. We have also cloned a full length cDNA coding for the activin Type IIB receptor from the goldfish ovary, suggesting that activin may have paracrine or autocrine actions on the ovarian functions. The identity of the cloned receptor was confirmed by specific binding of¹²⁵ I-activin on COS-1 cells transfected with the cloned Type IIB receptor.     

Evidence that 17α,20β,21-trihydroxy-4-pregnen-3-one is a maturation-inducing steroid in spotted seatrout

Ovarian steroidogenesis during final oocyte maturation (FOM) in the spotted seatrout (Cynoscion nebulosus) was investigated by incubating ovarian fragments with tritiated pregnenolone, followed by chromatographic separation of the radioactive products. The major tritiated steroid produced during FOM comigrated with 17α,20β,21-trihydroxy-4-pregnen-3-one (20β-dihydro-11-deoxycortisol, 20β-S) on HPLC and TLC. Only minor amounts of radioactive material coeluted with 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-P), 11-deoxycorticosterone (DOC), estradiol-17β and testosterone standards in the HPLC system. Additional chromatography by TLC confirmed the presence of radioactive estradiol-17β and testosterone but not 17α,20β-P and DOC. All the ovarian steroids producedin vitro during FOM were assayed for their ability to induce germinal vesicle breakdown (GVBD) of spotted seatrout oocytes. Twenty grams of ovarian tissue were incubated with human chorionic gonadotropin and exogenous pregnenolone. The steroidal products were purified by HPLC and TLC. Most of the maturation-inducing activity was confined to steroidal material which comigrated in these systems with 20β-S. This material was active at a concentration of 1 ng steroid/ml medium in the GVBD assay. Smaller amounts of material which coeluted with 11-deoxycortisol, DOC, 17α,20β-P and several minor unidentified fractions induced GVBD at concentrations of 10 ng steroid(s)/ml. The structure-activity relationships of authentic steroids in inducing GVBD of spotted seatrout oocytes was investigated. Hydroxylation at the 17α, 20β or 21 positions increased potency to induce GVBD. Steroids with multiple hydroxyl groups at the 17α and 20β positions (17α, 20β-P) and at the 17α, 20β, and 21 positions (20β-S) had maximum biological activity in the GVBD bioassay. The results suggest that 20β-S is a major maturation-inducing steroid in spotted seatrout.     

Fishing gear effects and ecology of the sea whip (Halipteris willemoesi (Cnidaria: Octocorallia: Pennatulacea)) in British Columbia,Canada: preliminary observations

• 1. We conducted a preliminary study of the effects of a shrimp beam trawl and prawn traps on sea whips (Halipteris willemoesi (Cnidaria: Octocorallia: Pennatulacea)) at two bays on Clio Channel, south central coast of British Columbia, Canada. Video and grabs surveys were conducted to estimate abundance and make observations on sea whips and their habitat.
• 2. No adult sea whips were caught in six beam trawls. In 600 prawn (Pandalus platyceros) trap sets at Turnour Bay, 30 sea whips were found entangled in the gear and of these 50% of the colonies were damaged. The length of broken sea whips ranged from 0.09 to 1.03 m. At Turnour Bay, the density of adults observed in video surveys ranged from 7.1 to 14.3 m^?2. Juvenile density in grab samples ranged from 53 to 123 m^?2 and a few adults were also caught. The lengths of intact adult sea whips caught in traps and grabs ranged from 0.20 to 1.98 m and juveniles from 0.3 to 0.7 cm.
• 3. Fewer adult sea whips (<0.1 m^?2) were seen in Bones Bay, where sediments were muddier (grain size <0.25 mm: 84.6 to 97.2%) relative to Turnour Bay (grain size <0.25 mm: 71.4 to 85.9%). Bottom water characteristics at the two bays were similar. Seasonal temperature range was 7.5–8.4°C, salinity 30.63–31.49 psu, and dissolved oxygen 2.76–5.97 mg L^?1. Differences in sediment characteristics and food availability may have been a factor influencing differences in abundance of sea whips between the two bays.
• 4. Further data are needed to investigate the effects of trawling and trapping on sea whips, as sample sizes were small in the present study and only two gear types were evaluated. Studies of effects of otter trawling and heavier traps, such as those used for Dungeness crab (Cancer magister), are warranted.

Copyright © 2005 John Wiley & Sons, Ltd.     

Histological examination of final oocyte maturation and atresia in wild and domesticated Penaeus monodon (Fabricius) broodstock

Oocyte maturation and gonadosomatic index (GSI) of eyestalk ablated Penaeus monodon females collected from the wild and from two first‐generation domesticated lines were assessed. Frequency and diameter of the different oocytes, and the intensity of oocyte atresia, were compared among groups through histological assessments of the sections of the middle ovarian lobe. Digitized images from ovary sections were used to record the frequency and diameter of different oocyte types. Spawning performance of the three groups were expressed in terms of the percentage of females that spawned at least once (productive females), time from eyestalk ablation to first spawning (latency period) and the number of spawnings per female stocked. Final ovarian maturation was attained in all groups, as indicated by the presence of mature oocytes with cortical rods (cortical oocytes), dark‐green ovarian colour and high GSI values (5.83–6.86%). However, domesticated groups presented significant larger immature oocyte types (previtellogenic and yolky oocytes) and smaller cortical oocytes compared with wild females, indicating a reduced vitellogenic activity during final oocyte maturation. Additionally, the frequency of atresia was comparatively higher for both domesticated groups, which could be related to their inferior spawning performance. The implications of these results on the reproductive potential and development of domesticated P. monodon are discussed.     

Contribution of the natural biota associated with substrates to the nutritional requirements of the post-larval shrimp, Penaeus esculentus (Haswell), inhigh-density rearing systems

The contribution of epiphytes associated with physical substrates to the nutritional requirements of post‐larval shrimp, Penaeus esculentus Haswell, was determined in high‐density rearing systems (3000, 6000 and 11 000 m^?3). Stable isotope signatures of epiphytes on polyethylene mesh substrate, AquaMats? and tank walls were compared with shrimp signatures. Two methods were used: the determination of carbon and nitrogen natural abundance ratios; and ¹⁵N‐nitrogen enrichment ratios after the addition of ¹⁵N‐ammonium to tanks. Using the natural abundance technique and a simple mixing model, epiphytes were found to contribute substantially to the carbon requirements of post‐larval shrimp (39–53%). This was despite the addition of formulated feed at satiation levels. There was no indication of a reduced contribution of carbon from epiphytes to shrimp nutrition at higher shrimp densities. The lack of a difference in the ¹⁵N/¹⁴N ratios of the two food sources meant that mixing models could not be used to calculate the contribution of nitrogen from epiphytes vs. artificial feed to shrimp nutrition. Using the ¹⁵N‐nitrogenenrichment method, the amount of nitrogen contributed by epiphytes to shrimp nutrition over 24 h could be determined. This method showed that nitrogen from epiphytes was assimilated by shrimp. ¹⁵N‐enrichment methods provided a more accurate alternative to natural abundance techniques, particularly when the stable isotope signals ofthe food sources are similar. This experiment hasshown the benefits in providing substrates for P.esculentus in high‐density rearing systems to provide an additional food source for shrimp.     

Factors affecting the persistence of traditional agricultural landscapes in Slovakia during the collectivization of agriculture

Collectivization of agriculture (1950s–1970s) was one of the most important periods in landscape development in Slovakia. Traditionally managed agricultural landscapes, that covered more than half of the Slovak territory, were transformed into large-scale fields and only fragments of traditional agricultural landscapes survived. We mapped the remaining traditional agricultural landscapes using aerial photos and historical maps. We then statistically analyzed the various geographical factors and their influence on the transformation process of traditional and collectivized fields, i.e., slope steepness, soil fertility, distance from settlements and isolation from regional capital cities. The comparison was performed using classification tree analysis. We constructed a set of decision rules that explain why fields were managed traditionally or collectivized. Our findings show that traditional agricultural fields were more likely to persist on steep terrain, less fertile soils, and on locations that were closer to the settlements, but more isolated from the regional capital cities. Steepness played the most important role: small-scale fields located on steep areas were not accessible to heavy machinery and therefore, frequently survived the collectivization. We show that the selected geographical factors are good explanatory variables for the collectivization of arable fields and orchards. For vineyards and grasslands, however, the explanatory power of the selected geographical factors is lower, and we suspect that other factors, not depicted in the analysis play an important role.     

Effects of Added Shelter and Stocking Density on Growth of Sleepy Cod Oxyeleotris lineolatus in Ponds

Sleepy cod Oxyeleotris lineolatus is a species of freshwater goby in demand in Australian markets by consumers of Asian origin. It is related to marble goby Oxyeleotris marmoratus , the most expensive freshwater food fish in Asia, which is cultured throughout southeast Asia in ponds and cages. The performance of sleepy cod in culture conditions was investigated to assess the viability of farming them in northern Australia. Sleepy cod fingerlings (62.8 ± 0.8 mm total length and 2.56 ± 0.095 g) were stocked into experimental ponds at 32,857 fish/ha, and grown out for 8 mo. Shelter was provided in each of three replicate ponds and was absent in three control ponds. The provision of shelter in juvenile growout was found to be of no benefit, although fish in ponds provided with shelter weighed slightly more per unit length than fish in ponds without shelter. Cannibalism was not a problem in growout, and survival was close to 100%. After the shelter trial was completed, fish were graded into large and small classes (three replicates of each), and grown out without shelter at the same density for 158 d. Following that, fish were again graded, and the largest 30% retained from growout at a density of 8,857 fish/ha (large, 198 ± 6.44 g) or 10,000 fish/ha (small, 48.9 ± 1.27 g). These were grown out for 188 d. Growth of selected stock at low densities was slower than earlier growth rates, although smaller fish gained weight more rapidly than larger fish. Growth rates were better than the only published data for marble goby. Further investigation into high density culture and different genotypes of sleepy cod needs to be undertaken to determine the viability of pond culture.     

The effect of temperature on the clearance of intraperitoneally-injected gonadotropin in the goldfish Carassius auratus

The serum half-disappearance time ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$), metabolic clearance rate and volume of distribution of intraperitoneally administered carp gonadotropin (cGtH) and endogenous GtH levels were determined in sexually mature male and female goldfish, Carassius auratus maintained at 12 ± 1°C or 20 ± 1°C. The results indicated that the rate of serum uptake of the injected cGtH from the peritoneal cavity was greater at 20°C than at 12°C in sexually mature male goldfish. Although increased endogenous serum GtH levels and decreased values of serum $\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ were associated with the elevated temperature, there was no difference in any of the parameters between sexually mature male and female goldfish acclimated to 12°C.     

Conservation of freshwater fish in the British Isles: The current status and biology of threatened species

1. The objective of this paper is to review the status of all freshwater fishes in the British Isles and to identify theatened species, races and communities which are in need of conservation. 2. Using objective criteria to assess the degree of threat, two of the 55 freshwater species known in the British Isles are thought to be extinct here and eight are believed to be under significant threat. 3. The biology of these ten species is reviewed. They are: Sturgeon Acipenser sturio, Allis Shad Alosa alosa, Twaite Shad Alosa fallax, Arctic Charr Salvelinus alpinus, Houting Coregonus oxyrinchus (extinct), Powan Coregonus lavaretus, Pollan Coregonus autumnalis, Vendace Coregonus albula, Smelt Osmerus eperlanus and Burbot Lota lota (extinct). 4. In addition, there are several races of commoner species which are believed to be worthy of special conservation measures. These fish include the purely freshwater race of River Lamprey Lampetra fluviatilis in Loch Lomond, dwarf Brook Lampreys Lampetra planeri in the Inner Hebrides, spineless Three-spined Sticklebacks Gasterosteus aculeatus in the Outer Hebrides and a number of important races of Brown Trout Salmo trutta in various waters in Ireland and Scotland. 5. There is also a range of fish communities in individual rivers and lakes which merit conservation attention. These are important for a variety of reasons, including high (and very low) diversity, unique assemblages, pristine stocks and highly characteristic communities. 6. The major threats to these species and communities include industrial and domestic pollution, acidification, land use changes, river barriers, drainage, fish farming, fishery management and the introduction of new species. 7. The identification of these threatened fishes and important freshwater sites provides a foundation for a review of existing protection in the British Isles and a major fish conservation management programme. These will be described in subsequent papers.     

Treatment of shrimp farm effluent with omnivorous finfish and artificial substrates

Long‐term environmental sustainability and community acceptance of the shrimp farming industry in Australia requires on‐going development of efficient cost‐effective effluent treatment options. In this study, we aimed to evaluate the effectiveness of a shrimp farm treatment system containing finfish and vertical artificial substrates (VAS). This was achieved by (1) quantifying the individual and collective effects of grey mullet (Mugil cephalus L.) and VASs on water and sediment quality, and (2) comparing the retention of N in treatment systems with and without the presence of finfish (M. cephalus and the siganid Siganus nebulosus (Quoy & Gaimard)), where light was selectively removed. Artificial substrates were found to significantly improve the settlement of particulate material, regardless of the presence of finfish. Mullet actively resuspended settled solids and reduced the production of nitrate when artificial substrates were absent. However, appreciable nitrification was observed when mullet were present together with artificial substrates. The total quantity of N retained by the mullet was found to be 1.8– 2.4% of the incoming pond effluent N. It was estimated that only 21% of the pond effluent N was available for mullet consumption. When S. nebulosus was added, total finfish N retention increased from 1.8% to 3.9%, N retention by mullet also improved (78±16 to 132±21‐mg N day^?1 before and after siganid addition respectively). Presence of filamentous macroalgae (Enteromorpha spp.) was found to improve the removal of N from pond effluent relative to treatments where light was excluded. Denitrification was also a significant sink for N (up to 24% N removed). Despite the absence of algal productivity and greater availability of nitrate, denitrification was not higher in treatments where light was excluded. Mullet were found to have no effect on the rates of denitrification but significantly reduced macroalgal growth on the surface of the water. When mullet were absent, excessive macroalgal growth led to reduced dissolved oxygen concentrations and nitrification. This study concludes that the culture of mullet alone in shrimp farm effluent treatment systems does not result in significant retention of N but can contribute to the control of macroalgal biomass. To improve N retention and removal, further work should focus on polyculturing a range of species and also on improving denitrification.