Band 62, Heft 1

In present study the systematic and taxonomic position of hymenolepidids parasitizing rodents of the family Gliridae from Europe and Central Asia is discussed. Hymenolepis myoxi is redescribed on the basis of the type material from the fat dormouse Glis glis deposited in the collection of the Museum für Naturkunde, Berlin, Germany. Significant corrections of morphologically distinctive characters attributable to H. myoxi include: (1) recognition of a rudimentary rostellar apparatus; (2) absence of rostellar hooks and spination on the suckers; and (3) tissues of the scolex and neck filled with numerous “firm elements”. Hymenolepis sulcata was recognised as a synonym of H. myoxi (sensu stricto) . The generic allocation of true H. myoxi and validity of the genus Armadolepis is clarified. Specimens from Eliomys quercinus originally designated as H. myoxi by Baer (1932) are described as a new species, Armadolepis jeanbaeri n. sp. The taxonomy has potentially been confused as Spassky (1954) designated misidentified specimens of H. myoxi sensu Baer (1932) as the type species of the genus Armadolepis . In the current article, this error is corrected and A. jeanbaeri n. sp. is fixed as the type of the genus Armadolepis . An additional new species of Armadolepis, A. tenorai n. sp., is described from Dryomys nitedula from Almaty Province, Kazakhstan. The generic diagnosis of Armadolepis is amended. Armadolepis (sensu stricto) is subdivided into two subgenera; the nominotypical subgenus includes species having well developed rostellar apparatus armed by rostellar hooks and A. (Bremserilepis) n. subgen. includes species with rudimentary and unarmed rostellar apparatus.

Multiple sclerosis is a chronic inflammatory central nervous system (CNS) disease, which affects about 1 in 1000 individuals in the western world. It has been suggested that this relatively high prevalence is linked to a high level of hygiene, i.e . a reduced exposure to various microorganisms, including parasites. Parasites are known to employ different immunomodulatory and antiinflammatory strategies, which enable them to evade destruction by the immune system. We have investigated the immunomodulation by the swine whipworm, Trichuris suis , by measuring the impact of oral administration of T. suis ova as well as of intraperitoneal administration of T. suis excretory/secretory products on the development and progression of experimental autoimmune encephalomyelitis – an animal model that shares clinical and pathological characteristics with multiple sclerosis. Intraperitoneal administration of excretory/secretory products before disease onset, resulted in a significant decrease in disease severity as well as markedly reduced T H 1 and T H 17 T-cell responses, centrally in the spinal cord as well as in the periphery, i.e . the spleen. Thus, parenteral administration of T. suis-derived products results in a skewing of the immune response with a significant impact on disease severity in a CNS inflammatory disease model.

Filarial parasites are complex mixtures of antigenic proteins and characterization of these antigenic molecules is essential to identify the diagnostically important filaria-specific antigens. In the present study, we have fractionated the somatic extracts from adults of Setaria cervi (bovine filarial parasite) on preparative SDS-polyacrylamide gel electrophoresis and tested the immunoreactivity of the separated gel fractions with polyclonal antibodies against filarial excretory-secretory antigens as well as filarial patients sera. The SDS-PAGE analysis of gel eluted fractions revealed 1 protein band in F-1 fraction, 2 protein bands in F-2 fraction and 2–3 protein bands in all other fractions (F3- F11). Seven gel eluted fractions (F1, F2, F3, F4, F5, F6 and F11) showed high ELISA reactivity with the polyclonal antibody (against excretory-secretory antigen) and four of these fractions (F-1, F-2, F3 and F6) exhibited high ELISA reactivity with antibodies present in filarial patient sera. The reactivities of the gel fractions (F1 and F2), recognized by filarial patients sera, were also tested with the monoclonal antibody (detecting the filarial circulating antigen). The F1 and F2 gel eluted fractions were found to have the target antigen of monoclonal antibody as evident by high reactivity with the monoclonal antibody in ELISA and immunoblotting. The S. cervi gel eluted F1 fraction (containing single antigen) could detect antibodies in filarial patients sera and not in non-filarial sera thereby suggesting its usefulness for specific serodiagnosis of human filariasis.

The relationships between water mite larvae parasitizing Coenagrion scitulum in core and edge populations were described. A total of 636 larvae of 7 water mite species were found on 143 C. scitulum adults (82 females and 61 males). C. scitulum was recorded for the first time as a host species for Arrenurus cuspidator, A. bruzelii, A. bicuspidator, A. tricuspidator, A. claviger and Hydryphantes octoporus . The degree of infestation by particular parasite species was typical for these species. In contrast, the parasites’ preferences for host body parts were not typical, as they preferred abdominal segments 2–4, which in earlier studies had been avoided by water mite larvae. No differences were found in degree of infestation of Coenagrion scitulum individuals between core and edge populations, with the exception of Hydryphantes octoporus , which parasitized damselflies only in core populations.

A new species of parasitic mites, Myocoptes (Comyocoptes) nesokia sp. nov. (Acariformes: Myocoptidae) is described from Nesokia indica (Gray, 1830) (Rodentia: Muridae) from Pakistan. Females of this species differ from the morphologically similar species, Myocoptes (Comyocoptes) striatus Fain, 1970, by the presence of a verrucose pattern in the middle part of the hysteronotum, longer setae cp (120–140 μm vs . 55–60 μm), and by the absence of scales on the opisthosomal cuticle posterior to the level of setal bases ps3 .

Two new digenean species are described from the intertidal blenny Scartichthys viridis (Valenciennes) (Blenniidae) collected off the coasts of Chile. The digenean Monorchimacradena viridis n. sp. (Lecithasteridae: Macradenininae) differs from the only known species described in the genus, M. acanthuri Nahhas and Cable 1964, in the presence of Drüsenmagen in the caeca, the location of the seminal vesicle between the testis and ovary (anterior to the testis in M. viridis n. sp.), and the pre-ovarian vitellarium. Megasolena littoralis n. sp. (Haploporidae), which is also reported from Scartichthys gigas (Steindachner), differs from the five valid species of Megasolena in that the post-caecal region (from the posterior edge of the caeca to the end of the body) is larger in M. viridis n. sp., meaning that the caeca are shortest in this species. Also, M. littoralis n. sp. is distinguished, from the other congeneric species, in a combination of characteristics, e.g., body length, suckers, pharynx, testes, hermaphroditic sac and sucker-length ratio. Sequences of ITS2 rDNA were generated for specimens of both species from various localities, indicating that it was invariant within the species. This study describes species of Monorchimacradena and Megasolena in Chilean waters, here reported for the first time off the Pacific Coast of South America.

Enterocytozoon bieneusi ( E. bieneusi ), infecting renal transplant (RT) recipients may be transmitted anthroponotically or zoonotically. Accordingly, we aimed to, a) evaluate genotypes of E. bieneusi ínfecting RT recipients, and b) infer phylogenetic interpretation on transmission of different genotypes among infected hosts. Stool samples of 22 RT recipients infected with microsporidia (identified using modified trichrome staining) were subjected to species identification. All E. bieneusi positive samples were subjected to genotyping. The phylogenetic tree was constructed using Mega 5 software. Of 22 microsporidia infected RT recipients, 21 (95.5%) had E. bieneusi . ITS sequences of 21 E. bieneusi were classified into eight genotypes (Ind1 to Ind8). Among them, 4 (Ind5 to Ind8) were novel. Genotypes Ind2, Ind3, Ind4, Ind7 and Ind8 showed close sequence similarity to genotypes reported exclusively from humans. Phylogenetic analysis further supported their anthroponotic transmission. Genotypes Ind1, Ind5 and Ind6 showed close sequence similarity to genotypes reported from both animals and humans. Phylogenetic analysis further supported their zoonotic transmission. Anthroponotic transmission of E. bieneusi was more common among males (11, 100% vs. 7/10, 70%; P = 0.05), presenting with diarrhea (11, 100% vs. 6/10, 60%; P = 0.02) and watery stool (10/11, 91% vs. 5/10, 50%; P = 0.03).

The microthrix pattern of common forms of the trypanorhynch cestode Prochristianella clarkeae Beveridge, 1990 on which the microscopically visible microtriches almost extended to the posterior end of the scolex was described using SEM and compared to other forms for which this pattern was different. The proximal bothrial surface and the scolex peduncle of all the examined specimens were covered with palmate spinitriches and papilliform filitriches. Unlike the common specimens of P. clarkeae , on the surface of those specimens with unusual distributions of microtriches as well as those on which the microscopically visible microtriches were not observable, palmate spinitriches extended to the mid-pars vaginalis and not to the posterior end of the scolex. Statistical tests did not detect any significant difference in the measurements of the specimens possessing different microthrix patterns. Furthermore, the partial 28S rDNA confirmed that the different forms were conspecific and that P. clarkeae possesses an intraspecific variation in the distribution of palmate spinitriches. Pastinachus sephen as a new host record was also established for Prochristianella clarkeae reported for the first time from the Persian Gulf.

The astigmatid family Hypoderatidae includes over 80 mite species exhibiting peculiar life cycle. Deutonymphs are parasitic instars inhabiting subcutaneous or visceral tissues of birds and rodents, while all other instars are nidicolous forms. In this study we investigated genetic diversity of deutonymphs of two hypoderatid species, Neottialges evansi and Phalacrodectes gaudi , collected from 16 individuals of the great cormorant Phalacrocorax carbo in SW Poland during host post-breeding dispersal. The initial alternative hypotheses were: (1) populations of deutonymphs in both mite species found in tissues of particular bird individuals form genetically distinct populations, (2) mites are panmictic among hosts. The topologies of NJ phylogenetic trees and median-joining haplotype networks reconstructed for COI haplotypes revealed lack of hostdependent genetic structure in populations of N. evansi and P. gaudi . Furthermore, high haplotype diversity (Hd) and low nucleotide diversity (Pi) prove high genetic differentiation of both mite species. We concluded, that unlimited dispersal of mites among cormorant specimens could be explained by host specific breeding behavior: free mating between cormorants originating from different European populations and low contribution of reunited pairs in breeding colonies in subsequent breeding seasons, reuse of the same nest material by different members of the colony, and host behavior called prospecting.

Parasitic nematodes of Oesophagostomum spp ., commonly known as ‘nodular worms’ are one of the most widely distributed and prevalent emerging zoonotic nematodes. However, little is known about the prevalence and gene characteristics of those parasites in Tibetan pigs. Therefore, a study was carried out to investigate the prevalence, isolation and identification of Oesophagostomum spp from Tibetan pigs by genetic markers of nad1 , cox3 and ITS1 for the first time. The results revealed that the infection rate of O. dentatum and O. quadrispinulatum by genetic markers of nad1 was 81.13%; 35 (66.04%); the O. dentatum and O. quadrispinulatum by genetic markers cox3 was 66.04%, and O. dentatum and O. stephanostomum by genetic markers ITS1 was found to be 77.36%. Interestingly, the O. stephanostomum specie was identified and isolated from 90.48% stomach and 69.23% colon samples by genetic markers of ITS1 . The present study, for the first time has described the presence and genetic characterization of Oesophagostomun spp of O. dentatum , O. quadrispinulatum and especially O. stephanostomum in Tibetan pigs from the high and remote Tibetan plateau. A public concern should be raised in terms of economical losses and severe public health problem.

Anisakiosis is nowadays one of the nematodoses more prevalent in Spain, with rates that oscillate between 0.43% in Galicia (N.W. Spain), and 15.7% and 22.1% in inland and southern regions, respectively. Likewise, it has been proved that Anisakis larvae have developed mechanisms to modulate the dichotomy of the host immune response for their own benefit. The experimental hypothesis of the present study was that Anisakis sp. larval products can be mediators of immune suppression and induce changes on the populations of αβ+ and γδ+ T cells. In the present study we determined the levels of anti- Anisakis antibodies in the serum of healthy people, and their relationship with the B and T cell subsets. Levels of anti- Anisakis antibodies (Ig’s, IgG, IgM, IgA and IgE) were measured by ELISA, while B and T cell subsets were studied by flow cytometry. Cells were labelled with monoclonal antibodies against CD45, CD4, CD8, CD56, CD3, CD19, TCRαβ and TCRγδ. All the specific isotypes studied were negatively correlated with NKT cell rates with the exception of IgG. A previous contact with Anisakis was related to a decrease in CD56+αβ+ and all γδ+ T cell subsets. The CD3+γδ+ population was lower in the group of subjects that showed IgA anti- Anisakis . We observed an inverse correlation among αβ-γδ NKT cells and anti- Anisakis sp. antibodies. CD3+CD56+ cells showed a significant decrease in the group of anti- Anisakis positive subjects. This fact was especially significant with CD3+CD56+γδ+ cells in the case of the anti- Anisakis IgA positive group.

Potentially pathogenic Free-Living Amoebae include members belonging to Naegleria genus. The species N. fowleri is known worldwide as the causative agent of the lethal Primary Amoebic Meningoencephalitis (PAM). Only one clinical case of N. fowleri has been reported in Iran. Several species of Naegleria have been reported to be natural carriers of other potentially pathogenic microbial agents. The thermotolerance properties of this genus facilitates their presence in geothermal water sources including hot springs and spas. In the current study water samples were collected from 22 therapeutic hot springs, Northern Iran and investigated for the presence of Naegleria spp. using morphological keys and PCR/DNA sequencing based methods. Incubation of collected samples were done at both 30°C and 45°C in order to detect Naegleria spp. and N. fowleri , respectively. Thermotolerance assay and flagellation tests were also performed. The obtained results revealed that 54% of the investigated water samples were positive for Naegleria spp. including N. australiensis, N. americana, N. dobsoni, N. pagei, N. polaris and N. fultoni . The pathogenic N. fowleri was not detected. The most detected Naegleria was belonged to N. australiensis . This is the first report on the Naegleria spp. occurrence in hot springs in Northern Iran showing that most of the surveyed hot spring sources were contaminated with non-pathogenic Naegleria spp. However, due to the recent report of PAM in the country, further studies to investigate the presence of pathogenic N. fowleri in the environment and clinical samples is needed in the region and worldwide.

Spermatological characters of Dictyocotyle coeliaca Nybelin, a unique endoparasitic monogenean from the body cavity of the ray Amblyraja radiata (Elasmobranchii: Rajidae), was investigated by means of scanning and transmission electron microscopy for the first time. The process of the spermatozoon formation begins with the appearance of the differentiation zone which contains two centrioles. Subsequently developed two free flagella rotate to lie parallel before their fusing. After fusion, both the mitochondrion and nucleus migrate alongside the axonemes. Spermatids are formed within parallel cytoplasmic canals. During late spermiogenesis, an electron-dense spiral end-piece is formed proximally to the centriolar region of some spermatids. At the end of spermiogenesis, this end-piece is separated by a plasma membrane from the late spermatid and remains in the residual cytoplasm when spermatozoa are released into the testis lumen. The mature spermatozoon of D. coeliaca contains two parallel axonemes with the 9 + “1” trepaxonematan pattern, mitochondrion, nucleus, parallel cortical microtubules, and electron-dense granules. Especially the presence of electron-dense granules, occurring frequenly in the spermatozoa of endoparasitic flatworms, may indicate an adaptation to endoparasitic lifestyle in D . coeliaca . The anterior extremity of the male gamete contains one centriolar derivative, a small component of the second centriolar derivative and anterior extremity of the mitochondrion. The posterior extremity of the mature spematozoon exhibits tubular elements of the disorganized axoneme. Variations of the spermatozoon ultrastructural characters within monocotylid monogeneans and possible evolutionary significance of glycogen in sperm to the endoparasitic habit are discussed.

The involvement of oligochaetes in the life cycles of fresh water myxozoan parasites in Brazil was investigated. Of 333 oligochaetes collected in a fish farm in the State of São Paulo, three (0.9%) released Aurantiactinomyxon type spores. From 86 worms collected in a fish farm in Mato Grosso do Sul State, 1 (0.9%) released actinospores with a novel morphology for which we propose the name Seisactinomyxon. Infected oligochaetes were identified by morphology: all belonged to family Naididae, with Pristina americana the host for Aurantiactinomyxon and Slavina evelinae the host of Seisactinomyxon. This is the first report of the involvement these two species of oligochaetes in the life cycle of myxozoans. Small subunit rDNA sequences of the Aurantiactinomyxon (1204 nt) and Seisactinomyxon (1877 nt) did not match any previously sequenced myxozoan. Phylogenetic analyses showed that both actinospore types fell in a clade formed by six Myxobolus spp. that parasitize Characiformes fishes.

An outbreak of severe parasitic pneumonia caused by Dictyocaulus viviparus was diagnosed in adult dairy cows in the municipality of Arabutã, Southern Brazil. The total morbidity in the herd was 71.9%, and the morbidity amongst adult lactating cattle was 100%. The main clinical signs observed were dyspnea, tachypnea, nasal discharge, decreased milk production, and cough. A necropsy was conducted on one animal in order to establish the diagnosis. The herd had been treated previously with levamisole; however, clinical signs persisted and became worse. After treatment with eprinomectin the severity of clinical signs decreased, and the respiratory condition subsequently disappeared. It is believed that the high morbidity presented in this outbreak is related to epidemiological factors, such as increased rainfall in 2014 and 2015, associated with low immunity of the herd. This is the first report of dictyocaulosis in adult dairy cattle in Brazil. Furthermore, it describes an outbreak presenting very high morbidity.

Trypanosoma evansi , the aetiological agent of Surra affects a wide range of livestock and wild animals in India. In the present study, we studied intra- and inter species genetic variability in the transferrin receptor encoding gene regions (ESAG6/7 gene region) of T. evansi isolates by cloning, sequencing and phylogenetic study collected from camel, cattle, donkeys and ponies from North-Western and Central India. The nucleotide sequence variation of ESAG6/7 gene region between Indian T. evansi isolates was up to 17.7% and amino acid sequence variation was up to 31%. Twenty nine clones from six T. evansi isolates from geographical regions of India were included into Clade 1, 5, 6, 7 and 9 consisting of ESAG6 variants reported among T. evansi isolates from South-east Asia and South America. The cladogram indicated a relation between the host species and the genetic variability in the hyper-variable region of ESAG6 gene. Analysis of the Indian ESAG6 variants and their respective Clade positions presented a host specific distribution indicating homogenous parasite population in their respective animal hosts.

A single nucleotide polymorphism on chromosome 4 (SNP TXNIP) has been reported to be associated with roundworm (Ascaris suum) burden in pigs. The objective of the present study was to analyse the immune response to A. suum mounted by pigs with genotype AA (n = 24) and AB (n = 23) at the TXNIP locus. The pigs were repeatedly infected with A. suum from eight weeks of age until necropsy eight weeks later. An uninfected control group (AA; n = 5 and AB; n = 5) was also included. At post mortem, we collected mesenteric lymph nodes and measured the expression of 28 selected immune-related genes. Recordings of worm burdens confirmed our previous results that pigs of the AA genotype were more resistant to infection than AB pigs. We estimated the genotype difference in relative expression levels in infected and uninfected animals. No significant change in expression levels between the two genotypes due to infection was observed for any of the genes, although IL-13 approached significance (P = 0.08; P unadjusted = 0.003). Furthermore, statistical analysis testing for the effect of infection separately in each genotype showed significant up-regulation of IL-13 (P<0.05) and CCL17 (P<0.05) following A. suum infection in the ‘resistant’ AA genotype and not in the ‘susceptible’ AB genotype. Pigs of genotype AB had higher expression of the high-affinity IgG receptor (FCGR1A) than AA pigs in both infected and non-infected animals (P = 1.85*10 -11 ).

Chinese Leishmania isolate MHOM/CN/90/SC10H2 ( L . H2), which was obtained from the spinal cords of patients from the Sichuan province of China, is an uncharacterized, pathogenic species closely related to Leishmania tarentolae . The in vitro transformation rate of L . H2 promastigotes into amastigotes has not been studied. This study is the first to successfully define the in vitro life cycle of L . H2 by investigating the percent conversion of L .H2 promastigotes to amastigotes in vitro under 216 different culture conditions. The highest proportion of L . H2 amastigotes observed (94%) was significantly higher than that previously reported. After conversion, the axenic amastigotes remained viable as verified by the levels of stage-specific genes (Gp46, A2 and β-tubulin) detected by RT-PCR. Meanwhile, morphological and protein characterizations of these axenic amastigotes were carried out in order to confirm the successful conversion. Specific antibodies were only able to detect 46 kDa, 52 kDa and 75 kDa proteins in samples isolated from axenic amastigotes. Afterward, these converted axenic amastigotes were transformed into the promastigote form by altering the culture condition. These converted axenic promastigotes still have the ability to infect macrophages, and their morphology changed back to the amastigote form following infection. These findings will assist further investigations into the biological characteristics of the host-parasite relationship and the process of pathogenesis.

Recombinant human TNFα (rhTNFα) has previously been shown to reduce fecundity in Schistosoma mansoni adult females maintained in vitro without males, and adversely affect the uptake of [ 14 C]-tyrosine, an amino acid required for schistosome vitellogenesis. Here we report on the effect of rhTNFα on [ 14 C]-methionine uptake in both separated and paired females, and the effect of three different preparations of rTNFα on schistosome oviposition in vitro . In the absence of rhTNFa, separated females incorporated only 30% of the [ 14 C]-methionine incorporated by paired females in a dose and time-dependent manner, suggesting low metabolic activity of females in the absence of males. Separated females and worm-pairs were treated with increasing doses of rhTNFα for 2 or 4 hr and then incubated in RPMI 1640 containing 10% fetal calf serum (FCS) and 5 µCi ml −1 [ 14 C]-methionine for 1 hr. Separated females treated with rhTNFa for 4 hr incorporated less methionine than those treated for 2 hr. In contrast, paired females treated with rhTNFa incorporated significantly smaller amounts of [ 14 C]-methionine in a TNFa dose-dependent but time-independent manner [2 hr (P = 0.001) or 4 hr (P = 0.027) One-Way ANOVA]. Worm-pairs maintained in RPMI 1640 containing 10% FCS and 100 ng ml −1 of any of the three rTNFa preparations laid significantly fewer eggs than the worms cultured without rTNFα(P = 0.001; Kruskal-Wallis Test). We also observed that among rTNFα-treated worm-pairs, females were sluggish and tended to separate from their male partners. These observations suggest that TNFa inhibits [ 14 C]-methionine uptake and reduces fecundity in females paired with males. Since paired females incorporate substantially greater amounts of [ 14 C]-methionine, the role of males in stimulating metabolic activity in females is affirmed. Reduced amino acid uptake, and possibly other nutrients, may contribute to the diminished fecundity observed in TNFa-treated females.

A new feather mite species, Neodectes pilgrimi sp. n. (Proctophyllodidae: Pterodectinae), is described from three species of New Zealand wrens (Passeriformes: Acanthisittidae): Xenicus gilviventris Pelzeln, 1867 (type host), X. longipes (Gmelin, 1789) (extinct species), and Acanthisitta chloris (Sparrman, 1787). Based on known host associations of the genus Neodectes , it is hypothesized that Neodectes pilgrimi sp. n. has a secondary origin on New Zealand wrens and was probably transferred onto the ancestor of its hosts from honeyeaters (Passeriformes: Meliphagidae).

The schistosome oviposition and granuloma constitution in the peritoneal cavity of AKR/J mice were evaluated. Groups of mice intraperitoneally infected with cercariae of Schistosoma mansoni were weekly euthanized during the acute (56 to 84 days post–infection (DPI)) and chronic (147 to 175 DPI) phase of infection. Schistosome developmental stages obtained via peritoneal lavage and perfusion of the portal system were inspected, counted and fixed, and peritoneal granulomata were then processed for histology. The morphological characterization and quantitative analysis of peritoneal schistosome eggs and granulomata were for the first time performed, such as the demonstration of the viability of miracidia obtained there from. Eutopic and ectopic mature schistosomes and normal pattern of worm oviposition were observed in all periods studied. However, the size of schistosome eggs from peritoneal cavity was smaller than observed for eggs laid by female worms from the portal system. The numbers of S. mansoni eggs and/or granulomata recovered from the peritoneal cavity was higher in chronic than acute infection, while the mean diameter of peritoneal chronic granulomata was smaller than for peritoneal acute granulomata. The constitution and evolution of these cellular reactions at histology were similar to that of hepatic granuloma, and peritoneal granulomata were subject to the host immunomodulation. In addition to the standardization of this experimental approach, which allows the obtaining of free schistosomal granulomata from peritoneal cavity of AKR/J mice, the potential use of these granulomata in ex vivo and in vivo studies is discussed.

Amoebiasis is a human disease produced by Entamoeba histolytica which causes widespread mortality and morbidity worldwide through diarrheal disease and abscess establishment in parenchymal tissues such as liver, lung, and brain. The true prevalence of infection is unknown for most areas of the world due to the difficulty to characterise E ntamoeba histolytica versus other non-pathogenic amoebas with identical morphology, as Entamoeba dispar , and Entamoeba moshkovskii . To overcome microscopy misidentification issues, we tested a nested multiplex polymerase chain reaction (PCR) and a real-time PCR on 194 stool samples collected from incoming dysentery patients in Cairo hospitals diagnosed with E. histolytica by microscopy. Nested PCR showed only 20 (10.3%) samples positive to E. histolytica and 17 (8.7%) to E. dispar . The real-time PCR detected only 19 and 11 samples positive to E. histolytica and E. dispar respectively, showing less sensitivity than the nested PCR. The data show that prevalence of E. histolytica in Cairo is lower when specific diagnosis methods are used instead of traditional microscopy, allowing to differentiate between morphologically identical human amoebas species.

Adequate knowledge of the genetic diversity among Babesia species infecting dogs is necessary for a better understanding of the epidemiology and control of canine babesiosis. Hence, this study determined the genetic diversity among the Babesia rossi detected in dogs presented for routine examination in Veterinary Hospitals in Abeokuta, Nigeria. Blood were randomly collected from 209 dogs. Field-stained thin smears were made and DNA extracted from the blood. Partial region of the 18S small subunit ribosomal RNA (rRNA) gene was amplified, sequenced and analysed. Babesia species was detected in 16 (7.7%) of the dogs by microscopy. Electrophoresed PCR products from 39 (18.66%) dogs revealed band size of 450 bp and 2 (0.95%) dogs had band size of 430 bp. The sequences obtained from 450 bp amplicon displayed homology of 99.74% (387/388) with partial sequences of 18S rRNA gene of Babesia rossi in the GeneBank. Of the two sequences that had 430 bp amplicon, one was identified as T. annulata and second as T. ovis . A significantly (p<0.05) higher prevalence of B. rossi was detected by PCR compared to microscopy. The mean PCV of Babesia infected dogs was significantly (p<0.05) lower than non-infected dogs. Phylogenetic analysis revealed minimal diversity among B. rossi with the exception of one sequence that was greatly divergent from the others. This study suggests that more than one genotype of B. rossi may be in circulation among the dog population in the study area and this may have potential implication on clinical outcome of canine babesiosis.

The detailed morphology of Cucullanus bourdini Petter et Le Bel, 1992 was further studied using both light and scanning electron microscopy, based on specimens collected from the yellowback fusilier Caesio xanthonota Bleeker (Perciformes: Caesionidae) from off the Taiwan Strait. In addition, C. bourdini was characterized genetically for the first time using molecular approaches by sequencing and analysing ribosomal [small ribosomal subunit (18S rDNA) and internal transcribed spacer 2 (ITS-2)] and mitochondrial [cytochrome c oxidase subunit 1 (cox1)] target regions. The molecular analysis of C. bourdini displayed no nucleotide differences in the 18S sequences, but detected 0.20–2.82% and 0.52–1.04% nucleotide divergence in the ITS-2 and cox1 regions, respectively. The new morphological and genetic data obtained herein would enable us to identify this hitherto little known species more rapidly and accurately.

Equine piroplasmosis (EP) is a hemoprotozoan tick-borne disease with worldwide distribution that is caused by Theileria equi and Babesia caballi . There are studies reporting the presence of equine piroplasmosis in Turkey but the situation in Erzurum is unknown. The aim of the current study was to determine the situation of equine piroplasmosis in jeered horses in Erzurum. Between April and August 2015, a total of 125 Arabian horse were examined and blood samples were collected. At the time of sampling, animals were also examined for tick infestations and clinical signs. Besides microscopic examination of Giemsastained blood smears, multiplex PCR performed with species specific primers partially amplifying the 18S rRNA gene of B. caballi and T. equi . During the microscopic examination of blood smears, T. equi piroplasms were found in 6 (4.8%) samples. In total, 11 (8.8%) T. equi DNA were detected with multiplex PCR. B. caballi piroplasms or DNA were not obtained. BLAST analysis of the sequenced T. equi samples (GenBank: KU921661-KU921667) indicated 98.8–100% identity to each other, and 100% similarity to T. equi isolates in South Africa, Iran, China, Sudan, India, Mongolia, Trinidad, Kenya, Spain, Serbia, Bosnia and Herzegovina and Turkey (Bursa). The results of our study indicate that T. equi occurs more frequently than B. caballi in the study area. To the authors’ knowledge, this is the first report of the molecular detection of equine piroplasmosis in jeered horses in Erzurum, Turkey.

In the production and management of beef and dairy cattle, controlling diarrhea is one of the important concerns. Pathogenic agents of the disease, protozoan parasites including Cryptosporidium spp., are difficult to control, making prevention, diagnoses, and treatment of diarrhea. In the present study, we investigated a farm with a history of calf deaths over a period of 10 years in order to determine the cause of disease and to clarify the detailed distribution of the pathogens. In four examined calves that were reared in calf pens, all were positive with Cryptosporidium and/or Giardia , while the other breeding stock and adult cattle were negative. Molecular analyses revealed that the isolates from calves were C. parvum subtype IIaA15G2R1 as a zoonotic and G. intestinalis assemblage E. Other pathogenic bacteria and diarrhea-causing viruses were not detected. After treating the calf pens with boiling water and milk of lime (Ca[OH] 2 ), oocysts of C. parvum and cysts of G. intestinalis were not found and no additional calves died. This is the first report to describe the mixed infection of both parasites in Japan.

During post-mortem examination of lungs and heart of a 7-month-old female French bulldog, 158 worms were collected from the lung vessels and they proved Angiostrongylus vasorum by their morphological and genetic identification by PCR. The histopathological investigation found a multifocal interstitial inflammation characterized by numerous lymphocytes and a smaller number of plasma cells and eosinophils whilst L1 stage larvae could be seen inside dilated alveoli. We suggested a lethal angiostrongylosis supposed to lead to a fatal effect. Our report attracts attention to the presence of the nematode A. vasorum as causative agent of canine cardiopulmonary disorder in the south-western region of Hungary.

Although the infection by Toxoplasma gondii in horses is usually asymptomatic, it may be characterized by the presence of tissue cysts and maintenance of detectable antibody titers. The aim of this study was to detect anti- T.gondii antibodies and to determine risk factors associated in sport horses from Paraíba State, Northeastern Brazil. A total of 138 apparently healthy sport horses of different age and gender were sampled. Anti-Toxoplasma gondii antibodies were detected in 16/138 (11.6%; 95% CI: 6.77-18.14%) horses by indirect immunofluorescence antibody assay (IFA). A significant association was found between the water source and seropositivity to T. gondii (p<0.05). The present study highlights the importance of providing good quality water to the animals.