Volume 58, Issue 4

Five new species and one new genus of the subfamily Harpirhynchinae (Acariformes: Harpirhynchidae) are described from birds in Canada: Harpyrhynchoides heatherae sp. nov. from Junco hyemalis (Passeriformes: Emberizidae), H. botaurus sp. nov. from Botaurus lentiginosus (Pelecaniformes: Ardeidae), H. phalaropus sp. nov. from Phalaropus lobatus (Charadriiformes: Scolopacidae), Neharpyrhynchus loxia sp. nov. from Loxia curvirostra (Passeriformes: Fringillidae), and Fainharpirhynchus contopus gen. nov., sp. nov. from Contopus cooperi (Passeriformes: Tyrannidae). Additionally, 3 species were recorded in Canada (Manitoba) for the first time: Harpyrhynchoides tracheatus (Fritsch, 1954) from Buteo jamaicensis (Accipitriformes: Accipitridae) (new host), H. modestus (Fain, 1976) from Columba livia (Columbiformes: Columbidae) (new host), and Neharpyrhynchus pilirostris (Berlese et Trouessart, 1889) from Passer domesticus (Passeriformes: Passeridae).

In the present study, transmission electron microscopy (TEM) has been used to study the ultrastructure of the digestive system, namely the pharynx, oesophageal glands and intestine, of the monogenean skin and fin parasite Macrogyrodactylus congolensis. The pharynx consists of an anterior highly muscular region and a posterior mainly glandular syncytial region. The anterior region is provided with six pharyngeal papillae, the centre of each of which is occupied by electron dense secretory bodies, identical with those in the posterior region of the pharynx. The intestine has an uninterrupted syncytial gastrodermis and the luminal surface is provided with many unbranched lamellae. The intestine of living specimens contains large and small granules which give it a reddish brown colour. Large particles, presumed to be lipid droplets, and small granules, presumed to be melanin granules, were found in the gastrodermis and in the intestinal lumen. Parasites were induced to feed and then preserved for TEM at the following intervals: just after feeding, 30 min after feeding, 1 h 30 min after feeding and 2 h after feeding. The specimens were then processed for TEM and sections cut through the intestine of each specimen were examined with the transmission electron microscope. Three types of vacuoles (V1, V2, V3) were detected in the gastrodermis. Vacuoles V1 have thick walls and are likely to be endocytotic, enclosing luminal contents at the surface of the gastrodermis. V2 vacuoles may be lysosomes that fuse with V1 vacuoles. V3 vacuoles may serve to dispose of residual digestive material into the lumen.

Alloglossidium demshini sp. nov. is described based on specimens from leeches Haemopis grandis collected in northwestern Minnesota. The new species is morphologically closest to Alloglossidium schmidti. The two species can be readily differentiated based on several morphological characters. The cirrus sac in A. schmidti is almost entirely situated anterior to the ventral sucker while in A. demshini sp. nov. it is situated dorsal to the ventral sucker and its proximal end almost reaches the posterior end of ventral sucker or extends posterior to it. The new species has a prepharynx that is substantially longer than the esophagus while in A. schmidti the situation is the opposite. The two species also differ in the position of the ovary and the position of the testes and vitelline fields in relation to the ends of the ceca. Hirudineatrema oschmarini described from leeches in Eastern Palaearctic and Alloglossidium richardsoni described in North America demonstrate great morphological similarity. Nevertheless, Hirudineatrema cannot be synonymized with Alloglossidium at this point because of several peculiar morphological features of H. oschmarini such as a V-shaped excretory bladder and the apparent presence of a true seminal receptacle in the latter species. These features need to be re-evaluated before any taxonomic decision can be made.

Hymenolepidid cestodes in Myodes glareolus from Lithuania and additional specimens originally attributed to Arostrilepis horrida from the Republic of Belarus are now referred to A. tenuicirrosa. Our study includes the first records of A. tenuicirrosa from the European (western) region of the Palearctic, and contributes to the recognition of A. horrida (sensu lato) as a complex of cryptic species distributed broadly across the Holarctic. Specimens of A. tenuicirrosa from Lithuania were compared to cestodes representing apparently disjunct populations in the eastern Palearctic based on structural characters of adult parasites and molecular sequence data from nuclear (ITS2) and mitochondrial (cytochrome b) genes. Morphological and molecular data revealed low levels of divergence between eastern and western populations. Phylogeographic relationships among populations and host biogeographic history suggests that limited intraspecific diversity within A. tenuicirrosa may reflect a Late Pleistocene transcontinental range expansion from an East Asian point of origin.

Fasciola hepatica is a liver fluke that infects 2.4 million of people and causes great economical loss in animal production. To date a 100% effective vaccine has not been developed and the disease is controlled by drug therapy. Great efforts are put into development of effective vaccine against parasite what is difficult since Fasciola spp. (like other helmints) during evolutionary process has developed sophisticated and efficient methods to evade immune response. During preliminary experiments it is convenient to use cell lines which are relatively cheap and allow for reproducible comparison of results between laboratories. We stimulated BOMA (bovine monocyte/macrophage cell line) and BOMAC (bovine macrophage cell line) with native or recombinant antigens of Fasciola hepatica and assessed IFN-γ, IL-4 and TNF-α level upon stimulation. We observed diminished secretion of proinflammatory TNF-α in LPS activated BOMA cells stimulated with Excretory/Secretory products of adult fluke (Fh-ES). We also observed greater changes in gene expression in LPS activated BOMA cells than in non activated BOMA cells upon stimulation using Fh-ES. The results show possibility of using cell lines for in vitro research of bovine immune response against liver fluke, although this model still requires validation and further characterization.

The course of anti-Neospora antibody kinetics during two consecutive pregnancies has been evaluated in five chronically infected dairy cows. The blood samples of cows were collected monthly. Anti-Neospora antibodies in blood sera were detected by indirect ELISA (ID-VET, France). During whole period of the study cows remained seropositive; with S/P% values (iELISA) ranging from 94 to 214%. The antibody kinetics determined by iELISA showed a significant increase (P<0.0059) of specific IgG antibodies in the third trimester of both pregnancies. The monitored cows gave birth to 10 healthy calves (4 steers and 6 heifers). To confirm the occurrence of vertical transmission of Neospora from mothers to offspring in the herd, dam-daughter serology was performed. Anti-Neospora antibodies was found in 4 from 6 heifers (>6 months old). Study presents original data reporting on the very similar persistent pattern of anti-Neospora antibody levels during the third trimester of pregnancies in all five dairies. Based on the high seropositivity of female offspring, the reactivation of a latent infection of cows rather than a re-infection can be supposed.

Although multiple vaccine strategy for lymphatic filariasis has provided tremendous hope, the choice of antigens used in combination has determined its success in the previous studies. Multiple antigens comprising key vaccine candidates from different life cycle stages would provide a promising strategy if the antigenic combination is chosen by careful screening. In order to analyze one such combination, we have used a chimeric construct carrying the well studied B. malayi antigens thioredoxin (BmTRX) and venom allergen homologue (BmVAH) as a fusion protein (TV) and evaluated its immune responses in mice model. The efficacy of fusion protein vaccine was explored in comparison with the single antigen vaccines and their cocktail. In mice, TV induced significantly high antibody titer of 1,28,000 compared to cocktail vaccine TRX+VAH (50,000) and single antigen vaccine TRX (16,000) or VAH (50,000). Furthermore, TV elicited higher level of cellular proliferative response together with elevated levels of IFN-γ, IL-4 and IL-5 indicating a Th1/Th2 balanced response. The isotype antibody profile showed significantly high level of IgG1 and IgG2b confirming the balanced response elicited by TV. Immunization with TV antigen induced high levels of both humoral and cellular immune responses compared to either cocktail or antigen given alone. The result suggests that TV is highly immunogenic in mice and hence the combination needs to be evaluated for its prophylactic potential.

The tegument of the microphallid digenean Maritrema feliui, examined by means of TEM, is described as a syncytial epithelium organised into two layers. The outer layer is an external anucleate, cytoplasmic region connected to a second region composed of nucleate perikarya (cytons) deeply embedded in the surrounding cortical parenchyma. The surface layer of the tegument is covered by a plasma membrane with many deep invaginations, which are apparently pinocytotic. This layer also bears numerous large, electron-dense spines of two types, which are intracellular and attached to the basal plasma membrane. Its cytoplasm is rich in free ribosomes, contains numerous mitochondria, disc-shaped granules frequently arranged in a rouleau, and several large, moderately electron-dense, membranous bodies. The subtegumentary perikarya and their nuclei, which are both flattened, are described in detail, as are their connections with the surface tegument. These perikarya appear to be the source of the disc-shaped granules and some of the other inclusions present in the surface layer. The main characteristics of the tegumental structure of M. feliui are commented upon in relation to the findings of previous publications and their suggested functions.

Larvae of a vast majority of water mite species are parasites of aquatic insects. Owing to this, they migrate to new localities, and are able to survive unfavourable environmental conditions. This also concerns species from subgenus Arrenurus s. str., parasites of dragonflies. The detailed analysis of this phenomenon, however, has only been possible in the last several years, since the key to the identification of larvae from genus Arrenurus Dug. was published. In 2010, the parasitism of Arrenurus s. str. larvae on dragonflies in the Lake Świdwie reserve (NW Poland) was analysed. Larvae of 9 species of water mites were recorded on 107 imagines of dragonflies from 8 species. The following were identified as hosts of water mites for the first time: Anax imperator, Libellula quadrimaculata, and Leucorrhinia caudalis. The highest prevalence occurred in the case of: Erythromma najas and Lestes dryas (100%), Coenagrion pulchellum (96.5%), and C. puella (80.0%). Coenagrion pulchellum was infested by 9 species of parasites, C. puella by 6, and Erythromma najas and Lestes dryas by three species. The highest number of host species occurred in the case of Arrenurus maculator (5); followed by A. cuspidator, A. batillifer cf., A. bicuspidator, and A. tetracyphus (3 each); A. papillator, A. tricuspidator, and A. bruzelii (2 each), and A. claviger (1). Differentiation of preferences of particular parasites towards various parts of the host body was observed, probably related to the coevolution of parasites and hosts, and competition between the host species. The body sizes of the parasites suggest that approximately 50% of body size growth of water mites from subgenus Arrenurus s. str. occurs at the stage of parasitic larva.

Two species of parasitic nematodes, Heliconema ahiri Karve, 1941 (Physalopteridae) and Procamallanus (Spirocamallanus) anguillae Moravec et al., 2006 (Camallanidae), were recorded from the Indonesian shortfin eel Anguilla bicolor bicolor Mc-Clelland in the Chalakkudi River and Veli Lake, respectively, both Kerala, India. The former species (H. ahiri), rediscovered from eels in India after 82 years from its original description, has been redescribed and revalidated based on its specific feature (presence of groups of minute denticles in the mouth) distinguishing it from other congeners; its finding in A. b. bicolor represents a new host record. The latter species (P. anguillae) has been recorded in India for the first time. SEM examination of its first-stage larvae from uterus has shown that P. anguillae is another species of Procamallanus whose larvae possess a crown of digit-like processes at the tail tip.

We conducted a parasitological study to identify nematodes infecting the gonads of bigeye barracuda Sphyraena forsteri Cuvier (Sphyraenidae) caught in the West Philippine Sea, off the Mariveles coast, Bataan Province, the Philippine archipelago. Males and very tiny nongravid female philometrid nematodes were collected from the ovaries of the fish. Based on light and scanning electron microscope observations, and supported by molecular analyses of the partial nucleotide sequences of the 18S small subunit ribosomal DNA, the philometrid nematode was determined to be a new species belonging to the genus Philometra Costa, 1845 in the family Philometridae (Nematoda: Dracunculoidea), and was named P. philippinensis sp. nov. This new species was clearly distinguishable morphologically from other congeners that parasitize the gonads of marine fishes. Males have unique sickle-shaped spicules and gubernaculum, and a vulva-like cloacal opening. Following Dentirumai philippinensis Quiazon et Moravec, 2013 and Philometra robusta Moravec, Möller et Heeger, 1992, this is the third philometrid species and the first gonad-infecting philometrid from the Philippine archipelago.

Ashworthius sidemi, a nematode belonging to the family of Trichostrongylidae, is a primary parasite of the Asian deer, mainly sika deer (Cervus nippon), with which it was introduced to Ukraine, as well as Slovakia, the Czech Republic and France. Migrating red deer carried this parasite from neighboring countries to Polish territory. Until now, in Poland, this parasite has been recorded in European bison, red deer, roe deer and fallow deer. As a result of post-mortem examinations of 10 elk, 2 of them from the Augustów Forest and Biebrza Marshes, A. sidemi were found in abomasa for the first time in Poland. The intensity of the invasions was 120 and 7 specimens, respectively. This finding of Ashworthius sidemi in elk indicates a further expansion of the focus of ashworthiosis in BiaŁowieża towards the north into the Biebrza Marshes and the Augustowska Forest. The growth of the elk population and their tendency for long distance migrations can contribute to the spread of the parasitosis in much greater distances than deer. On the basis of our own research and data from the literature, the current spread of ashworthiosis in Poland is discussed.

Sexually transmitted diseases are a major cause of acute disease worldwide, and trichomoniasis is the most common and curable disease, generating more than 170 million cases annually worldwide. Trichomonas vaginalis is the causal agent of trichomoniasis and has the ability to destroy in vitro cell monolayers of the vaginal mucosa, where the phospholipases A2 (PLA2) have been reported as potential virulence factors. These enzymes have been partially characterized from the subcellular fraction S30 of pathogenic T. vaginalis strains. The main objective of this study was to purify a phospholipase A2 from T. vaginalis, make a partial characterization, obtain a partial amino acid sequence, and determine its enzymatic participation as hemolytic factor causing lysis of erythrocytes. Trichomonas S30, RF30 and UFF30 sub-fractions from GT-15 strain have the capacity to hydrolyze [2-14C-PA]-PC at pH 6.0. Proteins from the UFF30 sub-fraction were separated by affinity chromatography into two eluted fractions with detectable PLA A2 activity. The EDTA-eluted fraction was analyzed by HPLC using on-line HPLC-tandem mass spectrometry and two protein peaks were observed at 8.2 and 13 kDa. Peptide sequences were identified from the proteins present in the eluted EDTA UFF30 fraction; bioinformatic analysis using Protein Link Global Server charged with T. vaginalis protein database suggests that eluted peptides correspond a putative ubiquitin protein in the 8.2 kDa fraction and a phospholipase preserved in the 13 kDa fraction. The EDTA-eluted fraction hydrolyzed [2-14C-PA]-PC lyses erythrocytes from Sprague-Dawley in a time and dose-dependent manner. The acidic hemolytic activity decreased by 84% with the addition of 100 μM of Rosenthal’s inhibitor.

Despite the importance of buffalo farming in Iran, little is known in this country about the abundance and distribution of Eimeria spp. in the animal species. The present study was designed to investigate the prevalence and species characterization of Eimeria oocysts in river buffalo calves of Khuzestan province, southwest of Iran. Of the total 108 fecal samples examined for Eimeria, 108 (100%) were found infected with 11 species of the parasite. Among the identified species of Eimeria, E. bovis was found to be the predominant etiological agent (76.85%), followed in order by E. canadensis (62.96%), E. zuernii (47.2%), E. ellipsoidalis (26.85%), E. subspherica (25.92%), E. brasiliensis (19.4%), E. auburnensis (18.51%), E. alabamensis (14.81%), E. pellita (11.1%), E. illinoisensis (5.5%) and E. bukidnonensis (2.7%). In most calves multiple infections with three species were present. While, 20.7% of calves showed heavy infection, 50.4 and 24.8% of calves showed weak and moderate infection, respectively. There was no significant difference in the OPG values between the calves of different localities. There was also no significant difference between the prevalence rate of infection in males and females. A total of 16.6% of all faecal samples were found to be diarrheic. A highly significant relationship could be identified between the occurrence of diarrhea and the level of E. bovis and E. zuernii oocysts excretion. Considering the pervasive occurrence and negative effects of the infection on the health condition and the growth performance of buffalo calves, infections should receive increased attention by both farmers and veterinarians.

Juvenile golden grey mullet, Liza aurata were collected from Sarıkum Lagoon Lake which connected to the Black Sea at Sinop, Turkey and examined for parasitic fauna. A total of 219 fish were investigated throughout a 1-year period. Parasite species recovered were Trichodina lepsii, T. puytoraci, Gyrodactylus sp., Ligophorus cephali, Ligophorus mediterraneus, Solostamenides mugilis, Ascocotyle sp. (metacercaria) and Ergasilus lizae. Overall infection prevalence (%) and mean intensity values were 95.9% and 412.65 ± 85.31 parasites per infected fish, respectively. Infection prevalence and mean intensity values for each parasite species in relation to season and fish size were also determined and discussed. While Ligophorus cephali and L. mediterraneus are new records for Turkish parasite fauna, the juvenile Liza aurata is a new host record for Ligophorus cephali and L. mediterraneus.

Parasitic diseases of livestock together with poor welfare conditions can negatively affect the health status and production of small ruminants. Protozoan parasites and tick-borne infectious agents are common threat of livestock including small ruminants mostly during the pasture season. Therefore the priority of the study was to analyse the circulation and presence of two protozoan parasites Toxoplasma gondii and Neospora caninum as well as tick-transmitted bacterium Anaplasma phagocytophilum in one selected goat farm in Eastern Slovakia. Throughout a three-year study period we have repeatedly screened the sera and blood of goats and dogs from monitored farm. In total, 343 blood serum samples from 116 goats were examined by ELISA. The mean seropositivity for T. gondii was 56.9% (66/116, CI (95%) = 48–66.0) and 15.5% (18/116, CI (95%) = 9.3–22.7) for N. caninum. The permanent occurrence of anti-Toxoplasma and anti-Neospora antibodies was detected in repeatedly examined goats during the whole monitored period. The presence of both parasites in the flock was analysed by PCR. DNA of T. gondii was confirmed in 12 out of 25 Toxoplasma-seropositive goats and N. caninum in 14 samples out of 18 Neospora-seropositive animals; four goats were co-infected with both pathogens. The risk of endogenous transmission of both parasites was pursued by examination of 41 kid’s sera, where seropositivity for toxoplasmosis was 31.7% and for neosporosis 14.6%. In dogs 61.1% seropositivity for T. gondii and 38.9% for N. caninum was found, however, their faeces were negative for coccidian oocysts. Eight out of 108 tested animals were infected with A. phagocytophilum, the causative agent of tick-borne fever. Seven of them were simultaneously infected with T. gondii and A. phagocytophilum, out of which four goats were concurrently infected with all three pathogens.

New data on the infection with Acanthocephala of 33 fish caught in Admiralty Bay in November 2007 to January 2008 are given. These fish belong to 5 species: Notothenia rossii (22 immature specimens), Lindbergichthys nudifrons (n = 7), Trematomus bernacchii (n = 1), T. newnesi (n = 1) and Harpagifer antarcticus (n = 2). Three species of Echinorhynchida: Aspersentis megarhynchus, Metacanthocephalus dalmori and M. johnstoni and four species of Polymorphida: Corynosoma arctocephali, C. bullosum, C. hamanni and C. pseudohamanni, were found. Prevalence of N. rossii and L. nudifrons was 100%. The mean abundance of infection of N. rossii (125.09) was larger than that of Notothenia coriiceps (82.93). Data of infection of N. rossii in 2007 was almost identical with that in 1979 (mean abundance 118.66). The most abundant in this host were A. megarhynchus, M. johnstoni, C. hamanni and C. pseudohamanni (mean abundances 36.36, 29.77, 13.86 and 44.73, respectively). In total Echinorhynchida were more abundant than Polymorphida in 2007/08 (66.18 versus 58.91). Reverse situation was in 1979 (mean abundance 47.36 for Echinorhynchida and 71.3 for Polymorphida. Only 7 L. nudifrons were examined in 2007/08 and Echinorhynchida were more numerous in this host (mean abundance 26.71) than Polymorphida (10.29). Single specimens of other fish were infected with a few Acanthocephala belonging to species recorded in the same hosts with those found in 1978/79.

Tumor-associated immune system cells secrete protease and cytokines that can inhibit the immune response. In particular, T-cell effector functions could be inhibited, potentially causing an increase in parasitic infestations. Demodex species are common inhabitants of normal hair follicles. Humans are the specific host for two species Demodex folliculorum and D. brevis. The aim of this study was to investigate the incidence and infestation of D. folliculorum and D. brevis in patients with cancer. In the present study, 101 patients with cancer were selected from among patients who were diagnosed and treated for cancer. The cancer patients were divided into four groups according to cancer type. Slides were examined for parasites using light microscopy at magnifications of ×40 and ×100. Infestation was defined as having at least five living parasites/cm2 of skin. The ages of the patients with cancer ranged between 38 and 82 years, with a mean of 65.5±10.1 years. It was determined that 77 of the 101 (76.2%) cancer patients were positive for Demodex species. Infestation was positive in 18 (47.4%) of the 38 cases in the breast cancer group, 7 (29.2%) of the 24 cases in the lung cancer group, 5 (18.5%) of the 27 cases in the gastrointestinal system cancer group, and 2 (16.7%) of the 12 cases in the urogenital system cancer group. Results showed that the rate of Demodex species infestation was higher in patients with breast cancer. Thus, cancer — and particularly breast cancer — is a risk factor for Demodex species infestation.

A total of 71 bat flies belonging to families Nycteribiidae and Streblidae, and 37 mites were collected on 12 species of bats (Phyllostomidae and Vespertilionidae) from the Chapada do Abanador (Minas Gerais, Brazil), between July 2009 and April 2010. Two new occurrences of ectoparasites were recorded on Histiotus velatus (bat fly Basilia producta) and on Carollia perspicillata (mite Parichoronyssus bakeri). Five new occurrences were recorded for the state of Minas Gerais, increasing the range for bat flies Anatrichobius passosi, Paraeuctenodes similis, Basilia juquiensis, Basilia producta and for mite Periglischrus vargasi. Moreover, two new species of mites were recorded for Brazil (P. bakeri and Macronyssus aff. leislerianus). With regard to infracommunities, the most frequent association was between Anastrebla modestini and Exastinion clovisi on bat Anoura geoffroyi. This study contributed to characterize the fauna of bat ectoparasites in representative but poorly-sampled environments of the Atlantic Forest, the campos de altitude (high altitude grasslands) and cloud forests of southern Minas Gerais.

Malaria is wide spread in poor world and its burden has been assessed by the enumeration of malarial parasites in blood of patients. This study was designed to find a relationship between social structure, and spread of malaria in Khyber agency. The average parasite density was 2050 parasite/μl in Khyber Agency. Due to economic and social setup most of the people have habit of sleeping in open air thus playing role in high malaria prevalence and Plasmodium vivax remains the prevalent species. Genetic study performed on 110 Blood samples showed less genetic diversity for both Plasmodium vivax and Plasmodium falciparum. Eight alleles were distinguished both for Pvmsp 3α and Pvmsp 3β in total of 20 and 39 amplified samples of P. vivax respectively. Out of 17 samples amplified for P. falciparum 11 showed genotype K1 and 10 for MAD at Pfmsp-1 while 14 alleles were identified for 3D7/1C and two for FC27 of corresponding families of Pfmsp-2 gene. This shows that Plasmodium parasites are not genetically diverse in Khyber agency.

Red foxes can be infected with diverse range of parasite species that can be transmitted to humans and pet animals, and the differential diagnosis is essential for veterinary and zoonotic risk assessment. In the present study, faecal flotation and microscopy of parasite eggs was done on 31 foxes originating from two distant localities in Denmark, the city of Copenhagen in the north east part of the island Zealand and from the southern part of the peninsular Jutland. In total, eggs of Eucoleus boehmi were recovered from a surprisingly high number of foxes (n = 22 samples; 71%). The findings represent the first observations of E. boehmi in Denmark, which is likely an overlooked infection of the upper respiratory tract of red foxes, which can be also found in other canids. Several morphological features of the eggs of E. boehmi can be used to distinguish it from other the closely related trichuroid eggs. Detecting cardiopulmonary parasites by faecal examination can be indicative of the presence of cardiopulmonary parasites based on the more sensitive post mortem analysis.

The importance of small-scale heterogeneity in local factors which overrides that of larger-scales factors, suggest that local factors play a major role in determining the richness and prevalence of larval digeneans in intertidal gastropods. The restricted distribution of the snail Heleobia australis along a 500 m transect in Cangrejo creek (Mar Chiquita, Argentina) provides a good opportunity to test the assumption that a study at spatial scale of 100s meters can detect spatiotemporal fluctuations of a larval digenean assemblage. To analyze that, 3600 specimens of H. australis were collected seasonally during the year 2011. A quantitative variation and a space-time interaction between sampling points and seasons of the year for the total prevalence of larval digeneans and snail’s densities were found, as well as a positive correlation with abiotic factors. These results revealed that the fluctuations in the community of larval digeneans of the snail H. australis can be detected at small spatial scale, using its natural distribution of 500 m. This study also highlights the importance of seasonality as a factor that must be considered in studies focused on the search for patterns structuring the communities of larval digeneans, at medium and large scales.

This study presents data from a molecular survey of the species of the genus Culicoides from the region of Kalimok Field Station (NE Bulgaria) and haemosporidian parasites occurring in them in order to investigate the host-parasite specificity of haemosporidians to their dipteran vectors. The identification of Culicoides spp. was carried out by morphological and molecular-genetic methods. We collected and analysed 230 individuals of the genus Culicoides. Nine species were found. Eight species were identified morphologically; Culicoides obsoletus, C. riethi, C. newsteadi, C. circumscriptus, C. festivipennis, C. punctatus, C. pictipennis and C. puncticollis. The ninth species might be classified as either of C. nubeculosus or C. riethi and its identification needs additional investigations. The total prevalence of Haemoproteus in the examined biting midges was 2.17%. Three individuals of C. pictipennis were infected with the Haemoproteus lineage TURDUS2 (prevalence 16.67%), a common parasite of thrushes (Turdidae). Two individuals of C. circumscriptus contained Haemoproteus lineages (prevalence 2.78%); these were the lineage HAWF2 (previously reported from Coccothraustes coccothraustes) and a new lineage CULCIR1 not previously reported in the literature.

The filarial nematode Dirofilaria repens is currently considered to be one of the most extensively spreading human and animal parasites in Europe. In Ukraine, reporting cases of dirofilariasis has been mandatory since 1975, and the disease was included in the national surveillance system for notifiable diseases. Up until December 31st 2012, a total of 1533 cases have been registered, with 1465 cases occurring within the previous 16 years. Most of the cases of dirofilariasis were registered in 6 regions: Kyiv, and the Donetsk, Zaporizhzhya, Dnipropetrovsk, Kherson and Chernihiv oblasts. In the years 1997–2002 the highest incidence rate was noted in the Kherson oblast in the south of the country (9.79 per 100 000 people), and the lowest in western Ukraine (0.07–1.68 per 100 000 people). D. repens infections were registered in all oblasts. Parasitic lesions were most often located in the head, the subconjunctival tissue and around the eyes. D. repens lesions were also found in the limbs, torso, male sexual organs, and female mammary glands. Dirofilariasis was diagnosed in persons aged from 11 months to 90 years old, most often among people between 21–40 years of age. Most patients had only one parasitic skin lesion; the majority of isolated nematodes were female. The results of our analysis point to a constant increase in D. repens dirofilariasis incidence in humans in Ukraine. Despite educational efforts, infections have become more frequent and the territory in which the disease occurs has enlarged to encompass the whole of Ukraine. Nevertheless, the Ukrainian sanitary-epidemiological services managed to achieve some measure of success, e.g. by creating a registration system for D. repens infections and establishing proper diagnostics for the disease.

Dicyemid mesozoan parasites, microscopic organisms found with high intensities in the renal appendages of benthic cephalopods, have a complex, partially unknown life cycle. It is uncertain at which host life cycle stage (i.e. eggs, juvenile, adult) new infection by the dispersive infusoriform embryo occurs. As adult cephalopods have a short lifespan and die shortly after reproducing only once, and juveniles are fast-moving, we hypothesize that the eggs are the life cycle stage where new infection occurs. Eggs are abundant and sessile, allowing a huge number of new individuals to be infected with low energy costs, and they also provide dicyemids with the maximum amount of time for survival compared with infection of juvenile and adult stages. In our study we collected giant Australian cuttlefish (Sepia apama) eggs at different stages of development and filtered seawater samples from the S. apama mass breeding aggregation area in South Australia, Australia, and tested these samples for the presence of dicyemid DNA. We did not recover dicyemid parasite cytochrome c oxidase subunit I (COI) nucleotide sequences from any of the samples, suggesting eggs are not the stage where new infection occurs. To resolve this unknown in the dicyemid life cycle, we believe experimental infection is needed.

During a routine inspection of the mosquito fauna in the Košická Basin (Eastern Slovakia), in one of the monitored locations we have caught 4 females of the invasive mosquito Aedes albopictus, using the CO2 baited CDC traps. Occurrence of this particular mosquito has already been reported in many European countries; in Slovakia, however, this is the first finding ever. The finding of Ae. albopictus extends the list of the mosquito fauna in Slovakia to 50 species and Slovakia thus ranks among other 20 European countries where this mosquito was observed. The presence of Ae. albopictus increased the probability of transmission of canine and human dirofilariosis in urban environment.

Water frogs, Pelophylax perezi, that are introduced in the Azores, were screened for parasites using PCR primers known to amplify Apicomplexa parasites, and using nematode-specific primers. With the former, three different organisms were detected: Hepatozoon, a trichodinid protozoan ciliate and a possible Stramenopile. Using the latter set of primers, a single unknown spirurid nematode was also detected. Phylogenetic analyses indicate that Hepatozoon detected within amphibian hosts appear to form a clade, although relationships of these parasites do not match the vertebrate intermediate host phylogeny. Regarding the possible Stramenopile, it is unclear whether this organism was actually present on the amphibian or in the water on the surface of the tissue sample. Our findings highlight that many different organisms can be detected with these primers and that they can be used to screen introduced host populations to detect parasites that have been brought with them.

Dirofilariosis, caused by Dirofilaria immitis and D. repens, is (re-) emerging worldwide. Dogs are the main reservoirs, while human infection has recently become an important focus of interest and attention. In Argentina, canine D. immitis infection has been described in eastern and northern subtropical and temperate humid regions, but never reported in mid-western arid regions so far. In this research note we report for the first time the occurrence of autochthonous human and canine D. immitis infection in the region.

The effects of a 5 versus 25 miracidia exposure of Echinostoma caproni on the lipid composition of Biomphalaria glabrata was studied using high performance thin layer chromatography (HPTLC)-densitometry. A 50 miracidia dose was not used because such a high level of exposure caused severe snail mortality by 3 weeks post-exposure (PE). Lipids were determined in the digestive-gland gonad complex (DGG) of the exposed snails and in the uninfected matched controls at 2 and 4 weeks PE. Extraction of lipids from DGGs was carried out by the Folch method with chloroform-methanol (2:1), and extracts were analyzed on Analtech HPTLC-HLF pre-adsorbent silica gel plates with measurement of separated bands using a CAMAG Scanner 3. For neutral lipids the mobile phase was petroleum ether-diethyl ether-glacial acetic acid (80:20:1) and the detection reagent was 5% ethanolic phosphoric acid, and for polar lipids chloroform-methanol-deionized water (65:25:4) mobile phase and 10% cupric sulfate in 8% phosphoric acid detection reagent were used. No significant differences in the concentrations of free sterols, free fatty acids, triacylglycerols, phosphatidylcholine, and phosphatidylethanolamine were seen at 2 weeks PE in any of the groups. At 4 weeks PE, the free fatty acid concentration increased significantly in the snails exposed to 25 miracidia compared to that of the 5 miracidia/snail group or the controls. Elevation of the free fatty acid fraction in the high dose snail group suggested that some changes occurred in the lipid metabolism of the snails in that group as a function of miracidia dose.

A heterologous prime-boost vaccination regime with DNA and recombinant vaccinia virus (rvv) vectors expressing relevant antigens has been shown to induce effective immune responses against several infectious pathogens. In this study, we describe the effectiveness of the prime-boost strategy by immunizing dogs with a recombinant plasmid followed by vaccinia virus, both of which expressed the glutamic acid-rich protein (BgGARP) of Babesia gibsoni. The dogs immunized with the prime-boost regime developed a significantly high level of specific antibodies against BgGARP when compared with the control groups. The antibody level was strongly increased after a booster immunization with a recombinant vaccinia virus. Two weeks after the booster immunization with a recombinant vaccinia virus expressing BgGARP, the dogs were challenged with B. gibsoni parasite. The dogs immunized with the prime-boost regime showed partial protection, manifested as a significantly low level of parasitemia. These results indicated that this type of DNA/rvv prime-boost immunization approach may have use against B. gibsoni infection in dogs.