Volume 61, Issue 2

Trichinellosis, a parasitic zoonosis with world-wide distribution, causes serious health problems in humans and is also of economic importance. In Slovakia the most frequent species is T. britovi , causing disease mainly in wild life species. T. spiralis occurs less frequently and T. pseudospiralis only sporadically. The paper describes the epidemiology of six human Trichinella infection outbreaks recorded in Slovakia between 1980 and 2008. Before 1990 wild boar meat was the main source of infection. Later, risk farm practices, especially feeding of pigs with the wild animal´s offal contributed to the formation of synanthropic cycle and pig meat caused the epidemics in 1990, 2001 and 2008. Sausages prepared from pork and T. britovi infected dog meat and offered as a local food specialty on traditional folk festival in 1998 (Brezno district, Central Slovakia) were the source of the largest human outbreak recorded in Slovakia. The anti- Trichinella antibodies were detected in 336 event visitors. The main reason of repeated human epidemics in Slovakia has been the permanent circulation of Trichinella spp. in sylvatic cycle, especially in red foxes and wild boars. High population density of both animal species, persistent prevalence of trichinellosis in wild boars and even increasing positivity of red foxes suggest that the risk of human outbreaks in Slovakia persists.

Immune responses of rats and sheep following vaccination with cDNA encoding phosphoglycerate kinase of Fasciola hepatica (cDNA-FhPGK/pCMV) and F. hepatica infection were investigated in the present study. cDNA-FhPGK/pCMV vaccinated female Sprague-Dawley rats were better protected by vaccination than their male counterparts – 48% reduction in fluke burden for females and no protection for males when compared with appropriate infection control groups. Moreover, male rats developed marked leukocytosis during the study with higher neutrophil, eosinophil and monocyte responses than females. Additionally, dynamics of eosinophil and monocyte responses varied between sexes. Increased titres of anti-FhPGK IgG1 and IgG2a correlated with the protective effect of vaccination that was observed among female rats. In the case of male sheep, no differences in worm burdens and in the course of the immune response were observed following vaccination. Titres of specific antibodies detected were low, and cellular responses were not significant. Apparently, sheep immune responses induced by cDNA-FhPGK/pCMV vaccination are not effective at controlling F. hepatica infection. Poor immunogenicity of DNA vaccines in large animals is still a major obstacle of this technology that has to be overcome.

Free-living amoebae (FLA) are widely distributed in soil and water. A few number of them are implicated in human disease: Acanthamoeba spp., Naegleria fowleri , Balamuthia mandrillaris and Sappinia diploidea . Species of Acanthamoeba can cause keratitis and brain infections. In this study, 72 water samples were taken from both hot tubs and thermal swimming pools in the city of Porto Alegre, RS, Brazil, to determine the presence of Acanthamoeba in the water as well as perform the phenotypic and genotypic characterization of the isolates. The identification of the isolates was based on the cysts morphology and PCR amplification using genus-specific oligonucleotides. When the isolates were submitted to PCR reaction only 8 were confirmed as belonging to the genus Acanthamoeba . The sequences analysis when compared to the sequences in the GenBank, showed genotype distribution in group T3 (12,5%), T5 (12,5%), T4 (25%) and T15 (50%). The results of this study confirmed the presence of potentially pathogenic isolates of free living amoebae in hot swimming pool and spas which can present risks to human health.

The present study describes the first isolation and characterization of Vairimorpha plodiae , a microsporidian pathogen of Plodia interpunctella (Lepidoptera: Pyralidae), from Turkey. We present characteristic light and electron microscopical features of the spores. Fresh binucleate spores are oval and measure 4.48 ± 0.23 (4.01–4.84) µm in length and 2.21 ± 0.15 (1.91–2.48) µm in width. Ultrastructural studies showed that the spore wall measures 150 to 200 nm and consists of a clear endospore (125–150 nm) and an electron-dense, uniform, thin exospore (30–50 nm). The polar filament is isofilar and with 10–12 coils. The well-developed polaroplast consists of two parts with thin lamellae anteriorly and thick, irregularly arranged lamellae posteriorly. The identity of our isolate is discussed.

Lymphatic filariasis is a debilitating diseases caused by filarial parasitic nematodes. The infection may be acquired in childhood but the symptoms become apparent only in later life. To evaluate the success of any intervention, sensitive diagnostics were used to identify infection among endemic normals that are likely to develop microfilaremia in due course of time. Capture assay was standardized using the recombinant protein Brugia malayi Abundant Larval Transcript-2 (ALT-2) specific monoclonal and poly-clonal antibodies and evaluated with serum samples of clinical groups from high and low filarial infection area individuals (HIA/LIA), Endemic Normal (EN, n = 478), microfilaeremics (MF, n = 77), chronic pathology (CP, n = 57) and non endemic normal (NEN, n = 20). In order to assess stage-specific infection, ALT-2 capture assay was compared with the early reported Venom allergen homologue (VAH) and microfilariae specific SXP-1 capture assays. Of the 632 serum samples tested, ALT-2 and VAH capture assays detected circulating filarial antigen (CFA) in 57% and 52% of HIA-EN individuals, respectively. As expected, the VAH and SXP-1 capture assays were positive for 100 % of MF individuals. The described capture assays can be useful for the detection of early and stage-specific filarial infections in endemic regions of developing countries.

The American cockroach Periplaneta americana (Linnaeus, 1758) (Blattodea: Blattidae) has been spreading worldwide by commerce and has successfully adjusted to living with humans. There are many reports of thelastomatid parasitic nematode isolated from P. americana in many countries including USA, Canada, India, Argentina, Bulgaria, but not in Japan. We have investigated the parasitic nematodes in P. americana lab strain and field-captured individuals in Japan and found that Thelastoma bulhoesi (de Magalhães, 1900) (Oxyuridomorpha: Thelastomatidae) parasitizes with high infection rates. We described morphological, molecular, and developmental characters of the parasitic nematode because such information was missing despite it has been discovered more than one hundred years ago. We described morphometrics with DIC microscopy and fine structure of male and female adult with SEM observation. We also reveal the embryonic and postembryonic development of this nematode. This is the first report of a thelastomatid nematode isolated from American cockroach in Japan, and the data showed here is also very useful and fundamental for further analysis of the cockroach and parasite relations.

In Central Europe the wild boar population is permanently growing and consequently Cf foodborne infections. In this study serological and molecular detection of Toxoplasma gondii and Neospora caninum in wild boars was evaluated. Moreover, same samples were screened for the presence and genetic variability of tick-borne bacterium Anaplasma phagocytophilum . Blood samples collected from 113 wild boars from Southern Slovakia were examined for antibodies to T. gondii by indirect and to N. caninum by competitive ELISA. The presence of parasitic DNA in blood samples was determined by standard or real time PCR techniques. Antibodies against T. gondii and N. caninum were detected in 45 (39.8%) and 38 (33.6%) animals, respectively. Females were more frequently infected for both pathogens than males. The high seropositivity against both coccidia indicates a permanent occurrence of these pathogens in the studied locality. T. gondii DNA was confirmed in five seropositive boars (4.4%) and N. caninum in 23 blood samples (20.4%). Three out of 23 N. caninum PCR positive animals did not show seropositivity. Three out of 113 blood samples of wild boars were positive for A. phagocytophilum (2.7%). The obtained A. phagocytophilum sequences were 100% identical with GenBank TM isolates from Slovak dog (KC985242); German horse (JF893938) or wild boar (EF143810) and red deer (EF143808) from Poland. Coinfections of T. gondii with N. caninum and N. caninum with A. phagocytophilum were detected in single cases. Results suggest a potential zoonotic risk of toxoplasmosis transmission to humans and the spread of neosporosis to farm animals.

In this study, prevalence of the protozoan parasites from the genus Giardia spp , with zoonotic potential and worldwide dissemination, was accessed in young dogs, which are reported as having higher prevalence rates. With that purpose, 49 animals from the Grupo de Intervenção Cinotécnico of the Guarda Nacional Republicana (Portuguese Gendarmerie Canine Unit) were chosen. They were housed individually in areas with a high number of kennels (up to 100), with ages ragging from newborns to 10 years old. Dogs were divided in four groups, according their age: under 6 months (n = 16), 6–12 months (n = 6), 12–18 months (n = 13) and 18–24 months (n = 14), comprising 22 females and 27 males. Fecal samples were collected from every animal and all were submitted to two different diagnostic tests, a passive flotation technique with a ZnSO 4 solution and a detection of fecal antigen using a commercially available ELISA test (Witness® Giardia – Zoetis). From the 49 samples, 5 (10.2%) were considered positive with ZnSO 4 flotation technique and 6 (12.24%) with the Witness® Giardia test. When considering the combination of both tests, 5 animals (10.2%) were considered positive. Of these, 3 (60%) were from the group under 6 months old, 1 (20%) from the 6–12 months and 1 from the 18–24 (20%) months. Within each group, in the under 6 months group 18.75% (n = 16) were considered positive, 16.67% in the 6–12 month group (n = 6), 0% in the 12–18 month group (n=13) and 7.14% in 18–24 month group (n = 14). None of the animals had clinical signs and no significant differences were found when comparing prevalence according to age, breed or gender. A combination of fecal flotation and antigen ELISA tests have good sensitivity and are easy to perform in practice and, therefore, could be a good choice to perform a diagnostic and small animal veterinarians should have this possible diagnostic in mind when in the present of clinical signs, particularly in young dogs.

Fascioliasis is an important food-borne parasitic zoonosis caused by two trematode species, Fasciola hepatica and Fasciola gigantica . The characterisation and differentiation of Fasciola populations is crucial to control the disease, given the different transmission, epidemiology and pathology characteristics of the two species. Lineal biometric features of adult liver flukes infecting livestock have been studied to characterise and discriminate fasciolids from Bangladesh. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected bovines (cattle, buffaloes) throughout the country. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardised measurements and the logistic model of the body growth and development of fasciolids in the different host groups. Since it is the first ever comprehensive study of this kind undertaken in Bangladesh, the results are compared to pure fasciolid populations of F. hepatica from the European Mediterranean area and F. gigantica from Burkina Faso, geographical areas where both species do not co-exist. Principal component analysis showed that the biometric characteristics of fasciolids from Bangladesh are situated between F. hepatica and F. gigantica standard populations, indicating the presence of phenotypes of intermediate forms in Bangladesh. These results are analysed by considering the present emergence of animal fascioliasis, the local lymnaeid fauna, the impact of climate change, and the risk of human infection in the country.

Two nematode species, Pseudoproleptus notopteri (Karve et Naik, 1951) and Spinitectus notopteri Karve et Naik, 1951 (both Cystidicolidae), are redescribed based on light and scanning electron microscopical studies of specimens collected from the digestive tract of the freshwater fish Notopterus notopterus (Pallas) (Notopteridae, Osteoglossiformes) in Thailand. Some new important morphological features, such as a detailed structure of the cephalic end and the presence of bifurcate deirids and a ventral median caudal protuberance in male, are reported for the former species ( P . notopteri ), which is provisionally assigned to Pseudoproleptus Khera, 1955; Notopteroides notopteri Chakravarty et Majumdar, 1962, Pseudoproleptus satendri Sahay, 1967, P . lamyi Le-Van-Hoa et Bui-Thi Lien-Huong, 1969, P . gomtii Gupta et Bakshi, 1984. P . sprenti Gupta et Masoodi, 1986 and P . thapari Gupta et Naiyer, 1992 are considered its junior synonyms. The first study of S . notopteri by SEM showed its morphological similarity with S . mastacembeli Karve et Naik, 1951, from which it clearly differs by the structure of eggs; Spinitectus alii Kalyankar, 1970, S . bengalensis Chakravarty, Sain et Majumdar, 1961, S . gomalensis Siddiqui et Kattak, 1984 and S . thapari Ali, 1957 are considered to be junior synonyms of S . notopteri . Pseudoproleptus notopteri and Spinitectus notopteri are reported from Thailand for the first time.

A new species of monogenean diplozoid, Paradiplozoon iraqensis n. sp., from the gills of Cyprinion macrostomum Heckel from the Tigris River in Tikrit, Iraq is described. P. iraqensis n. sp. is distinguished from other species of the genus by the size of plicae on the fore- and hind body, the number of the branches of the intestinal caecum in fore body, intestinal caecum not extending to haptor, the size of the clamps and the morphology of the clamp sclerites, and by the length of the central hooks. Details of the shape and size of eggs, and of the reproductive system and prohaptoral region are provided. The present study raises the number of species of Paradiplozoon recorded in Iraq to 15.

Diplomonadida are primitive flagellate protozoa, among which both commensals and pathogens have been recorded. To date, members of the genera Hexamita and Spironucleus have been reported in the digestive system of fish in the Baikal region. We determined the genetic diversity of Diplomonadida in fish of the genus Coregonus from south-eastern Siberia using molecular-genetic methods. Fish for analysis were caught in Lake Baikal and in the Barguzin, Nepa, Chechuy, and Kirenga rivers from 2010 to 2013. Gall bladders, hindguts and foreguts of 120 specimens of Coregonus migratorius representing three morpho-ecological groups, 25 specimens of Coregonus lavaretus baicalensis , 25 specimens of Coregonus tugun and 30 specimens of Coregonus lavaretus pidschian were analysed via amplification with primers specifically designed for eukaryotes. Amplicons positive for Diplomonadida were sequenced. A phylogenetic analysis revealed that diplomonad flagellates of whitefish from Southeastern Siberia belong to Spironucleus barkhanus . Positive Diplomonadida DNA samples were analysed with primers designed in the present study for the amplification of small subunits of ribosomal DNA fragments of S. barkhanus (about 1,430 bp) and sequenced. Phylogenetic analysis revealed inside the clade of S. barkhanus besides the cosmopolitan genotype from European salmon that was detected earlier in Baikalian grayling, a new genotype unique to the fish of the genus Coregonus from Lake Baikal.

The dynamics of the emergence, duration, and decline phases in epizootic cycles are well known for humans and some crops, but they are poorly understood for host–parasite systems in the wild. Parasites may be particularly insidious as they are often introduced unintentionally, simultaneously with their hosts, and later transferred to species in the new location. Here we investigate the epizootic dynamics of the tapeworm Ligula intestinalis in the Hamiz reservoir, Algeria, and explore its effects on the cyprinid fish Barbus callensis . Regular sampling was conducted from October 2005 to February 2008 with intermittent surveys carried out until 2010. Five percent of the 566 specimens of B. callensis that were caught were infected, with the maximum number of parasites found in spring. There was no obvious difference in weight between uninfected fish and infected ones, and infection did not affect fish condition. However, infected fish were significantly longer than uninfected fish and had inhibited gonad development. The proportion of infected fish caught was significantly higher in year 1 and by the second winter, infection collapsed to zero. The Ligula infection thus appeared to have minimal ecological effects and be of a temporary nature, thus exhibiting an epizootic cycle. Taken together, our data indicates that this infection declined or even failed during our study period. Failure may be due to the specific genetic strain of Ligula , but invasive carp may also have been influential in both the introduction and subsequent decline of this parasite.

Toxoplasma gondii is an obligate intracellular apicomplexan parasite, and can infect warmblooded animals and humans all over the world. In the past years, ROP family genes encoding particular proteins of T. gondii had made a great contribution to toxoplasmosis . In this study, we used multiple bioinformatics approaches to predict the physical and chemical characteristics, transmembrane domain, epitope, and topological structure of the rhoptry protein 48 (ROP48). The results indicated that ROP48 protein was mainly located in the membrane and had several positive linear-B cell epitopes and Th-cell epitopes, which suggested that ROP48 is a potential DNA vaccine candidate against toxoplasmosis. Then the PCR product amplified from the ROP48 cDNA was inserted into a pEASY-T1 vector to build a recombinant cloning plasmid. After sequencing, ROP48 was subcloned into a eukaryotic expression plasmid pEGFP-C1 to obtain pEGFP-C1-ROP48 (pROP48). After identification by PCR and restriction enzyme digestion, the recombinant plasmid pROP48 was transfected into HEK 293-T cell and identified by RT-PCR. The results showed that the eukaryotic expression plasmid pROP48 was constructed and transfected to the cells of HEK 293-T successfully. Western blotting showed that the expressed proteins can be recognized by anti-STAg mouse sera.

In the present study, a total of 14 (73.68%) cases of abortions and two (100%) cases of still births were detected positive for Trypanosoma evansi infection by wet and dry blood smear examination and fetal tissue PCR in camels of an organized farm. The abortions in infected dams were recorded from 8 to 11.5 month of gestation, however majority occurred during 9 th to 10 th month. The important laboratorial findings in infected dams were anemia, hypoglycemia, hyperproteinemia and leukocytosis. At necropsy the T. evansi infected aborted and still born fetuses showed subcutaneous edema, presence of moderate amount of dark red hemolysed blood in thoracic and abdominal cavity, bronchopneumonia, hepatic necrosis and acute congestion in all vital organs. Microscopically, there was severe congestion, thickening of bronchial and alveolar wall and mononuclear infiltration in the fetal lung, necrotic and degenerative changes in the liver, nephritis along with severe congestion and tubular necrosis in the kidneys and necrotic and degenerative changes and congestion of capillaries in the brain. The T. evansi DNA was detected by PCR from blood, lung, spleen, liver, kidney and brain of all the infected aborted and still born fetuses. The results of the study indicated that T. evansi can cross placental barrier and cause pathological events in the fetus resulting into abortion or still birth in pregnant camels.

Equine piroplasmosis is significant tick-borne disease with wide distribution. The prevalence of equine piroplasmosis in Serbia, Montenegro and Bosnia and Herzegovina is unknown. In aim to obtain a first insight into the prevalence we performed molecular epidemiological study which included 142 horses, on seven locations in these three countries. We first performed PCR for the detection of a 450bp long section of the 18S rRNA of piroplasma-specific region. For all positive samples we have done multiplex PCR for the species detection. Species determination was further confirmed by sequencing PCR products of 10 randomly selected Theileria equi and all Babesia caballi samples. The overall prevalence rates in analysed region for T. equi and B. caballi were 22.5% and 2.1%, respectively. Possible risk factors (such as location, age, sex and activity) associated with PCR positivity were evaluated. Marked differences were found in prevalence between geographic areas. There was no significant association between positivity and age group. T. equi was more prevalent in females and farming horses. This is the first report on the molecular survey of T. equi and B. caballi in central Balkan. Further prevalence studies on definitive host and vectors in this region are necessary.

Two new species of scale-mites parasitizing lizards of the family Agamidae (Sauria) are described: Pterygosoma blandfordi n. sp. from Psammophilus blanfordanus (Stoliczka) (Agamidae: Draconinae) from South India and Pterygosoma balochistani n. sp. from Laudakia nupta nupta (De Filippi) (Agamidae: Agaminae) from Pakistan. Pterygosoma blandfordi n. sp. (female) differs from P. foliosetis Jack, 1961 by the shape of the idiosoma which is much wider than long (vs. rounded idiosoma in P. foliosetis ), the presence of 110–139 pairs of the dorsal anterolateral setae (vs. presence of about 35 pairs of these setae), 20–26 pairs of the peripheral setae (vs. 10–19 pairs), 3 pairs of the genital setae (vs. 1 pair), 6 pairs of the pseudoanal setae (vs. 4 pairs), the absence of leg setae vGII and presence of setae vGIV (vs. presence of setae vGII and absence of setae vGIV ). P. balochistani n. sp. (female) differs from P. persicum Hirst, 1917 by the chelicerae 325–350 long (vs. 190–230 long in P. persicum ), the fixed cheliceral digit bearing small tines (vs. spinous fixed cheliceral digit), presence of subcapitular setae n (vs. absence of setae n ), serrate peripheral setae (vs. smooth peripheral setae), presence of leg setae vGII–III (vs. absence of setae vGII–III ), 4 pairs of the genital setae (vs. 3 pairs) and 7 pairs of the pseudoanal serrate setae (vs. 9–11 pairs of filiform setae ps ).

Spauligodon geckonis sp. nov. from the intestines of Ptychozoon kuhli (Gekkonidae) from East Malaysia is described and illustrated. Spauligodon geckonis sp. nov. represents the 52 nd species assigned to the genus and the third species from the Oriental Region. The new species is separated from congeners by the unique combination of aspinose filamentous tail and spicule in the male and spinose filamentous tail and fusiform, two knobbed eggs in the female. Gravid individuals 3 species of Nematoda, Meteterakis singaporensis, Physalopteroides grismeri , and Skrjabinelazia machidai , as well as larvae assignable to the Ascaridae were also found.

Polyopisthocotylean monogenean parasites of fishes are highly host specific and have been used as an appropriate model to study the host-parasite co-evolution. In the present study, eight monogeneans of the order Mazocraeidea were characterized by nuclear 28S rDNA sequences and their phylogenetic relationship with other polyopisthocotylean species was investigated. Neighbour-joining, maximum parsimony, maximum likelihood and Bayesian Inference methods were used for phylogenetic reconstruction. The topology sustained by high bootstrap was: (((Hexabothriidae (Mazocraeidae (Discocotylidae (Diplozoidae (Diclidophoridae (Plectanocotylidae (Heteromicrocotylidae (Microcotylidae (Heteraxinidae), (Thoracocotylidae, Gotocotylidae (Gastrocoylidae (Allodiscocotylidae: Protomicrocotylidae))). In addition, we have also developed DNA barcodes (COI sequences) for six species and the barcodes clearly discriminated all the species. The polytomy within Protomicrocotylidae family is resolved in this study for the first time and it appears that within this family, Bilaterocotyloides species are basal compared to Neomicrocotyle and Lethacotyle species while the latter is the more derived.

Anisakis and other parasites belonging to the Anisakidae family are organisms of interest for human health, because of their high zoonotic potential. Parasites belonging to this family can cause Anisakiasis, a parasitological disease caused by the ingestion of raw, infested fish products. Furthermore, evidence from the EFSA (European Food Safety Authority; EFSA 2010) has highlighted the allergological potential of nematodes belonging to the Anisakis genre. The detection and identification of Anisakidae larvae in fish products requires an initial visual inspection of the fish sample, as well as other techniques such as candling, UV illumination and artificial digestion. The digestion method consists of the simulation of digestive mechanics, which is made possible by the utilization of HCl and pepsin, according to EC Regulation 2075/2005. In this study, a new Anisakidae larvae detection method using a mechanical digestion system called Trichineasy ® was developed. A total of 142 fish samples, belonging to 14 different species, were examined to validate the method. A reaction mixture with 100 g of sample, 10 g of pepsin (1:10000 NF) and 50 ml of 10% HCl at 36 ± 1°C for 20 minutes was evaluated to be the best condition for the digestion of fish samples. These parameters have also allowed the detection of viable larvae after digestion. The results confirm this instrumentation as a valuable and safe tool for the detection of Anisakidae larvae in fishery products.

Trichomonosis, caused by the protozoan parasite Trichomonas vaginalis , is a curable sexually transmitted disease that is most commonly encountered worldwide. Increasing importance of trichomoniasis and emerging of resistance against metronidazole lead to search for alternative drugs with different mode of activity. The purpose of this study was to determine in vitro activity of ceragenins (CSA-13, CSA-44, CSA-13, and CSA-138) against the metronidazole-susceptible (ATCC 30001) and metronidazole-resistant (ATCC 50138) strains of T. vaginalis . The effective concentrations were evaluated using two strains of T. vaginalis with different metronidazole susceptibilities (ATCC 30001 and ATCC 50138) in the presence of dilution series of ceragenins in 24-well microtitre assays. Overall, all the ceragenins killed the metronidazole-susceptible (ATCC 30001) and metronidazole-resistant (ATCC 50138) strains of T. vaginalis (p>0.05). With regard to the their effects against the studied strains of T. vaginalis , in order of effectiveness, overall, the ceragenins ordered as CSA-13 (the most effective), CSA-131 and CSA-138 (effective similarly), and CSA-44 (the least effective) (p<0.05). All of the ceragenins reduced the trophozoite numbers of both of studied strains of T. vaginalis with a time- and dose- dependent manner (p<0.05). Although all of the study ceragenins, CSA-13, CSA-44, CSA-13, and CSA-138, killed the studied strains of T. vaginalis . CSA-13 is the leading ceragenin as the most effective anti-trichomonas compound, followed by CSA-131 and CSA-138. They have a potential to have a place in the armemantarium of gynecologic and urologic practice for the management of sexually transmitted diseases.

Enterocytozoon bieneusi is one of the most important zoonotic pathogen that can infect almost all animals, including humans. However, little information is available regarding prevalence and genotypes of E. bieneusi in sika deer. In the present study, the prevalence of E. bieneusi infection in sika deer in Jilin province, Northeastern China was examined using PCR amplification of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene. 23 (7.06%) of 326 samples were tested E. bieneusi -positive, and the risk factor significantly associated with E. bieneusi prevalence was the age of sika deer. Sequence analysis of the ITS rRNA gene suggested that 8 genotypes of E. bieneusi were found in this study, with five known genotypes, namely J ( n = 11), BEB6 ( n = 4), EbpC ( n = 1), CHN-DC1 ( n = 1), KIN-1 ( n = 1) and three novel genotypes, namely JLD-1 ( n = 2), JLD-2 ( n = 2) and JLD-3 ( n = 1). Phylogenetic analysis indicated that genotypes CHN-DC-1, KIN-1, EbpC, JLD-2 and JLD-3 fell into group 1, while other three genotypes (genotypes J, BEB6 and JLD-1) were clustered into group 2 (so-called bovine-specific groups). These findings indicated the presence of zoonotic E. bieneusi in Jilin province, Northeastern China. Effective strategies should be performed to control E. bieneusi infection in sika deer, other animals and humans.

Wild canides have a high epizootiological – epidemiological significance, considering that they are hosts for some parasites which spread vector born diseases. Increased frequency of certain interactions between domestic and wild canides increases the risk of occurrence, spreading and maintaining the infection of parasitic etiology in domestic canides. The research was conducted in 232 wild canides (172 red foxes and 60 golden jackals). The examined material was sampled from foxes and jackals, which were hunted down between 2010 and 2014, from 8 epizootiological areas of Serbia (North-Bačka, West-Bačka, Southern-Banat, Moravički, Zlatiborski, Raški, Rasinski and Zaječarski district). On completing the parasitological dissection and the coprological diagnostics, in wild canides protozoa from the genus Isospora were identified, 3 species of trematoda ( Alaria alata , Pseudamphistomum truncatum and Metagonimus yokogawai ), cestods from the genus Taenia and 5 species of nematodes ( Toxocara canis , Ancylostomatidae , Trichuris vulpis and Capillaria aerophila ). The finding of M. yokogawai in golden jackals were, to the best of our knowledge, one of the first diagnosed cases of metagonimosis in golden jackals in Serbia. The continued monitoring of the parasitic fauna of wild canides is needed to establish the widespread of the zoonoses in different regions of Serbia, because they present the reservoirs and/or sources of these infections.

Giardia duodenalis is one of the most common enteric parasites of humans and animals, including companion animals, livestock and wildlife. To date, the information about the prevalence and molecular characterization of G. duodenalis infection in white yaks was limited. In the present study, a total of 208 white yak fecal samples were collected from Tianzhu Tibetan Autonomous County (TTAC) between September 2013 and March 2014. Of the 208 white yak fecal samples, four samples (1.92%, all collected in March 2014) tested G. duodenalis -positive by PCR amplification of triosephosphate isomerase (tpi) gene. Sequence analysis confirmed the presence of G. duodenalis assemblage E. The present study revealed the presence and genotype of G. duodenalis in white yaks for the first time, and extended the host range of G. duodenalis . These results provided useful information for further genotyping or subtyping studies of G. duodenalis in white yaks.

The aim of the present study was to investigate the prevalence of antibodies against Toxoplasma gondii in turkeys and chickens on family farms in the semi-arid region of the state of Pernambuco, Brazil. In order to do so, 204 sera samples from turkeys were analyzed using the Modified Agglutination Test (MAT) and 322 sera samples from chickens were assessed using the Immunofluorescence Assay (IFA). Twenty-eight properties contained turkeys and 23 of these properties also contained chickens. The relative frequency of positive turkeys was 11% (21/204), with 46.6% (13/28) of the properties containing at least one positive turkey. The frequency of positive chickens was 25.8% (83/322), with 95.6% (22/23) of the properties containing at least one positive chicken. Based on the results of the present study, it was possible to conclude that turkeys can serve as an indicator of environmental contamination by oocysts of T. gondii . However, they are less effective than chickens bred in the same conditions. The increasing demand from consumers for naturally produced products should worry local sanitary authorities due to the high prevalence of antibodies against T. gondii found in this type of rearing system, particularly among chickens.

Cystoisospora suis is a pathogen that causes diarrhea in pigs and can lead to serious disease. Species identification, especially by histopathological examination, is often difficult because of morphologically similar parasites such as Eimeria species. In this study, we used histopathological, bacteriological, virological, and parasitological methods to identify the cause of the disease in two piglets with severe diarrhea. Villous atrophy, diffuse necrosis, and flattening of mucosal epithelial cells were found in the ilea of examined piglets, and coccidian parasites were found in the cytoplasm of the epithelial cells. In some merozoites in the meronts, the presence of two nuclei indicated type 1 merozoites, characteristic of C. suis . According to Cystoisospora- specific PCR targeting the rRNA internal transcribed spacer 1 (ITS1) gene, the sequences of the products were 98.5% similar to those of C. suis . Escherichia coli (O149 serogroup) exhibiting a virulence factor profile (LT, STb, and EAST1 as toxins and F4 as a colonization factor) was detected in one piglet. No other bacteria or significant enteric viruses were found. Co-infection with C. suis and E. coli could imply aggravation of the disease, although further study is needed to assess the pathogenicity of this interaction. This study is the first to clarify by molecular analysis the sequences of C. suis detected in piglets in Japan.

Procalcitonin (PCT) and C-reactive protein (CRP) may be useful to predict complicated forms of malaria. A total of 30 consecutive travelers diagnosed with Plasmodium falciparum malaria over a two-year period were included in the study. Patients with complicated Plasmodium falciparum malaria showed higher levels of parasitemia (P = 0.0001), PCT (P = 0.0018), CRP (P = 0.0005), bilirubinemia (P = 0.004), and a lower platelet count (P<0.0001) compared with patients with uncomplicated forms. PCT levels above 5 ng/mL showed the highest value of specificity (0.86) and positive predictive factor (0.67) among other parameters, and equal sensitivity (0.67) was displayed by CRP levels above 150 mg/dl. None of the patients with complicated malaria showed PCT levels within normal limits (<0.5 ng/ml). Both PCT and CRP correlated with parasitemia (P<0.001) and showed areas under ROC curve of 0.83. At multivariate analysis, only PCT was associated with an increased risk of complicated malaria (OR 8.2, IC 95% 1.2–57.2, P = 0.03). The determination of PCT on admission showed better results compared to CRP, platelet count, and bilirubinemia and can be useful in non-endemic areas for the initial clinical assessment of disease severity in travelers with Plasmodium falciparum malaria.

Theirelia orientalis is a tick-borne haemoprotozoan parasite, and infection with this parasite is one of the most important diseases for grazing cattle. Co-infection of cattle with different genotypes of T. orientalis often occurs. In this study, we investigated the temporal dynamics of genotypes in cattle in a year-round grazing system in Japan. Genotype-specific PCR assays to determine major piroplasm surface protein (MPSP) genotypes (types 1 to 5) of T. orientalis were performed by using time-course blood samples collected from grazing cattle and ticks in a pasture. All 20 cattle investigated in this study were infected with T. orientalis . By using genotype-specific PCR, we detected the combination of genotypes of T. orientalis (types 1 to 5) from each cattle. These multiple genotypes of T. orientalis were also confirmed in ticks. Notably, each genotype of T. orientalis in cattle was temporally detected from cattle and more variable genotypes were found in summer. The observed temporal dynamics of the MPSP genotypes of T. orientalis in cattle could be explained by host immunity against the parasites or genetic recombination of parasite in ticks.

A new isosporoid coccidian species (Protozoa: Apicomplexa: Eimeriidae) is reported from the southern house wren Troglodytes musculus , a very well distributed species in South and Central America. Isospora corruirae sp. nov. oocysts are subspherical to ovoidal, 24.1 × 21.4 μm, with smooth, bilayered wall. Micropyle and oocyst residuum are absent, but small spherules and splinter-like granules are frequently present. Sporocysts are ovoidal to piriform, 14.0 × 9.5 μm. Stieda body is prominent knob-like and substieda body is delicate. Sporocyst residuum is composed of scattered fragments of different sizes. Sporozoites are vermiform with posterior refractile bodies, anterior striations and a nucleus. This is the second description of an isosporoid coccidium infecting a New World wren.

Hymenolepis diminuta is an important model species in studies of therapeutics, biochemical processes, immune responses and other aspects of cestodiasis. The parasite produces numerous excretory-secretory (E-S) proteins and a glycocalyx covering its body. Our study focused on the mass spectrometry analysis of the E-S material with an objective to determine if E-S contains any new proteins, in particular those that can be identified as: antigens, vaccine candidates and drug targets. These proteins might engage directly in host-parasite interactions. Adult parasites collected from experimentally infected rats were cultured in vitro for 5 and 18h. Immunoblotting was used to verify which E-S protein bands separated in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) react with specific antibodies from sera of infected rats. We identified thirty-nine proteins by LC-MS/MS (liquid chromatography mass spectrometry). Results indicated the presence of proteins that have never been identified in cestode E-S material. Immunoblotting showed the immunogenicity of E-S products of H. diminuta , most probably associated with the presence of proteins known as antigens in other flatworm species. Among identified proteins are those engaged in immunomodulatory processes (eg. HSP), in response to oxidative stress (peroxidasin) or metabolism (eg. GAPDH). The predominant functions are associated with metabolism and catalytic activity. This is the first study identifying E-S-proteins in adult tapeworms, thus providing information for better understanding host-parasite interrelationships, and may point out potential targets for vaccines or drug discovery studies, as among the proteins observed in our study are those known to be antigens.