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The study lays stress on the morphological variability of metacercariae of Diplostomum pseudospathaceum under the influence of four factors: species and size of the host and density and age of the parasites. A total of 22 experiments were made, using 208 fish belonging to 10 species (three aquarium, seven native Polish species). Ranges of variations found in body size, number of excretory bodies, oral sucker and acetabulum, pharynx, holdfast organ and values of 9 biometric indices are wide and far exceed the ranges of variation given in the literature for this species of metacercaria.
The variability of metacercariae of Diplostomum pseudospathaceum under the influence of host species was studied experimentally. Four fish groups - two of Cyprinus carpio and two of Poecilia reticulata - were infected with the cercariae of D. pseudospathaceum originating from two snail species Lymnaea stagnalis and Galba palustris. For analysis of variability 14 characters of 152 metacercariae were measured and 8 indices were calculated. The morphometric data of the four experimental groups were examined statistically using a two way analysis of variance. The metacercariae of the four examined groups show highly significant variation in almost all analyzed characters. For particular characters a highly significant fish and snail host influence on the variation was noted. The source of variation was analyzed and the variability of previously investigated D. paracaudum was compared.
Metacercariae of Diplostomum pseudospathaceum and D. paracaudum were examined for genetic differences by random amplified polymorphic DNA analysis. Eleven different oligonucleotide decamers with arbitrary DNA sequence were tested as primers to amplify DNA from D. pseudospathaceum and D. paracaudum. Depending on the primer, the number of reproducible amplified fragments varied from 7 to 15, ranging in size from 0.2 to 2.0 kb DNA. Reproducible differences in patterns allowed differentiation of the two species with each primer. RAPD patterns unambiguously discriminated between D. pseudospathaceum and D. paracaudum, allowing good species identification.
The ultrastructure of the proto- and paranephridial excretory systems in the metacercaria of D. pseudospathaceum is described. Basically, the protonephridial system in the metacercaria, although being more complex, does not differ in ultrastructure from that in the cercaria. All protonephridial canals have nuclei and septate desmosomes between neighbouring cells as well as desmosomes closing the canals. Contrary to the protonephridial system, the paranephridial one has a syncytial structure with few cell nuclei in the wall. No septate desmosomes were visible in cross- and longitudinal sections of the paranephridial canals and terminal pockets. The heterocellular gap junctions (nexus) between tegumental cytons and paranephridial canal walls have not previously been observed and described in diplostomid metacercaria. The possible function of these heterocellular gap junctions is discussed.
The structure of the nervous system in the metacercaria of Clinostomum giganticum has been studied, using the methods of Gomori 1952 and Holt and Withers 1952 for the detection of acetylcholinesterase and non-specific esterase activities respectively. The nervous system of the metacercaria possess paired cerebral ganglia connected by the cerebral commissure, and three anterior and three posterior pairs of longitudinal nerve strands. The anterior and posterior nerves of corresponding pairs are connected by transverse commissures. There are twenty seven commissures in the whole body. The commissures are joined by numerous additional anastomoses giving the appearance of a dense net of nerve fibres.
The infection patterns of parasites are often tied to host behavior. Although most studies have investigated definitive hosts and their parasites, intermediate host behavior may play a role in shaping the distribution and accumulation of parasites, particularly the larval stages. In an attempt to answer this question, more than 4,500 pulmonate snails were collected from 11 states in the mid-Atlantic and Midwestern United States in the summer of 2012. These snails were necropsied and echinostome metecercariae were commonly observed infecting the snails as 2nd intermediate hosts (20.0%). The snails included species of 3 genera with distinct differences in the infection patterns of Echinostoma spp. metacercariae among them. Physa spp. (comprising of P. acuta and P. gyrina) snails exhibited a significantly higher prevalence of infection (23.5%) than both Lymnaea columella (11.6%) and Helisoma spp. (comprising of H. anceps and H. trivolvis) (14.2%; P < 0.05), with no difference in prevalence observed between the latter 2 genera (P > 0.05). The intensity of metacercariae within the snail hosts was significantly different between the 3 genera (P < 0.05), with L. columella having the highest intensity (24.3 ± 5.6), followed by Physa spp. (15.2 ± 1.5) and Helisoma spp. (5.0 ± 0.9). Differences in prevalence and intensity were also observed when the different snail families co-habited the same body of water. The disparities in infection patterns are likely due to distinct differences in the behavioral and feeding ecology of the snail hosts.
Ornamental fish, Xiphophorus maculatus, were imported from Singapore to Denmark for distribution to local aquarists. Importers observed lethargic and erratic swimming patterns among fish and forwarded a total of 30 fish for pathological examination to a university diagnostic service. All fish were diagnosed infected with encysted Centrocestus sp. metacercariae in gills (prevalence of 100% and mean intensity of 454.5 ± 161.9 parasites per fish). Metacercariae were identified by morphological and molecular methods. Cysts (mean length 163.3 ± 13.7 μm and mean width 113.3 ± 10.6 μm) contained a bent metacercaria with an X-shaped excretory bladder. PCR amplification of a rDNA region (5.8S rRNA gene, ITS-2, 28S rRNA gene) and subsequent sequencing confirmed the diagnosis. Metacercariae were found in gill filaments adjacent to the cartilage associated with cartilage hypertrophy, epithelial and mucous cell hyperplasia, clubbing and lamellar fusion. Host cell encapsulation of cysts comprised several layers of leucocytes, chondroblast-like and fibroblast-like cells. The observations raise concerns with regard to veterinary inspection and quarantine procedures. The zoonotic potential of these trematodes and a possible spread of the parasites in natural habitats in Europe should be regarded as a public health issue. So far, several cases of human infections have been reported only in Asia, but the potential intermediate host snail, Melanoides tuberculata, has been recorded in Germany. Accordingly, establishment of the parasite in Europe with climate changes should be considered a risk.
The distribution of the digenean Posthodiplostomum cuticola in cyprinids caught in the Polish part of the Vistula Lagoon is described. The metacercariae were found to be present in carp bream (Abramis brama), roach (Rutilus rutilus), Prussian carp (Carassius gibelio), white bream (Abramis bjoerkna), ziege (Pelecus cultratus), and common dace (Leuciscus leuciscus). Of the 1287 metacercariae found, 1175 occurred on the body surface, including 780 in the fins, 232 on the scales, and 163 in the skin underneath the scales. The remaining metacercariae occurred in the mouth cavity, gills, and eye sockets. Most of the parasites (1033) were located on the surface of the body below the longitudo caudalis. As is shown by the study, the digenea attack primarily those parts on the fish body that are closest to the bottom. The fins, sites that are more amenable to attachment and subsequently to penetration into the host's tissues, turned out to be the preferred site of the parasite (60.6% of the metacercariae).
The infection of metacercariae of Posthodiplostomum cuticola (v. Nordmann, 1832), the agent of black-spot disease, was investigated in 0+juvenile fish from 19 localities in south-east Moravia, Czech Republic. Of the 30 fish species examined, only cyprinids of 12 species were found to be infected. Five species of Leuciscinae, Rutilus rutilus, Leuciscus idus, Scardinius erythrophthalmus, Abramis bjoerkna and Abramis brama, fish with similar behaviour and habitat preference in juvenile period, were the most susceptible fish hosts for P. cuticola metacercariae. Especially high infection parameters were found in intergeneric hybrids of the studied leuciscine species. Environmental conditions were an important factor for parasite infection. The fish from flooded borrow pits, inhabiting lentic water bodies with dense vegetation in the littoral zone, showed high prevalence and abundance of P. cuticola infection contrary to their conspecifics in water reservoirs with steep banks and fish from rivers.
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