Population biology of Contracaecum rudolphii sensu lato (Nematoda) in the great cormorant (Phalacrocorax carbo) from northeastern Poland

Parameters related to the occurrence, aggregation, and population structure of the nematode Contracaecum rudolphii Hartwich, 1964 , in the great cormorant ( Phalacrocorax carbo ) from northeastern Poland were analyzed. A total of 491 birds of different ages (adults, immatures, and nestlings) was examined; the cormorants studied were taken from both fresh- and brackish water habitats. Contracaecum rudolphii were found in stomachs of 454 birds (92.5%); the 46,244 nematodes included third- (L3) and fourth-stage larvae (L4), and sub-adult and adult females and males. The distribution of nematodes in the host population were highly aggregated. The occurrence of C. rudolphii was significantly dependent on the host's age and habitat, as well as on season; the proportion of development stages differed significantly depending on birds' age and season (the latter only in adult birds). The infrapopulations of C. rudolphii in the adult cormorants showed distinct density-dependent correlations: that is, as the infrapopulation size increased, the number of adult females C. rudolphii decreased, and the proportion of larvae and sub-adult females increased. A higher proportion of larvae and sub-adult females characterized the component population structure of the nematodes in the cormorant nestlings, compared with adult birds, probably because of immune system deficiency in the immature birds, coupled with the development of the nematode population. Seasonal changes in the C. rudolphii population, observed in the adult cormorants, were not related to seasonality of the L3 occurrence in food; instead, the changes are believed to have resulted from independent processes of elimination of the oldest nematodes and their replacement by larval stages that subsequently matured.     

Prevalence and pathology of helminths of ciconiiform birds from the Brazilian swamplands

The prevalence of helminths recovered from 108 birds representing eight species of Ciconiiformes from the Brazilian west-central region are presented. The digeneans Ascocotyle (Phagicola) longa, Clinostomum marginatum, Cotylotretus grandis, Ithyoclinostomum dimorphum, the nematodes Contracaecum multipapillatum, Desmidocercella ardeae, Eustrongylides ignotus, and the cestode Valipora mutabilis were identified. Contracaecum multipapillatum was the most prevalent species and E. ignotus the most pathogenic. Gross lesions due to infections with C. multipapillatum were characterized by ulcerative processes and hyperemia of the mucosa whereas those caused by E. ignotus consisted of perforations of the gastric mucosa and fibrotic tubular lesions in the gastric serosa. Histopathological examinations revealed necrosis and mixed leucocyte infiltrations and discrete compression of the mucosa in C. multipapillatum infections. Destruction of the mucosa and submucosa with the presence of fibrous capsules were observed in E. ignotus infections. Reports of accidental human infections, with severe clinical signs induced by these parasites, indicate the necessity of a proper evaluation of the pathogenicity of helminths of aquatic birds.     

Anisakid infection in the European shag Phalacrocorax aristotelis aristotelis

A total of 100 specimens of the European shag Phalacrocorax aristotelis aristotelis were examined for anisakid infections in the south-east North Atlantic. The taxonomic status of the anisakid nematodes was studied by structural analysis using light and scanning electron microscopy. Two species, Contracaecum septentrionale and C. rudolphii A (Nematoda: Ascaridoidea), occurred in the European shag with high values of prevalence and mean intensities of infection. These constitute new host records for both parasites, widening their known geographical distribution. Although the histopathology associated with the infections indicated that the anisakids did not have the potential to cause bird death, the parasites are capable of contributing to and hastening avian mortality in the case of heavily-parasitized cormorants and when stressed by xenobiotics and/or through a synergetic effect by other bioagressors.     

Molecular identification of two strains of third-stage larvae of Contracaecum rudolphii sensu lato (Nematoda: Anisakidae) from fish in Poland

Contracaecum sp. larvae (L3) from fish were identified using nucleotide sequences of the internal transcribed spacers ITS-1 and ITS-2 of the ribosomal DNA. The nematode larvae originated from fish in a freshwater situation (crucian carp Carassius carassius, from Selment Wielki Lake in Mazury, northeastern Poland) and a brackish-water region (Caspian round goby Neogobius melanostomus from the Baltic Sea, Gdafisk Bay at the Polish coast). Two strains (Contracaecum rudolphii A and B) of Contracaecum rudolphii senso lato, a parasite common at the adult stage in fish-eating birds, were identified. In fish from the freshwater site, only the strain temporarily designated C. rudolphii B was identified; in the brackish-water region, both strains were found, suggesting that fish serve as paratenic host for both genotypes. Contracaecum rudolphii sensu lato has been recorded in several species of fish-eating birds in Poland, particularly in the great cormorant, Phalacrocorax carbo, in which the abundance is highest. The results, although based on a restricted number of larvae, suggest that the life cycles of both genotypes can be completed in the Polish region and that at least one of them, C. rudolphii B, can develop both in fresh and brackish water.     

A new anisakid species parasitizing the imperial cormorant Phalacrocorax atriceps from the North Patagonian Coast, Argentina

The anisakid species Contracaecum chubutensis n. sp. was found parasitizing the imperial cormorant Phalacrocorax atriceps at 2 different localities, Bahía Bustamante (45 degrees 11'S, 66 degrees 30'W) and Puerto Madryn (42 degrees 47'S, 65 degrees 02'W) on the Patagonian coast. Morphometrical analysis and further studies of adult specimens of C. chubutensis n. sp. and fourth-stage larvae of Contracaecum sp. were done using light and scanning electron microscopy. The possession and disposition of 7 pairs of tail papillae differentiate this new species from C. osculatum, C. radiatum, C. plagiaticium, and C. mirounga. Double-sized spicules separate it from C. variegatum and C. pelagicum. Moreover, the paracloacal papillae disposition and tail shape differentiate this new species from C. travassosi. The presence of bifurcated interlabia separate it from C. ogmorhini and C. margolisi. Three well-marked lip notches, a sharp tail shape, and the of the transversal paracloacal papillae disposition distinguishes C. chubutensis n. sp. from C. magnipapillatum and C. septentrionale. The species here described can be differentiated from C. caballeroi by its bifurcated interlabia, longer spicules, and for possessing more precloacal papillae pairs. It can be separated from C. rudolphii by a thinner interlabia at the base, deeper lip notches, a sharper tail, the disposition of transversal paracloacal papillae, and greater distance between both papillae of the first subventral distal papillae pair. In addition, fourth-stage larvae recovered from the same host and localities were identified and described as Contracaecum sp. Prevalence of adult parasites was 66.7 %, mean intensity was 4.0, and mean abundance was 7.1.     

<Emphasis Type="Italic">Contracaecum bioccai</Emphasis> n. sp. from the brown pelican <Emphasis Type="Italic">Pelecanus occidentalis</Emphasis> (L.) in Colombia (Nematoda: Anisakidae): morphology,molecular evidence and its genetic relationship with congeners from fish-eating birds

Contracaecum bioccai n. sp. is described from the brown pelican Pelecanus occidentalis (L.) in northern Colombia (Totumo Marsh) based on 20 enzyme loci studied using multilocus allozyme electrophoresis. Moreover, genetic relationships between the new taxon and related congeners are presented based on allozyme data-sets and sequence analyses (519 bp) of the mtDNA-cox2 gene. Fixed allele differences were found at some of the allozyme loci analysed in comparison with other Contracaecum spp. from pelicans and cormorants [i.e. the sibling species of the C. rudolphii Hartwich, 1964 complex, C. septentrionale Kreis, 1955, C. micropapillatum (Stossich, 1890), C. microcephalum (Rudolphi, 1809) and C. pelagicum Johnston & Mawson, 1942]. The genetic distance, at the allozyme level, between C. bioccai n. sp. and its congeners ranged from D ( Nei ) = 0.80 versus C. septentrionale to D ( Nei ) = 1.40 versus C. micropapillatum. The genetic distance at the mtDNA cox-2 level ranged, on average, from K-2P = 0.12 versus the C. rudolphii species complex to K-2P = 0.15 versus C. micropapillatum. An overall concordant tree topology, obtained from UPGMA and NJ tree analyses inferred from allozyme data, as well as from MP, UPGMA and NJ inferred from mtDNA-cox2 sequence analysis, showed C. bioccai n. sp. as a separated lineage to the other Contracaecum spp. A concordant result was also obtained by PCA analysis based on both the allozyme and mtDNA cox-2 data-sets. All of the tree topologies, derived from the phylogenetic analysis inferred from both allozymes and mtDNA data-sets, were in substantial agreement and depicted C. bioccai as closely related to the sibling species of the C. rudolphii complex (C. rudolphii A and C. rudolphii B) and C. septentrionale. Morphological analysis and a differential diagnosis based on male specimens of C. bioccai, which had been genetically characterised by both allozyme markers and mtDNA sequences analysis with respect to morphologically related congeners, enabled the detection of differences in a numbers of characters, including spicule length, the morphology of the distal end of the spicule and the distribution patterns of the distal caudal papillae.     

Male-biased parasitism by common helminths is not explained by sex differences in body size or spleen mass of breeding cormorants Phalacrocorax auritus

In vertebrates, males are often more parasitised than conspecific females. This bias in parasitism might result from sex differences in parasite exposure and/or susceptibility to infection. Such information is important for testing hypotheses about allocation of resources to life histories of males and females and for testing hypotheses about factors thought to influence parasite fitness and parasite dynamics. We tested whether double-crested cormorants Phalacrocorax auritus exhibit male-biased parasitism by gut helminths. The prevalence of nematode Contracaecum spp. and trematode Drepanocaphalus spathans infections was ∼90% and 39%, respectively. Cestode, primarily Paradilepis caballeroi and acanthocephalan Andracantha gravida infections were less common (<10%). Male and female cormorants did not differ in prevalence of infection by any helminth species. However, males had twice the abundance and intensity of Contracaecum spp. infections and twice the intensity of D. spathans infections than found in females. For common parasites showing male-biased parasitism, degree of parasitism was also unrelated to body size or mass in either sex. Males and females did not differ in spleen mass and spleen mass was unrelated to abundance of common parasites. Furthermore, abundance of trematodes and nematodes was not correlated. At present, male biases in parasitism by nematodes and trematodes in cormorants are independent patterns that remain unexplained, but are most likely attributable to sex differences in exposure and/or immunological differences not yet assessed.     

Infection by Contracaecum rudolphii (Nematoda: Anisakidae) in the Neotropic cormorant Phalacrocorax brasilianus,and fishes from the estuary of the Valdivia river,Chile

Seasonal parameters of infection by the nematode Contracaecum rudolphii in the Neotropic cormorant Phalacrocorax brasilianus (Phalacrocoracidae) and their relation with feeding and infection in fishes from River Valdivia, Chile, were determined. The prevalences of infection in birds were similar during seasons, whereas mean intensity and percentages of adult gravid females were higher in spring and summer, respectively. For fishes no seasonal differences were found in infection. Cormorant diet varied seasonally in relation to fish prey consumed and this can be related to seasonal differences in infection parameters of birds. The high infection in birds should be considered as a potential risk for salmoniculture.     

Gastrointestinal helminths of fish-eating birds from Chiloe Island, Chile

Of four species of fish-eating birds from Chiloé Island (Chile), three were infected with eight species of helminths. Tetrabothrius sp. was found in Larus scoresbii. Tetrabothrius cylindraceus, Profilicollis antarcticus, Anomotaenia dominicanus, Stephanoprora denticulata, Capillaria sp. and P. antarcticus were found in Larus dominicanus. Contracaecum rudolphii and Corynosoma sp. infected Phalacrocorax olivaceus. With the exception of S. denticulata, C. rudolphii and Capillaria sp., the above helminths are reported for the first time from Chile.     

Contracaecum rudolphii B: gene content, arrangement and composition of its complete mitochondrial genome compared with Anisakis simplex s.l

In the present study, we sequenced the complete mt genome (14,022 bp) of parasitic nematode Contracaecum rudolphii B and its structure and organization compared with Anisakis simplex s.l. The mt genome of C. rudolphii B is slightly longer than that of A. simplex s.l. (13,916 bp). C. rudolphii B mt genome is circular, and consists of 36 genes, including 12 genes for proteins, 2 genes for rRNA and 22 genes for tRNA. This genome contains a high A+T (70.5%) content. The mt gene order for C. rudolphii B is the same as those for A. simplex s.l., but it is distinctly different from other nematodes compared. The start codons inferred in the mt genome of C. rudolphii B are TTG and ATT. Six protein-coding genes use TAA as a stop codon whereas five genes use T and one genes use TAG as a termination codon. This pattern of codon usage reflects the strong bias for A and T in the mt genome of C. rudolphii B. Phylogenetic analyses using concatenated amino acid sequences of the 12 protein-coding genes, with three different computational algorithms (Bayes, ML and MP), all revealed distinct groups with high statistical support, indicating that C. rudolphii B and A. simplex s.l. is distinct but closely related species. These data provide additional novel mtDNA markers for studying the molecular epidemiology and population genetics of the C. rudolphii B, and should have implications for the molecular diagnosis, prevention and control of anisakidosis in humans and animals.     

Coexistence and genetic variability of Contracaecum rudolphii A and Contracaecum rudolphii B (Nematoda: Anisakidae) in cormorants, Phalacrocorax carbo sinensis, in the Baltic region

Although numerous genetically isolated entities within Contracaecum rudolphii sensu lato are presently defined, information on the distribution and ecology of these groups is nonetheless in demand. In the present study, information based upon DNA sequence data (restriction analysis and sequencing of rDNA) on the distribution of the species C. rudolphii A and C. rudolphii B in the cormorant Phalacrocorax carbo sinensis from the Baltic region (Poland and Finland) is provided. These data corroborate previous work that identified C. rudolphii A in brackish water regions and C. rudolphii B in fresh water sites, although mixed infections also occur. The 2 species may inhabit the same host specimen. One rare heterozygote of the species was recorded.     

Effect of selected anthelmintics on three common helminths in the brown pelican (Pelecanus occidentalis)

The effect of selected anthelmintics (albendazole, fenbendazole, piperazine dihydrochloride and clorsulon) against three major helminths (Contracaecum multipapillatum, Mesostephanus appendiculatoides, and Phagicola longus) were studied in 29 brown pelicans (Pelecanus occidentalis). Albendazole and fenbendazole were highly effective against all three parasites. Clorsulon had moderate effect against M. appendiculatoides and poor effect against C. multipapillatum and P. longus. Piperazine dihydrochloride had no effect against these helminths.     

The development of Eustrongylides tubifex (Nematoda: Dioctophymatoidea) in oligochaetes

Egg development of Eustrongylides tubifex (Nitzsch in Rudolphi, 1819) J?gerski?ld, 1909, was studied at 5, 10, 15, 20, 25, and 30 C. Eggs developed at 20, 25, and 30 C. Development ceased at 0, 5, 10, and 15 C but resumed when temperatures were raised. Eggs contained first-stage larvae in 23-26 days at 25 C. Seven species of aquatic oligochaetes were exposed experimentally to eggs of E. tubifex containing first-stage larvae. Third-stage larvae of E. tubifex developed in the aquatic oligochaetes, Limnodrilus hoffmeisteri Claparède, 1862, and Tubifex tubifex (Müller, 1774). Larvae developed in the ventral blood vessel of oligochaetes. Second-stage larvae were seen first in oligochaetes 30 days postinfection and third-stage larvae were seen 70-109 days postinfection at 25 C. Third-stage larvae in oligochaetes retained the cuticle of the first and second molt.     

Gastric nematodes of the Paraguayan caiman, Caiman yacare (Alligatoridae).

Examination of the stomach contents of 115 Caiman yacare Daudin, 1802, from Paraguay revealed the presence of the nematodes Brevimulticaecum baylisi (Travassos, 1933), Brevimulticaecum stekhoveni (Baylis, 1947), Contracaecum sp. of Railliet and Henry, 1912, Dujardinascaris paulista (Travassos, 1933), Eustrongylides sp. of Jagerskiold, 1909, Micropleura vazi Travassos, 1933, and Ortleppascaris alata (Baylis, 1947). Nematode prevalence was 28%; greatest species prevalence (14%) was found for B. baylisi. Greatest mean intensity, 25.4, was for Contracaecum sp. All occurrences are new host and distribution records.     

Sexual differences in the diet of great cormorants <Emphasis Type="Italic">Phalacrocorax carbo sinensis</Emphasis> wintering in Greece

Sexual differences in the diet of the great cormorant, Phalacrocorax carbo sinensis, were studied in four Greek wintering areas, the Amvrakikos Gulf, the Axios and Evros Deltas and the Messolonghi Lagoon, through the analysis of stomach contents. Great cormorants are birds sexually dimorphic in size, with males being generally larger than females. Although similar prey species were found in the stomachs of both sexes in all the studied areas, significant differences were observed with respect to the proportion of species taken. Male birds ate higher proportions of large fish species such as grey mullets, European sea bass, Dicentrarchus labrax, and Prussian carp, Carassius gibelio, while female birds took higher proportions of smaller species such as big-scale sand smelt, Atherina boyeri, and black goby, Gobius niger. As a consequence, male great cormorants were found to feed on significantly larger prey than did females by means of fish standard length and body mass. There was no significant difference between the sexes in the mass of food found in stomachs.     

Molecular phylogenetics and diagnosis of Anisakis, Pseudoterranova, and Contracaecum from northern Pacific marine mammals

Individual specimens of Anisakis, Pseudoterranova, and Contracaecum collected from marine mammals inhabiting northern Pacific waters were used for comparative diagnostic and molecular phylogenetic analyses. Forty-eight new sequences were obtained for this study of 14 Anisakis taxa, 8 Pseudoterranova taxa, 4 Contracaecum taxa, and 4 outgroup species. Partial 28S (LSU) and complete internal transcribed spacer (ITS-1, 5.8S, ITS-2) ribosomal DNA was amplified by the polymerase chain reaction and sequenced. Sequences of ITS indicated that Pseudoterranova specimens from Zalophus californianus (California sea lion), Mirounga angustirostris (northern elephant seal), Phoca vitulina (harbor seal), Enhydra lutris (sea otter), and Eumetopias jubatus (Steller's sea lion) exactly matched P. decipiens s. str., extending the host and geographic range of this species. Anisakis from northern Pacific marine mammals were most closely related to members of the A. simplex species complex. Comparison of Anisakis ITS sequences diagnosed the presence of A. simplex C in 2 M. angustirostris hosts, which is a new host record. Anisakis specimens from Phocoena phocoena (harbor porpoise), Lissodelphis borealis (Pacific rightwhale porpoise), and E. jubatus included 3 ITS sequences that did not match any known species. Contracaecum adults obtained from Z. californianus were most closely related to C. ogmorhini s.l. and C. rudolphii, but ITS sequences of these Contracaecum specimens did not match C. ogmorhini s. str. or C. margolisi. These novel Anisakis and Contracaecum ITS sequences may represent previously uncharacterized species. Phylogenetic analysis of LSU sequences revealed strong support for the monophyly of Anisakinae, Contracaecum plus Phocascaris, Pseudoterranova, and Anisakis. Phylogenetic trees inferred from ITS sequences yielded robustly supported relationships for Pseudoterranova and Anisakis species that are primarily consistent with previously published phenograms based on multilocus electrophoretic data.