Pleistophora mirandellae Vaney & Conte, 1901, a microsporidian parasite in cyprinid fish of rivers in Herault: taxonomy and histopathology

Abstract. Cyprinid fish from the Lergue and Lez rivers in Herault, southern France, were found to be parasitized by the microsporidian Pleistophora mirandellae Vaney & Conte, 1901. The parasite was found only in the gonads, forming cysts in the connective tissue between the seminiferous tubules of testes or occurring in the oocytes of the ovaries. Two types of uninuclear spores are produced. The synonomy of microsporidia parasitizing the gonads of cyprinid fish is discussed.     

Gonadal histology and some biochemical characteristics of <Emphasis Type="Italic">Chalcalburnus tarichi</Emphasis> (Pallas, 1811) having abnormal gonads

The gonad histology, gonado-somatic index (GSI), 17β-estradiol (E₂) levels and acetylcholinesterase (AChE) activity in the carp species Chalcalburnus tarichi from Lake Van and the Karasu river, eastern Turkey, have been investigated. Fish between 5 and 7 years old were sampled from November 2003 to February 2004. The ratio of female fish caught in Lake Van with abnormal ovaries (AbOF) was 43.3%, but the fork length and body weight of these fish were not correlated with this abnormality. The weight of the ovaries and the GSI values of AbOF were very low (P < 0.05). Histological observations on the samples caught each month revealed that the oocytes had degenerated in the perinucleolus and early cortical alveolus stages and that the ovaries were full of somatic stromal tissue. In addition, the seminiferous tubules of male fish with abnormal testes did not contain male reproductive cells at any stage. The ovaries of the fish caught from the Karasu river were also full of oocytes in the perinucleolus and early cortical alveolus stages, but there were fewer atretic follicles. Furthermore, apoptosis was observed in the ovary cells of these fish, in particular in the follicular cells, and the plasma E₂ levels of the AbOF was very low (P < 0.05). AChE activity was inhibited only in liver (P < 0.05). We conclude that our sample of C. tarichi must have been exposed to various polluting chemicals or another unknown factors (such as global warming) and that these factors have irreversibly impaired oocyte development in a high percentage of fish.     

Histological observations of seasonal reproductive and lunar-related spawning cycles in the female honeycomb grouper Epinephelus merra in Okinawan waters

Seasonal reproductive and lunar-related spawning cycles of the female honeycomb grouper Epinephelus merra inhabiting Okinawan waters were examined by histological observation of ovaries. Gonadosomatic index (GSI) increased beginning in May and peaked in June. Histological observations revealed that many oocytes laden with yolk were present in the ovaries from May to August. From September to October, ovaries were occupied by immature oocytes. These results suggest that the reproductive season of E.   merra lasts for 4 months from May through August in Okinawan waters. When the fish were collected according to the lunar cycle, GSI increased with the approach of the full moon. Oocytes at various development stages were observed from the first quarter to the full moon. Fresh ovulatory follicles were present in the ovaries around the last quarter moon. These results suggest that E.   merra has a lunar spawning cycle and spawns between the full moon and the last quarter moon. Ovarian features of the fish collected around the last quarter moon were different among individuals; some fish had many oocytes at the tertiary yolk stage in the ovaries, while the ovaries of the others were occupied by the oocytes at the peri-nucleolus and the oil droplet stages. This observation suggests that a minor release of eggs occurs in this species before or after a major spawning lunar day. Oocytes at the migratory nucleus and the maturation stages were not observed in any ovarian samples. This may mean that maturation of oocytes is related to the spawning behavior of this species and makes rapid progress in the process of aggregation at the spawning sites.     

Infestation parameters for Lepeophtheirus salmonis (Kroyer) (Copepoda: Caligidae) parasitic on sea trout, Salmo trutta L., off the west coast of Ireland during 1990 and 1991

Abstract. Infestation parameters for the calígíd copepod Lepeophtheirus salmonis (Kroyer) infesting sea trout, Salmo trutta L., were established for a number of locations off the west coast of Ireland during 1990 and 1991. Based on these parameters sites were classified into two groups in 1990 and three in 1991. Median parasitic intensity in these groups was 11·6 and 77 in 1990 and 9·5, 29·5 and 55 in 1991. Fish were parasitized mainly by chalimus stages of the parasite which attached preferentially to the fins. Heaviest infestations were invariably due to chalimus stages and early pre-adult lice and there was a progressive decrease in the correlation between parasitic intensity and the number of each successive stage in the life cycle. This suggested either host or parasite mortality as the parasite matured. Large numbers of lice-infested fish returned prematurely from the sea to estuarine areas at a number of sites. Extensive morphological damage, especially to the fins, was apparent on a proportion of these fish as a result of the infestation. Mortality of heavily infested fish was directly observed.     

Diurnal rhythm of serum steroid hormone levels in the Japanese whiting,Sillago japonica,a daily-spawning teleost

Ovarian developmental stages and serum steroid hormone levels were examined at six different times of day (0100, 0600, 1000, 1300, 1600, 2000 h) in a marine teleost, the Japanese whiting Sillago japonica, which has an asynchronous-type ovary containing oocytes at various stages of development and spawns every day during a period ranging up to three months. The largest oocytes in the ovaries at the active vitellogenic or post-vitellogenic stages were found between 0100 and 1300 h. Oocyte maturation indicated by germinal vesicle breakdown (GVBD) occurred at 1600 h, and ovulated oocytes were observed in the ovaries collected at 2000 h. These processes were accompanied by a significant daily change in serum steroid hormone levels. The serum level of estradiol-17β showed a peak in fish with mature oocytes sampled at 1600 h. In these fish, the second-largest oocytes in the ovaries were at the initial stage of vigorous vitellogenesis, the secondary yolk stage. Therefore the highest level of serum estradiol-17β was considered to be due to the second-largest oocytes. Testosterone levels remained low and constant throughout the experimental period. The serum levels of 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-diOHprog) peaked at 1600 h at which time all fish had mature oocytes. These results indicate that the Japanese whiting possesses a diurnal rhythm of oocyte development including vitellogenesis, oocyte maturation and ovulation, and further suggest that daily cycles in oocyte growth and maturation which simultaneously take place in an ovary are regulated by diurnal secretions of estradiol-17β and the maturation-inducing steroid, 17α,20β-diOHprog.     

One cryptic species of grey mullet (Mugil cephalus mitotype: NWP3) from Taiwan’s waters is worth cultivating for large roes using aquaculture

The grey mullet, Mugil cephalus, is an important aquaculture species in Taiwan. They were thought to be a single fish species in terms of the morphology‐based taxonomy; however, three cryptic species of grey mullet (mitotype: NWP1, NWP2 and NWP3) have been identified within the Taiwan waters. In general, the NWP2 females are known to have small body sizes with large developed ovaries regardless of the cultivation time, whereas the NWP1 females have large body sizes with small ovaries. In this study, the NWP3 females are found to have significantly larger body and ovary weights than do the NWP2 females under the same aquaculture conditions. Therefore, even though the NWP2 females are good fish for producing large ovaries, this study indicates that the NWP3 females have even more potential to produce huge ovaries than do the NWP2 females.     

Distribution and structure of the population of sea lice, Lepeophtheirus salmonis Krøyer, on Atlantic salmon, Salmo salar L., under typical rearing conditions

Abstract. Samples of Atlantic salmon, Salmo salar L., held in sea cages and tanks with running water were examined for presence of sea lice, Lepeophtheirus salmonis. The number of lice, their sex stage (only post-chalimus stages) and location on the fish body were recorded. A new method has been used to express parasitic intensity in fish of different size. Effects of different factors on the degree and character of infestation (structure of the lice population), such as fish size, habitat and chemical treatment, have been studied. Different lice categories have shown different susceptibility to delousing with dichlorvos. Second pre-adults are most susceptible, and males generally more than females. Running water has been found to'flush out'the lice from the fish body. Pre-adults are flushed out easier than adults, and males generally easier than females. Adult males show high tolerance to water of lowered salinity and great mobility manifested in occasional transfer between hosts. The higher activity level of adult males may partly explain a drop in proportion of males after delousing and in running water. Frequency of mating has been found to increase with parasitic intensity.     

Out-of-season spawning of cultured pikeperch [Sander lucioperca (L.)]

The aim of this study was to determine the effectiveness of out‐of‐season spawning of cultivated pikeperch (fish that were reared from the larval stage in re‐circulating systems and fed commercial feed exclusively) stimulated hormonally with human chorionic gonadotropin (hCG). The impact of fish age (2+ and 3+) and hormone dosage [200 or 400 IU hCG kg⁻¹ body weight (BW)] on spawning was analysed and expressed as the share of stripped females, commercial fecundity (% BW) and survival of embryos until the eyed‐egg stage (EES index). The possibility of utilizing changes in female pikeperch body weight (CBW index) that are observed following hormone injections as an additional indicator for determining maturity was also investigated. The age and hormone dosage were not noted to have a significant impact on the number of stripped females (≥80% on all groups), the latency period (90–100 h), commercial fecundity (11.3–13.3% BW) or the values of the EES index (61–73%; P>0.05). The mean value of EES from the 3+ age group females was higher than that in the 2+ females, and the interaction between the tested factors (fish age and hormone dosage) wasstatistically significant (P<0.05). In the fish from the control group (injected with a 0.9% NaCl solution), no progress was noted in the maturation of oocytes and no eggs were obtained from any female. It was noted that these females lost BW over the course of the subsequent 24 h of the measurements (P<0.05). In the groups of females that were stimulated hormonally, the opposite phenomenon was observed; in these groups, the CBW index increased significantly between 48 and 96 h following hormone injection. The value of the CBW index was not noted to have been statistically significantly determined by either hormone dose or fish age (P>0.05). The regression equations that described the dependence between CBW and the oocyte maturity stage were highly significant statistically, and the determination coefficient R² assumed a value of 0.76. The most significant increase in BW was related to the oocytes achieving maturity stage III. The BW of pikeperch females with oocytes in this stage was 103% higher than the initial BW. This might be a valuable and useful tool for determining maturity in females of this fish species.     

Modulation of the response of rainbow trout (Oncorhynchus mykiss Walbaum) to confinement, by an ectoparasitic (Argulus foliaceus L.) infestation and cortisol feeding

The effects of a 21 day infestation with the ectoparasite Argulus foliaceus on the response of rainbow trout Oncorhynchus mykiss to an acute stressor (confinement) were investigated. Fish were divided into a control and two parasitized groups, with one parasitized group having received cortisol feeding 72 and 24 h before infection. Prior to confinement only control fish showed a significant increase in body mass and condition factor. Following 4 h confinement, plasma glucose levels in the normal- fed/infested group were significantly higher than in the controls, while the cortisol-fed/infested group had significantly lower plasma levels of adrenocorticotropic hormone (ACTH) and glucose. Both infested groups, but not controls, had higher plasma ACTH and cortisol levels 48 h after confinement when compared with their pre-confinement levels. Stressor-induced immunomodulation was more evident in the parasitized fish after confinement, as was concluded from decreased lysozyme activity, alternative complement activity and oxygen radical production either immediately after, or 48 h post 4 h confinement. Circulating lymphocyte numbers were significantly reduced in the control and normal-fed/infested fish. Results indicate that low ectoparasitic infestations can, for a prolonged period, lead to a stress experience with effects that impair the response to subsequent stressors. Such effects may be further confounded by a previous elevation of plasma cortisol levels.     

Long-afterglow phosphorescent pigment is a potent tool for manipulation of reproductive performance in the tropical damselfish <Emphasis Type="Italic">Chrysiptera cyanea</Emphasis>

Photoperiod plays a role in the regulation of reproduction in many fish. We examined the effectiveness of the long-afterglow phosphorescent pigment LumiNova on the reproductive performance of the sapphire devil Chrysiptera cyanea, a tropical damselfish that reproduces under long-day conditions. During the breeding season, the experimental fish, but not the control fish, were reared in aquaria covered with LumiNova sheets for 75 days under natural photoperiod and temperature. At days 60 and 75 after onset of the experiment, the gonadosomatic index of experimental fish was higher than that of control fish. Histological observations at 60 and 75 days revealed that the ovaries of experimental fish contained large oocytes laden with yolk, whereas the control fish had few yolky oocytes at 60 days and mostly immature oocytes at 75 days. Spawning occurred repeatedly in experimental fish, but not in control fish, throughout the study period. Because LumiNova sheets continue to emit green light for some time after sunset, the extended light conditions probably contributed to active reproduction in experimental fish. In conclusion, long-afterglow phosphorescent pigment can be used for energy-efficient aquaculture to regulate the reproduction of fish, although its effect needs to be evaluated in different species.     

The physiological response of Atlantic salmon, Salmo salar L., to a single experimental challenge with sea lice, Lepeophtheirus salmonis

The sea louse, Lepeophtheirus salmonis , is an ectoparasitic copepod of Atlantic salmon, Salmo salar L., capable of causing severe damage. This study was conducted to examine the physiological response of salmon to the stress of sea lice infestation. Smoltified salmon were acclimatized in 30‰ saltwater and exposed to high levels of lice infestation. The number of copepods per fish ranged from 15 to 285, with a mean of 106. The infested salmon were sampled six times over the 29-d experimental duration and examined for alterations in the primary and secondary stress indicators, including plasma concentrations of cortisol, glucose, electrolytes, thyroid hormones T3 and T4, as well as the haematocrit level. The results were examined for correlations between the stress indicators, the number of copepods per fish and the life stage of the copepods. The presence of L. salmonis elevated stress indicators in relation to the specific sea lice stage. By day 21, both cortisol (mean 63.1 nmol L−1 controls: 179.8 nmol L−1 for parasitized) and glucose (mean 3.545 mmol L−1 controls: 4.567 mmol L−1 for parasitized) levels were significantly increased due to the presence of the lice. This was believed to be a direct result of the sea lice development into the larger life stages, thus increasing the level of host damage.     

Induced Spawning of Wild American Shad Alosa sapidissima Using Sustained Administration of Gonadotropin-Releasing Hormone Analog (GnRHa)

American shad Alosa supidissima broodstock were collected from the Susquehanna River during their spawning migration. Mean volume of expressible milt (± standard deviation) was 2.5 (±1.7) mL/kg body weight; mean spermatozoid count was 66.2 ± 10⁹ (±163 ± 10⁹) spermntozoa/mL; and duration of 50% motility was 36.5 (±10.3) see. Ovarian biopsies indicated the presence of oocytes of various sizes (200–2,000 μm in diameter) and stages of development. Fish were implanted with a delivery system loaded with gonadotropin-releasing hormone analog (GnRHa) and started spawning 2 d after treatment. Fertile eggs were collected daily for the next 9 d, for a total of 50,100 eggs/kg body weight with a mean fertilization success of 62%. Upon cessation of spawning, the ovaries of all females still contained large numbers of oocytes at various stages of development, as at the beginning of the experiment, but with a greater number of atretic oacytes. Our observations show that American shad have an asynchronous ovarian development, and treatment with a GnRHa delivery system is effective in inducing several successive spawns of fertile eggs.     

Characterization of glycans in the developmental stages of Myxobolus cerebralis (Myxozoa), the causative agent of whirling disease

Glycans and sugar-binding molecules (lectins) form an interactive recognition system, which may enable parasitic organisms to adhere to host cells and migrate into target tissues. The aim of the present study was to analyse surface-associated glycans in the developmental stages of Myxobolus cerebralis (Hofer), the causative agent of whirling disease. A panel of biotin-labelled plant lectins was used to detect a broad spectrum of glycan motifs with high specificity. Binding sites were detected histochemically in the tissue sections of infected rainbow trout, Oncorhynchus mykiss (Walbaum), and infected Tubifex tubifex (Müller), and were characterized by light, fluorescence and transmission electron microscopy. With mannose-specific lectins [Lens culinaris agglutinin, Pisum sativum agglutinin, Canavalia ensiformis agglutinin (LCA, PSA, CanA)] mannose-containing glycans were detected in all the developmental stages and host tissues. No binding sites for galactose-specific lectins were present in M. cerebralis spores but reactivity with host tissues occurred. Diversity in glycans was detected by N-acetyl-D-galactosamine-specific lectins in sporoplasm cells of M. cerebralis and triactinomyxon spores. In the group of lectins with monosaccharide-specificity for N-acetyl-D-glucosamine (GlcNAc), the reactivity of Datura stramonium agglutinin (DSA), Lycopersicon esculentum agglutinin (LEA) and Solanum tuberosum agglutinin (STA) was restricted to polar capsules whereas Griffonia simplicifolia agglutinin II (GSA II) also bound to sporoplasm cells of stages in the fish host but not in those present in infected T. tubifex. Moreover, Triticum vulgaris (wheat germ) agglutinin (WGA) and succinylated WGA indicated the presence of N-acetyl-D-glucosamine polymers in polar capsules. No specificity for spores was observed concerning 'bisected'N-glycans and no reactivity in parasitic stages was observed with the fucose-binding lectin Ulex europaeus agglutinin (UEA) I, Sambucus nigra agglutinin (SNA) (specific for alpha2,6-sialylated glycans) and Maackia amurensis agglutinin (MAAI) (specific for alpha2,3-sialylated glycans). Arachis hypogaea (peanut) agglutinin (PNA), Erythrina cristagalli agglutinin (ECA), GSA I, Sophora japonica agglutinin (SJA), Dolichos biflorus agglutinin (DBA) and GSA II detected reactive sites solely confined to the developmental stages of M. cerebralis and were not reactive in the fish host. These parasite-specific glycans may play a role in the adhesion process of the parasite to fish epidermis prior to infection, but may provide protection to the host by activating the complement system, or stimulating an adaptive immune response as putative antigens.     

Spawning frequency,fecundity, egg size and ovarian histology in groups of Tilapia zillii maintained upon two distinct food ration sizes from first-feeding to sexual maturity

Duplicate groups of first-feeding (4-5-d-old) Tilapia zillii (Gervais) were maintained on two feeding regimes (high and low ration size) for a total period of 550 d (~ 17 months) in environmentally-controlled aquaria. Differences in mean individual fish weight were evident between fish fed the different ration sizes after just 3 months, though these differences were not statistically significant until day 180. Thereafter, fish maintained upon the higher ration were significantly larger than fish fed a reduced ration. Mortality in high ration fish over the course of the experiment was ~60 % compared to ~88 % in low ration fish. Mortality in both treatment groups was most pronounced during the first three months (particularly during the first 30 d) but was much reduced there-after. Spawning trials undertaken from day 450 onwards indicated that there was no significant difference between the two ration groups in terms of spawning periodicity over a discrete time period of 60 d. Mean total fecundity was significantly higher in fish on high rations though mean egg diameter and gonadosomatic index remained unchanged. Adjustment of spawning data to a common maternal fish size using one-factor ANCOVA failed to detect significant differences between the two ration levels in terms of either mean total fecundity or mean egg diameter. Stereological analysis of plastic-embedded ovarian tissue revealed that the ovaries of low ration fish possessed significantly more stage 2 and 3 (early and late perinucleolar) oocytes and significantly less stage 6/7 (late-vitellogenic/maturing) oocytes than the ovaries of fish fed the higher ration. No significant differences were detected between the two rations in terms of ovarian atresia. It is suggested that under prolonged food restriction, female T. zillii sacrifice somatic growth such that reproductive investment can be maintained.     

Effect of temperature and photoperiod on the reproductive condition and performance of a tropical damselfish <Emphasis Type="Italic">Chrysiptera cyanea</Emphasis> during different phases of the reproductive season

Effects of temperature and photoperiod on the reproductive activity of a reef associated tropical damselfish Chrysiptera cyanea were evaluated under three phases with different environmental patterns, phase I (April–May; increasing water temperature and photoperiod), phase II (June–July; increasing water temperature and peak/decreasing photoperiod), and phase III (August–September; peak/decreasing water temperature and decreasing photoperiod). When the fish were reared at 20, 25, or 30°C under natural photoperiod, the reproductive conditions differed within and among the phases depending on experimental temperature and environmental patterns. From phases I through III, ovaries with vitellogenic oocytes were notable only at 25°C, whereas regressing and immature oocytes were noticed at 20 and 30°C. The fish underwent active spawning at 25°C, whereas no or few spawnings were observed at other temperatures. In phase III, there was a resultant prevention of decrease in the gonadosomatic index and disappearance of vitellogenic oocytes in the ovaries of fish under a long photoperiod (LD14:10) compared to those under a short photoperiod (LD10:14). These results indicate that a long photoperiod with a suitable range of water temperature is a principal determinant in continuity of reproductive activity and performance, and that a high temperature has a negative impact on their ovarian development.     

Histological evidence of lunar-synchronized ovarian development and spawning in the spiny rabbitfish Siganus spinus (Linnaeus) around the Ryukyus

ABSTRACT: This study clarifies the annual reproductive cycle and the lunar-synchronized spawning of the spiny rabbitfish ( Siganus spinus ) that inhabit the Okinawan waters. Annual and weekly changes in the gonadosomatic index (GSI) and the histological features of the ovaries were checked. Gonadosomatic index was high during the months of May to July, and yolk-laden oocytes were observed in the ovaries from March to July. Some of the ovaries collected during June and July contained oocytes at maturation stage or ovulatory follicles. These results suggest that the spiny rabbitfish undergo active vitellogenesis and spawning from May to July. During the reproductive season (May to July), collection of fish according to the lunar phase revealed that a high GSI occurred around the time of the new moon. Cyclic oocyte development with peaks around the time of the same moon phase was also observed, suggesting that, in Okinawan waters, this species is a lunar-synchronized spawner and spawns three times.     

Histopathology of Myxidium giardi Cepede, 1906 infection in European eels, Anguilla anguilla L., in Portugal

Infection of Myxidium giardi in skin, gills, cartilage, liver, spleen, kidney, urinary bladder and intestine of eels in different stages of development is described. Some 233 eels were examined by standard histological procedures. Histopathological and clinical findings demonstrated that pathological damage due to infection is severe only in the kidney of elvers. In older eels, histopathological findings indicate an established tolerance to infection. Findings of free spores and plasmodia with spores at different stages of differentiation in eels of all age groups suggest a continuous process of infestation among eels under culture conditions.     

Possible application of mibolerone for induced sex inversion of grouper Epinephelus coioides

SUMMARY: Thirty immature juvenile grouper Epinephelus coioides (19–168 g bodyweight, BW) were randomly stocked in four units 6 t tanks to determine if mibolerone can be used to induce sex inversion in groupers. After acclimatization and weaning to artificial feed, the feed given daily (4% BW/day) was supplemented with 0, 50, 100, and 200 μg mibolerone/kg feed for about 18 weeks. Thereafter, the hormone treatment was withdrawn and the experiment was terminated at Week 24. Ten fish were killed for gonad histology at stocking to serve as an initial control while about three to five fish were killed every 8 weeks. In general, ovaries of initial controls showed the presence of moderate stromal cells and gonia and few primary oocytes. At Weeks 8 and 16, ovaries of the control fish (0 μg/kg) were similar to that of the initial control except that primary oocytes increased at Week 24. Gonads of fish fed diets containing 100 and 200 μg/kg had none to moderate spermatocytes and few spermatids at Week 8 and 16, although spermatozoa were not observed, indicating that the fish were undergoing spermatogenesis. Spermatogenesis at 50 μg/kg was not as advanced since only few spermatocytes occurred at Weeks 8 followed by moderate gonia and no spermatocytes and spermatids at Week 16. However, the presence of few primary oocytes was observed when mibolerone was withdrawn suggesting that sex-inversed fish reverted back to a female condition. These results show that sex inversion in juvenile grouper can be induced by oral administration of mibolerone and may have possible application on mature females to produce functional males.     

长鳍篮子鱼繁殖季节性腺的组织学研究

采用常规组织切片法研究了长鳍篮子鱼繁殖季节各期卵巢和精巢的组织学结构特征。在所获得样本中,未发现Ⅰ期、Ⅱ期的卵巢和精巢。Ⅲ期卵巢以第Ⅲ时相卵母细胞为主,这时期液泡、卵黄颗粒出现,同时含有少量Ⅰ时相和Ⅱ时相的卵母细胞;Ⅳ期卵巢以第Ⅳ时相卵母细胞为主,Ⅳ时相晚期卵母细胞开始出现油球,细胞核偏移和变形,放射带明显;Ⅴ期卵巢的卵细胞游离,卵细胞的外层分别有胶膜、放射带和质膜;Ⅵ期卵巢主要由Ⅲ时相卵母细胞和大量的空滤泡外膜组成。长鳍篮子鱼精巢为辐射型,精巢内生殖细胞分为初级精母细胞、次级精母细胞、精子细胞和精子,各期生殖细胞和支持细胞组成了精小管,同一精小管中的生精细胞发育不同步。精子成熟后,充满整个精小管,完成发育过程。