The ultrastructure of the posterior gut and caecum inAlona affinis (Crustacea,Cladocera)

Summary The posterior midgut, the anterior hindgut and the caecum ofAlona affinis were studied by transmission electron microscopy. The caecum arises from the junction of the entodermal midgut and the ectodermal hindgut. It consists of gastrodermis and epidermis. Because of the ultrastructural similarity of the caecum with the posterior midgut and the anterior hindgut it is concluded that the caecum is a functional supplement of the latter gut parts. But the functional significance of these gut parts is poorly understood. Some ultrastructural features suggest that they contribute in excretion and salt regulation.Abbreviations Au autolysosome - Ba bacteria - Bl basal lamina - Ca canaliculi - Cae caecum - Crc crypt cell - Cu cuticle - Ec epicuticle - Ep epidermis - G Golgi complex - Hae hemolymph - Hc head cell - La basal labyrinth - Lu gut lumen - lMv long microvilli - Mg midgut - Mi mitochondria - mC mitochondria rich cell - nBc new border cell - Nc neck cell - Nu nucleus - oBc old border cell - Pc procuticle - pMe peritrophic membrane - pMg posterior midgut - Rm ring muscle - S secretion (? uncertain) - sMv short microvilli - vC vacuolated cell     

The anatomy and histology of the midgut and hindgut of Charybdis (Goniohellenus) truncata (Fabricius, 1798) (Decapoda: Brachyura)

The midgut of C. (G.) truncata accounts for half of the postgastric intestinal tract. The paired anterior midgut caeca arise just behind the pyloric stomach, on either side of the midgut. The unpaired posterior midgut caecum arises dorsally at the rear end of the midgut, where this joins the hindgut. The midgut and its caeca help in the digestive absorption of food. The hindgut is of ectodermal origin and is lined with chitin of a collagenous nature. The connective tissue of the anterior part of the hindgut is packed with tegumental glands whose secretion contains both sulphated and weakly acidic mucosubstances, which facilitate the passage of faecal matter and help to bind food particles. The digestive gland - the hepatopancreas - opens into the anterior part of the midgut, below the anterior midgut caeca. Histologically, its tubules contain three different types of cells - "F", "R" and "B" cells.     

Morphology and ultrastructure of the adult ovarian cycle in Mithracidae (Crustacea,Decapoda, Brachyura,Majoidea)

The ultrastructure of the ovary during development and yolk production is poorly known in Brachyura and Majoidea in particular. Here, we describe the histology, histochemistry and ultrastructure of the adult ovarian cycle in four Mithracidae species from three different genera: Mithrax hispidus, Mithrax tortugae, Mithraculus forceps and Omalacantha bicornuta. All species showed a similar pattern of ovarian development and vitellogenesis. Macroscopically, we detected three stages of ovarian development: rudimentary (RUD), developing (DE) and mature (MAT); however, in histological and ultrastructural analyses, we identified four stages of development. The oocytes of the RUD stage, during endogenous vitellogenesis, have basophilic cytoplasm filled with dilated rough endoplasmic reticulum. The reticulum lumen showed many granular to electron-dense materials among the different stages of development. The Golgi complexes were only observed in the RUD stage and are responsible for releasing vesicles that merge to the endogenous or immature yolk vesicles. At the early DE stage, the oolemma showed many coated and endocytic vesicles at the cortex. The endocytic vesicles merge with the endogenous yolk to form the exogenous or mature yolk vesicles, always surrounded by a membrane, characterizing exogenous vitellogenesis. The exogenous yolk vesicles comprise glycoproteins, showing only neutral polysaccharides. At the late DE stage, endocytosis still occurs, but the amount of endogenous yolk decreases while the exogenous yolk increases. The late DE stage is characterized by the beginning of chorion production among the microvilli. The MAT stage is similar to the late DE, but the endogenous yolk is restricted to a few cytoplasmic areas, the ooplasma is filled with exogenous yolk, and the oolemma has very few coated vesicles. In the MAT stage, the chorion is fully formed and shows two electron-dense layers. The ovarian development of the species studied has many similarities with the very little known Majoidea in terms of the composition, arrangement and increment of the yolk vesicles during oocyte maturation. The main differences are in the vitellogenesis process, where immature yolk formation occurs without the direct participation of the mitochondria but with the participation of the rough endoplasmic reticulum in the endogenous phase.     

The complete mitogenome of the hydrothermal vent crab Xenograpsus testudinatus (Decapoda, Brachyura) and comparison with brachyuran crabs

In this study, we analyzed the complete mitochondrial (mt) genome of a hydrothermal vent crab Xenograpsus testudinatus (Decapoda: Brachyura) obtained from the hydrothermal vents off Kueishantao Island, Taiwan, which extend from the deep sea Okinawa Trench. The mitogenome of X. testudinatus was 15,796 bp in length and contained the same 37 genes (e.g. 2 rRNAs, 22 tRNAs, and 13 PCGs) found in other metazoan mitogenomes. Analysis of the structural mt gene order in X. testudinatus revealed that the 13 PCGs, excluding a translocation of ND6-Cyt b cluster, were similarly ordered when compared to the pancrustacean ground pattern; however the tRNAs were severely rearranged. Phylogenetic analysis of decapod mitogenomes showed that the molecular taxonomy of the vent crab was in accordance with its morphological systematics. Together, these findings suggest that the vent crab studied here has little mitochondrial genetic variation when compared with morphologically defined conspecifics from other marine habitats.     

The Grapsidae and Ocypodidae (Decapoda: Brachyura) of Tanzania

Twenty-nine species of the Grapsidae and 15 species of the Ocypodidae were collected from the littoral zone in the area around Dar es Salaam. Nine species of Grapsidae and three of Ocypodidae were new records for the East African area, bringing the total of recorded species for the region to 35 and 26 respectively. The distribution of these crabs indicates a very limited endemism in the western Indian Ocean, with the majority of species forming an attenuated extension of the abundant Indo/West Pacific brachyuran fauna.
The habitat preference of each species is described. Within each subfamily there is a relative uniformity in general habitat preference, mode of life and manner of feeding. The species of each subfamily are then separated by substrate choice and preferred level on the shore. The greatest numbers of species occur in the most diverse habitats—the creek mangrove, the coastal mangrove and the reef flat.     

Cleavage and gastrulation in the shrimp Penaeus (Litopenaeus) vannamei (Malacostraca, Decapoda, Dendrobranchiata)

While most malacostracan crustaceans develop through superficial cleavage, in the Amphipoda, Euphausiacea, and Dendrobranchiata (Decapoda) cleavage is complete. Euphausiaceans and dendrobranchiate shrimp share a similar early cleavage pattern, early cleavage arrest and ingression of mesendoderm progenitor cells, a ring of crown cells (prospective naupliar mesoderm) around the blastopore, and hatching as a nauplius larva. Yet recent phylogenies do not support a close relationship between Euphausiacea and Decapoda. In addition, some variation is reported in the timing of mesendoderm cell arrest and number of crown cells for a number of dendrobranchiates. To determine the representative pattern of development in the Dendrobranchiata, embryos of the Pacific white shrimp Penaeus (Litopenaeus) vannamei were stained with Sytox Green to label chromosomes and nuclei and examined with confocal microscopy. The early cleavage pattern, mesendoblast arrest and subsequent ingression at the 32-cell stage, presence of 8 initial crown cells, and fates of the mesendoblasts are the same for P. vannamei (family Peneaeidae) and Sicyonia ingentis (family Sicyoniidae). The lineage of the primordial endoderm cells differs from that reported for P. kerathurus. These characters were discussed in the context of the evolution of development in the Dendrobranchiata and in comparison to the Euphausiacea.     

The ultrastructure of the midgut epithelium in millipedes (Myriapoda,Diplopoda)

The midgut epithelia of the millipedes Polyxenus lagurus, Archispirostreptus gigas and Julus scandinavius were analyzed under light and transmission electron microscopies. In order to detect the proliferation of regenerative cells, labeling with BrdU and antibodies against phosphohistone H3 were employed. A tube-shaped midgut of three millipedes examined spreads along the entire length of the middle region of the body. The epithelium is composed of digestive, secretory and regenerative cells. The digestive cells are responsible for the accumulation of metals and the reserve material as well as the synthesis of substances, which are then secreted into the midgut lumen. The secretions are of three types – merocrine, apocrine and microapocrine. The oval or pear-like shaped secretory cells do not come into contact with the midgut lumen and represent the closed type of secretory cells. They possess many electron-dense granules (J. scandinavius) or electron-dense granules and electron-lucent vesicles (A. gigas, P. lagurus), which are accompanied by cisterns of the rough endoplasmic reticulum. The regenerative cells are distributed individually among the basal regions of the digestive cells. The proliferation and differentiation of regenerative cells into the digestive cells occurred in J. scandinavius and A. gigas, while these processes were not observed in P. lagurus. As a result of the mitotic division of regenerative cells, one of the newly formed cells fulfills the role of a regenerative cell, while the second one differentiates into a digestive cell. We concluded that regenerative cells play the role of unipotent midgut stem cells.     

Growth and reproduction in the freshwater crab, Potamon fluviatile (Decapoda, Brachyura)

SUMMARY. 1. The patterns of relative and absolute growth and the reproductive biology of the freshwater crab Potamon fluviatile were studied in a natural population inhabiting a hill stream close to Florence, Italy, over an annual cycle. Periodical inspections of a stretch of the stream were made and morphological and anatomical analysis carried out.
2. As in other decapods, the females are smaller than the males (they can however occasionally reach larger dimensions). This may be determined by a lengthening of the moulting interval in the females, by their higher energetic cost of reproduction (since reproduction occurs simultaneously with moulting, at least in this habitat) and/or by a higher mortality rate resulting from the risks associated with carrying eggs and vagility.
3. The analysis of relative growth in secondary sexual characters (abdomen width and major chela length) with respect to carapace length, shows that the pre-puberty and puberty ecdysis occur at 15 and 35 mm carapace length respectively, which was also confirmed by the gonad weight, vas deferens in males, and the onset of vitellogencsis in females.
4. A delay between anatomical and functional (i.e. the ability to copulate successfully) maturity was observed in the males. Being larger may be an advantage in intra-sexual competition for mating, but larger males, being more vagile, are also more likely to meet receptive females. The females may mate before their puberty moult and store sperms in their seminal receptacles for when they attain full maturity; this could be adaptive since opportunities of encountering males are few and far between in their adult phase, characterized by their vagile and amphibious habits.     

The ultrastructure of plasma and thylakoid membrane vesicles from the fresh water cyanobacteriumAnacystis nidulans adapted to salinity

Summary Photoautotrophically growing cultures of the fresh water cyanobacteriumAnacystis nidulans adapted to the presence of 0.4–0.5 M NaCl (about sea water level) with a lag phase of two days after which time the growth rate reassumed 80–90% of the control. Plasma and thylakoid membranes were separated from cell-free extracts of French pressure cell treatedAnacystis nidulans by discontinuous sucrose density gradient centrifugation and purified by repeated recentrifugation on fresh gradients. Identity of the plasma and thylakoid membrane fractions was confirmed by labeling of intact cells with impermeant protein markers prior to breakage and membrane isolation. Electron microscopy revealed that each type of membrane was obtained in the form of closed and perfectly spherical vesicles. Major changes in structure and function of the plasma membranes (and, to a much lesser extent, of the thylakoid membranes) were found to accompany the adaptation process. On the average, diameters of plasma membrane vesicles from salt adapted cells were only one-third of the diameters of corresponding vesicles from control cells. By contrast, the diameters of thylakoid membrane vesicles were the same in both cases.Freeze-etching the cells and counting the number of membrane-intercalating particles on both protoplasmic and exoplasmic fracture faces of plasma and thylakoid membranes indicated a roughly 50% increase of the particle density in plasma membranes during the adaptation process while that in thylakoid membranes was unaffected. Comparison between particle densities on isolated membranes and those on corresponding whole cell membranes permitted an estimate as to the percentage of inside-out and right-side-out vesicles. Stereometric measurement of particle sizes suggested that two distinct sub-populations of the particles in the plasma membranes increased during the adaptation process, tentatively correlated to the cytochrome oxidase and sodium-proton antiporter, respectively. The effects of salt adaptation described in this paper were fully reversed upon withdrawal of the additional NaCl from the growth medium (deadaptation). Moreover, they were not observed when the NaCl was replaced by KCl.Abbreviations CM cytoplasmic or plasma membrane - ICM intracytoplasmic or thylakoid membrane - EF exoplasmic fracture face - PF protoplasmic fracture face - DABS diazobenzosulfonate; Hepes N-2-hydroxyethylpiperazine-N-2-ethane-sulfonate - PMSF phenylmethylsulfonylfluoride Dedicated to the memory of Professor Oswald Kiermayer     

Epithelial ultrastructure of the midgut of the horsefly, Hybomitra schineri (Tabanidae)

The study of the midgut of Hybomitra schineri by means of electron microscopy has shown that epithelial cells are of cylindrical shape. According to the electron density of cytoplasm the cells can be arranged into three subtypes. The apical surface of all the cells is covered by microvilli with filamentous glycocalyx. The peritrophic membrane of the 1st type is formed in females only during the blood digestion. Two types of secretion of digestive enzymes (merocrine and macroapocrine ones) were found, the latter being prevalent at the carbohydrate feeding.     

Male courtship cycles in three species of tropical Ilyoplax crabs (Decapoda,Brachyura, Ocypodidae)

The courtship behaviour and cycles of male courtship activity and colouration of Ilyoplax orientalis, I. delsmani and I. gangetica were studied in the field in Malaysia and Thailand. Each species had a distinctive chela waving or beckoning display. Depending on species, the chelipeds, carapaces, or both of waving males blanched to white in contrast to the cryptic colour of nonwaving males and females. All three of these tropical Ilyoplax exhibited semilunar cycles in male waving activity at the colony level. It was confirmed for I. orientalis that individual males cycled each semilunar period between waving and non-waving phases and exhibited different behaviour toward females during these two behavioural phases.     

Foregut morphology and ontogeny of the mud crab Dyspanopeus sayi (Smith, 1869) (Decapoda,Brachyura, Panopeidae)

The morphology of the foregut of the Say's mud crab Dyspanopeus sayi was described in adults and larvae. The ossicle system was illustrated based on a staining method with Alizarin-Red. The gastric teeth and cardio-pyloric valve were dissected and examined using optical and scanning electron microscopy. In the adults, the morphology of ossicles and gastric teeth of D. sayi is very similar to the related species Rhithropanopeus harrisii. The foregut of first zoea (ZI) presented a functional cardio-pyloric valve while the filter press was lacking. The filter press was observed in the pyloric chamber from ZII. The most significant changes in morphology take place after metamorphosis from ZIV to megalopa, including the occurrence of the gastric mill. The organization and morphology of many megalopal foregut ossicles are recognizable in the adult phase, although the morphology of the gastric teeth differs from the morphology of adults. A correlation of gastric mill structures with food preferences and their contribution to the phylogeny are briefly discussed.     

Phylogeny, biogeography and a new taxonomy for the Gecarcinucoidea Rathbun, 1904 (Decapoda: Brachyura)

Phylogenetic relationships of gecarcinucoid freshwater crabs were investigated, based on morphology of the male second gonopod. In addition, a comparison of sequences from the mitochondrial large subunit rRNA gene helped to resolve the phylogeny of this group and relationships to other Old World freshwater crabs. As a result, we recognise two sister groups within the Gecarcinucoidea, the African Deckeniidae and the Asian Gecarcinucidae. Deckeniidae includes three monophyletic clades, the Deckeniinae in East Africa and on the Seychelles, the West African Globonautinae and the Malagasy Hydrothelphusinae. Gecarcinucidae comprises two sister groups, the Gecarcinucinae with representatives in Sri Lanka, India and southeast Asia, and the Parathelphusinae in India, southeast Asia, the Sundaic Islands and Australia. Interpretation of our phylogenetic results leads us to propose a new biogeographic hypothesis for the Gecarcinucoidea. Most likely, the gecarcinucoid freshwater crabs have an African origin; their distribution can be explained by successive events of dispersal. This model can be correlated with palaeogeographical and palaeoclimatological data for the Cenozoic, suggesting a gecarcinucoid dispersal to Asia via the “Lemurian Stepping-Stones”, a chain of islands in the West-Indian Ocean that were emergent in times of low sea levels during the Oligocene.     

The Host Range ofEccritotarsus catarinensis(Heteroptera: Miridae), a Potential Agent for the Biological Control of Waterhyacinth (Eichhornia crassipes)

The host range ofEccritotarsus catarinensiswas determined using 33 plant species to assess the risk of using this insect, a native of South America, for the classical biological control of waterhyacinth (Eichhornia crassipes) in Australia and Papua New Guinea. The results, in conjunction with the results of Hillet al.(1999) who tested 67 species (mostly South African), strongly suggest thatE. catarinensisis restricted to the Pontederiaceae, a family of aquatic plants. All five species of Pontederiaceae in the Australian testing,E. crassipes, Pontederia cordata, Monochoria vaginalis, M. cyanea,andM. australasica,were suitable for insect development. Colonies persisted for at least four generations onE. crassipes, P. cordata,andM. vaginalis.Two-way choice and multiple-choice preference trials were conducted and discussed.E. catarinensisdid not exhibit a clear preference for waterhyacinth over other Pontederiaceae in these trials. Most oviposition occurred into the upper surface of the lamina during laboratory testing despite observations that the underside was preferred in the field. Although not considered suitable for release in Australia, this insect may be useful in other countries where more serious waterhyacinth problems occur and whereM. vaginalisis a serious weed, such as in Southeast Asia.     

Mycobiota associated with larval mines of Thrypticus truncatus and T. sagittatus (Diptera, Dolichopodidae) on waterhyacinth, Eichhornia crassipes, in Argentina

Thrypticus truncatus is a candidate agent for biocontrol of waterhyacinth; the larvae of this diptera mine in the petioles and feed on the phloem in the vascular bundles. The mycobiota associated with T. truncatus and T. sagittatus mines was investigated during two surveys undertaken in the spring and autumn in the Delta of the Paraná River, Argentina. Isolations were made from the mines and larval feeding points, as well as from the larvae, following dissection of the petioles, and plated onto agar. Young and upper parts of the petioles without Thrypticus mines were used as controls. Twenty eight fungal species were isolated from the mines. Pestalotiopsis guepinii, Mucor attenuatus, Phoma tropica, Achlya americana, Fusarium avenaceum, Cladosporium cladosporioides, Clonostachys rosea, Epicoccum purpurascens, Plectosporium tabacinum, Alternaria alternata, and Acremonium sp. were the most common fungi associated with mines and feeding points. Cladosporium cladosporioides, Cytospora sp., Mucor attenuatus, and Phoma tropica were associated with the larval body. The list of fungi in mines was compared with bibliographic information to determine if the species are known pathogens on waterhyacinth or other plant species. This is the first study on mycobiota associated with T. truncatus and T. sagittatus mines in waterhyacinth petioles in Argentina.     

Active NaCl absorption across split lamellae of posterior gills of Chinese crabs (Eriocheir sinensis) adapted to different salinities

Split lamellae of posterior gills of Eriocheir sinensis adapted to fresh water, brackish waters (9 or 18‰) or seawater (36‰) were mounted in Ussing chambers, and transepithelial short-circuit currents and conductances were measured with salines, containing approximately in vivo-like NaCl concentrations. Active sodium and chloride absorption (I_Na and I_Cl), the transcellular conductances and the leak conductance were identified with external amiloride and/or DIDS. Split gill lamellae of crabs adapted to fresh water displayed similar magnitudes of I_Na and I_Cl with 10 mmol l⁻¹ NaCl in the external medium (internally haemolymph-like NaCl saline). Augmenting external NaCl (50 mmol l⁻¹) resulted in an increase of I_Cl, whereas I_Na decreased. Split gill lamellae of crabs adapted to brackish waters (external NaCl of 125 and 225 mmol l⁻¹, respectively) showed lower currents than preparations of freshwater crabs (50 mmol l⁻¹ external NaCl). With split gill lamellae of seawater crabs no currents were detected (450 mmol l⁻¹ NaCl on both sides). The transcellular conductances showed similar changes as the currents. The leak conductance of split gill lamellae of crabs adapted to fresh or brackish waters was low (0.3–0.8 mS cm⁻²), whereas it was much higher (7 mS cm⁻²) with preparations of seawater crabs.     

The ultrastructure of the sinus gland of Carcinus maenas (Crustacea: Decapoda)

Summary The sinus gland of Carcinus maenas consists of the swollen axonal endings of the neurosecretory cells of the major ganglia and acts as a storage release centre for the membrane bound neurosecretory material. These neurosecretory granules fall into five different types based on size and electron density. Their contents are released by exocytosis of the primary granules or smaller units budded from the primary granules.I thank Professor E. Naylor for his constant advice and Professor E. W. Knight-Jones, Department of Zoology, University College, Swansea, for the provision of laboratory facilities. I am grateful to the Science Research Council for the financial support. Finally, I thank the Electron Microscope Unit, Southampton General Hospital, where the work was completed.     

Morphology and ultrastructure of the midgut in Piscicola geometra (Annelida, Hirudinea)

This paper presents information on the organization of the midgut and its epithelium ultrastructure in juvenile and adult specimens of Piscicola geometra (Annelida, Hirudinea), a species which is a widespread ectoparasite found on the body and gills and in the mouth of many types of fish. The analysis of juvenile nonfeeding specimens helped in the explanation of all alterations in the midgut epithelium which are connected with digestion. The endodermal portion (midgut) of the digestive system is composed of four regions: the esophagus, the crop, the posterior crop caecum, and the intestine. Their epithelia are formed by flat, cuboidal, or columnar digestive cells; however, single small cells which do not contact the midgut lumen were also observed. The ultrastructure of all of the regions of the midgut are described and discussed with a special emphasis on their functions in the digestion of blood. In P. geometra, the part of the midgut that is devoid of microvilli is responsible for the accumulation of blood, while the epithelium of the remaining part of the midgut, which has a distinct regionalization in the distribution of organelles, plays a role in its absorption and secretion. Glycogen granules in the intestinal epithelium indicate its role in the accumulation of sugar. The comparison of the ultrastructure of midgut epithelium in juvenile and adult specimens suggests that electron-dense granules observed in the apical cytoplasm of digestive cells take part in enzyme accumulation. Numerous microorganisms were observed in the mycetome, which is composed of two large oval diverticles that connect with the esophagus via thin ducts. Similar microorganisms also occurred in the cytoplasm of the epithelium in the esophagus, the crop, the intestine, and in their lumen. Microorganisms were observed both in fed adult and unfed juvenile specimens of P. geometra, which strongly suggests that vertical transmission occurs from parent to offspring.     

Descriptions of some planktonic larvae of the Calappidae (Crustacea: Decapoda; Brachyura)

Some unidentified planktonic larval stages have been assignedto the Calappidae using Rice's key of Brachyura families. Zoeastage I from the eastern region of the Algerian coast (westernMediterranean Sea) has been attributed, according to Manningand Holthuis (1980), to Calappa granulata (L.), the only recordedrepresentative of the family from the Mediterranean Sea. Zoealstages 1-lV from the Jordanian coast of the Gulf of Aqaba, NorthernRed Sea, have been reported to the subfamily Matutinae. Comparisonsof larvae of Calappa and of Hepatus and then, of larvae of Hepatusand of Matuta, show that Hepatus larvae are more closely relatedto those of Calappa, while those of Matuta are clearly distinct.It s suggested to place the genus Hepatus with the Calappinae.Guinot (1966. 1978) placed some genera, including Hepatus. ina 'unit parthenoxystomienne, lea Aethrinae'. Larval developmentis unknown for Aethra, Actaeomorpha. Hepatella andOsachila.However, comparison of Hepatus larvae with those of Parthenopidaeshows that larval characters, particularly the setation of theendopod of the first maxilliped, are not conclusive for supportingsuch a regrouping. Moreover, the Calappidae have a combinationof primitive and advanced features which suggest that theirgrouping in the Oxystomata seems more likely to be an adaptationto a similar way of life. The morphotypal taxon Oxystornatahas been definitively rejected by Guinot (1978).