Volume 56, Issue 2

The impact of in vitro isolation and molecular characterisation of Neospora caninum as well as sequence analyses was studied. The brain homogenate of a naturally Neospora-infected dairy cow (positive in ELISA and Western blot) was intraperitoneally inoculated into Mongolian gerbils (Meriones unguiculatus). The brain of gerbils on day 60 post-inoculation was homogenized, and, after trypsin-digestion, cultured on Vero cells. Neospora-like tachyzoites were first observed after 77 days of cultivation. The parasite was confirmed by polymerase chain reaction (PCR) using Neospora-specific primers Np21 and Np6. The PCR product of the first Slovak isolate (NC-SKB1) was subsequently sequenced and published in GenBank under accession number GU300774. Sequencing and BLAST search identified the isolate as N. caninum.

Demidospermus annulus sp. nov. (Platyhelminthes, Monogenea, Ancyrocephalidae) is described from the gills of the catfish Parapimelodus valenciennis Lütken collected in Samborombón Bay, Argentina. The new species differs from all congeneric species mainly by the structure of the accessory piece of the male copulatory organ, the sclerotized ring-shaped vaginal aperture and the dorsal bar articulation.

The musculature of the marginal hooklets of adult Macrogyrodactylus congolensis (Prudhoe, 1957) Yamaguti, 1963 has been studied. Each marginal hooklet of M. congolensis is associated with three pairs of muscles. The possible role of these muscles in the operation of the marginal hooklet is discussed. Transmission electron microscopy has been used for the first time to study the marginal hooklets of M. congolensis. The handle articulates with the blade in the region of the guard. Internally, the handle, the proximal region of the blade in the articulation region and the distal pointed region of the blade consist of three layers. Distal to the articulation region, the blade consists of four layers with differing electron densities. A cavity is associated with the distal region of the blade and the handle. A cyton containing secretory bodies of different sizes and shapes was found in association with each marginal hooklet. The possible function of these secretions is discussed.

The ultrastructure of the uterus proper of the aspidogastrean Aspidogaster limacoides Diesing, 1835 and two digenean species, Phyllodistomum angulatum Linstow, 1907 (Plagiorchiida, Gorgoderidae) and Azygia lucii (Müller, 1776) (Strigeida, Azygiidae), was examined by transmission electron microscopy (TEM). The uterine epithelial lining of these species is thin, except for the perinuclear region of the epithelial cells. Septate junctions occur between adjacent epithelial cells within the uterine wall. The luminal surface of the cells is elevated into microlamellae, which project into the uterine lumen and cover the entire epithelial lining. Basally the uterine epithelium is attached to a basal matrix, and its supporting layers of muscle fibres are weak and composed of scattered circular muscles. Despite the marked similarity in the ultrastructural pattern of the aspidogastrean and two digeneans studied, there is some degree of variation in the secretory activity of their uterine epithelium. The high level of vesicular exocytotic activity in the epithelial cytoplasm of A. lucii may be associated with differences in egg emission and the subsequent life cycle involving a non-ciliated, non-swimming and non-free-living miracidium, as opposed to the free-swimming miracidium of P. angulatum. The similar nature of the uterine epithelium in all three species studied represents an ultrastructural marker possibly supporting a close phylogenetic relationship between the Aspidogastrea and the Digenea.

A new bucephalid species, Prosorhynchoides carvajali sp. nov. is described. This parasite was found in three marine fish, Auchenionchus microcirrhis (type-host), A. variolosus and Sicyases sanguineus (other-hosts), collected from the intertidal rocky zones of central Chile. P. carvajali sp. nov. is characterized by a pharynx in a post-equatorial position, a large cirrus sac length (half of the total worm length) and rounded caecum extending dorsally and anteriorly from pharynx. Although Prosorhynchoides carvajali sp. nov. closely resembles P. labiata; the latter has an elongated, narrow and inverted-U-shape caecum, contrasting to P. carvajali sp. nov. which has a larger rounded caecum, directed anteriorly. To our knowledge this is the first known report of Prosorhynchoides on the South American Pacific coast.

The natural infection with parasitic helminths is common in wild rodent populations. Once such interactions are better understood in the laboratory, it will be more feasible to extend the findings to infected hosts in nature. The flukes recovered from laboratory-infected Akodon cursor at 63 days post-infection were stained with hydrochloric carmine and individually mounted on glass slide as whole-mounts. Light and laser scanning confocal microscopy studies of adult male and female Schistosoma mansoni are reported. The parasites were examined morphologically and biometrically, which was obtained in a digital system for image analysis. Parameters used were: tegument thickness, digestive, excretory and reproductive systems. The overall conclusion of this experiment is that the morphological features of adult worm were similar to laboratory mice. It has been confirmed that the grass mouse is a permissive host to S. mansoni infection.

Alloglossidium fonti sp. nov. is described based on specimens from black bullheads (Ameiurus melas) collected from several localities in northwestern Minnesota. The new species is morphologically closest to A. corti and A. geminum. These two species were also found in the same region as A. fonti sp. nov. and sometimes in the same individual fish. Examination of numerous genetically marked specimens has demonstrated that the most reliable distinguishing feature among the three species is the position of anterior margin of the vitelline fields. In the new species they are situated at the level of anterior margin of ventral sucker while in A. corti it is at the level of intestinal bifurcation or even further anteriorly, and in A. geminum it is at the level of posterior margin of ventral sucker or further posteriorly. The new species has a relatively smaller cirrus sac than A. geminum. Despite rather minor, albeit stable morphological differences, the molecular data strongly supported the status of A. fonti sp. nov. as a new species. Moreover, molecular data and morphological examination of new material suggested that A. kenti, earlier synonymized with A. corti, is a valid species that is resurrected here. Comparison of 2420 base long sequences of nuclear rDNA (partial 18S, complete ITS region, and partial 28S) obtained from multiple specimens of all 4 species collected in Minnesota, North Dakota, Nebraska and Mississippi, showed essentially no intraspecific variability (except for 1 base difference in the ITS2 region of A. fonti sp. nov.), but has demonstrated pronounced interspecific differences. The new species was not found in the four examined catfish species from the Red River of the North (on the border between North Dakota and Minnesota) and neighbouring North Dakota.

Over a two-years period, a survey was carried out in order to increase the knowledge of digeneans parasitising the commonest intertidal gastropods on the Patagonian coast, Southwestern Atlantic Ocean. A total of 4,725 gastropods were examined. Six species of digenean parasitising four snail species were found; four of them were registered for first time: Maritrema sp. 1 (Microphallidae) in Crepidula dilatata (Calyptraeidae), Parorchis sp. (Philophtalmidae) and sporocyst of Renicolidae in Trophon geversianus (Muricidae), and Diphterostomum sp. (Zoogonidae) in Buccinanops globulosus (Nassariidae). Two other species were found in Siphonaria lessoni (Siphonariidae): Maritrema sp. 2 and Hemiuridae. One snail species, Tegula patagonica (Trochidae) was not parasitised. These gastropods act as first intermediate host, and C. dilatata, S. lessoni and B. globulosus also frequently host metacercariae within the sporocyst. Overall prevalences varied from 0.16% of Diphterostomum in the intertidal population of B. globulosus to 33.45% of Maritrema sp. 1 in C. dilatata.

The first description of vitellogenesis in the Diphyllidea is presented in this paper. Though the type of vitellogenesis and mature vitellocyte in Echinobothrium euterpes appear to be unique among the Eucestoda, however, they somewhat resemble that observed in the two orders of the lower cestodes, Tetraphyllidea and Proteocephalidea. Vitellocyte maturation is characterized by: (1) an increase in cell volume; (2) extensive development of short, parallel, frequently concentric cisternae of GER that produce dense proteinaceous granules; (3) development of Golgi complexes engaged in packaging this material; (4) progressive formation of saturated lipid droplets; their continuous enlargement and fusion; (5) formation of small accumulations of glycogen particles scattered between and among lipid droplets in the cytoplasm of maturing vitellocytes; (6) concentration of dense proteinaceous granules in the peripheral layer of cytoplasm, around the cell plasma membrane; and (7) vacuolization of cytoplasm of mature vitellocytes accompanied by a rapid increase in its volume. A new, unreported type of dense proteinaceous granules, situated around the limiting plasma membranes of mature vitellocytes, is described. Vitellogenesis evidently differs from that with typical shell-globules and shell-globule clusters previously reported in other taxa of lower cestodes. Cytochemical staining with periodic acidthiosemicarbazide-silver proteinate for glycogen indicates a strongly positive reaction for glycogen particles between and around large unsaturated lipid droplets of the maturing and mature vitellocytes. Some hypotheses concerning the interrelationship between this pattern of vitellogenesis, possible mode of egg formation, embryonic development and diphyllidean life cycle, and their phylogenetic implications are drawn and discussed.

We describe the spermiogenesis process and the ultrastructural characters of the spermatozoon of Acanthobothrium crassicolle by means of transmission electron microscopy, including cytochemical analysis for glycogen. Spermiogenesis in A. crassicolle begins with the formation of the differentiation zone that contains two centrioles associated with striated rootlets and an intercentriolar body. The latter is formed by one electron-dense layer. The centrioles develop into two free flagella that first grow orthogonally to a median cytoplasmic process and then undergo flagellar rotation becoming parallel to that median cytoplasmic process. After flagellar rotation only one of the flagella completes its growth and both short and long flagella undergo proximodistal fusion with the median cytoplasmic process. In the final stages of spermiogenesis, the nucleus becomes filiform and migrates into the spermatid body. Later, the ring of arched membranes constricts and the spermatozoon is liberated from the residual cytoplasm. The ultrastructural organization of the spermatozoon of A. crassicolle follows the general pattern of spermatozoa of the other Tetraphyllidea-Onchobothriidae species, but exhibits some differences. It is filiform, tapered at both extremities and lacks mitochondrion. It contains two axonemes of unequal length showing the 9 + “1“ pattern of Trepaxonemata, a nucleus, parallel cortical microtubules and electron-dense granules of glycogen. The anterior extremity of the male gamete contains a single crested body surrounding a thin and long apical cone. This type of apical cone has never been described in a tetraphyllidean spermatozoon. Another particularity is the presence of a single electron-dense microtubule at the vertex of the crested body.

Cosmocerca kalesari sp. nov. from the large intestine of frogs, Euphlyctis cyanophlyctis, collected in the Kalesar Wildlife Sanctuary, India is described and photomicrographed. It is most similar to those species bearing 5 pairs of plectanes, i.e., C. acanthurum, C. banyulensis, C. cruzi, C. japonica, C. ornata, C. novaeguineae, C. paraguayensis, C. parva, C. podicipinus and C. travassosi, but differs from these species with the exception of C. cruzi and C. japonica in the length of spicules and length of gubernaculum. The new species can be separated from C. cruzi and C. japonica by the arrangement of postanal mammiliform papillae: 6 postanal pairs in C. kalesari sp. nov.; 4 postanal pairs in C. cruzi; no postanal mammiliform papillae in C. japonica. Cosmocerca kalesari sp. nov. represents the 25th species assigned to the genus, but only the second species from the Oriental region.

Falcaustra sinensis sp. nov. collected from the feces of Indotestudo elongata (Blyth) is described and illustrated. The new species can be distinguished from the congeners by the arrangement of caudal papillae of the male (6 pairs precloacal, 4 pairs postcloacal, 1 median), length of spicules (3.84–4.74 mm) and the presence of pseudosucker.

Two new species of Camallanus are described from Australian freshwater turtles. Camallanus beveridgei sp. nov. is reported from Elseya dentata in Northern Territory. It differs from other species of the genus parasitic in turtles by several characters including the shape of the median ridge in the buccal capsule and the position of the anterior pair of caudal papillae in males. Camallanus sprenti sp. nov. is reported from Elseya latisternum (type host) and Emydura krefftii in northern Queensland. It is closely related to Camallanus tuckeri, and differs from the latter species in possessing a shorter oesophagus. We summarize data on morphology, distribution and specificity of 5 known Camallanus spp. from Australian turtles and provide a key for their identification. Sequence comparison of more than 500 base pairs at the 5′ end of the nuclear 28S rDNA gene confirms the status of C. sprenti and C. beveridgei as new species. Camallanus sprenti differs from the other 4 species of Camallanus from Australian turtles by 16–59 bases (3.1–11.5%) while C. beveridgei differed from the other 4 species by 23–60 bases (4.5–11.6%). Phylogenetic analysis demonstrates close interrelationships among C. tuckeri, C. sprenti and C. beveridgei, the three species with most similar buccal capsules.

As an adult, Eustrongylides ignotus, recognized as a zoonotic parasite, is a parasitic nematode found in the gastrointestinal tract of fish-eating birds. As larvae, they are found in the connective tissue or body cavities of freshwater fish. In Japan, E. ignotus and E. tubifex have been found in three avian species, but their distribution in intermediate and paratenic hosts remains unclear. For this study, 16 commercial raw Japanese smelts, Hypomesus transpacificus nipponensis, were used for parasitological examination. A pinkish red worm collected from the abdominal wall of one smelt (6.3%) was identified as E. ignotus based on the high sequence similarity (99.9%) of the partial 18S ribosomal RNA gene and the morphology of its cephalic and caudal ends. Furthermore, a Raphidascaris-like nematode was found in the intestines of five smelts examined (31.3%) and was identified as R. gigi in view of the absence of lateral cuticular alae. Eustrongylides ignotus and R. gigi have been reported in various freshwater fish, but not in smelts. Therefore, this finding of E. ignotus and R. gigi in H. transpacificus nipponensis represents a new host record.

In Europe, Serbia ranks among countries with a high prevalence of Trichinella infection in pigs, which continues to be a serious human health problem. While in some Balkan countries, more than one Trichinella species/genotype has been described in both the sylvatic and domestic cycles, these data are lacking for Serbia. To date, only a few Serbian isolates of Trichinella have been genetically specified, and all were classified as T. spiralis. Although transmission of Trichinella from domestic pigs to wildlife could be assumed, neither the infection status nor the species of Trichinella circulating among wildlife in Serbia has been investigated. This study shows the presence of two Trichinella species, T. spiralis and T. britovi, in wild animals originating from five districts in Serbia, where Trichinella infections in domestic pigs and humans have been recorded. Trichinella spiralis was detected in jackals (n = 3), red foxes (n = 2) and a wild cat (n = 1). We also established that wolves (n = 4) and red foxes (n = 2) serve as sylvatic reservoirs for T. britovi. This is the first report on the presence of T. britovi in Serbia.

The occurrence of the dracunculoid nematode Philometra chilensis Moravec, Chávez et Oliva, 2011 (Philometridae) infecting the ovary of the red cusk-eel, Genypterus chilensis (Guichenot), off the Pacific coast of northern Chile is reported. A sample of 266 fish specimens (total body length 34–96 cm) revealed the nematode overall prevalence (April 2007) of this parasite to be 18.5% and the mean intensity of infection 17.9 (SD = 30.8). All infected fish were mature females. The present results suggest that infection by the nematode shows seasonality and coincides with specific reproductive stages of the red cusk-eel, a partial spawner, and as a result the infection can affect the reproductive success of the host.