Glycogen and degeneration in the pyloric gland of Dendrodoa grossularia (Ascidiacea,Tunicata)

Summary Degeneration is observed in cells of the pyloric gland of Dendrodoa grossularia in which glycogen storage occurs. The ultrastructure of four phases of the degeneration cycle is described. Natural senescence seems to be the cause of the degeneration. Glycogen storage might be the result of metabolic disturbance, but its presence reveals the importance of glycogen in the function of the organ. The role of the pyloric gland is discussed.     

On the Presence of a Transverse System in Tunicate Muscle

Abstract Ultrastructural investigation of the caudal muscle cells in the tadpole larva of the ascidian Dendrodoa grossularia has shown that in this species, a transverse tubular system is present; diad and triad couplings are formed with the sarcoplasmic reticulum. Peripheral couplings of the sarcoplasmic reticulum are infrequent and have only been observed at the longitudinally apposed faces of the caudal muscle cells.     

Ultrastructure of the non-germinal cells in the testes of ascidians (Urochordata) and their role in the phagocytosis of sperm

 In three species of the Enterogona (Clavelina lepadiformis, Ciona intestinalis and Ascidiella aspersa) and three species of the Pleurogona (Dendrodoa grossularia, Styela clava and Molgula manhattensis) the testis was found to be invested by an epithelium separating the germ cells from the surrounding connective tissue or haemal sinuses. Each epithelial cell probably bears a single cilium, which in C. lepadiformis has a rootlet. Cilia are absent in S. clava. Lipid droplets are common and glycogen-rosettes occur in C. lepadiformis and D. grossularia. The basal plasmalemma varies from smooth to very irregular and in A. aspersa is anchored with hemidesmosomes. Except in S. clava, desmosome-like junctions occur between adjacent cells. Elimination of waste sperm following the reproductive season was observed to be undertaken by the epithelial wall cells in all species except C. lepadiformis. In C. intestinalis, D. grossularia, S. clava and M. manhattensis many of these cells detach and migrate to the interior of the testis where they continue and complete the phagocytosis of sperm. In C. lepadiformis, the non-germinal epithelium plays no role in the elimination of superfluous sperm which is probably phagocytosed, together with the rest of the body, by wandering trophocytes. Within the Urochordata the effectiveness of the testis epithelium as a blood-testis barrier varies, but is not correlated to modes of reproduction as postulated for other taxa. Accepted: 7 March 1997     

Microanatomy of the digestive tract and accessory organs of the Japanese flathead (Inegocia japonica Cuvier, 1829) (Scorpaeniformes,Platycephalidae)

The Japanese flathead, Inegocia japonica Cuvier, 1829 is a commercially important fish in small-scale coastal fisheries in Thailand; however, an explanation of its digestive biology is missing. This study describes the digestive tract and accessory organs of I. japonica, using morphological and histological methods. The fish (10 individual fish, 24.5 ± 0.98 cm in total length) were obtained from Libong Island, Thailand. Integrated morphological and histological data showed that the digestive tract was composed of oesophagus, stomach, pyloric caeca and intestine, with accessory organs. All digestive tracts consisted of four layers, including mucosa, submucosa, muscularis and serosa. Two stomach regions were identified (cardiac and pyloric stomachs). Several clusters of gastric glands were identified in the cardiac stomach. Each gland was a unicellular structure. The apical surface of this gland contained the vacuolar cell. The intestine was lined with a simple columnar structure with goblet cells that was similar to pyloric caecum. Goblet cells were rare in the anterior intestine, in contrast to the posterior intestine where goblet cells were abundant. The numerous of hepatocyte was mostly observed in the liver, whereas an exocrine acinar cell of pancreas was also identified. The results of our observations provided the first information of the digestive tract of I. japonica and can be applied to advanced study, such as physiology and histopathology.     

Evolution of digestion of carbohydrates in the separate parts of the digestive tract of the edible snail Helix lucorum (Gastropoda: Pulmonata: Stylommatophora) during a complete 24-hour cycle and the first days of starvation

In the present study we examined carbohydrase activities during a complete 24-h cycle and during the first days of starvation in both adult and juvenile snails. The results indicated the predominant role of the digestive gland in the secretions of the enzymes responsible for degradation of most of the carbohydrates tested. Salivary glands secreted some digestive enzymes but in amounts lower than secreted by the digestive gland. Enzymatic activities fluctuated during the first hours of digestion and also after the digestive tract was empty. The relatively high enzymatic activities recorded 24 h after the intake of food and during starvation could be due to the circadian rhythm of this species and/or to the participation of an existing microflora in the digestive tract of Helix lucorum. The double origin (exogenous and endogenous) of some digestive enzymes such as cellulases is discussed.Abbreviations CMC Carboxymethyl cellulose - LSD-test least significant difference test - PNP p-nitrophenyl - SA specific activity - U units     

Nitric oxide (NO) synthase immunoreactivity in the starfish Marthasterias glacialis

The neuroendocrine system of the starfish Marthasterias glacialis was investigated immunocytochemically using antisera specific for rat neuronal, bovine aortic endothelial, and mouse macrophage, nitric oxide (NO) synthases. Immunoreactivity was detected only with the antibodies specific for the neural enzyme, in the ectoneural and hyponeural tissues of the radial nerve cords and in the basiepithelial plexus and endocrine cells of the digestive tract. The pyloric stomach showed more immunoreactive structures than the other digestive organs, with the rectal caeca showing the least activity. Immunoreactive endocrine cells were located in the cardiac and pyloric stomachs and in the pyloric caeca. Co-localization of the enzyme immunoreactivity, and the staining for NADPH-diaphorase, demonstrate the presence of NO synthase in echinoderms. These results provide further evidence that NO is a neuronal messenger of early phylogenetic origin which has been conserved throughout evolution.     

Studies from the “UWL Helgoland” on the macrobenthic fauna of rocks and boulders in Lübeck Bay (western Baltic Sea)

During a fortnight's saturation mission with the Underwater Laboratory (UWL) Helgoland in May and June 1975, samples of the macrobenthic fauna of rocks and boulders at 15 m depth in the Lübeck Bay were collected from the same area as in October 1974, using a diver-operated suction sampler. Salinity varied from 11.1 to 15.0 S and water temperature from 7.5° C to 11.5° C. Visibility was usually about 3 to 4 m. On average, 10.449 individuals/m² were recorded in 1975 compared to 30.474 individuals/m² in 1974. Two species, the ascidianDendrodoa grossularia and the polychaetePolydora ciliata account for 90.8% of the total number of individuals sampled. Five species (Dendrodoa grossularia, Nereimyra punctata, Neoamphitrite figulus, Asterias rubens, Facelina drummondi) were significantly less abundant than in 1974, while two species and one group of species (Anaitides maculata, Corophium bonelli, Nemertini indet.) were more abundant in 1975. As in 1974, the angle of inclination of the substrate showed no relationship with species composition. In 1974, four motile benthic invertebrates,Nereimyra punctata, Idothea baltica, Diastylis rathkei, andFacelina drummondi were observed swarming. In 1975, onlyI. baltica was observed swarming and the individuals seemed to be less active than in 1974, perhaps related to the lower water temperatures recorded in May and June 1975 compared to those in October 1974.     

石磺消化系统的组织学观察

对石磺消化系统各部分结构进行组织学观察.石磺的消化系统由消化道和消化腺两部分组成.消化道包括口、食道、贲门胃、幽门胃、中肠和后肠,不具吻;消化腺包括肝胰腺、唾液腺和肛门腺.在光学显微镜下,消化道由粘膜层、粘膜下层、肌层和外膜4层组成;肌层主要为环肌,粘膜层主要为柱状细胞.肝胰腺甚为发达,组织结构显示肝胰腺由很多分支的腺管组成,腺管由腺细胞、分泌细胞等组成.唾液腺和肛门腺发达.     

Zur feinstruktur der Chorda dorsalis niederer Chordaten [Dendrodoa grossularia (v. Beneden) und Oikopleura dioica Fol]

Zusammenfassung Der Aufbau der Chorda dorsalis wurde bei zwei Tunikaten — Dendrodoa grossularia und Oikopleura dioica — elektronenmikroskopisch untersucht.Die Chorda von Dendrodoa besteht aus einem Strang solider, mit Dottereinschlüssen gefüllter, glykogenreicher Zellen. Sie sind von einer Paserschicht umhüllt, an der die quergestreifte Muskulatur des Schwanzes unmittelbar ansetzt.Die Chorda von Oikopleura besteht aus peripher gelegenen epithelartigen Zellen, die eine mit elastischem Material gefüllte, langgestreckte Vakuole umschließen. Ihr Cytoplasma enthält locker verteilt Golgi-Apparat, glattes und granuliertes endoplasmatisches Reticulum, Mitochondrien vom tubulären Typ und optisch leere Vakuolen. An der Faserhülle der Chorda setzen lateral die beiden Stränge quergestreifter Muskulatur an. Am Schwanz beider Formen werden ventrale und dorsale Flossen auf unterschiedliche Weise ausgebildet.

---

On the fine structure of the notochord of lower chordates (Dendrodoa grossularia and Oikopleura dioica)

Summary The fine structure of the notochord of two tunicate species — Dendrodoa grossularia and Oikopleura diowa — was investigated with the electron microscope.In Dendrodoa, the notochord consists of a cord of solid glycogen-rich cells, which contain numerous big yolk droplets. These cells, which are otherwise poor in cytoplasmic organelles, are surrounded by a layer of fibrils, to which the muscle cells of the tail are firmly attached.In Oikopleura the notochord consists of peripheral flat epitheliumlike cells, which surround a single central rodlike vacuole filled with elastic material. The cytoplasm of the notochordal cells contains loosely distributed Golgi apparatus, smooth and granulated endoplasmic reticulum, mitochondria of the tubular type and empty-appearing vacuoles. The two muscle tracts of the tail are laterally attached to the fibrous sheath of the notochord. The dorsal and ventral fin of the tail exhibit in both animals a completely different ultrastructure.

---

---

Die Untersuchungen wurden mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.

---

Herrn Prof. Dr. A. Remane danken wir für Hinweise zur Diskussion.

---

Herrn Prof. Dr. W. Bargmann danke ich für Überlassung eines Arbeitsplatzes im Anatomischen Institut Kiel.     

Histological structure of the digestive tract,liver, and pancreas of Ambassis nalua (Hamilton, 1822) with ultrastructural details of the gastric gland

The scalloped perchlet Ambassis nalua is one of the dominant fishes in the Estuarine Pranburi River, Thailand. It is suggested that this fish is in the secondary trophic level with a carnivorous nature. Studies on digestive system will help us further identify the niche of this species in the food web/food chain. The present study therefore aimed to report the detailed structure and ultrastructure of A. nalua digestive system. Fish samples (n = 30) with a total length of 5.7 ± 0.5 cm were obtained using beach seines from the Estuarine Pranburi River. Their digestive tract length and intestine coeficient were 3.6 ± 0.07 cm and 0.91, respectively. Light microscopic observation showed that the digestive wall comprised four layers, namely mucosa, submucosa, muscularis, and serosa. The prominent mucous-secreting cells were found in the mucosal oesophagus. The stomach had many gastric folds, with height and width being 649.76 ± 85.15 and 370.30 ± 68.56 μm, respectively. Gastric glands were found in the anterior stomach but not in the posterior stomach. Each gastric gland was made up of a single type of columnar cells. The gastric cells were ultrastructurally characterized by numerous mitochondria and well-developed secretory granules of varying sizes. A few small vacuoles were also identified in the apical area of the gastric cells. The intestine had two regions (anterior and posterior intestines), and pyloric caecum was absent. The density of the goblet cell was significantly higher in the posterior intestine. These results provide basic knowledge of the digestive system of A. nalua, and the low intestine coefficient and the absence of pyloric caecum suggest the carnivorous feeding habit of this species.     

野生与养殖黄鳍鲷消化道形态组织结构

采用常规石蜡组织切片的方法对野生和养殖黄鳍鲷(Sparus latus)消化道的形态组织结构进行了比较观察。结果表明,野生和养殖黄鳍鲷的消化道存在一定差异。(1)形态学研究表明,食道粗而短,胃呈V形,分为贲门部、胃体部和幽门部,胃与肠的连接处有4条幽门盲囊,肠道在体腔内迂回两个回折。野生黄鳍鲷牙齿更为坚硬锋利,体腔中脂肪较少,消化道更为粗短。野生和养殖黄鳍鲷的肠道系数分别为0.71±0.03和0.94±0.12。(2)组织学研究表明,食道黏膜上皮由扁平细胞层和杯状细胞层组成,杯状细胞发达。胃黏膜由单层柱状上皮组成,无杯状细胞,贲门部和胃体部胃腺发达。幽门盲囊组织学特征与肠相似,上皮为柱状上皮,其中的杯状细胞少于肠。肠中,前肠杯状细胞最多,中肠次之,后肠最少。直肠杯状细胞多于肠。野生与养殖黄鳍鲷组织学的区别在于,消化道相同部位养殖鱼的杯状细胞多于野生鱼,野生鱼的肌层厚度大于养殖鱼。黄鳍鲷消化道的形态组织结构与其生活环境和食物是相关的。     

Nutritional indices of juvenile Caribbean spiny lobsters in a Mexican reef lagoon: Are changes over a 10-year span related to the emergence of Panulirus argus Virus 1 (PaV1)?

We compared two indices of nutritional condition, the relative weight of the digestive gland (RWDG) and the ratio of weight to carapace length (weight/CL ratio), of the first three benthic phases, algal juveniles (7–25 mm CL), postalgal juveniles (25–45 mm CL), and subadults (45–79 mm CL) of Caribbean spiny lobster (Panulirus argus) dwelling in a shelter-poor reef lagoon before (1995) and after (2005) deployment of large artificial shelters (“casitas”) in 1998. To provide an experimental baseline, in 2005 we also conducted a laboratory experiment to assess the effect of starvation on both nutritional indices. Through 25-day trials, starvation significantly decreased the mean RWDG of all three juvenile phases, but did not affect their weight/CL ratios. In the field, casitas significantly enhanced juvenile lobsters relative to control sites and to prior values, but between 1995 and 2005 there was a significant decrease in condition of algal juveniles (both indices) and postalgal juveniles (only the RWDG), but not of subadults. These results would appear to counter previous findings that, for P. argus, food is not limiting and nutritional condition is not density-dependent. However, in 2000, a new pathogenic virus that has a distinct predilection for small juveniles, Panulirus argus Virus 1 (PaV1), emerged in the reef lagoon. Among other effects, this lethal virus causes a marked atrophy of the digestive gland. Through the laboratory trials and in the field, prevalence of visibly diseased individuals was highest in algal juveniles and lowest in subadults. These individuals were excluded from statistical analyses, but the disease progression may affect the digestive gland progressively given that newly infected lobsters take several weeks to become visibly diseased. Up to 2006, PaV1 had not significantly impacted lobster density on casita sites, but because P. argus is highly gregarious and transmission of PaV1 appears to be mainly by contact, futher investigation is required. These issues are relevant to enhancement of spiny lobsters and assessment of ecological impacts of marine diseases.     

Effects of complexing agents on the distribution of Hg(II) species provided by food to starfish Leptasterias polaris

Abstract

Mature starfish Leptasterias polaris were exposed to labelled mercury (II) species via food contaminated at a level of 5.0 μg g^?1. The distribution of inorganic Hg and methylmercury (MeHg) in starfish organs and tissues and the effect of a series of complexing agents on mercury translocation between organs and tissues were examined over a 24-h period. The distribution of mercury species in coelomic fluid components, ammonia excretion rate and mercury excretion were also measured. The highest concentrations were observed in the stomach (the source organ) and in pyloric caecum (up to 0.32 μg g^?1 wet weight for inorganic Hg and 0.22 μg g^?1 for MeHg). Concentrations of MeHg in gonads ranged from ≤ 0.01 to 0.08 μg g^?1 whereas concentrations of inorganic Hg never exceeded 0.06 μg g^?1. In all studied cases, mercury concentration was very low the coelomic fluid (≤ 0.01 μg g^?1). The short-term distribution of Hg species via contaminated food in starfish L. polaris seems to be controlled by the haemal system, a primitive circulatory system responsible for the transport of soluble nutrients from the digestive track towards organs and tissues, but a possible role of the coelomic fluid can not be excluded. Very low Hg contents were observed in gonads and in the coelomic fluid which fills the general cavity. Except for mercaptoethanol (merOH) and dimercaptosuccinic acid (DMSA), the addition of complexing agents to the food had little effect on the distribution of Hg species. MerOH appeared as an efficient carrier for methylmercury transport through the digestive system. DMSA enhanced the translocation of inorganic mercury from stomach and pyloric caecum toward external tissues and markedly increased its excretion.     

A comparative study of chitinase activity in digestive tract of Serranus cabrilla and Serranus scriba

Chitinolytic activity was searched for in oesophagus, stomach, intestine, pyloric caeca and liver of two sympatric closely related species of Serranidae (Teleostei, Perciformes): Serranus cabrilla (L.) and Serranus scriba (L.) living near the rocky shore of Calvi (Corsica, Mediterranean Sea). The chitinase activity was much higher in stomach than in other parts of the digestive tract and liver in both species. In extracts of gastric mucosa, the chitinase activity was more important in S. cabrilla than in S. scriba; an appointed situation was found in the extracts of intestine and pyloric caeca. These findings are discussed as aspects of the ability of these species to share very near ecological niches.     

Morphology and histochemical analysis of glycoproteins in the digestive tract of Dabry's sturgeon

The aim of this study is to explore the morphology and histochemistry of the digestive tract of Acipenser dabryanus (Duméril, 1869). The digestive system of the Dabry's sturgeon contained an oropharyngeal cavity, esophagus, stomach, pyloric caeca, duodenum, valvula intestine, and rectum. Both the pyloric caeca, and the valvula intestine, which belonged to elasmobranch, were present simultaneously in the digestive system of the Dabry's sturgeon, which indicated that it was an archaic species. According to the results, we found four types of mucous cells exist in the digestive tracts of the Dabry's sturgeon. The statistical results showed that there were various types and densities of mucous cells in different parts. In comparison, large quantities of mucous cells existed in intestines. The difference in type and distribution of mucous cells are closely related to the functions of the organs where they are found; 5‐hydroxytryptamine were used to identify the endocrine cells in the digestive tract of the Dabry's sturgeon.The 5‐hydroxytryptamine immunoreactive cells were distributed throughout the digestive tract except the esophagus.     

Pathophysiologic alterations in Biomphalaria glabrata infected with Angiostrongylus costaricensis

Hemolymph glucose, alkaline phosphatase, lactic dehydrogenase, and creatine phosphokinase in Biomphalaria glabrata infected with Angiostrongylus costaricensis were significantly higher on day 27 postinfection (PI) than in uninfected snails. Hemolymph total calcium from infected snails was less on days 6, 12, and 27 PI than that from controls. Total hemolymph protein was similar for controls and infected animals during the entire study. Throughout the study the mean number of amoebocytes/mm³ hemolymph from infected snails was significantly less than that for controls. Mean total wet weights of digestive gland and foot muscle from infected and uninfected snails was similar throughout the study. Mean μg glycogen/mg wet weight of digestive gland from infected snails was significantly greater on days 24, 27, and 28 PI than that from controls. Mean μg glycogen/mg wet weight of foot muscle from infected snails was significantly reduced between days 12 and 28 PI from that of uninfected snails. It is suggested that hemolymph glucose and digestive gland glycogen in infected snails are augmented by glycogen breakdown in the foot muscle of parasitized animals. Elevations in hemolymph enzymes are due to tissue destruction by larvae emerging from the foot muscle of infected snails. Parasite-induced derangements in shell metabolism underlie observed changes in hemolymph calcium in infected snails.     

Cloning of cDNAs and hybridization analysis of lysozymes from two oyster species, Crassostrea gigas and Ostrea edulis

In bivalve molluscs including oysters, lysozymes play an important role in the host defense mechanisms against invading microbes. However, it remains unclear in which sites/cells the lysozyme genes are expressed and which subsequently produced the enzyme. This study cloned lysozyme cDNAs from the digestive organs of Pacific oyster Crassostrea gigas and European flat oyster Ostrea edulis. Both complete sequences of two oysters' lysozymes were composed of 137 amino acids. Two translated proteins present a high content in cysteine residues. Phylogenetic analyses showed that these oysters' lysozymes clustered with the invertebrate-type lysozymes of other bivalve species. In the Pacific oyster, lysozyme mRNA was expressed in all tissues except for those of the adductor muscle. In situ hybridization analyses revealed that lysozyme mRNA was expressed strongly in basophil cells in the digestive gland tubule of C. gigas, but not in digestive cells. Results indicated that the basophil cells of the oyster digestive gland are the sites of lysozyme synthesis.     

The hypervariable domain of the mitochondrial control region in Atlantic spiny lobsters and its potential as a marker for investigating phylogeographic structuring

Atlantic spiny lobsters support major fisheries in northeastern Brazilian waters and in the Caribbean Sea. To avoid reduction in diversity and elimination of distinct stocks, understanding their population dynamics, including structuring of populations and genetic diversity, is critical. We here explore the potential of using the hypervariable domain in the control region of the mitochondrial DNA as a genetic marker to characterize population subdivision in spiny lobsters, using Panulirus argus as the species model. The primers designed on the neighboring conserved genes have amplified the entire control region (approx. 780 bases) of P. argus and other closely related species. Average nucleotide and haplotype diversity within P. argus were found to be high, and population structuring was hypothesized. The data suggest a division of P. argus into genetically different phylogeographic groups. The hypervariable domain seems to be useful for determining genetic differentiation of geographically distinct stocks of P. argus and other Atlantic spiny lobsters.     

The suitability of gallium as a substitute for aluminum in tracing experiments

Aluminum is a toxic metal whose complex aquatic chemistry, mechanisms of toxicity and trophic transfer are not fully understood. The only isotope of Al suitable for tracing experiments in organisms—²⁶Al—is a rare, costly radioisotope with a low emission energy, making its use difficult. Gallium shares a similar chemistry with Al and was therefore investigated as a potential substitute for Al for use in aquatic organisms. The freshwater snail, Lymnaea stagnalis was exposed to either Al or Ga (0.0135 mM) under identical conditions for up to 40 days. Behavioural toxicity, metal accumulation in the tissues, and sub-cellular partitioning of the metals were determined. Al was more toxic than Ga and accumulated to significantly higher levels in the soft tissues (P < 0.05). The proportion of Al in the digestive gland (DG; detoxificatory organ) relative to other tissues was significantly lower than that of Ga (P < 0.05) from day 14 onwards. There were also differences in the proportions of Al and Ga associated with heat stable proteins (HSPs) in the digestive gland, with significantly more HSP present in the DGs of snails exposed to Al, but significantly less Al than Ga associated with the HSP per unit mass protein present. From this evidence, we conclude that Ga may be of limited use as a tracer for Al in animal systems.     

DNA damage (comet assay) and 8-oxodGuo (HPLC-EC) in relation to oxidative stress in the freshwater bivalve Unio tumidus

The relationships between DNA damage and oxidative stress in the digestive gland, gills and haemocytes of the freshwater bivalve Unio tumidus were investigated. Two markers of genotoxicity were measured: DNA breaks by means of the comet assay, and oxidative DNA lesions by means of 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo) measured using high-performance liquid chromatography (HPLC) coupled to electrochemical detection. Lipid peroxidation was evaluated by measuring malondialdehyde (MDA) tissue levels. Effects were studied after exposure of bivalves for 6 days to benzo[a]pyrene (B[a]P) (50 and 100 μg?l^?1) and ferric iron (20 and 40 mg?l^?1), applied alone or in combination. Lipid peroxidation in the digestive gland and gills resulted from exposure to Fe³⁺ or B[a]P whatever the concentrations tested. DNA oxidatively formed lesions were induced in the two tissues at a higher level after B[a]P exposure than after Fe³⁺ treatment. No significant dose–response relationship was found with the two compounds and no synergistic effect was observed between Fe³⁺ and B[a]P. The gills appeared less sensitive than the digestive gland to DNA lesions expressed as 8-oxodGuo and comet results. Good correlations were noted between 8-oxodGuo and comet. MDA and DNA damage did not correlate as well, although it was stronger in the digestive gland than in the gills. Production of mucus by the gills likely served to prevent lesions by reducing the bioavailability of the chemicals tested, which could explain that dose–effect relationships and synergistic effects were not observed.