Cryopreservation of male and female gonial cells by vitrification in the critically endangered cyprinid honmoroko <Emphasis Type="Italic">Gnathopogon caerulescens</Emphasis>

We investigated the feasibility of cryopreservation of spermatogonia and oogonia in the critically endangered cyprinid honmoroko Gnathopogon caerulescens using slow-cooling (freezing) and rapid-cooling (vitrification) methods. Initially, we examined the testicular cell toxicities and glass-forming properties of the five cryoprotectants: ethylene glycol (EG), glycerol (GC), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (BG), and we determined cryoprotectant concentrations that are suitable for freezing and vitrification solutions, respectively. Subsequently, we prepared the freezing solutions of EG, GC, DMSO, PG, and BG at 3, 2, 3, 2, and 2 M and vitrification solutions at 7, 6, 5, 5, and 4 M, respectively. Following the cryopreservation of the testicular cells mainly containing early-stage spermatogenic cells (e.g., spermatogonia and primary spermatocytes), cells were cultured for 7 days and immunochemically stained against germ cell marker protein Vasa. Areas occupied by Vasa-positive cells indicated that vitrification led to better survival of germ cells than the freezing method, and the best result was obtained with 5 M PG, about 50% recovery of germ cells following vitrification. In the case of ovarian cells containing oogonia and stage I, II, and IIIa oocytes, vitrification with 5 M DMSO resulted the best survival of oogonia, with equivalent cell numbers to those cultured without vitrification. The present data suggest that male and female gonial cells of the endangered species G. caerulescens can be efficiently cryopreserved using suitable cryoprotectants for spermatogonia and oogonia, respectively.     

Bioremediation of organically enriched sediment deposited below fish farms with artificially mass-cultured colonies of a deposit-feeding polychaete <Emphasis Type="Italic">Capitella</Emphasis> sp. I

ABSTRACT:   For bioremediation of organically enriched sediment deposited below fish farms, the extremely high potential for population growth of a deposit-feeding polychaete, Capitella sp. I, in the organically enriched sediment, and the effect on decomposition of organic matter in the sediment, were examined. A mass-culturing technique was conducted for this species. Bioremediation experiments were conducted on the organically enriched sediment in a fish farm in Kusuura Bay, Japan in 2003–2006. Approximately 1.7 million individuals of the worms were placed on the sediment below one net pen in December 2003, 9.3 million individuals in November 2004, and 2.2 million individuals in November 2005. After the worms were spread on the sediment, they rapidly increased in number and reached the highest densities of approximately 134 000 inds/m² in February 2004, 527 000 inds/m² in March 2005 and 103 000 inds/m² in January 2006. In the process of rapid population growth, the decomposition of the organic matter of the sediment was enhanced markedly. Our results demonstrate that the promotion of population growth by spreading cultured colonies of Capitella can enhance the decomposition rate of organic matter markedly in organically enriched sediment below fish farms. This method is promising for minimization of the negative effects of fish farms.     

Maturity and reproduction of goneplacid crab <Emphasis Type="Italic">Carcinoplax vestita</Emphasis> (Decapoda,Brachyura) in Tokyo Bay

ABSTRACT:     Sexual maturity, morphological sexual dimorphism, and reproduction of the goneplacid crab Carcinoplax vestita were investigated in Tokyo Bay, Japan, from November 2002 to October 2003. The puberty molt in males was evidenced by changes in the relative size of the chelipeds and merus of the walking legs, and was estimated to occur at a size range of 13.20–18.85 mm carapace length. Post-pubertal females were identified by the relative size of the abdomen and puberty was estimated to occur at a size range of 12.81–15.46 mm carapace length. Sexual dimorphism in C. vestita was observed in all features that showed secondary sexual characteristics. Monthly changes of gonad index in males, and of seminal receptacle index and occurrence of sperm plugs in females were synchronized, and indicated that mating was intense in spring. Ovaries began developing in March. Ovigerous females were found in all months except December, but were clearly more abundant between August and October. Fecundity ranged from 7800 to 57 000 mature oocytes per female per batch and was highly correlated with body size. The results suggest that some females may spawn more than one batch per year.     

Aspects of the reproductive biology of the sharpsnout seabream Diplodus puntazzo (Cetti, 1777). I. Gametogenesis and gonadal cycle in captivity during the third year of life

A histological study was made of the reproductive cycle of captive sharpsnout seabream Diplodus puntazzo during the 3rd year of life. All specimens contained both ovarian and testicular tissue. Ovarian tissue prevailed in 64 out of 120 fish sampled throughout one year. The ovary underwent a long resting phase from October to August, during which time it contained oogonia, primary growth phase (PGP) and occasional yolk vesicle stage oocytes. Intense ovarian development took place in September leading to the formation of advanced vitellogenic oocytes. Final maturation and spawning were not achieved in any of the fish. The testicular area in the predominantly female gonads contained only spermatogonia and was found to be histologically normal in 76% of individuals, but severely regressed in the remaining fish. The testicular portion was found to be dominant in 56 out of 120 fish sampled throughout one year. These underwent a resting phase from November to July, a maturing/pre-spawning phase in August, and a spawning phase with spontaneous sperm release in September to October, as in the natural environment. The ovarian portion in the functional male gonads contained only oogonia and oocytes which did not develop beyond the perinucleolus stage at any point during the reproductive cycle. Apart from the pre-spawning/spawning phase, during which the gonadosomatic index (GSI) was comparable in the two groups, the GSI values were approximately ten-fold higher in those fish in which the ovary predominated than in those in which the testis predominated.     

An insulin-like system involved in the control of Pacific oyster Crassostrea gigas reproduction: hrIGF-1 effect on germinal cell proliferation and maturation associated with expression of an homologous insulin receptor-related receptor

The putative involvement of insulin-like peptides in the control of the reproduction of the Pacific oyster Crassostrea gigas was investigated using different approaches. In conjunction with a monthly histological analysis of the oyster reproductive cycle, in vitro biological effects of the human recombinant IGF-1 (hrIGF-1) on dissociated germinal cells were mesured over 1 year using [³H]-thymidine and [¹⁴C]-amino acid mixture as tracers for DNA and protein synthesis. DNA synthesis was stimulated by hrIGF-1 in November (114 ± 11% for 10^− 7M), December (46 ± 6% for 10^− 7 M) and January, which was identified as the highest gonial mitosis period. A clear dose-effect was observed in January with a maximum activation of 68 ± 7% for 10^− 12 M. Germinal cell protein synthesis was also stimulated in March (20 ± 1% for 10^− 10 M), April (41 ± 5% for 10^− 13 M), May (25 ± 4% for 10^− 13 M), and by almost all of hrIGF-1 doses in June (21.5 ± 2% for 10^− 13 M) and July (34 ± 1% for 10^− 13 M). This suggests the involvement of insulin-like substances in gonadal tubule rebuilding (December), as well as in the development of germinal cells (March, April), and in the summer maturation of gametes (May, June, July). These insulin-like effects conform with the expression pattern of the recently identified C. gigas insulin receptor-related receptor (CIR): It appeared highly expressed in the gonadal area during gonial mitosis phase, but also in maturating oocytes, suggesting the involvement of an insulin-like system in gonial proliferation and maturation. Moreover, CIR showed differential expression during embryogenesis and larval developmental stages. The expression of maternal CIR during the embryonic and early larval development, followed by the increasing zygotic CIR expression from D larvae to 11-day-old veliger larvae, then a decrease until metamorphosis, also suggest that insulin-like peptide is involved in organogenesis.     

Testicular development and serum profiles of steroid hormone levels in captive male Pacific herring Clupea pallasii during their first maturational cycle

The present study investigates the relationship between testicular development and serum steroid hormone levels in captive Pacific herring Clupea pallasii during the first reproductive cycle. The maturity of the testis was divided into five periods based on histological observation. These are early spermatogenic stage (April to July), mid-spermatogenic stage (August to November), late spermatogenic stage (December to March), functional maturation stage (early April) and spent stage (late April). The pattern of seasonal change in gonadosomatic index (GSI) clearly reflected testicular maturity. 11-Ketotestosterone (11-KT) levels increased from October to a peak level (6.58 ± 1.87 ng/mL) in January, and were maintained at this level until March. In contrast, testosterone levels were consistently low, less than 1 ng/mL, at all times. These results suggest that 11-KT is the predominant androgen that controls spermatogenesis in this species. 17,20β-Dihydroxy-4-pregnen-3-one (DHP) showed a single sharp peak (3.38 ± 0.35 ng/mL) in early April of the second year, suggesting that milt production is induced by DHP as in some other teleost species.     

Scp3 expression in relation to the ovarian differentiation in the protogynous hermaphroditic ricefield eel <Emphasis Type="Italic">Monopterus albus</Emphasis>

Synaptonemal complex protein 3 (Scp3), which is encoded by scp3, is a meiotic marker commonly used to trace the timing of gonadal differentiation in vertebrates. In the present study, the ricefield eel scp3 cDNA was cloned, and a fragment encoding amino acids 49 to 244 was overexpressed. The recombinant Scp3 polypeptide was purified and used to generate a rabbit anti-Scp3 polyclonal antiserum. In adult ricefield eels, scp3 mRNA was predominantly detected in the gonads and faintly detected in discrete brain areas. In the gonads, Scp3 immunoreactivities were shown to be localized to the germ cells, including meiotic primary growth oocytes, spermatocytes, and pre-meiotic spermatogonia. During early ovarian differentiation, immunoreactive Scp3 was not detected in the gonads of ricefield eels at 6 days post-hatching (dph) but was found to be abundantly localized in the cytoplasm of some oogonia at 7 dph, coinciding with the appearance of the ovarian cavity and ovarian differentiation. At 14 dph, strong Scp3 immunostaining was detected on one side of the nucleus with a distinct polarity in some germ cells, presumably at the leptotene stage. Consistent with these results, the expression of scp3 mRNA was faintly detected in the gonads of ricefield eels at 6 dph, increased at 8 dph, and then remained relatively high thereafter. Taken together, these results suggest that the appearance of immunoreactive Scp3 in oogonia could be a marker for early ovarian differentiation in ricefield eels. The translocation of the Scp3 protein from the cytoplasm to the nucleus in the oogonium of ricefield eels appears to be a controlled process that warrants further study.     

Endocrine and gonadal changes during the annual reproductive cycle of the freshwater teleost,Stizostedion vitreum

The annual reproductive cycle of walleye (Stizostedion vitreum) was characterized by documenting changes in gonadal development and serum levels of estradiol-17β (E₂), testosterone (T), 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P), and 11-ketotestosterone (11-KT) in wild fish captured from upper midwestern lakes and rivers throughout the year. Fish from the populations used in this study spawn annually in early- to mid-April. Walleye showed group synchronous ovarian development with exogenous vitellogenesis beginning in autumn. Oocyte diameters increased rapidly from ∼ 200 μm in October to ∼ 1,000 μm in November, and reached a maximum of 1,500 μm just prior to spawning. Changes in gonadosomatic indices (GSIs) paralleled changes in oocyte diameters. Serum E₂ levels in females increased rapidly from low values in October (< 0.1 ng ml⁻¹) to peak levels of 3.7 ng ml⁻¹ in November, coinciding with the period of the most rapid ovarian growth. Subsequently, E₂ levels decreased from December through spawning. Serum T levels exhibited a bimodal pattern, increasing to 1.6 ng ml⁻¹ in November, and peaking again at 3.3 ng ml⁻¹ just prior to spawning. We detected 11-KT in the serum of some females at concentrations up to 5.6 ng ml⁻¹, but no seasonal pattern was apparent. In this study (unlike our results in a related study) 17,20-P was not detected. In males, differentiation of spermatogonia began in late August, and by January the testes were filled (> 95% of germ cells) with spermatozoa. Mature spermatozoa could be expressed from males from January through April. GSIs ranged from 0.2% (post-spawn) to 3.2% (pre-spawn). Serum T levels rose from undetectable levels in post-spawn males to 1.6 ng ml⁻¹ by November, remained elevated throughout the winter, and peaked at 2.8 ng ml⁻¹ I prior to spawning. Levels of 11-KT in males remained low (< 10 ng ml⁻¹, from post-spawning through January, then increased significantly by March and peaked just prior to spawning at 39.7 ng ml⁻¹. Our results indicate that vitellogenesis and spermatogenesis are complete or nearly so, in walleye by early winter, and suggest that it may be possible to induce spawning in this species several months prior to the normal spawning season by subjecting fish to relatively simple environmental and hormonal treatments.     

Ovarian development and induced oviposition of the overwintering swimming crab Portunus trituberculatus (Brachyura: Portunidae) reared in the laboratory

ABSTRACT:   Females of the swimming crab Portunus trituberculatus were cultured under natural temperature and high temperature (21°C) conditions to examine ovarian development and oviposition from autumn (October) to the beginning of breeding season in the following spring (May). Ovaries developed because of vitellogenesis of oocytes from October to December and their developmental state did not change during the overwintering period from December to March. In spring, from late March to mid May, some ovaries reached prematuration and maturation stages and ovipositions began from mid–late April under natural temperature conditions. Females reared in tanks of high temperature regimes oviposited earlier than those reared in tanks of natural temperatures. The number of days to oviposition tended to decrease with advancement of the beginning times (early February to late April) of culture of females under high temperature regimes; induced ovipositions were achieved more easily with increasing photoperiod from c . 12 h in early February to c . 14 h in late April. Therefore, it is inferred that the temperature and photoperiod are important environmental factors controlling ovarian development and oviposition.     

Endocrine growth regulation of adult Atlantic salmon in seawater: The effects of light regime on plasma growth hormone, insulin-like growth factor-I, and insulin levels

Plasma growth hormone (GH), insulin-like growth factor-I (IGF-I), and insulin were measured in two groups of Salmo salar L. during a one-year study. The fish were reared under either a simulated natural photoperiod (SNP) from January to December or a regime of continuous light from January to June, followed by SNP until December (LL/SNP). Plasma GH levels during spring were low, and lower in the LL/SNP fish (< 0.9 ng ml^− 1) than in the SNP fish (> 1.9 ng ml^− 1), although the LL/SNP grew better (0.8% per day) than the SNP fish (0.5% per day). Plasma IGF-I levels increased transiently from January (64.7 ng ml^− 1) to maximum in late September in the LL/SNP (85.8 ng ml^− 1) and in November in the SNP group (87.3 ng ml^− 1). The ratio GH:IGF-I was lower in the LL/SNP group during spring when this group grew better than the SNP group.     

Classification of differentiating oocytes during ovarian cycle in the giant freshwater prawn, Macrobrachium rosenbergii de man

Based on the light microscopic observations of cells' sizes, chromatin patterns, amount of lipid droplets and yolk granules, the female germ cells could be classified into four different phases, which include 1) oogonia (Oog), 2) primary oocytes (pOc), 3) secondary oocytes (sOc), and 4) mature oocyte (mOc). Oog are small oval-shaped cells with irregular-shaped nuclei sizing 4–6 μm in diameter. They rest on the connective tissue germinal cord at the tip of each ovarian pouch (lobule). Oogonia increase their number through mitotic division, and the daughter cells move into ovarian pouch where they undergo first meiotic division to become primary oocytes, which have various steps of 1st meiotic prophase accumulating at the innermost zone of the ovarian pouch. The primary oocytes are small oval-shaped cells (8.5–10 μm in diameter) with large nuclei containing chromatin in various states of condensation that finally transform into chromatids. Their nuclei are surrounded by thin rim of faint blue-stained cytoplasm. The secondary oocytes derived from 2nd meiosis and comprise five steps: Oc1 and Oc2, classified as previtellogenic oocytes, Oc3 and Oc4, classified as vitellogenic oocytes, and mature oocyte (mOc) The zones of ovarian pouch are defined based on the accumulation of various steps of developing oocytes, namely, oogenic, previtellogenic, vitellogenic and mature zones, respectively. The ovarian cycle is divided into five stages based on the number and types of oocytes present in each stage. Stage 0 and I are spawn and spent stages. Stage II and III are proliferative and premature stages, while stage IV is mature stage. During ovarian stage I, each ovarian pouch contains primarily oogonia, primary oocytes, Oc1 and a few Oc2. In stage II, the pouch contains mainly Oc2 and Oc3, while in stage III the predominant cells are Oc4. Mature oocytes appear synchronously, in stage IV. The ovulating mature oocytes pass through the thin disrupted wall of ovarian pouch into subcapsular space, that leads into the oviduct situated on the ventro-lateral side of the ovarian lobe. At spawning, the ovarian pouches break down and only connective sheaths and hemolymph sinuses remain. The germinal cords and islets of oogonia remain in the central area of stage 0 ovary. The ovarian capsule, including the muscular layer, becomes attenuated as the ovary progresses from stage 0 to IV. The hemolymph vessels become highly convoluted in the central area of the ovary, and they branch radially into smaller hemolymph sinuses around each oogenic pouch.     

Reproductive biology of <Emphasis Type="Italic">Sillago aeolus</Emphasis> in Okinawa Island,Japan

ABSTRACT:   A total of 414 fish (female, 59.3–275.0 mm and male, 61.0–220.0 mm standard length) were collected from December 2000 to March 2002 around Okinawa Island, Japan, for the determination of sex, spawning season, maturity and fecundity. Monthly changes in the gonadosomatic index (GSI) exhibited similar trends for both sexes and the mean GSI maintained a high level between February and May. Furthermore, the percentage frequency of hydrated oocytes and the spermatozoic activity suggested that main spawning occurred between February and May. Some hydrated oocytes were found throughout the year, with a GSI value greater than 1.0, suggesting that sporadic spawning occurred. Length at first maturity was determined for females and males as 120 and 113 mm standard length, respectively. Almost 50% and 100% males were matured at the end of their 0 and 1 year of age group, respectively. In contrast, no females were found to be mature at 0+ age group, and almost 60% of females were mature at 1+ age group. At ages over 2 years, all males and females were found to be mature. Batch fecundity (BF) of 33 females was related to standard length, and the relationship between standard length and BF was expressed by the exponential equation: BF = 269.5e^0.020954SL.     

Autumn conditioning of the oyster Crassostrea gigas: a new approach

Gametogenesis, emitted gametes and larval yield (D larvae) of the oyster, Crassostrea gigas (Thunberg), were simultaneously examined in laboratory under two different conditions of temperature and photoperiod. One batch (“natural” cycle, NC) followed the equivalent cycle of average conditions of these parameters measured in La Tremblade (Charente-Maritime, France) during the last 15 years. The second batch (“accelerated” cycle, AC) was maintained under a variation two times faster of these parameters. Three conditioning experiences were performed during October, November and December 1998 with oysters from both NC and AC conditions. “Accelerated individuals” produced oocytes in growing stage from October and mature oocytes were noticed from December. In contrast, oysters from NC produced growing oocytes until January. Animals in AC produced 42% (October), 56% (November) and 96% (December) of mature oocytes after conditioning. Only the “natural” lot conditioned in December showed growing (23%) and mature oocytes (8%). Higher Walne–Mann index (WMI) values were recorded for “accelerated oysters” and significant differences (P<0.05) between treatments were noticed during October and November, suggesting nutrient accumulation before conditioning in the “accelerated” condition. Gamete emission and D larval yield results were equivalent to those reported in literature during early spring for the same species. The effect of temperature and photoperiod is discussed in order to understand their relationship with gametogenesis under these particular experimental conditions. It was demonstrated that internal clocks regulating gametogenesis can be change if stimulating factors (environmental cues) also change.     

Identification of overwintering vegetative cells of the bivalve-killing dinoflagellate <Emphasis Type="Italic">Heterocapsa circularisquama</Emphasis> in Uranouchi Inlet,Kochi Prefecture,Japan

ABSTRACT:   Red tides of Heterocapsa circularisquama have led to serious damage of bivalve aquacultures in western coastal areas of Japan. To understand the whole picture regarding the ecology of this species, it is essential to clarify its overwintering mechanisms. In this study, the population dynamics of H. circularisquama were investigated from February 2004 to November 2005, and overwintering cells were identified for the first time in water columns of Uranouchi Inlet, Kochi Prefecture, Japan. Heterocapsa circularisquama cells were detected by the indirect fluorescent antibody technique using monoclonal antibodies that specifically recognize and react to this species. Vegetative cells were almost always detected from the first observation in February 2004 to November 2005 with temperatures of 10.5–30.6°C. During the period from winter to spring, this species survived in areas with a temperature higher than 10°C. The overwintering cells of H. circularisquama were isolated in March 2004, and identification was made via observation of the morphology and body scales of the cultured cells. These overwintering cells were identified as H. circularisquama and reacted to the monoclonal antibody. These results indicate that H. circularisquama can overwinter and survive throughout the year in a vegetative cell state in Uranouchi Inlet.     

Interspecific germ cell transplantation: a new light in the conservation of valuable Balkan trout genetic resources?

Interspecific transplantation of germ cells from the brown trout Salmo trutta m. fario and the European grayling Thymallus thymallus into rainbow trout Oncorhynchus mykiss recipients was carried out in order to improve current practices in conservation of genetic resources of endangered salmonid species in the Balkan Peninsula. Current conservation methods mainly include in situ efforts such as the maintenance of purebred individuals in isolated streams and restocking with purebred fingerlings; however, additional ex situ strategies such as surrogate production are needed. Steps required for transplantation such as isolation of high number of viable germ cells and fluorescent labeling of germ cells which are to be transplanted have been optimized. Isolated and labeled brown trout and grayling germ cells were intraperitoneally transplanted into 3 to 5 days post hatch rainbow trout larvae. Survival of the injected larvae was comparable to the controls. Sixty days after transplantation, fluorescently labeled donor cells were detected within the recipient gonads indicating successful incorporation of germ cells (brown trout spermatogonia and oogonia—27%; grayling spermatogonia—28%; grayling oogonia—23%). PCR amplification of donor mtDNA CR fragments within the recipient gonads additionally corroborated the success of incorporation. Overall, the transplantation method demonstrated in this study presents the first step and a possible onset of the application of the germ cell transplantation technology in conservation and revitalization of genetic resources of endangered and endemic species or populations of salmonid fish and thus give rise to new or improved management strategies for such species.     

The reproductive biology of the baby clam, Marcia opima, from two geographically separated areas of India

A study of the reproductive cycle of the baby clam, Marcia opima (Gmelin) was conducted at two sites along the southeast (Tuticorin Bay) and southwest (Ashtamudi estuary) coast of India from December 1998 to January 2000. Histological evidence showed that baby clam from both the sites showed two spawning seasons. First spawning season was recorded from May to July in the southeast coast. In the southwest coast, the first spawning season was observed through March to May. A second spawning season from September to December was recorded in both the coasts. In the southeast coast, minimum and maximum condition indices were obtained in January 2000 and April 1999 respectively and decreased from May to June and November to January, signifying the two spawning periods. Most of the animals were either in spent or indeterminate gonadal stage from May to August and November to January indicating the active spawning during that months in the southeast coast.     

Seasonal distribution, age, growth, and reproductive biology of marbled sole Pleuronectes yokohamae in Tokyo Bay, Japan

ABSTRACT:   The seasonal distribution, age, growth and reproductive biology of marbled sole Pleuronectes yokohamae (Günther) from Tokyo Bay, Japan were studied. A clear seasonal distribution trend was observed, with P. yokohamae limited to the south of the bay in summer and expanding almost throughout the bay in other seasons. The formation and dynamics of hypoxic areas in the bay limited the northward distribution of P. yokohamae in summer. Age was determined by counting growth rings on otoliths; maximum ages were found to be 5 years for males and 6 years for females. The mean back-calculated length for females at each estimated age was greater than that for males. Growth of males and females was shown by the von Bertalanffy growth equations. Reproductive biology was studied on the basis of gonadosomatic indices and histological sections of gonads. The spawning season lasted from November to March, peaking in December and January. Sixty per cent of males at 1 year and all males ≥ 2 years had mature testes, whereas 6.7% of females at 1 year, 58.3% of females at 2 years, and all females ≥ 3 years had attained maturity.     

Life cycle of <Emphasis Type="Italic">Dilophus okamurae</Emphasis> (Phaeophyceae) and its associated invertebrate fauna in Onagawa Bay,Japan

ABSTRACT:   The life cycle of Dilophus okamurae and the associated benthic invertebrate fauna were studied monthly or bimonthly from June 1998 to September 1999 at a depth of 4–7 m in Onagawa Bay along the Pacific coast of north-eastern Honshu, Japan (38°28'N, 149°29'E). Dry biomass ranged from 230 g/m² in June to 10 g/m² in October. Sporophytes with tetrasporangia appeared during July to September 1998. Gametophytes with oogonia or antheridia were observed for the first time in May 1999 during observation. The absence of mature sporophytes in 1999 suggests that the life cycle is completed over two years. The sympatric gastropods Lirularia iridescens , Homalopoma sangrarense, Conotalopia minima and Cantharidus callichroa synchronized with seasonal changes in biomass of this algae. In contrast, Barleeia angustata and B. trifasciata fluctuated independently. The juvenile sea urchin Strongylocentrotus nudus appeared in the algal population starting in October at 0.7 mm mean test diameter at high density, and disappeared after March.     

Growth of the fourspine sculpin <Emphasis Type="Italic">Cottus kazika</Emphasis> in the Gonokawa River,Japan, and effects of water temperature on growth

ABSTRACT:   The fourspine sculpin Cottus kazika is indigenous to Japan and found in Honshu except for the waters facing the Seto Inland Sea, and was also found in southern Shikoku and eastern Kyushu. This species has a catadromous lifestyle and migrates as juveniles from the sea to the middle reaches of rivers to grow. The growth pattern of this fish was investigated by a mark-and-recapture method from July 1994 to December 1996, in the Nigorikawa River, a tributary of the Gonokawa River system, Shimane Prefecture. 0-year-old fish of 50–70 mm total length ( TL ) occurred in the study area from June to July, grew to 90–140 mm  TL by the following April, and attained 160–210 mm  TL by December. This fish grew rapidly in September–November and April–July, almost ceasing to grow in July–September. It seems that this stagnant growth phase in summer is a characteristic of the seasonal growth pattern of C. kazika . A rearing experiment indicated that the growth rate of C. kazika was higher at 16–22°C than at 12–14 and 24–26°C. This result supports the field evidence of a stagnant growth phase in summer in the Nigorikawa River.     

Horizontal movement and emigration of juvenile spotted halibut <Emphasis Type="Italic">Verasper variegatus</Emphasis> released in a shallow brackish lagoon: Matsukawa-ura,northeastern Japan,revealed by acoustic telemetry

To elucidate the post-release movement and emigration of juvenile spotted halibut Verasper variegatus, ten hatchery-reared age-0 individuals (10 months old, 17.0–19.7 cm total length) surgically implanted with acoustic transmitters were released near the center of Matsukawa-ura, a shallow brackish lagoon, on 25 November 2009 (water temperature 13.8 °C). They were monitored by ten acoustic receivers for 5 months. Of ten individuals, eight left the release site during December–March. Three of these emigrated to the outer ocean on 17 and 29 January and 30 March 2010, respectively. Juveniles probably started wintering migration in December and January when water temperatures decreased considerably in the lagoon (mean 9.5 and 6.0 °C, respectively). They stopped their migration in the coldest month, February (mean 5.7 °C), and restarted it in March when water temperatures frequently exceeded a plausible threshold for movement (≥6 °C). Statistical analyses revealed that the fish started migration significantly more frequently at nighttime. The migration track of an individual recorded from 11 to 30 March showed gradual nocturnal movements and a slow migration speed (estimated maximum speed 2.2 m/min). Our results revealed that water temperature primarily governed the seasonal timing of nocturnal migratory movements of juvenile spotted halibut.