Effect of the extract of Thespesia populnea leaves on mice testis

Male swiss mice were administered with the leaf extract of Thespesia populnea at a daily dose of 400 mg/kg body weight for 15 days and the testis were subjected to structural analysis. The structure of the seminiferous tubule in the testis of treated animal was elongated. Sertoli cells were enlarged in its structure and spermatids became round and disintegrated. It is suggested that the extract of T. populnea treatment leads to pathological changes in the seminiferous tubules, Sertoli cells and spermatids of the testis.     

Reproduction of the commercial sea cucumber Holothuria whitmaei [Holothuroidea: Aspidochirotida] in the Indian and Pacific Ocean regions of Australia

Seasonal trends in the gonad index (GI) of two widely separated populations of black teatfish, Holothuria whitmaei (formerly included in Holothuria nobilis), were investigated between Pacific (Great Barrier Reef) and Indian Ocean (Ningaloo Reef) coral reefs of Australia. Reproductive activity followed a similar annual trend, with the GI of both populations peaking typically between April and June. Macroscopic and histological analysis of Ningaloo Reef specimens revealed that large germinal tubules, positioned centrally on the gonad basis, progressed through four maturity stages: growing (II); mature (III); partly spawned (IV); and spent (V). Growing tubules dominated the central region of the gonad basis between January and March, followed by an increase in the number of mature tubules throughout the GI peak (April–June). The progressive appearance of partly spawned and spent tubules between June and October suggests that spawning in H. whitmaei continues intermittently over an extended period throughout the austral winter. The examination of the gonad structure of sexually mature male and female specimens identified five tubule size classes in total (C₁ to C₅), each of differing physical and gametogenic status. In females, smaller C₁ tubules located at the anterior edge of the gonad basis contained pre- to early vitellogenic oocytes. Larger C₂ and C₃ tubule cohorts, positioned centrally on the gonad basis, contained mid- to late-stage vitellogenic oocytes. Smaller C₄ and C₅ tubules, located at the posterior edge, contained only relict oocytes. Similar physical and gametogenic differences were apparent between tubule cohorts in male specimens. We propose that these results, together with evidence of incomplete gonad resorption over the austral summer, indicate that gonad development in H. whitmaei conforms to the predictions of the Tubule Recruitment Model (TRM). The TRM is reported rarely among tropical aspidochirotes, and results presented here (1) provide the first direct evidence of this model in H. whitmaei, and (2) confirm that this species is one of the few winter-spawning tropical invertebrates.     

原尾蜥虎(Hemidactylus bowringii)雄性生殖周期的研究

对福州产的原尾蜥虎（Ｈｅｍｉｄａｃｔｙｌｕｓｂｏｗｒｉｎｇｉｉ） 雄性生殖周期的研究结果表明：１０ 月份精子发生开始新的周期性复生，精原细胞发育进入增殖期，中旬曲细精管除精原细胞外尚有较多的初级精母细胞；１１ 月初已有较多的精细胞，曲细精管管腔中开始出现少量的精子；１２ 月份，生精活动非常活跃，输精管始见有成熟的精子，１２ 月下旬进入冬眠期，成熟的精子释放到输精管中过冬，冬眠期间生精活动趋于停滞，表现为曲细精管管径和生精上皮厚度都有所下降；次年３ 月底４ 月初陆续出蛰，生精活动又恢复并逐渐增强，４、５ 月份精子发生相当活跃，６ 月份是精子发生的高峰期，也是交配繁殖的高峰期；原尾蜥虎一年只繁殖一次，繁殖期过后，７ ～９ 月份，生精活动逐渐减弱，曲细精管的管腔逐渐消失，初级、次级精母细胞、精细胞逐渐减少、退化，精子逐渐退化、消失，到９ 月下旬曲细精管和输精管都见不到精子，此时的曲细精管主要由精原细胞组成，一个周期到此结束．     

Ultrastructure of the spirocyst tubule in black corals (Coelenterata: Antipatharia) and its taxonomic implications

Spirocyst tubules in five shallow-water antipatharian species Antipathes pennacea, Antipathes sp. cf. salix, A. fiordensis, A. galapagensis, and Cirrhipathes luetkeni contain an electron-opaque, helically arranged structure, composed of four distinct and separate bundles of microfibril-like material. Each bundle appears to be solid, rather than hollow, and is enveloped by a sheath of lesser electron-opacity that is continuous with the pleats. Tubules from a more distantly related, deep-water antipatharian, are similarly configured but are composed of only three distinct bundles, separated from one another by electron-opaque sheath material. The bundles are electron-lucent; their substructure was not preserved because this specimen was prepared for microscopy after preservation in alcohol for 27 yr, apparently without initial fixation. Most other details of the spirocyst tubule and capsule can be determined from this museum specimen. The arrangement within the tubule of multiple bundles, clearly defined by sheaths, is structurally distinct from other zoantharian spirocysts, and appears to be unique to the Antipatharia. The tubule structure of the anemone Aiptasia sp., examined for comparison, is consistent with that of spirocysts from other actiniarians and zoantharians generally, including the appearance of electron-opaque rodlets within the tubule lumen. No substructural information was obtained from Aiptasia sp. rodlets. It is unclear whether the microfibrillar bundles of antipatharian spirocysts differ from those of other zoantharians by being more amenable to resolution, or if their tubule substructure is fundamentally distinct from those of more typical zoantharians. In terms of overall structure, Aiptasia sp. spirocyst tubules contain a single bundle of rodlets without a clearly defined sheath, in contrast to antipatharians which have three or four bundles, each clearly defined. These structural differences may be useful in determining relationships both within the Antipatharia, and between antipatharians and allied orders.     

虎纹蛙和大树蛙肾细胞超微结构及其与环境适应性的关系

用透射电镜比较了大树蛙和虎纹蛙肾细胞的超微结构，大树蛙近曲小管上皮细胞具发达的质膜内褶迷路，虎纹蛙近曲小管上皮细胞无比结构；小树蛙远曲小管上皮细胞含丰富的质膜内褶，虎纹蛙远曲小管上皮细胞的质膜内褶较少；两种蛙的集合管上皮细胞都有质膜内褶迷路；结果表明，虎纹蛙和大树蛙之间肾细胞超微结构特征的差异是与它们不同的栖息环境和生态习性相适应的。     

Effect of sampling procedures,transportation stress and laboratory maintenance on the structure and function of the digestive gland epithelium of the mussel<Emphasis Type="Italic"> Mytilus galloprovincialis</Emphasis>

The digestive glands of four groups of mussels (Mytilus galloprovincialis) were examined under light and electron microscopes. Individuals of the first group were fixed immediately after their extraction from seawater; those of the second, after having been kept out of water for 2 h; while those of the third and fourth groups, after maintenance in the aquarium for 1 day and 1 week, respectively. The digestive gland cells of the mussels kept out of the water for 2 h presented, in many cases, extensive morphological disorganization, expressed mainly by cytoplasmic extrusions into the tubule lumen. In contrast, the mussels kept in the aquarium for 1 day showed no disintegration phenomena in their digestive cells, but presented a decrease in the height of the tubule epithelium and increased size of the tubule lumen. Similarly, the mussels kept in the aquarium for 1 week presented cells characterized by increased height compared to those of the mussels of the previous group, as well as a quite inactive epithelium, with no extrusion of cytoplasmic materials into the lumen. The values of the "lysosomal membrane stability" test (LMS), a bioindicator of general stress, were also evaluated. The mussels kept in the aquarium for 1 day showed decreased LMS values compared to the values for other mussel groups, which indicated that general stress had increased. Results of the present study show that care should be taken in sampling and handling of mussels for use in biomonitoring studies.     

An electron microscope study on the mineralization of the skeleton of the staghorn coral Acropora hebes

The skeleton formation in the reef-building coral Acropora hebes (Dana) was ultrastructurally investigated by observing the skeletogenic tissue and the skeleton in the apical portion of the branches. The skeletogenic tissue was made of a layer of tall calicoblastic cells which displayed high exocytotic activities. A number of hollow spherules and tiny vesicles were found in the sub-epithelial space between the calicoblastic cell layer and the skeletal plate. These organic materials appeared to occur in the perinuclear Golgi vesicles in the cells. The energy-dispersive X-ray analysis revealed a calcium element in the osmiophilic granules of the calicoblastic cells, but not in any other cell organelles. As the granules showed no diffraction pattern, it was suggested that they could be a Ca-reservoir in the cells. Crystalline particles were found to deposit on the periphery of the sub-epithelial spherules. The spherules developed to spherular crystals by depositing granulated crystalline particles. Two of the spherules appeared to fuse with each other to form a spindle-shaped crystal. The spherular and spindle-shaped crystals accumulated on the thecal ridge and the lateral side surface of the thecal plate, and seemed to contribute to the elongation and thickening of the thecal plate. The thecal plate exhibited a porous structure which probably originated from an aggregation of the central cores of these crystals. On the surface of the thecal plate more than about 5 m thick, scale-like structures composed of spherular crystal substructures were observed. These observations suggest that mineralization in A. hebes occurs in the extracellular space by elabolating the spherular and spindle-shaped precursor structures and that growth of the corallite is brought about by an aggregation and coalescence of these crystals.     

Association between the mollusc bivalve Loripes lucinalis and a Chlamydia-like organism,with comments on its pathogenic impact,life cycle and possible mode of transmission

Loripes lucinalis is a littoral bivalve which has already been confirmed to harbour endo-cellular sulfur-oxidizing bacteria within its gills. Examination of the digestive gland of L. lucinalis collected from the Moulin Blanc Beach in the Bay of Brest (Brittany, France) revealed the existence of an additional association involving a Chlamydia-like organism. Three different forms of Chlamydia-like bacteria were observed: reticulate rod-shaped cells, electron-dense cells and enlarged cells. The reticulate rod-shaped cells and the electron-dense bodies are thought to represent the germinal initial body and infectious form of the bacteria, respectively. The enlarged cells were always associated with what are believed to be spherical or icosahedral phages. Initial infestation seems to occur by phagocytosis at the apical pole of the digestive cells of the tubule and duct epithelia. Within the host cell, the bacteria undergo binary fission and budding, forming an inclusion which gradually fills up the cell. Inclusions are generally between 15 and 30 m in size, and > 85% of all individuals examined possessed inclusion bodies. The level of infestation varied between individuals, some being heavily colonized, but did not seem to be related to season. Histological and ultrastructural observations suggest that, once developed, the colony has three possible fates: (1) the cells will degenerate due to phage infection; (2) colony overcrowding will occur, causing the development of electron-dense bodies that will be released into the lumen; (3) the entire membrane-bound inclusion will be released into the lumen and subsequently into the pallial cavity. Inclusions within the pallial cavity may be ingested by the host or may even be phagocytized by bacteriocyte cells of the gill. It is proposed that this association could be a form of symbiosis and that L. lucinalis may, therefore, be a rare example of an organism adapted to harbour two very different symbioses.     

Ultrastructural changes in the lysosomal-vacuolar system in digestive cells of Mytilus edulis as a response to increased salinity

The effects of increasing salinity on the ultrastructural morphology of the lysosomal-vacuolar system in digestive cells of the common mussel Mytilus edulis were investigated in order to relate structural changes to previous biochemical and cytochemical observations. After 3 h of increased salinity, from 21 to 35%., the digestive cells showed an increase in numbers of heterolysosomes. There was some evidence of digestive cell breakdown, the contents forming membrane-bound bodies and being released into the tubule lumen. After 12 h of increased salinity, heterolysosomes were prevalent in the digestive cells. There was increased evidence for digestive-cell breakdown, many of the tubule lumina being packed with membrane-bound bodies. It is concluded that increasing salinity from 21 to 35%. stimulates the lysosomal-vacuolar system, as a result of autophagocytosis or apoptosis; this is consistent with the hypothesis that intracellular, lysosomally-mediated, catabolism of proteins is a source for free amino acids during the adaptation of mussels to increased salinity.     

The collagenous nature of problematical ligaments in crinoids (Echinodermata)

A major adaptive problem among comatulid crinoids is the need for a system of articulation and muscularization of the skeleton which provides rigidity in the cirri, arms, and pinnules for maintaining position and feeding in moving water masses, and also the ability to undergo occasional rapid flexions. Most skeletal ossicles of comatulids are united by so-called ligament fibers, while arm articulations possess masses of muscle as well as ligament. Light and electron microscopic work shows that the ligament consists primarily of collagen fibers. Similarity of this dual system of crinoid arm musculature to the muscles and collagenous catch apparatus of echinoid spines suggests that the crinoid ligaments may provide the needed rigidity to the crinoid skeleton, in a manner similar to that by which the echinoid catch apparatus can hold the spine tightly to the test in one position.     

Digestion in a subtidal population of Mercenaria mercenaria (Bivalvia)

The digestive processes of Mercenaria mercenaria (Linn.) were studied in a natural subtidal population. Hourly sampling of 4 to 5 quahogs was conducted over one 25 h period in August, 1978, at Woods Hole, Ma., USA. Crystalline style length did not vary significantly with time, tide, particulate C, N or suspended material in the water. Changes in secretory activity of the style sac epithelium were not evident from histological sections. Based on cellular morphological differences, digestive tubules were individually classified into one of 4 categories, indicative of the state of intracellular digestion within the digestive cells. All 4 tubule types were present in individuals from each sample hour, signifying that intracellular digestion occurs continually within the digestive gland. A 3 h time lag is evident between times of peak C, N or suspended materials and apparent peak levels of absorptive (Type II) tubules.Contribution No. 76 Marine Science Institute, Northeastern University; Nahant, MA 01908, USA     

Metabolic dysregulation and inhibition of spermatogenesis by gibberellic acid in rat testicular cells

Enzymatic and histological change in the testicular cells of rats treated orally and intradermally for 45 days with gibberellic acid (GBA) in independent studies is reported. Assay of hexokinase (HK), acid phosphatase (AcP) and alkaline phosphatase (AkP) in rat testicular tissue homogenate preparations yielded results that suggested changes in these enzyme activities relative to their respective controls. Histological studies showed loss of germ cells, derangement of the germinal cells, and reduction in the size of the seminiferous tubules and dystrophy of Leydig cells. More importantly decreased sperm count in the lumen was observed. A dysregulatory role is thus established for GBA in rat testicular cell function. This compound may serve as an inhibitor of testicular cell function.     

Defensive mechanisms and ecology of some tropical holothurians

The ecology, defensive behavior and toxicity of three species of reef flat holothurians (Actinopyga mauritiana, Holothuria atra and Holothuria difficilis) were studied at Eniwetok Atoll, Marshall Islands. The average diurnal population density of H. difficilis ranged from 1.4 to 32 holothurians/900 cm²; resting respiratory rates (0.05 ml O₂/g wet wt/h) were comparable during day and night; nourishment in H. difficilis may be primarily from bacteria and foraminifera in which about 2% of the dry weight of sediment consumed is utilized, and the species probably passes at least 3 g dry wt of sediment/m²/day (>1 kg/m²/year). A fundamental difference in energy flow is suggested: considerably more energy is passed from benthic algae to grazing and browsing fishes to predatory fishes on coral reefs whereas, in extra-tropical latitudes, more energy is shunted from benthic algae to invertebrates to predators. The effects of holothurin leading to death in fishes are irreversible. Holothuria difficilis is best protected from predation. Its body wall is toxic and it can accurately eject Cuvierian tubules, which are also toxic. The discharge of tubules was regulated by a circadian rhythm in May. Studies on holothurians and sponges suggest that many exposed coral reef invertebrates have evolved effective defensive mechanisms in association with high intensity predation.Supported by A.E.C. Contract AT (29-2)-226 with the University of Hawaii.     

Microarchitecture and function of the lophophore in the bryozoan Cryptosula pallasiana

The architecture and function of the lophophore of the marine bryozoan Cryptosula pallasiana (Moll) are described, including some new features not previously discovered in bryozoans. The nature of fluid movements within the lophophoral coelom during feeding activities is postulated on the basis of the arrangements of epithelia and muscles. Epithelial cells at the tentacle bases are blastemic in nature, and there is a ciliated pit of unknown function in the angle between every pair of tentacles. There are 6 nerves in each tentacle, including a pair of single-axon subperitoneal nerves. Neurosecretory-like vesicles and glycogen occur in some neurons of the ganglion. The basal lamina collagen has a diameter smaller than that previously recorded for an invertebrate. Filament dimensions are given for several different muscles. Tentacle muscles and lophophore retractor muscles are smooth. Thick paramyosin-like filaments up to 75 nm diameter occur in two muscle types. A new set of muscles is described: the basal transverse muscles of the tentacles.     

The effect of vitamin E supplementation on ovary and testis histopathology in Oreochromis niloticus exposed to deltamethrin

The effects of deltamethrin on the histology of Oreochromis niloticus testis and ovary and the protective effect of vitamin E supplementation were investigated. The fish were divided into four groups, i.e. a control group, a vitamin-E-treated group, a deltamethrin-treated group and a vitamin E+deltamethrin-treated group. In the control and vitamin-E-treated groups, the ovaries and testes were normal at the end of 7, 14, and 21 days. In the deltamethrin-treated group, deformed oocytes, atretic oocytes, melanomacrophage centers, and focal necrotic areas were noted in the ovaries. Deltamethrin also caused cell necrosis, nuclear pycnosis, decreasing number of spermatogenic cells, decreasing spermatozoa within the lumen of tubule, decreasing number of spermatocyte cells, hypertrophy and degeneration of spermatogonia cells, and increasing number of macrophages in testis. Vitamin E decreased some histopathological changes induced by deltamethrin, but did not confer complete protection.     

Effect of FSH and LH on testis during nonbreeding season in Calotes versicolor (Daud.)

Histological and biochemical studies carried out on the male reproductive organs of the Indian garden lizard, Calotes versicolor after the treatment with pituitary gonadotrophins (FSH and LH), showed a significant increase in the weight, protein content and diameter of testis, but decrease in its cholesterol. The spermatogonia, spermatocytes and spermatids increased significantly in the germinal epithelium of seminiferous tubule, and spermatozoa appeared in its lumen. The Leydig and Sertoli cells were hypertrophied with increase in their nuclear diameter. The epidymal weight, diameter and protein content also increased after gonadotrophins treatment. There was a significant decrease in the testicular cholesterol indicating the utilization of cholesterol for steroid hormone synthesis. The combined gonadotrophin (FSH + LH) treatment was more effective than the individual gonadotrophin treatment.     

Naked Dictyocha speculum — a new type of phytoplankton bloom in the Western Baltic

Chlamydomonas pulsatilla Wollenweber, a euryhaline, marine flagellate (isolated from rockpool at St. Andrew's, New Brunswick, Canada in 1980 by J. A. Hellebust), shows decreasing rates of activity of its four contractile vacuoles in the salinity range of 0 to 15% artificial seawater (ASW). Electron microscopy shows that the contractile vacuole complex persists as a spongiome (collection of small vesicles or tubules) in cells grown at salinities above the range for operation of contractile vacuoles. From calculations of rates of water expulsion, based on size and frequency of contraction of individual vacuoles determined by light microscopy, the time necessary to empty one cell volume increased from ca 20 min at 1% ASW to ca 600 min at 15% ASW. Analysis of inorganic and organic solute contents of cells grown at 1 and 5% ASW allowed the calculation of internal osmotic pressures. Estimates of hydraulic conductivities based on rates of water expulsion via contractile vacuoles and differences in internal and external osmotic pressures resulted in values ranging from 1.1 to 1.4×10^-14 m s^-1 Pa^-1 for individual cells. Growth experiments at low photon flux densities over a salinity range of 1 to 15% ASW, over which contractile vacuole activity varied by a factor of 30, showed little difference in growth rates. This indicates that the cost for operation of contractile vacuoles must be very low. The secretion of large molecular weight organic substances does not appear to be associated with the functioning of contractile vacuoles.     

Reproductive biology of the commercial sea cucumber Holothuria fuscogilva in the Solomon Islands

 Reproduction of Holothuria fuscogilva (Selenka, 1867) in the Solomon Islands was investigated over a 4 yr period (1994 to 1998) by macroscopic and microscopic examination of the gonad tubules, the gonad index (GI) method, histological examination of gametogenesis, and spawning-induction trials. The gonad consisted of numerous tubules that dominated the coelom of gravid specimens. New tubules appeared in March, and grew in size and extent of branching until they reached their maximum size and maturity in August. Spawning occurred from August to October, with the majority of gametes released during October, although it was only partial in many individuals. After spawning, the tubules appeared wrinkled and resorbed into the gonad basis. A five-stage gonad maturity scale based on the macroscopic appearance of the gonad tubules corresponded with discrete stages of gametogenesis identified by histology. Gametogenesis was initiated in mid-March, with oogenesis and spermatogenesis occurring in parallel, followed by the growing stage (May to July) which was marked by active gamete development. Successful induction of spawning during the breeding period corroborated the GI and histological data. The uniform growth of gonad tubules indicated that H. fuscogilva in the Solomon Islands does not conform to the progressive tubule recruitment model described for other holothurians. An important application of this study is that the appearance of gonad tubules, removed by biopsy, can be used to determine the gonad condition of wild adults or captive broodstock. Received: 28 June 1999 / Accepted: 8 March 2000     

Benzene-induced histopathological changes and germ cell population dynamics in testes of Sprague Dawley rats

Benzene has been considered as an occupational hematotoxin and leukemogen. The present study was conducted to determine the effects of oral administration of benzene on reproductive organs and testicular spermatogenesis in rats. Adult rats were divided into three weight matched groups (Gr. I-III) containing 6 each. Gr. I rats received vehicle only and served as control. Rats in Gr. II and III were fed orally with 0.5 and 1 ml kg(-1) dose of benzene for 14 and 9 days, respectively and autopsy was done on 15th and 10th day. Food and water intake and gross behavioral changes were recorded daily during the entire treatment. Results showed no significant change in reproductive organ weights viz. testis, epididymis and ventral prostate in benzene-treated (0.5 or 1 ml kg(-1)) rats than that in controls. But, caused a significant decrease (p < 0.005) in weights of seminal vesicles in rats treated with both 0.5 and 1 ml kg(-1) doses compared to control. In contrast, at higher dose (1 ml kg(-1)) of benzene, significant (p < 0.001) decline in body weight and 100% mortality was observed on day 10 of autopsy. In treated rats, testicular cytotoxicity was marked by multinucleated giant cells formation, cytoplasmic vacuolization, pyknosis of nuclei, chromatolysis, desquamation and dissolution of germ cells in tubular lumen. The quantitative analysis of spermatogenesis showed a significant (p < 0.001) decrease in number ofA-spermatogonia (in 1 ml kg(-1) dose only), primary spermatocytes (non-pachytene and pachytene) and spermatids (round and elongated) in treated as compared to control rats. The diameters of testicular tubules and Leydig cells nuclei were also significantly (p < 0.001) reduced in treated rats. A steady loss in food and water intake recorded and signs of ill health were observed in treated (0.5 or 1 ml kg(-1)) rats. Results of the study indicated antitesticular lantispermatogenic effects of benzene at 0.5 and 1 ml kg(-1) dose in rats.     

Comparative effects of trivalent and hexavalent chromium on spermatogenesis of the mouse

It has been repeatedly stressed that the biological effectiveness of chromium depends entirely on its oxidation state. The present work compares the histological effects of ingested trivalent and hexavalent chromium compounds on the mouse testes. The results showed comparable data of food consumption and gain in body weight. Both compounds produced ambiguous levels of degeneration in the outmost cellular layers of a number of seminiferous tubules, reduced the number of spermatogonia per tubule and the sperm count, and caused significant increases in the percentage of morphologically abnormal sperms. Generally, all these effects were more pronounced in animals which ingested the hexavalent chromium.