We examined the in vitro and in vivo efficacy of plant cysteine proteinases (CPs) derived from pineapple (Ananas comosus) and kiwi fruit (Actinidia deliciosa), and compared their efficacy as anthelmintics to the known effects of CPs from the latex of papaya (Carica papaya) against the rodent intestinal nematode, Heligmosomoides bakeri. Both fruit bromelain and stem bromelain had significant in vitro detrimental effects on H. bakeri but in comparison, actinidain from kiwi fruit had very little effect. However, in vivo trials indicated far less efficacy of stem bromelain and fruit bromelain than that expected from the in vitro experiments (24.5% and 22.4% reduction in worm burdens, respectively) against H. bakeri. Scanning electron microscopy revealed signs of cuticular damage on worms incubated in fruit bromelain, stem bromelain and actinidain, but this was far less extensive than on those incubated in papaya latex supernatant. We conclude that, on the basis of presently available data, CPs derived from pineapples and kiwi fruits are not suitable for development as novel anthelmintics for intestinal nematode infections.

Aspidogastreans are commonly found infecting freshwater and marine molluscs, teleosts fishes and freshwater turtles. The subclass comprises four families – Rugogastridae Schell 1973, Stichocotylidae Faust & Tang 1936, Multicalycidae Gibson & Chinabut 1984 and Aspidogastridae Poche 1907 – and it is characterized by the presence of a ventral adhesive disc divided into rows of alveoli. In the current work, using light and scanning electron microscopy and molecular approaches, a new species of Aspidogastridae of the genus Rohdella Gibson & Chinabut, 1984, is described as a parasite of Colomesus psittacus in Brazil. The new taxon is distinguishable by the presence of oesophageal glands, teguments covered by ciliated papillae, and the position and shape of the hermaphroditic duct. The present work describes the third species of the genus Rohdella, thereby adding new morphological and molecular data regarding Aspidogastridae.

Pontoporia blainvillei (Gervais and d'Orbigny, 1844) is an endangered small cetacean endemic to South America with four Franciscana Management Areas (FMA) recognized as different population stocks. The role of the intestinal parasite Synthesium pontoporiae (Digenea: Brachycladiidae) as a possible biological marker to differentiate P. blainvillei stocks was evaluated using nuclear and mitochondrial DNA markers. Internal transcribed sequence 1 and 2 (ITS1 and ITS2) regions of S. pontoporiae did not show intraspecific variability. The mitochondrial NADH dehydrogenase subunit 3 (ND3) and cytochrome oxidase subunit I (COI) gene sequences suggested lack of population structure in S. pontoporiae and population expansion. The apparent panmixia of S. pontoporiae may be due to the high mobility of one or more of its intermediary hosts. Alternatively, it may be due to the small sample size. This result is incongruent with the previously proposed FMA.

Morphological and molecular sequence data were combined with cross-hybridization studies and used to identify a new Steinernema sp. from Free State, South Africa. Molecular and morphological data indicate that the new species belongs to the ‘glaseri-group’ of Steinernema spp. Key morphological diagnostic characters for S. innovationi n. sp. include the morphometric features of the third-stage infective juveniles: total body length = 1054 (1000–1103) μm, tail length = 108 (97–117) μm, location of the excretory pore = 88 (82–91) μm, and D% = 58 (54–63), E% = 115 (104–137) and H% = 43 (37–46). Additionally, the morphology of the spicules and gubernaculum of the first-generation males are considered key diagnostic traits. Steinernema innovationi n. sp. was also characterized by analysis of both rDNA and mitochondrial gene sequence data, which further indicate the uniqueness of this Steinernema species.

The aim of this study was to develop a low-cost antifilarial immunoglobulin (Ig) G4 detection kit for the diagnosis of lymphatic filariasis. The kit was designed to be used by minimally trained personnel without the constraints of expensive laboratory equipment. We provide a description of the development and validation of a single-serum-dilution based enzyme-linked immunosorbent assay (ELISA) kit with ready-to-use reagents for measuring antifilarial IgG4 antibodies. The kit was tested on residents in Brugia malayi-endemic areas in southern Thailand. Detection was performed by naked-eye observation of the resultant colour of the immunological reactivity. The coefficient of variation (CV) was used to assess the reproducibility of the results. Long-term stability was measured over a 6-month period. Sensitivity of the test kit was 97% when compared with microfilariae detection in thick blood smears. Specificity was 98.7% based on the sera of 57 patients living outside the endemic areas who were infected with other parasites and 100 parasite-free subjects. All positive CVs were < 10%. The test kit was remarkably stable over 6 months. Field validation was performed by the detection of antifilarial IgG4 in 4365 serum samples collected from residents of brugian filariasis-endemic areas and compared with outcome colours of the test samples by the naked eye. Subsequent ELISA evaluation of these results using an ELISA reader indicated high agreement by the kappa statistic. These results demonstrate that the test kit is efficient and useful for public health laboratories as an alternative tool for the diagnosis of lymphatic filarial infection.

A cornerstone in parasitology is why some species or populations are more parasitized than others. Here we examine the influence of host characteristics and habitat on parasite prevalence. We studied the helminths parasitizing the Pyrenean brook newt Calotriton asper (n= 167), paying special attention to the relationship between parasites and ecological factors such as habitat, sex, ontogeny, body size and age of the host. We detected two species of parasites, Megalobatrachonema terdentatum (Nematoda: Kathlaniidae) and Brachycoelium salamandrae (Trematoda: Brachycoeliidae), with a prevalence of 5.99% and 1.2%, respectively. Marginally significant differences were found in the prevalence between sexes, with females being more parasitized than males. The present results show significant differences in the body length of paedomorphic and metamorphic individuals, the former being smaller. Nevertheless, no significant correlations between parasite prevalence and either newt body length, ontogenetic stage or age were found. In comparison with other Salamandridae living in ponds, prevalence and diversity values were low. This may be due to a long hibernation period, the species' lotic habitat and its reophilous lifestyle, which probably do not allow for a high parasite load.

During the past decade, golden jackal populations have substantially increased, yet little is known of their potential for transmitting parasites within animal and human hosts. In the present study, between 2005 and 2010, 447 jackals from six localities in Serbia were examined for intestinal parasites. Two species of trematodes (Alaria alata, Pseudamphistomum truncatum), three nematodes (Toxocara canis, Ancylostoma caninum, Gongylonema sp.), and seven cestodes (Taenia pisiformis, Taenia hydatigena, Multiceps multiceps, Multiceps serialis, Mesocestoides lineatus, Mesocestoides litteratus, Dipylidium caninum) were identified. Pseudamphistomum truncatum and M. serialis species were recorded for the first time. The overall prevalence of parasitic infection was 10.3%. No significant differences were found in the prevalence of infection between males and females (P>0.817), between localities (P>0.502), or with regard to annual cycles (P>0.502). In the infected jackal population, 65% harboured multiple infections and one individual was a host to five different types of parasite species, the highest number of parasites we recorded in a single host. These findings indicate that although the prevalence of gastrointestinal helminths in the jackal population in Serbia is significantly lower than expected from earlier studies, further monitoring is required given the jackal's rapid population increase.

This study investigated the epidemiological factors that contribute to the seroprevalence of Toxoplasma gondii and Toxocara spp. in children from Paraná state, Brazil. Immunoglobulin G (IgG) antibodies to T. gondii were detected using indirect immunofluorescence, and IgG antibodies to Toxocara were detected using an enzyme-linked immunosorbent assay. For each individual, a questionnaire was completed that contained epidemiological and clinical data. The data analysis was performed using multiple logistic regression. Of the 544 children investigated, 3.2% presented co-infection with T. gondii and Toxocara spp. Of this total, 7.4% were positive for antibodies to T. gondii, and 25% were positive for antibodies to Toxocara spp. The presence of antibodies to Toxocara spp. increased the risk of T. gondii infection (P= 0.029). Children who were 1–8 years of age were less infected by T. gondii than those who were 9–12 years of age. The variables that influenced positivity for anti-Toxocara spp. were the origin of the children and contact with sand. Children with positive serology for Toxocara spp. presented more eosinophilia compared with those with non-reactive serology. Infection with both parasites reveals the need for preventive measures, such as guidance about modes of infection, parasite control and monitoring recreational areas.

Morphometric analysis of Schistosoma mansoni male worms obtained from AKR/J and Swiss mice was carried out. Rodents infected by the intraperitoneal route with 80 cercariae of the schistosome (LE strain) were killed by cervical dislocation at 45 and 60 days post-infection and both peritoneal lavage and perfusion of the portal system were performed for the recovery of adult worms. Characteristics including total body length, the distance between oral and ventral suckers, extension of testicular mass and the number of testes were considered in the morphological analysis. Changes that occurred in S. mansoni recovered from the peritoneal cavity or from the portal system of AKR/J and Swiss mice included total body length and reproductive characteristics. Significant morphometric alterations were also observed when worms recovered from the portal system of both strains of mice were compared with the schistosomes obtained from hamsters (Mesocricetus auratus), the vertebrate host in which the LE strain had been adapted and maintained by successive passages for more than four decades. The present results reinforce the idea that S. mansoni has high plastic potential and adaptive capacity.

Human cystic echinococcosis (CE) is known to be endemic in the Tibet Autonomous Region (TAR), China; however, there is relatively little data from hospital records or community prevalence studies, and the situation regarding occurrence of human alveolar echinococcosis (AE) is unclear. Here we review the available reports about human echinococcosis in the seven prefectures of TAR. In addition, two pilot studies by mass screening using ultrasound (with serology) were undertaken (2006/7) in Dangxiong County of Lhasa Prefecture (north central TAR) and Dingqing County of Changdu Prefecture (eastern TAR). In Dangxiong County a prevalence of 9.9% (55/557) for human CE was obtained but no human AE cases were detected. By contrast, in Dingqing County (N= 232 persons screened), 11 CE cases (4.7%) and 12 AE cases (5.2%) (including one mixed CE and AE case) were diagnosed by ultrasound. Hospital records and published reports indicated that CE cases were recorded in all of seven prefectures in Tibet Autonomous Region, and AE cases in four prefectures. Incidence rates of human CE were estimated to range from 1.9 to 155 per 100,000 across the seven prefectures of TAR, with a regional incidence of 45.1 per 100,000. Incidence of AE was estimated to be between 0.6 and 2.8 cases per 100,000. Overall for TAR, human AE prevalence appeared relatively low; however, the pilot mass screening in Dingqing in eastern TAR indicated that human AE disease is a potential public health problem, possibly similar to that already well described in Tibetan communities bordering TAR in north-west Sichuan and south-west Qinghai provinces.

Plagioscion squamosissimus (Heckel, 1840) (Osteichthyes: Sciaenidae) is considered piscivorous and is a generalist species endemic to the Amazon region. This fish is an important part of the natural ecosystems in which it occurs and provides basic functional components in the food web. The genus Neoechinorhynchus Stiles & Hassall, 1905 is distributed worldwide and parasitizes fish and turtles, but there are few reports of parasites of this genus in South America, due to the high diversity of fish that can be found in this region. A new species of thorny-headed worm (Acanthocephala: Neoechinorhynchidae) is described from P. squamosissimus from Guajará Bay, Belém, Pará, Brazil. In general, the unique characteristics of the hooks on the anterior end of the proboscis and the length-to-width ratio relationship separate this new species from other described species in the genus Neoechinorhynchus. Although the species in this genus are mostly found in North America, the dearth of species known from the neotropics may be due to the lack of studies in this region.

Paragonimus heterotremus, which is an important pathogen for human paragonimiasis in Asia, is recognized as having the smallest metacercariae (maximum diameter < 300 μm) of any previously reported Paragonimus species. Recently, P. pseudoheterotremus has been described from Thailand as a new species having metacercariae (about 200 μm) slightly smaller than those of Thai P. heterotremus. In fact, the small size of P. pseudoheterotremus metacercariae is compatible with those of P. heterotremus from India and China. In this study in Vietnam, we found variably sized small metacercariae which are expected to consist of both P. heterotremus and P. pseudoheterotremus. Contrary to expectation, the adult flukes obtained by separate infection of experimental cats with different sized metacercariae were all identified as P. heterotremus, using both morphological and molecular characteristics. The molecular analyses of an extensive collection of P. heterotremus/P. pseudoheterotremus isolates from Asian countries also indicated that genetic distances between different populations of P. heterotremus are even larger than that between P. pseudoheterotremus and P. heterotremus. The haplotype network showed that all P. heterotremus and P. pseudoheterotremus isolates formed a P. heterotremus complex consisting of three groups with strong geographical origins. In addition, the Indian P. heterotremus group is the root of the other P. heterotremus and P. pseudoheterotremus populations. Based on the observed metacercarial polymorphisms and genetic variation in P. heterotremus,P. pseudoheterotremus should be considered a geographically isolated population of the P. heterotremus complex.

This review presents the historical and current situation of echinococcoses in Mongolia. Since the collapse of the Soviet Union in 1991, Mongolia's health surveillance infrastructure has been very poor, especially as it pertains to chronic diseases, including neglected zoonotic diseases (NZDs). Although there is anecdotal evidence of people dying from hepatic disease due to infection with the larval stage of Echinococcus spp., there are very few published reports. All confirmed cases of echinococcoses in Mongolia are from hospitals located in the capital city of Ulaanbaatar. Cases of cystic echinococcosis (CE), caused by either Echinococcus granulosus sensu stricto or Echinococcus canadensis are believed to be relatively common throughout Mongolia. In contrast, cases of alveolar echinococcosis (AE), caused by Echinococcus multilocularis, are believed to be rare. Recent wild-animal surveys have revealed that wolves (Canis lupus) are the major definitive hosts of E. canadensis, whereas both wolves and red foxes (Vulpes vulpes) are the primary definitive hosts of E. multilocularis. Although wild-animal surveys have begun to elucidate the transmission of Echinococcus spp. in Mongolia, there have yet to be large-scale studies conducted in domestic dogs and livestock. Therefore, further epidemiological studies, in addition to education-based control campaigns, are needed to help combat this NZD.

Sparganosis is a serious food-borne parasitic zoonosis caused by infection with Spirometra spargana. The prevalence of sparganum infection in wild frogs (Rana nigromaculata, R. limmochari, R. temporaria and Bufo gargarizans) was investigated in Henan Province of central China during 2008–2012. Of 3482 caught wild frogs, 565 (16.23%) were found to be infected with plerocercoids (spargana) of the genus Spirometra. Spargana were found in 14.85% (320/2155) of R. nigromaculata, 20.82% (233/1119) of R. limmochari and 10.91% (12/110) of R. temporaria frogs. However, no sparganum was found in B. gargarizans. To investigate the phylogenetic position of collected spargana, three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunits 1 and 3 (cox1 and cox3), and NADH dehydrogenase subunit 4 (nad4), were amplified, sequenced and analysed. Sequences of cox1, cox3 and pnad4 were 417, 390 and 578 bp in length, respectively. The base composition of cox1, cox3 and pnad4 were generally AT rich with a mean of 63.5%, 68.3% and 67% AT, respectively. Phylogenetic analysis showed that all the sparganum isolates in Henan Province represented Spirometra erinaceieuropaei and were a well-supported clade. These findings demonstrated clearly the usefulness of the three mtDNA sequences for molecular identification and population genetics studies of S. erinaceieuropaei spargana of human and animal health significance.

Neurocysticercosis (NCC) caused by Taenia solium cysts is a frequent but neglected parasitic disease of the central nervous system (CNS) worldwide. The aim of this study was to describe anatomical locations of cysts in the CNS and the corresponding inflammation. A total of 17 naturally infected pigs were used to evaluate the distribution of cysts and, of these, seven were used to evaluate the corresponding inflammation further, through histopathology. Clinical signs in the pigs included dullness, sluggishness, somnolence, apathy and loss of consciousness. Cysts were distributed in all cerebral lobes, i.e. 39.7% in the frontal lobe, 20.3% in the parietal lobe, 20.0% in the occipital lobe and 19.7% in the temporal lobe, and only 0.4% in the cerebellum. No cysts were found in the spinal cord. Cysts were localized as follows: 47.9% in the dorsal subarachnoid, 46.9% in the parenchyma, 4.4% in the subarachnoid base and 0.9% in the ventricles. The results of the histopathology revealed lesions in an early inflammatory stage, i.e. stage I, in all anatomical locations except for two, which showed more of an inflammatory reaction, stage III, in one pig. It was concluded that clinical signs in pigs were neither pathognomonic nor consistent. These signs, therefore, cannot be used as a reliable indicator of porcine NCC. Furthermore, T. solium cysts were found to be in abundance in all cerebral lobes, and only a few were found in the cerebellum. Regarding the inflammatory response, no significant differences were found in the location and total number of cysts. Thus, further studies are needed to explain the determinants of cyst distribution in the CNS and assess in detail clinical signs associated with porcine NCC.

To better evaluate the usefulness of female Haemonchus specimens for specific identification, we undertook a detailed analysis of the morphology of a collection of worms obtained from cattle and sheep in shared pastures. Based on the results, we also more precisely evaluated the host-specificity of Haemonchus contortus, H. placei and H. similis occurring sympatrically in a farm located in the western region of São Paulo State, Brazil. A synlophe analysis was employed to identify the three species of Haemonchus. In cattle, the predominant species was H. similis (90.9%), followed by H. placei (9.1%). With the exception of one H. placei specimen, only H. contortus was found in sheep. The longest body length was found for H. placei specimens, followed by H. contortus and then H. similis. It was possible to distinguish H. similis females from H. contortus and H. placei on the basis of vulval structure. The synlophe analysis proved to be very useful for identification of H. contortus, H. placei and H. similis in epidemiological studies involving different species of ruminants in the same pastures. The finding that H. placei and H. similis were adapted to cattle and that H. contortus was adapted to sheep also confirmed the high host-specificity of the three nematodes species.

Canids, particularly dogs, constitute the major source of cystic echinococcosis (CE) infection to humans, with the majority of cases being caused by Echinococcus granulosus (G1 genotype). Canine echinococcosis is an asymptomatic disease caused by adult tapeworms of E. granulosus sensu lato (s.l.). Information on the population structure and genetic variation of adult E. granulosus is limited. Using sequenced data of the mitochondrial cytochrome c oxidase subunit 1 (cox1) we examined the genetic diversity and population structure of adult tapeworms of E. granulosus (G1 genotype) from canid definitive hosts originating from various geographical regions and compared it to that reported for the larval metacestode stage from sheep and human hosts. Echinococcus granulosus (s.s) was identified from adult tapeworm isolates from Kenya, Libya, Tunisia, Australia, China, Kazakhstan, United Kingdom and Peru, including the first known molecular confirmation from Gaza and the Falkland Islands. Haplotype analysis showed a star-shaped network with a centrally positioned common haplotype previously described for the metacestode stage from sheep and humans, and the neutrality indices indicated population expansion. Low Fst values suggested that populations of adult E. granulosus were not genetically differentiated. Haplotype and nucleotide diversities for E. granulosus isolates from sheep and human origin were twice as high as those reported from canid hosts. This may be related to self-fertilization of E. granulosus and/or to the longevity of the parasite in the respective intermediate and definitive hosts. Improved nuclear single loci are required to investigate the discrepancies in genetic variation seen in this study.

New data relating to the fauna of Haploporidae (Digenea) in mugilid fish from Primorsky Region of the south of the Russian Far East are presented. In the most recent revision of the family Haploporidae Nicoll, 1914, Parasaccocoelium Zhukov, 1971 was synonymized with Pseudohapladena Yamaguti, 1952 (Overstreet & Curran, 2005). Here, we restore the genus Parasaccocoelium. The species described by Zhukov can be distinguished from the species Pseudohapladena sensu Yamaguti, 1952 using a combination of the following features: (1) diffuse or absent eye-spot pigment; (2) subterminal or terminal oral sucker; (3) present or absent genital atrium; and (4) vitellarium with two large and compact vitelline masses, or vitelline follicles tubular and divided into two groups. For the species of Pseudohapladena, i.e. P. scatophagi, Yamaguti noted that the vitelline follicles occupy almost the entire post-testicular region, but in the present specimens, the vitelline follicles do not occupy the post-testicular region. Based on the diagnosis of Pseudohapladena by Overstreet & Curran (2005), the genus Parasaccocoelium differs in the following features: (1) a single testis that is longitudinal, longitudinally oval, spherical, transversally oval, irregular, bilobed or V-shaped, or a testis that is subspherical to irregular to elongate; (2) the vitellarium in the lateral fields formed from large compact follicles that have an irregular form and extend between the anterior margin of the ventral sucker or pharynx and posterior margin of the testis, and the vitelline follicles do not occupy post-testicular regions; or the vitellarium consists of elongate groups of follicles that usually extend at least one-half of a body length and extend to the posterior end of the body in some and to the posterior margin of the testis in others; (3) the eggs are operculate, unembryonated and developed when laid, or the eggs in some cases are only operculate; and (4) a developed miracidium with or without an eye spot. New data have been added to the description of Parasaccocoelium mugili Zhukov, 1971 based on new material, and two new species of this genus, Pa. haematocheilum n. sp. from Liza haematocheila and Pa. polyovum n. sp. from L. haematocheila and Mugil cephalus, are described using morphological and molecular approaches. We support the taxonomic status of the genus Parasaccocoelium and its inclusion within the sub-family Waretrematinae, and we consider Pseudohapladena lizae Liu et Yang, 2002 to be a junior synonym of Parasaccocoelium mugili Zhukov, 1971.

The aim of the present study was to calculate the prevalence and intensity of intestinal helminths in the house mouse (Mus musculus) and the black rat (Rattus rattus) trapped in rural households of Yucatan, Mexico. Sampling was conducted during the rainy season from October to December 2011 and the dry season from January to March 2012. A total of 154 M. musculus and 46 R. rattus were examined, with 84.2% of M. musculus being infected with helminths compared with a significantly lower prevalence of 52.2% in R. rattus (P< 0.01). Adult M. musculus were more likely to be infected with helminths (89%) than subadults (63%) (P< 0.01). Four helminth species were identified: Taenia taeniaeformis larvae, Nippostrongylus brasiliensis, Syphacia muris and Trichuris muris. Nippostrongylusbrasiliensis was present more frequently in M. musculus than in R. rattus (P< 0.01) and in adult mice compared to subadults (P< 0.01). Trichuris muris was present only in adult mice. This is the first report of N. brasiliensis, S. muris and T. muris in Yucatan, Mexico, as well as the first to report the presence of N. brasiliensis in M. musculus from Mexico. The helminth fauna of commensal rodents present in households appears to constitute a low potential health risk to local inhabitants; however, it would be advisable to conduct further studies to better understand the public health risk posed by these rodent intestinal helminths.

The liver fluke, Opisthorchis viverrini, and the minute intestinal fluke, Haplorchis taichui, are prevalent in many Asian countries. This study analysed the patterns of infections of O. viverrini and H. taichui in Lahanam and Thakhamlien villages (Savannakhet Province, Lao PDR), in two cross-sectional investigations. Out of a total of 207 human participants, post-anthelmintic treatment positivity rates for expelled worms were 170 (82.1%) for H. taichui and 65 (31.4%) for O. viverrini. Both these species co-exist in the study villages. When each parasite was analysed separately, H. taichui infections reached a plateau among people aged >20 years. Opisthorchis viverrini infection rates were highest in the age group 21–30 years, with decreasing infection rates after the age of 30. Our findings indicated that fish-borne trematode infections were more prevalent among adults. Fish, common intermediate hosts, were acquired in the study area for analysis. The examination of 35 species of fish as intermediate hosts found O. viverrini metacercariae in only six species, and these were found mostly during the month of November. Many farmers who live on the rice fields obtain their food from their immediate environment, including these intermediate-host fish, potentially putting them at greater risk of O. viverrini infection. By contrast, H. taichui metacercariae were found in three species of fish obtained from the market, meaning that anyone could consume them and become infected. If people who work in rice fields limit the species of fish they consume, or avoid consuming raw fish during the month of November, they may reduce their risk of O. viverrini infection.