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Four new species of the genus Hatschekia Poche, 1902 (Copepoda: Siphonostomatoida: Hatschekiidae) are described based on female specimens collected from pufferfishes (Tetraodontiformes: Tetraodontidae) caught in coastal waters off the Ryukyu Islands, Japan: H. longiabdominalis sp. n. on Arothron hispidus (Linnaeus), H. geniculata sp. n. on A. hispidus (type host) and A. stellatus (Bloch et Schneider), H. ellipsocorpa sp. n. on A. mappa (Lesson), and H. boonah sp. n. on A. nigropunctatus (Bloch et Schneider) (type host) and A. meleagris (Schneider). Hatschekia longiabdominalis sp. n. and H. boonah sp. n. differ from all other congeners by sharing an unusual, projected abdomen and a fusiform trunk with posterior lobes; these two species are differentiated from each other by the shape of the dorsal chitinous frame on the cephalothorax. Hatschekia geniculata sp. n. can be distinguished by the combination of the following morphological characters: a rhomboidal cephalothorax with a pair of lateral conical protrusions, a cylindrical trunk with posterior lobes and a bent abdomen with a dorsal protrusion. Hatschekia ellipsocorpa sp. n. resembles H. pholas (Wilson, 1906) but can be distinguished from the latter by the possession of one distal and one inner setae on the terminal endopodal segment of legs 1 and 2. Hatschekia pholas is also redescribed based on female specimens from the tetraodontid A. stellatus. At present, 44 nominal species of the genus have been reported from Japan, including four new species described in this paper; 38 of them have been described originally from Japan.

Single-miracidium infections of Lymnaea cubensis (Pfeiffer) from Guadeloupe with the giant liver fluke Fascioloides magna (Bassi, 1875) (Digenea) were carried out during five successive snail generations to determine if this lymnaeid might sustain complete larval development of the parasite. Controls were constituted by a French population of Galba truncatula (Müller) (a single generation) infected according to the same protocol. It was recorded that prevalence and intensity of F. magna infection in L. cubensis progressively increased from F1 to F5 generations. Cercarial shedding of F. magna was noted only within F5 generation of L. cubensis. However, most measured parameters of infection in this species were significantly lower than those noted for G. truncatula and most L. cubensis died after a single shedding wave. Despite this, L. cubensis can be added to the list of potential intermediate hosts of F. magna.

A new nematode species, Comephoronema multipapillatum sp. n. (Cystidicolidae), is described from the anterior intestine and caecum of the squirrelfish Holocentrus adscensionis (Osbeck) (Beryciformes: Holocentridae) collected in Angra dos Reis, State of Rio de Janeiro, offshore Brazil. The new species was placed in Comephoronema Layman, 1933 by having an oval oral aperture, four submedian labia, four bilobed submedian sublabia, two narrow lateral pseudolabia and four single cephalic papillae, but mainly by numerous precloacal papillae in males (more than six pairs), in which it differs from species of the otherwise morphologically similar genus Ascarophis van Beneden, 1871. Comephoronema multipapillatum sp. n. can be easily distinguished from other congeners based on the high number of precloacal papillae in males (18 pairs + one unpaired) and also by the bidentate plate structure on the inner margin of pseudolabia, mature eggs with two long filaments on a single pole, body length of male (9.4-11.5 mm) and female (10.2-19.9 mm), left spicule size (222-278 µm) and length ratio of spicules (1 : 2.2-2.8). This is the fifth nominal species of Comephoronema, the first nematode registered parasitizing H. adscensionis and the first species of the genus in the Neotropical part of the Atlantic Ocean.

This study revises the originally monotypic genus Afrogyrodactylus Paperna, 1968 (Monogenea), the species of which infect alestid fish (Characiformes) in Africa, and includes new records of these parasites from three geographically distant countries, Senegal, Sudan and South Africa. Morphology of opisthaptoral hooks and bars and nuclear ribosomal DNA data revealed three Afrogyrodactylus species. Afrogyrodactylus girgifae sp. n. is described from the fins of the Sudanese nurse tetra, Brycinus nurse (Rüppell), and A. kingi sp. n. presents from the gill arches of the South African sharptooth tetra, Micralestes acutidens (Peters), whereas a previously undescribed Afrogyrodactylus sp. occurred on the fins of B. nurse from Senegal. All three species differ conspicuously from the only one known species of this genus, A. characinis Paperna, 1968, by the dimensions of their haptoral hard parts. Detailed morphological and molecular descriptions and comparisons are presented.

Moxidectin is a macrocyclic lactone belonging to milbemycin family closely related to ivermectin and is currently progressing towards Phase III clinical trial against human infection with the filaria Onchocerca volvulus (Leuckart, 1894). There is a single report on the microfilaricidal and embryostatic activity of moxidectin in case of the human lymphatic filarial parasite Brugia malayi (Brug, 1927) in Mastomys coucha (Smith) but without any adulticidal action. In the present study, the in vitro and in vivo antifilarial efficacy of moxidectin was evaluated on, B. malayi. In vitro moxidectin showed 100% reduction in adult female worm motility at 0.6 µM concentration within 7 days with 68% inhibition in the reduction of MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide dye) (which is used to detect viability of worms). A 50% inhibitory concentration (IC50) of moxidectin for adult female parasite was 0.242 µM, for male worm 0.186 µM and for microfilaria IC50 was 0.813 µM. In adult B. malayi-transplanted primary screening model (Meriones unguiculatus Milne-Edwards), moxidectin at a single optimal dose of 20 mg/kg by oral and subcutaneous route was found effective on both adult parasites and microfilariae. In secondary screening (M. coucha, subcutaneously inoculated with infective larvae), moxidectin at the same dose by subcutaneous route brought about death of 49% of adult worms besides causing sterilisation in 54% of the recovered live female worms. The treated animals exhibited a continuous and sustained reduction in peripheral blood microfilaraemia throughout the observation period of 90 days. The mechanism of action of moxidectin is suggested to be similar to avermectins. The in silico studies were also designed to explore the interaction of moxidectin with glutamate-gated chloride channels of B. malayi. The docking results revealed a close interaction of moxidectin with various GluCl ligand sites of B. malayi.

Cardicola langeli n. sp. (Digenea: Aporocotylidae) infects the heart of sheepshead, Archosargus probatocephalus (Walbaum, 1792) (Perciformes: Sparidae) in the northern Gulf of Mexico off Horn Island (type locality), Mississippi, USA. The new species is described herein using light and scanning electron microscopy of adult specimens and can be most easily distinguished from the other 24 accepted species of Cardicola Short, 1953 by the combination of having (i) an ovovitelline duct that extends anteriad and that (ii) is posterior to the ootype, (iii) a male genital pore that is lateral to the oviducal seminal receptacle and (iv) a female genital pore lateral to the ootype. The new species is the only member of Cardicola so-far reported to have tegumental spines that are distally flattened and broad, rather than pointed. The new species generally resembles the two other species of Cardicola that infect sparids, i.e. Cardicola cardiocolum (Manter, 1947) (type species) from jolthead porgy, Calamus bajonado (Block et Schneider), in the Gulf of Mexico and Cardicola aurata Holzer, Montero, Repullés, Sitja-Bobadilla, Alvarez-Pellitero, Zarza et Raga, 2008, from gilthead seabream, Sparus aurata Linnaeus, in the Mediterranean Sea, by having a spheroid anterior sucker with concentric rows of minute spines anterior to the mouth and by having a similar general arrangement of the vitellarium, gonads and genitalia. However, it differs from them by having the combination of the aforementioned five features plus asymmetrical posterior caeca and a dextral posterior caecum that extends beyond the posterior margin of the ovary. Probable eggs of C. langeli n. sp. that contain a ciliated miracidium infect gill epithelium and are spheroid. An updated list of hosts, infection sites and geographic localities for the 25 accepted species of Cardicola is provided.

Abstract: Three new species of dicyemid mesozoans are described for the first time from Australian octopus and cuttlefish species. Dicyemennea floscephalum sp. n. is described from Octopus berrima Stranks et Norman (southern keeled octopus) collected from Spencer Gulf and Gulf St. Vincent, South Australia, Australia and represents the first description of a species of Dicyemennea Whitman, 1883 from Australian waters. Dicyema papuceum sp. n. and D. furuyi sp. n. are described from Sepia papuensis Hoyle (Papuan cuttlefish) collected from Shark Bay, Western Australia, Australia. Dicyemennea floscephalum sp. n. is a medium to large species that reaches approximately 4.9 mm in length. The vermiform stages are characterised by having 23-28 peripheral cells, and a disc-shaped, flower-like calotte in larger individuals. An anterior abortive axial cell is absent in vermiform embryos and verruciform cells were not observed in nematogens and rhombogens. Infusoriform embryos comprise 37 cells; one nucleus is present in each urn cell. Dicyema papuceum sp. n. is a small species that reaches approximately 1.1 mm in length. The vermiform stages are characterised by having 30-33 peripheral cells and a relatively small, cap-shaped calotte. An anterior abortive axial cell is absent in vermiform embryos and verruciform cells were occasionally observed in nematogens. Infusoriform embryos comprise 37 cells; two nuclei are present in each urn cell. Dicyema furuyi sp. n. is a large species that reaches approximately 5.3 mm in length. The vermiform stages are characterised by having 22-24 peripheral cells and an elongate calotte. An anterior abortive axial cell is absent in vermiform embryos and verruciform cells were not observed in nematogens and rhombogens. Infusoriform embryos comprise 37 cells; one nucleus is present in each urn cell. Three secondary nematogens were also observed in the right renal appendages of two host individuals, confirming the occurrence of this form.

Blood smears prepared from the peripheral blood of 20 wild caught Amietia quecketti (Boulenger) from the North-West University Botanical Gardens, North West Province, South Africa, were examined for the presence of haemogregarines. A haemogregarine species comparative in morphology, host and geographical locality to that of Haemogregarina theileri Laveran, 1905 was detected. The original description of H. theileri was based solely on frog peripheral blood gamont stages. Later, further parasite stages, including trophozoites and merogonic liver stages, were recorded in a related Amietia sp. from equatorial Africa. This species was originally classified as a member of the genus Haemogregarina Danilewsky, 1885, but due to the close life cycle and morphological resemblance to those of Hepatozoon species, H. theileri was later transferred from Haemogregarina to Hepatozoon Miller, 1908. In the present study, meront and merozoite stages not described before, along with previously observed trophozoite, immature and mature gamont stages, are described from the peripheral blood of hosts. In addition, comparative phylogenetic analysis of the partial 18S rDNA sequence of Hepatozoon theileri to those of other haemogregarine species, including those of species of Hepatozoon and a Haemogregarina, support the taxonomic transfer of H. theileri to Hepatozoon, nesting H. theileri within a clade comprising species parasitising other amphibians. This is the first molecular and phylogenetic analysis of an African anuran species of Hepatozoon.

This study describes the proteocephalidean tapeworm Pseudocrepidobothrium chanaorum sp. n. (Proteocephalidae: Proteocephalinae), which was found in the intestine of Pseudoplatystoma reticulatum (Eigenmann et Eigenmann) from the Colastiné River, a tributary of the Paraná River. The new species differs from the two other species of the genus, P. eirasi (Rego et de Chambrier, 1995) and P. ludovici Ruedi et de Chambrier, 2012, parasites of Phractocephalus hemioliopterus (Bloch et Schneider) from the Amazon River in Brazil, in having fewer proglottides (4-8 without ventral appendages vs 7-12 with ventral appendages and 20-36 without ventral appendages, respectively), a smaller scolex (350-450 µm wide vs 495-990 µm and 515-1 020 µm wide, respectively), in the total number of testes (21-25 vs 21-51 and 37-79, respectively), a cirrus-sac usually directed anteriorly if the vagina is posterior to the cirrus-sac vs transversely situated in the known species. The study of the tegumental surface of Pseudocrepidobothrium spp. revealed the presence of four types of microtriches: papilliform, acicular and capilliform filitriches, and gladiate spinitriches. The three species have a similar microthrix pattern, with minor differences on the immature proglottis surface. Pseudocrepidobothrium chanaorum sp. n. is the ninth proteocephalid reported from P. reticulatum.

Histochemical techniques were applied to whole mounts, to study the distribution of the enzymes alkaline phosphatase, acid phosphatase, adenosine triphosphatase, 5'-nucleotidase and glucose-6-phosphatase in the organs and tissues of a viviparous monogenean, Macrogyrodactylus clarii Gussev, 1961, from the gills of the North African catfish Clarias gariepinus (Burchell) in Egypt. The following organs and tissues were studied: head region, anterior adhesive glands, mouth region, pharynx, intestine, testis, vesicula seminalis, male accessory gland, male accessory reservoir, copulatory organ, receptaculum seminis, egg-cell forming region, embryonic cells, excretory system, nerve cells, haptor, muscle fibres and subtegumental cell bodies (cytons). The enzymes showed marked differences in their activities among the studied organs and tissues. Alkaline phosphatase and acid phosphatase activities were detected in many organs and tissues, while the activities of adenosine triphosphatase, 5'-nucleotidase and glucose-6-phosphatase were restricted to a few organs. Although no positive reaction for any enzyme was observed in the anterior adhesive gland cells, a positive reaction for acid phosphatase was detected in the anterior adhesive areas. All enzymes showed marked activity in the digestive and excretory systems. The distribution of the enzymes in the tissues and organs of M. clarii is compared with those of other monogeneans, including other gyrodactylids parasitizing the same host fish. Some possible functions of the enzymes are discussed.

First step in developing an epitope-based vaccine is to predict peptide binding to the major histocompatibility complex (MHC) molecules. We performed computational analysis of unique available EgA31 sequence to locate appropriate antigenic propensity positions. T-cell epitopes with best binding affinity values of < 50% inhibitory concentration were selected using different available servers (Propred and IEDB). Peptides with 100% population coverage were selected. A DNA fragment corresponding to the furin linker enriched in Golgi apparatus was inserted sequentially between each epitope sequences in a synthetic DNA in order to cleave the chimeric protein into four separated peptides. Subsequently, the synthetic DNA was cloned into the pGEX4T-1 and pEGFP-N1 vectors and GST-ChEgA31 was expressed in E. coli strain BL21-DE3. The recombinant protein was detected by western blotting using an HRP-conjugated polyclonal anti-GST antibody. Fusion protein purified by affinity chromatography was used to raise antisera in rabbits. Results in agar gel immunodiffusion assay indicated induction of specific antibodies against multiepitope antigen in the tested rabbits. Cytokine assay was carried out in C57Bl/6 mice and the levels of cytokines were analyzed by sandwich ELISA. Interestingly, production of specific IFN-γ was prominently higher in mice immunized with GST-ChEgA31 and pEGFP-ChEgA31 (650-1 300 pg/ml) compared to control groups. No difference was observed in the level of IL-10 and IL-4 in immunized and GST control group. Challenge study with 500 live protoscolices of Echinococcus granulosus on immunized mice demonstrated protectivity level (50-60%). Based on our results, it appeared that the chimeric protein in the study was able to stimulate T-helper cell-1 (Th1) development and high level of cell mediated immunity in mice.

Two new species of two genera of the order Diphyllidea van Beneden in Carus, 1863, Halysioncum Caira, Marques, Jensen, Kuchta et Ivanov, 2013 and Echinobothrium van Beneden, 1849 sensu stricto are described from Aetomylaeus cf. nichofii (Bloch et Schneider) off the Iranian coast of the Persian Gulf. Halysioncum kishiense sp. n. differs from all other congeners in the number of apical hooks with the exception of H. hoffmanorum (Tyler, 2001) and H. pigmentatum (Ostrowski de Núñez, 1971). Halysioncum kishiense sp. n. can be easily differentiated from H. hoffmanorum and H. pigmentatum by the number of hooklets and testis numbers. Echinobothrium parsadrayaiense sp. n. is differentiated from all its congeners except for E. acanthinophyllum Rees, 1961 by its hook formula. The number of spines per column on the cephalic peduncle, the number of testes and possession of a thick-walled rather than thin-walled vagina distinguish E. parsadrayaiense sp. n. from E. acanthinophyllum. To date, with these two new species, five species of Diphyllidea have been reported from the Persian Gulf.

Two new species of feather mites are described from nightjars (Caprimulgiformes: Caprimulgidae) of Brazil: Hartingiella neotropica sp. n. (Xolalgidae) described from Hydropsalis parvula (Gould) and Paragabucinia brasiliensis sp. n. (Gabuciniidae) from H. albicollis (Gmelin). The former differs from the type species by having, in males, the anterior projections on epimerites III towards setae 3b and the adanal shield bearing setae ps3 present; in both sexes, a pair of small sclerites situated posterior to setae se have flat suprategumental processes. Paragabucinia brasiliensis sp. n. differs from P. petitoti (Gaud et Mouchet, 1959) by the smaller size of the incisions in the internal margins of opisthosomal lobes of males. These mites are the first representatives of corresponding genera described from the Neotropical region. The genus Hartingiella Gaud, 1980 was previously known solely from its type species. Keys to males and females of the genus Paragabucinia Gaud et Atyeo, 1975 are presented. In addition, all previous records of feather mites associated with birds of the order Caprimulgiformes of the world are summarised.

The trichostrongylid nematode Travassostrongylus scheibelorum sp. n. from the Linnaeus' mouse opossum, Marmosa murina (Linnaeus) (type host), and the woolly mouse opossum, Marmosa demerarae (Thomas), from French Guiana is described. The nematodes have a synlophe with ridges frontally oriented from right to left, six dorsal and six ventral, at midbody; seven dorsal and seven ventral posterior to the vulva, and two cuticular thickenings within the lateral spaces; a long dorsal ray and a pointed cuticular flap covering the vulva. This is the 12th species of Travassostrongylus Orloff, 1933, which includes species featuring ridges around the synlophe and a didelphic condition. These traits contrast with those in other genera in the Viannaiidae Neveu-Lemaire, 1934, which feature ventral ridges on the synlophe of adults and a monodelphic condition. Members of the family are chiefly Neotropical and are diagnosed based on the presence of a bursa of the type 2-2-1, 2-1-2 or irregular, and cuticle without ridges on the dorsal side (at least during one stage of their development). Herein, we present a reconstruction of the ancestral states of the didelphic/monodelphic condition and the cuticular ridges that form the synlophe in opossum-dwelling trichostrongyles, namely Travassostrongylus and Viannaia Travassos, 1914. Our investigations suggest they are not reciprocal sister taxa and that the change from didelphy to monodelphy and the loss of dorsal ridges, occurred in the common ancestor of species of Viannaia. These results suggest a synlophe with three ventral ridges is not plesiomorphic in the opossum dwelling trichostrongylids.

Apical membrane antigen-1 (AMA-1) of Plasmodium vivax Grassi et Feletti, 1890 is a promising malaria vaccine candidate. However, antigenic variation is a major problem to design a universal malaria vaccine. Hence, detailed understanding of the pvama-1 gene polymorphism can provide conductive information on this potential vaccine component. Therefore, this study investigated the extent of genetic polymorphisms at domain I (DI), DII and partial DIII of AMA-1 among Iranian P. vivax isolates. Out of 107 blood samples, 92 were analysed based on the quality of the sequencing data. The sequences were classified into 53 haplotypes. Amino acid changes were observed at 31 positions that 17 were located at DI, 11 were at DII and the rest of them (3 positions) were at DIII. Thus, codon polymorphisms at DI were found to be higher than DII. Also, five of these polymorphic codons (D242E, T374P, S389R, Y391F, I395F) were novel and have not been reported yet. Neutrality analysis by using the dN-dS difference (the difference between the rate of non-synonymous and synonymous mutations) showed a negative diversifying selection at DI, DII and across the length of both domains. The potential B-cell epitopes were found in 5 regions of the PvAMA-1 with 10 mutation sites (E145A, K188N, E189N/K/D, K190Q/E, P210S, E227V, D242E, R249H, G253E, K352E), whereas only one mutation (G288E) has been detected in intrinsically unstructured/disordered regions. Fixation index (Fst) estimation between Iranian and Indian isolates (0.0131) indicated a significant low genetic differentiation. Distribution of the polymorphic sites and IURs mapped on a three dimensional structure of PvAMA-1 showed that these regions were located at two opposite faces of the molecule. In conclusion, the results have significant value in the design and development of a malaria vaccine based on this antigen.

Six new species of coccidia are described from endangered Phelsuma spp. geckoes (Sauria: Gekkonidae) endemic to Mauritius, Indian Ocean. Five new species (3 Eimeria and 2 Isospora species) are described from Phelsuma rosagularis Vinson et Vinson; all lack a micropyle and an oocyst residuum, and all have a sporocyst residuum. Oocysts of Eimeria swinnertonae sp. n. are ellipsoidal, 22.2 × 17.8 (20.8-24.8 × 16.8-18.4) μm; SI 1.25; polar granule absent. Sporocysts are ellipsoidal, 8.8 × 7.0 (8.0-9.6 × 6.4-8.0) μm; SI 1.3; Stieda body absent. Oocysts of Eimeria stebbinsi sp. n. are ellipsoidal, 17.4 × 11.7 (16.0-19.2 × 11.2-12.8) μm; SI 1.5; polar granules present. Sporocysts are elongate-ellipsoidal, 7.7 × 4.0 (7.2-8.0 × 3.2-5.6) μm; SI 1.9; Stieda body present. Oocysts of Eimeria raleighi sp. n. are spheroidal to sub-spheroidal, 17.0 × 15.5 (16.0-19.2 × 14.4-16.8) μm; SI 1.1; polar granule present. Sporocysts are sub-spheroidal, 7.8 × 6.6 (7.2-8.0 × 6.4-7.2) μm; SI 1.2; Stieda body absent. Oocysts of Isospora cottinghamae sp. n. are ellipsoidal, 19.8 × 15.5 (17.6-21.6 × 14.4-17.6) μm; SI 1.3; polar granules present. Sporocysts are ellipsoidal, 10.8 × 6.9 (9.6-12.8 × 6.4-8.0) μm; SI 1.6; Stieda body present. Oocysts of Isospora pearlae sp. n. are ellipsoidal, 16.0 × 11.5 (15.2-17.6 × 9.6-12.8) μm; SI 1.4; polar granule present. Sporocysts are ellipsoidal, 8.8 × 5.4 (8.0-9.6 × 4.8-6.4) μm; SI 1.6; Stieda and substieda bodies present. One new Eimeria species is described from the blue-tailed day gecko, Phelsuma cepediana Merrem. Oocysts of Eimeria hartleyi sp. n. are sub-spheroidal to ellipsoidal, 18.2 × 14.5 (16.0-20.8 × 13.6-16.0) μm; SI 1.26; polar granules present. Sporocysts are ellipsoidal to cylindroidal, 7.5 × 5.3 (6.4-8.0 × 4.8-6.4) μm; SI 1.4; Stieda body present. We report the presence of tetrazoic spheroidal to sub-spheroidal oocysts or sporocysts 10.2 × 8.5 (9.9-10.4 × 8.3-8.8) μm; SI 1.2 from an individual of P. cepediana. These oocysts or sporocysts are significantly larger than the Cryptosporidium species so far described from reptiles, and likely represent excretion of spuriously ingested sporocysts of a Sarcocystis or Adelina coccidian.

Abstract: Rhabdias blommersiae sp. n. (Nematoda: Rhabdiasidae) is described from the lungs of Domergue's Madagascar frog, Blommersia domerguei (Guibé) (Amphibia: Mantellidae), in Madagascar. The new species differs from congeners parasitizing amphibians in having a smaller body and buccal capsule, six equal lips, large excretory glands of unequal length and a posteriorly inflated body vesicle. A combination of characters distinguishes it from Afromalagasy species of Rhabdias Stiles et Hassall, 1905. Rhabdias blommersiae is the third species of the genus described from amphibians in Madagascar. Close similarities in the number and shape of circumoral structures in two Rhabdias species described from mantellid hosts in Madagascar suggest a close relationship and common origin of the two species, with subsequent adaptation to separate hosts within the Mantellidae.

The intestinal protozoan parasite Giardia duodenalis (Lambl, 1859) Kofoid & Christiansen, 1915 [syn. Giardia intestinalis and Giardia lamblia] has emerged as a widespread enteric pathogen in humans and domestic animals. In recent years, G. duodenalis has been found in cattle worldwide and longitudinal studies have reported cumulative prevalence of 100% in some herds. In the present study, we determined the prevalence and genetic characterisation of G. duodenalis in 200 dairy cattle from 10 dairy farms in São Paulo state, Brazil. All faecal specimens were screened for the presence of G. duodenalis using microscopy examination, enzyme immunoassay (EIA) and polymerase chain reaction (PCR). DNA was extracted from faecal samples and G. duodenalis were identified by amplification of the small subunit ribosomal (SSU-rDNA) and glutamate dehydrogenase (GDH) genes followed by restriction fragment length polymorphism (RFLP) or sequencing analysis. Giardia was identified in eight farm locations (80% prevalence). Overall, 15/200 (7.5%) animals were positive for infection, only one of which was a cow. Giardia duodenalis genotype E was present in 14 of the animals tested. Zoonotic genotype AI was present in one positive sample. Genotype E and genotype A represented 93% and 7% of G. duodenalis infections, respectively. This study demonstrates that G. duodenalis infection was prevalent in dairy calves in São Paulo state and that the non-zoonotic genotype E predominates in cattle in this region. Nevertheless, calves naturally infected in Brazil can shed Giardia cysts that can potentially infect humans, and thus, they may represent a public health risk.

Abstract: Pseudocapillaria moraveci sp. n. (Nematoda, Capillariidae) is described from the stomach of Gobius paganellus Linnaeus (Perciformes, Gobiidae) from rocky shores of the Vigo estuary (Northwest Atlantic coast of Spain). Scanning electron microscopy reveals the absence of a dorsal cuticular membrane interconnecting the caudal lobes of male, thus confirming the assignment of this new species to the nominotypical subgenus Pseudocapillaria Freitas, 1959. Pseudocapillaria moraveci can be easily differentiated from other marine congeneric species belonging to subgenera Pseudocapillaria and Ichthyocapillaria Moravec, 1982 by its muscular oesophagus, which is distinctly thickened at the distal third, the terminal or almost terminal anus in female, the presence of a basal internal papilla on each ventrolateral caudal lobe in male, and its exclusive site preference for the stomach. In addition, the spicule, which measures 0.180-0.292 mm in length (4.9-7.9% of body length) and possesses a proximal end expanded and oblique in lateral view, a narrowed middle part, and a distal end almost pointed, also distinguishes this species. Pseudocapillaria moraveci is, together with P. tomentosa (Dujardin, 1843) Moravec, 1987, the only species in the genus Pseudocapillaria that has been reported to infect members of the family Gobiidae.

Abstract: Recent collections of cestode parasites from two species of the myliobatid genus Aetomylaeus Garman from several localities in the Pacific Ocean resulted in the discovery of two new species of Halysioncum Caira, Marques, Jensen, Kuchta et Ivanov, 2013. Halysioncum gibsoni sp. n. from Aetomylaeus maculatus (Gray) in the South China Sea off Borneo differs from all of its congeners in having the following combination of characters: 27 apical hooks (14 type A and 13 type B hooks), 11-12 lateral hooklets, 22-28 spines per column on the cephalic peduncle, testes distributed in a single column and an internal seminal vesicle. Halysioncum arafurense sp. n., recovered from Aetomylaeus cf. nichofii 2 (sensu Naylor et al. 2012b) in the Arafura Sea off the Wessel Islands, Northern Territory, Australia, can be distinguished from its congeners based on the following combination of characters: 23 apical hooks (12 type A and 11 type B hooks), the number of lateral hooklets (9-11), the number of spines per column on the cephalic peduncle (20-24), the number and distribution of the testes (13-15 testes in two irregular columns), and the distribution of vitelline follicles (interrupted dorsally at the level of the ovarian lobes). Both species represent the first verified records of diphyllideans from eagle rays of the genus Aetomylaeus and formally extend the host associations of diphyllideans to include a third genus of Myliobatiformes. The myliobatiforms are indeed an understudied group of available hosts for diphyllideans and represent interesting target hosts if the diversity of diphyllidean tapeworms is to be fully estimated and understood.

There are many strategies to control leishmaniasis, but majority of them are inadequate. Killed Leishmania vaccine (KLV) has been applied for its immunogenicity in human and mouse model. Bacillus Calmette-Guerin (BCG) as adjuvant is an immunemodulator inducing humoral and cellular immune responses during zoonotic cutaneous leishmaniasis (ZCL). Both KLV and BCG have been applied for their immune responses in hosts for controlling leishmaniasis. In this study, KLV and BCG were applied to inhibit replication and visceralization of Leishmania major in BALB/c mice. Mice were injected with KLV and BCG, followed by infection with promastigotes of L. major. Six weeks after infection, a small nodule appeared, which was followed by development of a large lesion and visceralization. Effects of KLV and BCG, physiopathological changes, lesion size, delay of lesion formation, proliferation of amastigotes inside macrophages and detection of amastigotes in target organs were studied. Results showed that the KLV had anti-leishmanial activity by reducing lesion size on late infection. In KLV and BCG group, the average number of amastigotes in macrophages was lower than in other groups. Significant reductions in number of amastigotes in both spleen and lymph node were observed, indicating lower visceralization of Leishmania parasites in these target organs. No significant changes were presented in body weights, survival rates and degrees of splenomegaly in test group. It can be concluded that application of KLV and BCG had acceptable efficacy in reduction of skin lesions size and proliferation of parasites, even though a few side-effects were observed. It is indicated that KLV/BSG may have ability to modulate host immune responses against Leishmania parasites and to reduce pathophysiology of the disease during infection.

Abstract: Nomasanguinicola canthoensis gen. et sp. n. infects the branchial vessels of bighead catfish, Clarias macrocephalus Günther (Siluriformes: Clariidae), in the Mekong River near Can Tho, southern Vietnam. Nomasanguinicola differs from all other genera of fish blood flukes (Digenea: Aporocotylidae) by the combination of lacking body spines and by having an anterior sucker with two flanking columns of large denticles, an intestine comprising several short papilla-like caeca, an inverse U-shaped uterus, and an ootype located near the separate genital pores. The new species has an ootype that is posterior to the level of the female genital pore. That feature most easily differentiates it from the only other putative aporocotylid species having an anterior sucker with two flanking columns of large denticles, Plehniella dentata Paperna, 1964 and Sanguinicola clarias Imam, Marzouk, Hassan et Itman, 1984, which have an ootype that is lateral (P. dentata) or anterior (S. clarias) to the level of the female genital pore. These two species apparently lack extant type materials, infect North African catfish, Clarias gariepinus (Burchell), and herein are considered incertae sedis, but likely comprise species of Nomasanguinicola. An updated list of hosts, sites of infection and geographic localities for the six species and three genera of blood flukes that mature in catfishes is provided. The new species is the first fish blood fluke recorded from Vietnam and only the third reported from a walking catfish (Clariidae).

Kudoid parasites are known to infect a large variety of fish. A significant proportion of Kudoa species have relatively low host specificity, with a single species able to infect multiple host species representing various host families even from different host orders. Since DNA sequences have been associated with myxosporean species characterisations, it has become far easier to determine host range of new species and validate host records from earlier descriptions. This study investigated the host specificity of a kudoid parasite, Kudoa thalassomi Adlard, Bryant, Whipps et Kent, 2005, from the Great Barrier Reef in Australia using DNA sequence analysis and morphology. The results revealed the host specificity to be broad, with K. thalassomi identified in 18 different fish species representing six different fish families. This study also compares current genetic information from different host isolates of Kudoa Meglitsch, 1947 to their host ranges recorded in existing literature. From this analysis, only half of the Kudoa species with multiple host records (27 Kudoa species) have half or more isolates that are genetically characterised, and thus specifically identified with a high confidence, from their known hosts. Only five kudoid species have genetically characterised isolates from all of their recorded hosts.

The liver fluke Clonorchis sinensis (Digenea) is a high-risk parasite that causes serious diseases such as cirrhosis, carcinogenic liver damage and clonorchiasis in East Asia. This study was conducted to evaluate the relationship between stress/endocrine hormones and inflammation induced by infection as well as the expression of heat shock proteins (hsp-27, hsp-90), cox-2 and cytokines in the livers of hamsters infected with C. sinensis. The average body weight of infected hamsters decreased up to 25% compared with that of the control group, and bile duct hyperplasia with inflammation, liver fibrosis and hepatic necrosis were observed in C. sinensis-infected livers. The expression of hsp-27, hsp-90, and cox-2 was significantly increased in the livers of C. sinensis-infected hamsters compared with the control group. Moreover, the expression levels of inflammatory cytokines (IL-1β, IL-2, TGF-β2 and IFN-α1) were markedly increased in the livers of the infected group compared with those of the control group. Consistently, plasma IL-3 and IL-6 levels gradually increased during the infection period, and the concentration levels of testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), corticosterone, and adrenocorticotropic hormone (ACTH) in C. sinensis-infected hamsters increased over 25%, compared with those of the uninfected normal group. These results demonstrate that C. sinensis infection may increase the expression of hsp27, hsp90 and cox-2 as well as it may cause periductal fibrosis, chronic inflammation and hepatic necrosis in the liver. Furthermore, the results indicate that C. sinensis infection induces not only stress-induced hormone imbalance but also the sustained secretion of inflammatory cytokines through chronic stress/stimuli.

Abstract: Therapeutic efficacy of sulfadoxine-pyrimethamine (SP), which is commonly used to treat falciparum malaria, was assessed in isolates of Plasmodium falciparum (Welch, 1897) and Plasmodium vivax (Grassi et Feletti, 1890) of Aligarh, Uttar Pradesh, North India and Taif, Saudi Arabia during 2011-2012. Both the species showed mutations in dihydrofolate reductase (DHFR) enzyme as they have common biochemical drug targets. Mutation rate for pfdhfr was higher compared to pvdhfr because the drug was mainly given to treat falciparum malaria. Since both the species coexist, P. vivax was also exposed to SP due to faulty species diagnosis or medication without specific diagnosis. Low level of mutations against SP in P. falciparum of Saudi isolates indicates that the SP combination is still effective for the treatment of falciparum malaria. Since SP is used as first-line of treatment because of high level of resistance against chloroquine (CQ), it may result in spread of higher level of mutations resulting in drug resistance and treatment failure in near future. Therefore, to avoid further higher mutations in the parasite, use of better treatment regimens such as artesunate combination therapy must be introduced against SP combination.

Abstract: Based on the study of type material, two new genera of cestodes (Cyclophyllidea: Anoplocephalidae) are proposed for Paranoplocephala Lühe, 1910 sensu lato species from African rodents. Afrojoyeuxia gen. n., proposed for A. gundii (Joyeux, 1923) comb. n. from Ctenodactylus gundi (Rothmann) (Hystricomorpha: Ctenodactylidae), is characterized by a high length/width ratio of mature proglottids, longitudinally extensive testicular field positioned anterior to the female glands, an ovoid or subspherical cirrus-sac and a thick, conical cirrus. Hunkeleriella gen. n., proposed for H. dasymidis (Hunkeler, 1972) comb. n. from Dasymys incomtus (Sundevall) (Myomorpha: Muridae), differs from related genera mainly by its short (10-20 mm) and wide strobila and neck, unilateral genital pores (exceptionally with a few changes per strobila), the position of the genital pores (slightly anterior to the middle of proglottid margin) and initially tube-like early uterus (later reticulated). Parandrya Gulyaev et Chechulin, 1996, earlier suggested to be a junior synonym of Paranoplocephala, is considered to be a valid, independent genus. Evidence of non-monophyly and need for a taxonomic revision of Paranoplocephala sensu lato, as well as the phylogenetic position of A. gundii and H. dasymidis are discussed.

Abstract: A myxosporean producing actinospores of the tetractinomyxon type in Hydroides norvegicus Gunnerus (Serpulidae) in Denmark was identified as a member of the family Parvicapsulidae based on small-subunit ribosomal DNA (SSU rDNA) sequences. Myxosporean samples from various Danish and Norwegian marine fishes were examined with primers that detect the novel myxosporean. Sprattus sprattus (Linnaeus) and Clupea harengus Linnaeus (Teleostei, Clupeidae) were found to be infected. The sequences of this parvicapsulid from these hosts were consistently slightly different (0.8% divergence), but both these genotypes were found in H. norvegicus. Disporic trophozoites and minute spores of a novel myxosporean type were observed in the renal tubules of some of the hosts found infected through PCR. The spores appear most similar to those of species of Gadimyxa Køie, Karlsbakk et Nylund, 2007, but are much smaller. The actinospores of the tetractinomyxon type from H. norvegicus have been described previously. In GenBank, the SSU rDNA sequences of Parvicapsulidae gen. sp. show highest identity (82%) with Parvicapsula minibicornis Kent, Whitaker et Dawe, 1997 infecting salmonids (Oncorhynchus spp.) in fresh water in the western North America. A phylogenetic analysis places P. minibicornis and Parvicapsulidae gen. sp. in a sister clade to the other parvicapsulids (Parvicapsula spp. and Gadimyxa spp.).

Abstract: Gill monogenean species of Ligophorus Euzet et Suriano, 1977 were studied from the teleost Mugil cephalus Linneaus (Mugilidae) from the Mediterranean and Atlantic coasts of Morocco. We report the presence of L. mediterraneus from both the Mediterranean and Atlantic coast and L. cephali and L. maroccanus sp. n. from the Atlantic coast only. The latter species, which is described herein as new, resembles L. guanduensis but differs from this species mainly in having a shorter penis compared to the accessory piece, a proportionally longer extremity of the accessory piece and a less developed heel. The utility of Ligophorus spp. as markers of cryptic species of the complex M. cephalus is discussed in the context of species diversity and geographical distribution of these monogeneans on this host around the world. Presence of different species of Ligophorus on M. cephalus sensu stricto from the Atlantic and Mediterranean coast of Morocco demonstrates the usefulness of these species as fine resolution markers of genetic populations of their host, which are known to inhabit those coasts.

The current work describes two new species of Gyrodactylus von Nordmann, 1832 collected from pipefish Syngnathus scovelli (Evermann et Kendall) and Syngnathus typhle L. during two separate gyrodactylosis episodes on fish held in a public aquarium located in northern Italy. The gyrodactylids collected from the skin, fins and gills of pipefish were subjected to a morphological analysis of the attachment hooks and the morphometric data were compared to the four species of Gyrodactylus previously described from syngnathid hosts, namely G. eyipayipi Vaughan, Christison, Hansen et Shinn, 2010, G. pisculentus Williams, Kritsky, Dunnigan, Lash et Klein, 2008, G. shorti Holliman, 1963 and G. syngnathi Appleby, 1996. Principal components analysis (PCA) of the morphological data indicated six clusters; two discrete groups among the specimens taken from the pipefish held in the Italian aquarium and four further groups representing G. eyipayipi, G. pisculentus, G. shorti and G. syngnathi. Molecular sequences of the ribosomal internal transcribed spacers (ITS1 and ITS2) and the 5.8S gene for the new species considered here were then compared with those available for other species in GenBank. The comparison did not reveal any identical match, supporting the morphological analysis that Gyrodactylus corleonis sp. n. from S. typhle and Gyrodactylus neretum sp. n. from S. scovelli represent distinct species. Both G. corleonis and G. neretum possess robust hamuli, marginal hook blades that curve smoothly from their sickle base to a point beyond the toe and, ventral bars with a broad median portion and a reduced membrane. Gyrodactylus corleonis, however, can be distinguished on the basis of its heart-shaped ventral bar; G. neretum has a 1:2 hamulus point:shaft ratio and a rectangular-shaped ventral bar. A redescription of the haptoral hard parts of the four species previously recorded on pipefish is also presented.

Trafficking of the rhoptry chimeric protein RhopH2-GFP, which contains RhopH2 signal peptide plus the downstream five amino acids, was dissected by treating parasites with Brefeldin A at three different time points. Twenty eight hrs-stage trophozoites accumulated the chimera within the parasite endoplasmic reticulum. In 32 hrs-stage schizonts, the chimera was distributed in the parasite cytoplasm but not in the parasitophorous vacuole. In 36 hrs stage-schizonts, the chimera was detected in the individual parasitophorous vacuoles of the developing merozoites and, in contrary to non-treated parasites, no immature rhoptry vesicles could be detected in the cytoplasm of immature merozoites. These data show that this chimera is trafficked to the rhoptries via Brefeldin A-sensitive pathway indicating that this trafficking is similar to that of the endogenous rhoptry proteins, and that the five amino acids downstream of the signal peptide cleavage site may contain the sorting signal required for rhoptry targeting.