A freeze-fracture electron microscope study of Trichomonas vaginalis Donné and Tritrichomonas foetus (Riedmüller)

Two strains of Trichomonas vaginalis, JH162A , with low pathogenicity, and Balt 44, with high pathogenicity, as well as one highly pathogenic strain, KV-1, of Tritrichomonas foetus were studied by freeze-fracture electron microscopy. The protoplasmic faces ( PFs ) of the cell membranes of all three strains of both species had similar numbers of intramembranous particles (IMPs); however, the particles in the external faces (EFs) of these membranes were least abundant in Trichomonas vaginalis strain Balt 44 and most numerous in those of strain JH162A of this species. In Tritrichomonas foetus strain KV-1 the number of IMPs in the EF was close to but somewhat lower than that in the mild strain of the human urogenital trichomonad . In both species, the anterior, but not the recurrent, flagella had rosette-like formations, consisting of approximately 9 to 12 IMPs on both the PFs and EFs. The numbers and distribution of the rosettes appeared to vary among different flagella and in different areas of individual flagella of a single organism belonging to either species. The freeze-fracture electron micrographs provided a more complete understanding of the fine structure of undulating membranes of Trichomonadinae , as represented by Trichomonas vaginalis, and of Tritrichomonadinae (the Tritrichomonas augusta -type), as exemplified by Tritrichomonas foetus, than was gained from previous transmission and scanning electron microscope studies. Typically three longitudinal rows of IMPs on the PF of the recurrent flagellum of Trichomonas vaginalis were noted in the area of attachment of this flagellum to the undulating membrane. The functional aspects of the various structures and differences between certain organelles revealed in the two trichomonad species by the freeze-fracture method are discussed.     

Cattle pathogen tritrichomonas foetus (Riedmüller, 1928) and pig commensal Tritrichomonas suis (Gruby & Delafond, 1843) belong to the same species

A number of reports suggest that the sexually transmitted pathogen of cattle, Tritrichomonasfoetus, and a gastrointestinal commensal of pigs, Tritrichomonas suis, are very similar and may be co-specific. A conclusive review of the taxonomic and nomenclatural status of these species has not been presented so far. Toward this end, we reexamined and compared porcine and bovine trichomonads with regard to their morphology, pathogenic potential, and DNA polymorphism. Using light and electron microscopy, no distinguishing features between T. foetus and T. suis strains were found in size, general morphology, and karyomastigont structure. Both bovine and porcine trichomonads showed pathogenic potential in the subcutaneous mouse assays and did not separate into distinct groups according to strain virulence. Three DNA fingerprinting methods (i.e. RFLP, RAPD, and PCR-based analysis of variable-length DNA repeats) that produce species-specific DNA fragment patterns did not distinguish between the bovine and porcine strains. Sequencing of a variable 502-bp DNA fragment as well as comparison of 16S rRNA gene sequences did not reveal species-specific differences between the cattle and porcine strains. Therefore, we conclude that T. foetus and T. suis belong to the same species. To prevent confusion that may arise from T. foetus-T. suis synonymy, we propose to suppress the older name suis and maintain its accustomed junior synonym foetus as a nomen protectum for both cattle and porcine trichomonads. The case has been submitted to the International Commission on Zoological Nomenclature for ruling under its plenary power.     

Are Tritrichomonas foetus and Tritrichomonas suis synonyms?

Tritrichomonas suis, a tritrichomonad of pigs, and the related species Tritrichomonas foetus, a tritrichomonad of cattle, are morphologically identical. The taxonomic relationship between these two tritrichomonads has been questioned ever since they were established as distinct species in 1843 and 1928, respectively. Here, we compare the similarities of morphology, ultrastructure, distribution, host specificity, characteristics of in vitro cultivation, immunology, biochemistry and analysis of molecular data from published sources between these two species. All data indicate that these two tritrichomonad species are identical. Thus, we propose that T. foetus and T. suis are synonyms.     

Life history of Lineus viridis (Müller, 1774) (Heteronemertea, Nemertea)

Lineus viridis is a common nemertean species of North-Atlantic intertidal sand flats. Its mating behaviour is peculiar insofar as this species is reported to be polyandric. However, detailed information on this topic is lacking. In order to get more data on the reproduction, oogenesis and life history of this species, a population in the Wadden Sea on the Isle of Sylt (North Sea) was studied between 2005 and 2011. We conducted regular surveys, during which we sampled, measured and recorded the sex status of 25–100 individuals at each sampling event; at least three individuals were fixed for histological studies at each sampling date. In addition, animals were kept in the laboratory for 3?years to complement field data on sexual identity. Lineus viridis was found to reproduce annually in several successive year; the females are significantly larger than the males. Oogenesis starts in spring, shortly after the preceding reproductive period, and continues until the end of December. Spermiogenesis starts in late autumn and also ends late in December. During mating, several males are generally found crawling on a single female, which forms a cocoon that encloses both the female and the associated males. Fertilization is internal. While females discharge all of their eggs during a single mating event and lose more than 40% of their wet weight, males only empty a few of their gonads, and are thus able to fertilize more than one female. Our study clearly shows that Lineus viridis is a perennial, iteroparous species with a pronounced sexual size dimorphism. During this long-term study, no evidence for sequential hermaphroditism has been found. The observed polyandric mating system in this species raises further questions regarding mate and sperm competition that deserve additional research.     

The population biology ofPlanaria torva (Müller) (Turbellaria,Tricladida)

Summary 1. A quantitative study of the population biology ofPlanaria torva (Müller) living in a productive lake, based on monthly samples over a period of 20 months, is presented. 2. Samples of triclads, their cocoons and other organisms were taken from bricks placed at a shallow depth in the lake. They provided relative information on changes in population size structure and demonstrated the temporal breeding of this triclad. 3. Cocoon production byP. torva began in January and reached a peak in March–April. This was followed by a peak in recently-hatched young during May–June. Adults reached a maximal proportion of the total population in January and most had died by July. 4. Changes in size structure suggest that intra-specific competition occurred during June–July following the recruitment of young. This hypothesis is supported by other independent evidence. 5. Attempts to measure fecundity based on cocoons and young gave very different results attributed to the sampling technique.     

A chromosome study ofSchistocephalus solidus (Müller, 1776) (Cestoda: Pseudophyllidea)

Analysis of Giemsa-stained metaphase plates of plerocercoids ofSchistocephalus solidus revealed the mode diploid number of chromosomes for this species as 2n=18. Chromosome pairs 1, 5, 6, 8 and 9 are metacentric, pairs 2, 4 and 7 are subtelocentric, and pair 3 is subtelocentric to acrocentric. The karyotype is characterised by mean absolute chromosome length between 1.9 and 5.9 m and a total chromosome length of the haploid complement of 32 m. A comparison with other known karyotypes of related species of pseudophyllidean cestodes was made.     

Spermatological characters of the pseudophyllidean cestode Bothriocephalus scorpii (Müller, 1776)

Spermiogenesis of Bothriocephalus scorpii (Cestoda, Pseudophyllidea) includes an orthogonal development of two flagella, followed by a flagellar rotation and a proximo-distal fusion with the median cytoplasmic process. The fusion occurs at the level of four attachment zones. The presence of dense material in the apical region of the differentiation zone in the early stage of spermiogenesis appears to be a characteristic feature for the Pseudophyllidea. The mature spermatozoon possesses two axonemes of 9+"1" pattern of the Trepaxonemata, nucleus, cortical microtubules, electron-dense granules and crested body. The anterior part of the gamete exhibits a centriole surrounded by electron-dense tubular structures arranged as incomplete spiral. When the crested body disappears, the electron-dense tubular structures are arranged into a ring encircling the axoneme. The electron-dense tubular structures and their arrangement appear to be a specific feature for the clade "Bothriocephalidea". The organization of the posterior extremity of the gamete with the nucleus is described for the first time in the Pseudophyllidea.     

Possible sites of ultrafiltration in Tubifex tubifex Müller (Annelida,Oligochaeta)

Summary The endothelia of Tubifex tubifex Müller consist of myoendothelial cells, chloragocytes, or podocytes. The latter seem to occur only as windows on the ventral vessel which has an endothelium of myoendothelial cells elsewhere. The podocytes are large cells, with several processes on the inner side which ramify into several pedicels. These are aligned upon the outside of the basement membrane which lines the inside of the endothelium. The gaps between adjacent pedicels are about 40 nm wide. In capillaries fenestrated endothelia occur with irregular spacings measuring up to 0.4–1 m. A diaphragm in podocytes or capillary fenestrations do not seem to exist. The basement membrane is the only continuous layer lining the blood vessels and capillaries of Tubifex with a rather uniform diameter in the range of 50 nm. It is the only permeability barrier between blood and coelomic fluid.     

The localization of inorganic elements,particularly vanadium and sulphur,in haemolymph from the ascidians Ascidia mentula (Müller) and Ascidiella aspersa (Müller)

Vanadium and sulphur were found by X-ray microanalysis in three different cell types, morula cells, signet-ring cells, and granular amoebocytes, from each of the ascidians Ascidia mentula (Müller) and Ascidiella aspersa (Müller). In addition to the cells containing vanadium and sulphur a few vacuolar cells containing high concentrations of iron, magnesium and calcium were also found. After fixation in the presence of strontium or barium chlorides another cell type containing large amounts of precipitated sulphur was also found, confirming our previous findings that many haemolymph cells in these species contain large amounts of sulphate only. The presence of these various cell types is discussed in relation to the question of the acidity of ascidian blood cells.     

A cytogenetics study of Hydrodroma despiciens (Müller, 1776) (Acari: Hydrachnellae: Hydrodromidae)

The karyotypes of water mites (Acari: Hydrachnellae: Hydrodromidae) are largely unknown. The present investigation is the first report of a study designed to characterize the chromosomes of water mites. The study was carried out with specimens of Hydrodroma despiciens collected from Eber Lake in Afyon, Turkey. Several different methods were tried to obtain chromosomes of this species. However, somatic cell culture proved to be the most effective for the preparation of chromosomes. In the present study, we determined the diploid chromosome number of Hydrodroma despiciens to be 2n = 16. However, a large metacentric chromosome was found in each metaphase, which we believed to be the X chromosome. We could not determine the sex chromosomes of this species. This study is the first approach to the cytogenetic characterization of this water mite group. Furthermore, these cytogenetic data will contribute to the understanding of the phylogenetic relationship among water mites. To our knowledge, this is the first report on the cytogenetics of water mites.     

Immunohistochemical analysis of adhesive papillae of Clavelina lepadiformis (Müller, 1776) and Clavelina phlegraea (Salfi, 1929) (Tunicata,Ascidiacea)

Almost all ascidian larvae bear three mucus secreting and sensory organs, the adhesive papillae, at the anterior end of the trunk, which play an important role during the settlement phase. The morphology and the cellular composition of these organs varies greatly in the different species. The larvae of the Clavelina genus bear simple bulbous papillae, which are considered to have only a secretory function. We analysed the adhesive papillae of two species belonging to this genus, C. lepadiformis and C. phlegraea, by histological sections and by immunolocalisation of β-tubulin and serotonin, in order to better clarify the cellular composition of these organs. We demonstrated that they contain at least two types of neurons: central neurons, bearing microvilli, and peripheral ciliated neurons. Peripheral neurons of C. lepadiformis contain serotonin. We suggest that these two neurons play different roles during settlement: the central ones may be chemo- or mechanoreceptors that sense the substratum, and the peripheral ones may be involved in the mechanism that triggers metamorphosis.Key words: Settlement, neurotransmitter, serotonin, β-tubulin, papillary nerves, metamorphosis.Ascidians (phylum Chordata; subphylum Tunicata) are sessile filter-feeding organisms that can be found in all benthic marine environments and develop through a swimming tadpole larva. Larvae of colonial ascidians have a short planktonic life that can vary from minutes to hours (Burighel and Cloney, 1997). Prior to metamorphosis, the larva attaches to the substratum by means of peculiar organs of ectodermic origin, located in the anterior region of the trunk. These organs, known as adhesive papillae, secrete sticky substances and effect primary adhesion of the larva to the substrate (Cloney, 1977). They have an important role in the initiation of settlement and metamorphosis and there is evidence that, at least in some species, they participate in substrate selection (Torrence and Cloney, 1983; Svane and Young, 1989; Groppelli et al., 2003). In many species, they are organised in a triangular field, whereas in others they are aligned along the mid-sagittal plane of the trunk. Adhesive papillae have been classified into two types: eversible papillae, typical of some colonial species, composed by several cell types and rapidly changing shape as they touch the substrate, and non-eversible papillae, typical of solitary species, which do not change shape after settlement (Burighel and Cloney, 1997).With few exceptions, all adhesive papillae are formed by elongated secretory and sensory cells, which are recognised as primary neurons (Cloney, 1977, 1979). It has been proposed that sensory cells may detect the chemical and physical characteristics of the substratum at potential sites for settlement and metamorphosis (Young and Braithwaite, 1980; Groppelli et al., 2003).The presence of primary neurons in the papillae has been reported in the larvae of several species such as Distaplia occidentalis, Diplosoma macdonaldi, Phallusia mammillata, Ciona intestinalis and Ascidia malaca (Cloney, 1977;Torrence and Cloney, 1983; Sotgia et al., 1998; Takamura, 1998; Gianguzza et al., 1999).These neurons have axons that join together to form the papillary nerves that enter the central nervous system at the level of the sensory vesicle (Imai and Meinertzhagen, 2007).Recently, different neurotransmitters have been localised in the sensory neurons of the papillae of different species. The presence of GABAergic neurons has been reported in the papillae of Ciona savygni (Brown et al., 2005) and of Ciona intestinalis (Zega et al., 2008), while serotonergic neurons have been localised in the papillae of Phallusia mammillata (Pennati et al., 2001) and Botrylloides leachi (Pennati et al., 2007). Moreover, it has been demonstrated that serotonin plays a role in the mechanism triggering metamorphosis in ascidians (Zega et al., 2005).After attachment, all papillae retract to draw the larva closer to the substratum. In the colonial ascidian Distaplia occidentalis, the process of retraction is reversibly inhibited by cytochalasin B, suggesting that microfilaments are involved in this process (Cloney, 1979).Clavelina lepadiformis is a colonial species, whose larvae bear non-eversible simple bulbous papillae.They have been described as being formed only by columnar glandular cells, whose secreted material is responsible for the sticky properties of the organ. These papillae do not contain sensory cells and were considered the simplest among those studied (Turon, 1991).In this work, the morphology of the adhesive papillae of C. lepadiformis and C. phlegraea was further investigated by histological analysis and immunolabelling techniques in order to clarify the actual cellular composition and function of these organs.     

Biological studies and phenology of the slug Deroceras reticulatum (Müller, 1774) (Pulmonata: Stylommatophora)

Summary

In the southeast of Buenos Aires Province (Argentina), slugs cause reductions in crop yield by killing seeds and seedlings, by destroying stems and growing points, and by reducing leaf area. Deroceras reticulatum (Müller, 1774) was introduced into Argentina a long time ago, but the biology and ecology of this species in this country are still unknown. The aim of this study was to determine how temperature affects growth rate, reproduction and survival of D. reticulatum and also to assess the seasonal phenology of this species. The life cycle of D. reticulatum was investigated at 12°C, 20°C and at a temperature which alternated between 12°C for 16 h and 20°C for 8 h. The light/dark regime during the experiment was LD: 8/16 h. Slug populations were sampled from grassland located at the Experimental Station of INTA Balcarce. Eggs hatched after 16.3 to 39.8 days and fertility varied between 83.9% and 91.1%. The mean time from hatching to oviposition was longer and fecundity lower at 20°C than 12°C and 12/20°C (p <0.05). The net reproductive rate was 1.49 to 70.53 and the mean generation time varied between the 27 and 30 weeks. In natural conditions, eggs of D. reticulatum were found from early winter until the end of the spring, when soil temperature was 8–15°C and humidity was above 15%. These results indicate that D. reticulatum has one generation per year, univoltine phenology, with peak densities of slugs from the beginning of winter to the end of spring, which coincides with the sowing and emergence of winter and summer crops.     

Reproductive organs of the giant looper,Boarmia selenaria Schiffermüller (Leopidotera : Geometridae)

With some exceptions, the internal and external reproductive organs of the female and male giant looper, Boarmia selenaria Schiffermüller (Lepidoptera : Geometridae) are similar to those of other Lepidoptera; the same is true for the spermatophore. However, there is no enlargement of the vas deferens before it enters the ductus ejaculatorius duplex, the accessory glands are not closely united, the claspers are of a complex type, transtilla, furca, socii and coremata are not present, and the 2 arms of the gnathos fuse distally, whereas in the Satyridae, Nymphalidae and Lycaenidae, the 2 halves remain separate.     

Conjugation in the spirotrich ciliate Halteria grandinella (Müller, 1773) Dujardin, 1841 (Protozoa, Ciliophora) and its phylogenetic implications

The conjugation of Halteria grandinella was studied in protargol preparations. The isogamontic conjugants fuse partially with their ventral sides to a homopolar pair. The first maturation division generates dramatic transformations: (i) the partners obtain an interlocking arrangement; (ii) the number of bristle kineties decreases from seven to four in each partner; and (iii) the right conjugant loses its buccal membranelles, the left the whole adoral zone. The remaining collar membranelles arrange around the pair's anterior end and are shared by both partners; finally, the couple resembles a vegetative specimen in size and outline. The vegetative macronucleus fragments before pycnosis. The micronucleus performs three maturation divisions, but only one derivative each performs the second and third division. The synkaryon divides twice, producing a micronucleus, a macronucleus anlage, and two disintegrating derivatives. Scattered somatic kinetids occur during conjugation, but disappear without reorganization. An incomplete oral primordium originates in both partners. The conjugation of Halteria grandinella resembles in several respects that of hypotrich spirotrichs; however, the majority of morphological, ontogenetical, and ultrastructural features still indicates an affiliation with the oligotrich and choreotrich spirotrichs. Accordingly, the cladistic analysis still contradicts the genealogy based on the sequences of the small subunit rRNA gene.     

The life-history of Haementeria costata (Fr. Müller) (Glossiphoniidae: Hirudinea)

The frequency of occurrence of Haementeria costata in the waters of north-eastern Poland is 66.5%. This leech is usually found in shallow water zones frequented by water fowl and mammals.The different numbers of individuals taking food and different amounts of food consumed at one meal during successive feedings resulted in individuals of a cohort reaching sexual maturity at the age of 1, 2, and 3 years and surviving for 1.5, 2.5 and 3.5 years, respectively. The differences in weight and age of individuals of populations in their natural environment and in the laboratory depend on the accessibility of food.     

Factors affecting feeding rate, reproduction and growth of an oligochaete Lumbriculus variegatus (Müller)

Worm density of a deposit feeding oligochaete (Lumbriculus variegatus) did not affect egestion whereas both temperature and sediment type had a significant influence. The worms egested less actively at the lowest temperature (6 °C). The egestion rate, expressed as mg dry feces produced, was highest in the sandy sediment and lowest in the sediment derived almost exclusively from decaying plant material. The amount of dry material and the volume of wet sediment passing through the worms varied between the test sediments; the results are probably dependent on the chosen unit of measure. Reproduction was significantly decreased in sandy sediment with low organic carbon content. Reproduction was also dependent on the worm size, larger worms reproducing more frequently. Most worms lost weight during the tests, but the loss was lowest in the sediment with the highest organic carbon content. This revised version was published online in July 2006 with corrections to the Cover Date.