Study on the reproductive biology of Pinctada fucata (Gould) (Mollusca: Pteriidae)

Annual cycle of gonad development and spawning in pearl oyster, Pinctada ficata (Gould) in Nakhiloo, Northeast Persian Gulf, was investigated over two years from August 1994 to June 1996. Gonadal condition was assessed by staging criteria to describe gametogenic development from histological preparations of randomly collected individuals of all sizes. A bimodal gametogenic pattern with summer and autumn spawning periods was evident throughout the study. Gametogensis commenced in November-December which proceeded by major gonadal maturation during February-April. Summer spawning was observed from April to July with major spawning at the latter end. During spawning peak in July, low level of gametogensis was noticed. Gametogenic activity was picked up again in August-September which proceeded by autumn spawning from September to December. Towards the end of spawning season, incidence of gonadal inactivation increased. Minimum level of gonadal activity was observed in November. Temperature regime appears to have influential role in regulation of gametogenic and spawning processes. Gonadal development and spawning trends were similar in both sexes. P. radiaata was found to be protandrous hermaphrodite which matured as a male at shell height greater than 20 mm. Biseivality was uncommon and the sex ratio was about 1:1. Ultrastructure of gametes were investigated in the Pictada fucata (Gould). "Auxiliary cells" closely accociated with developing oocytes were observed. Each oocyte seems to be associated with only one secretory cell. which is characterized by an abundant rough endoplasmic reticulum at the onset of vitellogenesis. Contact between this cell and a developing oocytes is maintained by a desmosome-like junction which can be observed when the vitelline coat is formed. these "auxiliary or nursing cells" seem to play a tropic role in vitellogenesis, and may be involved in the formation of the vitelline coat of the oocytes. Oocytic degeneration is observed in this species, it is a continuous phenomenon of varing intensity throughout the year. The ultrastructural changes resulting in lysis of the oocyte are described. Mature spermatozoa consist of a broad, cap-shaped acrosomal vesicle, subacrosomal material, a round nucleus, two triplet substructure centrioles surrounded by four spherical mitochondria, and a flagellum anchored to the distal centriole and plasma membrane. Spermatozoa of Plucata closley resemble to those of other investigated Pteriidae. Changes in proximate composition of soft tissue and gonadal cycle of Pinctada fucata was studied. Mobilization and utilization of stored reserves are apparent during gametogenesis and gonadal maturation. Protein reserves are utilized during spermatogenesis while reserved carbohydrates form the main energy donor in oogenesis. The role of lipid as am.: energy reserve is second to that of carbohydrate.

Morphological and ultrastructural changes in tobacco BY-2 cells exposed to microcystin-RR

Tobacco BY-2 cells were exposed to microcystin-RR (MC-RR) at two concentrations, 60 mu g mL(-1) and 120 mu g mL(-1), to study the changes in morphology and ultrastructure of cells as a result of the exposure. Exposure to the lower concentration for 5 d led to typical apoptotic morphological changes including condensation of nuclear chromatin, creation of a characteristic 'half moon' structure, and cytoplasm shrinkage and decreased cell volume, as revealed through light microscopy. fluorescence microscopy, and transmission electron microscopy, respectively. Exposure to the higher concentration, on the other hand, led to morphological and ultrastructural changes typical of necrosis, such as rupture of the plasma membrane and the nuclear membrane and a marked swelling of cells. The presence of many vacuoles containing unusual deposits points to the involvement of vacuoles in detoxifying MC-RR. Results of the present study indicate that exposure of tobacco BY-2 cells to MC-RR at a lower concentration (60 mu g mL(-1)) results in apoptosis and that to a higher concentration (120 mu g mL(-1)), in necrosis. (C) 2009 Elsevier Ltd. All rights reserved.

EFFECT OF ELEVATED BICARBONATE CONCENTRATION ON GROWTH, CHLOROPHYLL A FLUORESCENCE AND ULTRA-STRUCTURE OF Microcystis aeruginosa (CYANOBACTERIUM)

The influence of bicarbonate (HCO3-) on Microcystis aeruginosa FACHB 905 was assessed in this study. Growth curves, chlorophyll a fluorescence and ultrastructure were measured at two HCO3- concentrations, 2.3 mM and 12.4 mM. A treatment of sodium chloride (NaCl) was also conducted alongside to establish the influence level of sodium. It was found that upon treatment with elevated HCO3- concentrations of 2.3 mM and 12.4 mM, cell densities were 13% and 27% (respectively) higher than controls. In photosynthetic performance, elevated HCO3- concentration initially stimulated Fv/Fm at the prophase of culture and then subsequently inhibited it. The inhibition of 2.3mM was higher than that of 12.4mM HCO3-. The maximum relative electron transport rate (ETRmax) exhibited inhibition at elevated HCO3- concentrations. DI0/CS was decreased at 2.3 mM and increased at 12.4mM. In the case of both treatments. ABS/CSI TR0/CS, ET0/CS, RC/CS0 and RC/CSm were decreased by elevated HCO3- concentrations, which indicated damage to photosynthetic apparati and an inactivation of a fraction of reaction centers. This point was also proven by ultrastructural photos. High HCO3--exposed cells lost the characteristic photosynthetic membrane arrangement compared with the control and high salinity treated samples. At the 2.3mM concentration of HCO3-. damage to photosynthetic apparati caused decreased photosynthetic activity. These findings suggested that elevated HCO3- concentration stimulated the growth and photosynthesis of M. aeruginosa FACHB 905 in a short time. Exposure to high HCO3- concentrations for a longer period of time will damage photosynthetic apparatus. In addition, the ultrastructure indicated that elevated HCO3--concentration lead to photosynthetic apparati damage. In our experiment, it was observed that the inhibition effect of 2.3mM HCO3- was higher than that of 12.4mM HCO3-. We hypothesized that M. aeruginosa FACHB 905 induced a protective mechanism under high concentrations of HCO3-.

Microcystin-RR induces physiological stress and cell death in the cyanobacterium Aphanizomenon sp DC01 isolated from Lake Dianchi, China

The phytoplankton community in Lake Dianchi (Yunnan Province, Southwestern China) is dominated in April by a bloom of Aphanizomenon, that disappears Suddenly and is displaced by a Microcystis bloom in May. The reasons for the rapid bloom disappearance phenomenon and the temporal variability in the composition of phytoplankton assemblages are poorly understood. Cell growth, ultrastructure and physiological changes were examined in cultures of Aphanizomenon sp. DC01 isolated from Lake Dianchi exposed to different closes of rnicrocystin-RR (MC-RR) produced by the Microcystis bloom. MC-RR concentrations above 100 mu g L-1 markedly inhibited the pigment (chlorophyll-a, phycocyanin) synthesis and caused an increase of soluble carbohydrate and protein contents and nitrate reductase activity of toxin-treated blue-green algae. A drastic. reduction in photochemical efficiency of PSII (Fv/Fm) was also found. Morphological examinationn showed that the Aphanizomenon filaments disintegrated and file cells lysed gradually after 48 h Of toxin exposure. Transmission electron microscopy revealed that cellular inclusions of stressed cells almost leaked out completely and the cell membranes were grossly damaged. These findings demonstrate the allelopathic activity of Microcystis aeruginosa inducing physiological stress and cell death of Aphanizomenon sp. DC01 Although the active concentrations of microcystin were rather high, we propose that microcystin may function as allelopathic Substance due to inhomogeneous toxin concentrations close to Microcystis cells. Hence, it may play a role in species Succession of Aphanizomenon and Microcystis in Lake Dianchi.

In situ studies on physiological and biochemical responses of four fishes with different trophic levels to toxic cyanobacterial blooms in a large Chinese lake

Physiological and biochemical responses of four fishes with different trophic levels to toxic cyanobacterial blooms were studied in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. We sampled four fishes: the phytoplanktivorous Hypophthalmichthys molitrix and Aristichthys nobilis, the omnivorous Carassius auratus, and the carnivorous Culter ilishaeformis. Alterations of the antioxidant (GSH) and the major antioxidant enzymes (CAT, SOD, GPx, GST) in livers were monitored monthly, and the ultrastructures of livers were compared between the bloom and post-bloom periods. During the cyanobacterial blooms, the phytoplanktivorous fishes displayed only slight ultrastructural changes in liver, while the carnivorous fish presented the most serious injury as swollen endomembrane system and morphologically altered nuclei in hepatocytes. Biochemically, the phytoplanktivorous fishes possessed higher basal GSH concentrations and better correlations between the major antioxidant enzymes in liver, which might be responsible for their powerful resistance to MCs. This article provided physiological and toxicological evidences for the possible succession of fish communities following occurrence of toxic cyanobacterial blooms and also for the applicability of using phytoplanktivorous fish to counteract toxic cyanobacterial blooms in natural waters. (C) 2007 Elsevier Ltd. All rights reserved.

Electron microscopic examination of the viromatrix of Rana grylio virus in a fish cell line

Rana grylio virus (RGV), a Ranavirus belonging to the family Iridoviridae, assembles in the viromatrix which is a factory for viral genome replication and particle assembly. Ultrastructural studies of the viromatrix will clarify the pathway of assembly. The viromatrix and quantitative changes in RGV infected epithelipma papulosum cyprini (EPC) cells, one of fish cell lines, were studied by electron microscopy. It was shown that viromatrices were adjacent to the nucleus, and the electron density was lower than that of the surrounding cytoplasm. The viromatrix contained virus particles with different forms, electron-dense materials and amorphous structures which included tubules and membranous materials. Tubules were often observed in direct continuity with empty capsids. Several bundles of intermediate filaments were seen alongside the viromatrix and crystalline aggregates. Large clusters of mitochondria occurred in proximity to viromatrix. A total of 990 cells profiles were examined. The results showed that 394 cells contained viromatrix: 89.3% contained one, and 10.7% contained two to four viromatrices. The number of viromatrices increased gradually and reached a peak at 16 h p.i. The viromatrix area at 24 h p.i. increased up to 7.4 +/- 0.69 mu m(2) which was three-times lower than that at 6 h p.i. The number of empty capsids within viromatrix was generally more than that of "full" particles at different time points, and there was a strong positive correlation between them. (c) 2005 Elsevier B.V. All rights reserved.

In vivo studies on toxin accumulation in liver and ultrastructural changes of hepatocytes of the phytoplanktivorous bighead carp i.p.-injected with extracted microcystins

Phytoplanktivorous bighead carp were injected i.p. with extracted microcystins (mainly MC-RR and -LR) at two doses, 200 and 500 MC-LReq. mu g kg(-1) bw, and the changes in extractable MCs in liver and in the ultrastructure of hepatocytes were studied at 1, 3, 12, 24 and 48 h after injection. Quantitative and qualitative determinations of MCs in the liver were conducted by HPLC and LC-MS, respectively. MC concentration in the liver reached the maxima at 12 It (2.89 mu g MCs g(-1) dry weight at the lower dose) or at 3 h (5.43 mu g MCs g(-1) dry weight at the higher dose) post-injection, followed by sharp declines afterwards, whereas the ultrastructural changes of hepatocytes in both dose groups suggest progressive increases in severity toward the directions of apoptosis and necrosis from I to 24 h, respectively. There were two new findings in fish: widening of intercellular spaces was among the early ultrastructural changes induced by MCs and ultrastructural recovery of hepatocytes was evident at 48 h post-injection in both dose groups. Both the present and previous studies suggest that with in vivo or in vitro exposure to microcystins, hepatocyte damage in fish tends to proceed toward the direction of apoptosis at lower MC concentrations but toward the direction of necrosis at high MC concentrations. The temporal dynamics of MCs in the liver suggest that bighead carp may have a mechanism to degrade or bind MC-LR actively after it enters the blood system. (c) 2005 Elsevier Ltd. All rights reserved.

A survey of the gomphonemaceae and cymbellaceae (Bacillariophyta) from the Jolmolungma Mountain (Everest) region of China

Gomphonemaceae and Cymbellaceae from Jolmolungma Mountain region of China, comprised 79 taxa belonging to five genera. The dominant species were Gomphonema angustatum var. productum, G. gracile var. gracile, G. parvulum var. parvulum, G. hedini, Cymbella aequalis var. pisciculus, C. delicatula, C. cistula, C. cistula var. hebetata, C. cesatii, C. cymbiformis, C microcephala, C. minuta var. minuta., C. minuta var. silesiaca, C. pusilla, C. affinis. Some arctic and alpine forms also occurred, and the following taxa were unique to this region: Cymbella delicatula, C. naviculiformis, C. gracilis, C. minuta var. minuta, C. affinis, C. cistula, C. cymbiformis, C. cistula var. hebetata, C. arctica, Didymosphenia geminata, Gomphonema hedinii, G. olivaceum. Numbers of taxa in each samples varied greatly, It appears that species diversity of diatoms is related not only to macro-environments (e.g., geographic zonation) but also to microenvironments (e.g., microhabitats and microclimates). Cymbella spp. are not only comparatively widespread in the region but also outnumber Gomphonema spp. A large number of species found in the Jolmolungma Mountain region are also found in Tibet and the headwaters of Yangtze River.

Infection and propagation of lymphocystis virus isolated from the cultured flounder Paralichthys olivaceus in grass carp cell lines

The causative agent of lymphocystis disease that frequently occurs in cultured flounder Paralichthys olivaceus in China is lymphocystis virus (LV). In this study, 13 fish cell lines were tested for their susceptibility to LV. Of these, 2 cell lines derived from the freshwater grass carp Ctenopharyngodon idellus proved susceptible to the LV, and 1 cell line, GCO (grass carp ovary), was therefore used to replicate and propagate the virus. An obvious cytopathic effect (CPE) was first observed in cell monolayers at 1 d post-inoculation, and at 3 d this had extended to about 75% of the cell monolayer. However, no further CPE extension was observed after 4 d. Cytopathic characteristics induced by the LV were detected by Giemsa staining and fluorescence microscopic observation with Hoechst 33258 staining. The propagated virus particles were also observed by electron microscopy. Ultrastructure analysis revealed several distinct cellular changes, such as chromatin compaction and margination, vesicle formation, cell-surface convolution, nuclear fragmentation and the occurrence of characteristic 'blebs' and cell fusion. This study provides a detailed report of LV infection and propagation in a freshwater fish cell line, and presents direct electron microscopy evidence for propagation of the virus in infected cells. A possible process by which the CPEs are controlled is suggested.

Scanning electron microscopy of Aspidogaster ijimai Kawamura, 1913 and A-conchicola Baer, 1827 (Aspidogastrea, Aspidogastridae) with reference to their fish definitive-host specificity

Two species of aspidogastreans, namely Aspidogaster ijimai and A. conchicola, were studied by scanning electron microscopy. In nine lakes and an old river course, the Tian'ezhou oxbow, investigated in the flood plain of the Yangtze River, A. ijimai was obtained from the common carp (Cyprinus carpio) in three lakes, and A. conchicola from the black carp Mylopharyngodon piceus in three lakes and the oxbow. In none of the localities, however, were the two species found together. It is suggested that A. ijimai may be considered as a specialist parasite for the common carp, at least in the flood-plain lakes of the Yangtze River. The two parasites were similar in many aspects of their morphology. Their bodies can both be separated into a dorsal part and a ventral disc, with the body surface of the dorsal part elevated by transverse folds, and the disc subdivided into alveoli by transverse and longitudinal septa, although the number of alveoli was different in the two species. The depression on the ventral surface of the neck region was prominent for both species, and their ventral disc was covered densely with non-ciliated bulbous papillae. The position of mouth, osmo-regulatory pore and marginal organ was also similar for A. ijimai and A. conchicola. However, microridges in the trough of the folds in the neck region and numerous small pits on the upper part of the septa were found exclusively in A. ijimai, but uniciliated sensory papillae in A. conchicola.

棘蛙族(Tribe Paini)精子形态结构及精巢年周期研究

棘蛙族(Tribe Paini)隶两栖纲(Amphibia)、无尾目(Anura)、蛙科(Ranidae)、叉舌蛙亚科(Dicroglossinae)，由棘蛙属(Paa)、倭蛙属(Nanorana) 和沙巴蛙属(Chaparana)构成（Dubois,1992）。由于特殊的形态特征和染色体核型，棘蛙族受到国内外学者的广泛重视和研究，但是到目前为止，棘蛙族的系统发育关系尚未明晰，族下属种的分类和归属问题还有待进一步研究和新的证据出现。本文通过光学显微镜、电子显微镜和石蜡切片对棘蛙族10 物种的精子和精巢进行研究，旨在了解棘蛙族精子的形态、量度、超微结构特征及不同季节精巢结构的变化规律，同时为棘蛙族的系统研究提供新的依据，也为棘蛙族濒危物种的保护和经济物种的繁殖提供基础资料。研究结果表明：棘蛙族各属物种精子的形态基本相似，精子整体呈线形，由头部、中片和尾部构成。精子头部呈长条状，顶体呈锥状，位于头部顶端并向前伸出，中片较长，尾部波动弯曲。棘蛙族各属物种精子量度差异较大，将各属物种精子头部、中片、尾部、头宽、尾宽的量度数据进行聚类分析，结果表明棘蛙族10 物种可分为三类：第一类包括棘侧蛙、合江棘蛙、小棘蛙、棘腹蛙和棘胸蛙，特点是精子较短，全长在72.6～103.35µm 之间；第二类包括倭蛙、高山倭蛙、腹斑倭蛙，特点是精子较长，全长在107.74～129.75µm 之间；第三类包括隆肛蛙和双团棘胸蛙，特点是精子最长，全长在145.89～165.84µm 之间。棘蛙族各属精子超微结构基本相似：精子头部由顶体、细胞核构成；中片由中心粒、线粒体构成；尾部由单根轴丝构成。精子顶体横切呈圆环状，细胞核电子密度高；线粒体为卵圆形，呈环状围绕轴丝排列，线粒体数目较多，约30层；尾部轴丝为典型的9＋2结构，即由2根中央微管和9对外周微管组成。不同季节的倭蛙精巢结构变化表明倭蛙精巢每年只有一个生精周期，生精周期始于7 月，繁殖季节从5 月到6 月，生精高峰期为9 月；根据倭蛙不同季节精巢结构的变化，可将生精周期分为3 个阶段：第一阶段从7 月到9 月，为精子形成期；第二阶段从10 月到翌年4 月，为精子的贮存阶段，也即倭蛙的冬眠期；第三阶段从5 月到6 月，为精子的排放阶段，即倭蛙的繁殖期。不同季节的隆肛蛙精巢结构变化表明5 月为隆肛蛙的繁殖高峰期。根据棘蛙族各属精子的形态、量度和超微结构特征，结合已有的棘蛙族形态学、生态学、染色体核型及系统学研究成果，本文认为：1．基于精子数据对棘蛙族的划分和基于形态学及分子系统学数据对棘蛙族的划分均有相同之处，精子形态结构可为棘蛙族的系统研究提供新的证据。2． 棘蛙族各属精子的形态、量度及超微结构不仅与蛙科其他属种有明显差异，而且在无尾类中也较为特殊，精子学研究结果支持将棘蛙族从蛙科中分离出来，归隶于叉舌蛙科的叉舌蛙亚科的系统学修正。3. 精子的顶体、细胞核、中片的形态结构及量度可作为蛙科的分类指标。On the base of unique morphological and kyrotype characters, Dubois(1992)recognized three genera Paa, Narnorana, Chaparana as tribe Paini, which is amember of Dicroglossinae, Ranidae. In present study, the sperm shape, size andultrastructure of 10 paini species were investigated through the light and electronmicroscope, and testis structure of N. pleskei and F. quadrana was also studied. Wesuppose this study could offer some spermatological evidence to phylogeny andreproduction study of tribe Paini. The results were as follows:The sperm shape of tribe paini is homologically similar, the spermatozoa arefiliform, composed of elongate head, long mid-piece and waved tail. The acrosome isapically associated with the nucleus and extend anteriorly.The sperm length of tribe paini differ remarkably among genera. Cluster for thelength of sperm head, mid-piece, tail, total length, head-width, tail-width of ten painifrogs indicated the 10 species could be separated into three groups: GroupⅠcontainsP. shini, P. robertingeri, P. spinosa, P. exilispinosa, P. boulengeri, the spermatozoa ischaracterized with short in total length, ranging from 72.6µm to 103.35µm; GroupⅡcontains N. pleskei, N. parkeri, N. ventripunctata, the spermatozoa ischaracterized with relatively long in total length, ranging from 107.74µm to129.75µm; Group Ⅲ contains F. quadrana and P. yunnanensis, the spermatozoa is characterized with longest in total length, ranging from 145.89µm to 165.84µm. thethree groups based on spermatological data is partially match the classification basedon morphological and molecular data.The ultrastructure of spermatozoa in tribe paini is also basic similar, includingacrosome vescile, nuleus of the head proper, centriole, mitochondriol of themid-pieces, axoneme of the tail. The acrosome vescle is circle in TEM transversesection, the density of nucleus is high; The mitochondrions is oval, surrounding theaxial filament with about 30 layers of mitochondria; The axoneme has the typical 9+2pattern of microtubules.The seasonal changes in testis of N. pleskei indicates it has only onespermatogenesis circle, which begin in July, the reproduction season is from May toJune, the spermatogenesis is active in September. On the base of seasonal changes intestis, the spermatogenesis circle can be separated into three stages: In stageⅠfromJuly to September, spermatids are formed; In stage Ⅱ from October to April next year,the spermatozoa are stored in testis,which is the hibernated period; In stage Ⅲ fromMay to June, mature spermatozoa were released from the testis, which is thereproduction season of N. pleskei. As to F. quadrana, reproduction is active in May.With the previous study of morphology, ecology, karyotypes and phylogenyresearch of tribe Paini, the spermatological data in present study suggests:1. The spermatological classification of tribe paini is partially consistant with themorphological and molecular classification respectively.2.The sperm morphology and ultrustructure of tribe paini is unique not only inthe family Ranida but also in Anura, which suggest the tribe paini is monophyletic andmight be transfered from the family Ranida to the family Dicroglossidae based onmolecular evidence.3. The acrosome, nuleus, shape, length and ultrastructure of mid-piece can beused as an alternative taxonomic character in Anura.

Variations in morphology and PSII photosynthetic capabilities during the early development of tetraspores of Gracilaria vermiculophylla (Ohmi) Papenfuss (Gracilariales, Rhodophyta)

Background: Red algae are primitive photosynthetic eukaryotes, whose spores are ideal subjects for studies of photosynthesis and development. Although the development of red alga spores has received considerable research attention, few studies have focused on the detailed morphological and photosynthetic changes that occur during the early development of tetraspores of Gracilaria vermiculophylla (Ohmi) Papenfuss (Gracilariales, Rhodophyta). Herein, we documented these changes in this species of red algae. Results: In the tetraspores, we observed two types of division, cruciate and zonate, and both could develop into multicellular bodies (disks). During the first 84 hours, tetraspores divided several times, but the diameter of the disks changed very little; thereafter, the diameter increased significantly. Scanning electron microscopy observations and analysis of histological sections revealed that the natural shape of the disk remains tapered over time, and the erect frond grows from the central protrusion of the disk. Cultivation of tissue from excised disks demonstrated that the central protrusion of the disk is essential for initiation of the erect frond. Photosynthetic (i.e., PSII) activities were measured using chlorophyll fluorescence analysis. The results indicated that freshly released tetraspores retained limited PSII photosynthetic capabilities; when the tetraspores attached to a substrate, those capabilities increased significantly. In the disk, the PSII activity of both marginal and central cells was similar, although some degree of morphological polarity was present; the PSII photosynthetic capabilities in young germling exhibited an apico-basal gradient. Conclusions: Attachment of tetraspores to a substrate significantly enhanced their PSII photosynthetic capabilities, and triggered further development. The central protrusion of the disk is the growth point, may have transfer of nutritive material with the marginal cells. Within the young germling, the hetero-distribution of PSII photosynthetic capabilities might be due to the differences in cell functions.

Functional characterization of sll0659 from Synechocystis sp PCC 6803

Synchocystis sp. PCC 6803 lacks a gene for the any known types of lycopene cyclase. Recently, we reported that sll0659 (unknown for its function) from Synechocystis sp. PCC6803 shows similarity in sequence to a lycopene cyclase gene-CruA from Chlorobium tepidum. To test, whether Sll0659 encoded protein serves as lycopene cyclase, in this study, we investigated the carotenoids of the wild types ans mutants, In the sll0659 deleted mutant, there is no blockage at the lycopene cyclization step. Our results demonstrate that sll0659 does not affect lycopene cyclization. However, the ultrastructure of mutants suggests the involvement or necessity of sll0659 in the cell division.

Effect of ultraviolet irradiation on sperm of the left-eyed flounder, Paralichthys olivaceus

In this study, at proper dosage of ultraviolet (UV) irradiation (180 sec: 36,000 erg/mm(2)), sperm chromosomes of left-eyed flounder, Paralichthys olivaceus, were inactivated, while spermatozoa maintained ability to move and inseminate eggs. Gynogenetic haploids were detected by morphological observation, chromosome counting, and flow cytometer analysis. The ultrastructure of treated sperm was observed under scanning electronic microscope (SEM) and transmission electronic microscope (TEM). The results showed that after being irradiated at lower dosage of irradiation (0-180 sec: 0-36,000 erg/mm(2)), the surface structure of spermatozoa was not affected by UV irradiation, while the inner structures including membrane system and karyoplasm denseness of treated spermatozoa were little changed. However, obvious changes were observed in their membrane system, mitochondria, and nucleus if the dosage of irradiation increased to 240 sec: 48,000 erg/mm(2) or 300 sec: 60,000 erg/mm(2). The sperm survival rates did not change at the lower dosages of the UV irradiation (0-180 sec: 0-36,000 erg/mm(2)) but decreased as the irradiation dosage increased. The motility of treated sperm was lower than that of control group in general but did not change with UV irradiation dosage increasing at the certain range of 0-300 sec: 0-60,000 erg/mm(2).

Morphological and ultrastructural characterization of the coelomocytes in Apostichopus japonicus

The coelomocytes suspended in the coelomic fluid and occurring in the coelomic epithelial layer of the sea cucumber Apostichopus japonicus (Selenka) (Holothuroidea: Aspidochirota: Stichopodidae) function as mediators of the immune system, trephocytic cells and nutrient transport cells. Types of coelomocytes are characterized based on their morphological and ultrastructural features. Flow cytometry plus light and electron microscopic analyses were conducted in order to characterize the coelomocytes of A. japonicus. Six types of coelomocytes were identified: lymphocytes, morula cells, amoebocytes, crystal cells, fusiform cells and vibratile cells. Within these major categories, several distinctive cell types occurred that might represent developmental stages. The mean +/- SD coelomocyte concentration in the individuals (body length: 10 to 15 cm; weight: 100 to 150 g) was (3.79 +/- 0.65) X 10(6) cells ml(-1). The coelomic fluid contained mainly hyalinocytes (76.69%) and granulocytes (23.31 %).