Molecular and serological survey on taeniasis and cysticercosis in Kanchanaburi Province, Thailand.

A community-based field survey on taeniasis and cysticercosis was performed in two villages in Thong Pha Phum District, Kanchanaburi Province, central Thailand, where 3 Taenia species, T. solium, T. saginata and T. asiatica, are sympatrically occurring. Four (0.6%) out of 667 stool samples were egg-positive for Taenia sp. by Kato-Katz technique. Three out of those four persons and other three persons who were Taenia egg-negative but having a recent (<1 year) history of discharging worms in stool were treated with niclosamide. One Taenia egg-positive woman was not treated because of severe ascites. After treatment, three persons expelled long strobilae with scolices and two persons expelled strobilae without scolex. One Taenia egg-positive person did not expel any worms post-treatment. Among 5 persons, four expelled a single worm, whereas one expelled multiple worms, may be 6 worms but not confirmed by detection of scolices. One scolex was armed with hooklets, whereas 2 others did not. Multiplex PCR of 10 expelled proglottids (including 6 estimated worms from one patient) revealed that one sample was T. solium, one T. saginata, and 8 T. asiatica. A total of 159 residents agreed to receive a serological test for cysticercosis. By ELISA using partially purified glycoprotein antigen, 9 cases, 5 and 4 from villages A and B respectively, were found to be sero-positive. The five and an additional sample on the border line from village A were evaluated using confirmative immunoblot using recombinant chimeric antigen. Among the six samples, four including the border line sample were confirmed to be cysticercosis by immunoblotting. One of the 4 persons had neurological symptoms with nodular lesions in the brain by computed tomography. These 4 confirmed or suspected cysticercosis cases were free of T. solium worms, but two of them including confirmed NCC case had a past (>1 year) history of expelling proglottids in the stool.

[Occurrence of nematodes in horses from the Northern region, State of Tocantins, Brazil]. / Ocorrência de nematóides em equídeos na região Norte do Estado do Tocantins, Brasil.

The occurrence of nematodes in horses of Northern Region, State of Tocantins was studied. Fecal samples were collected from 70 horses and 31 donkeys, in a total of 101 animals of different ages, in the municipalities of Nova Olinda, Araguanã, Babaçulândia and Santa Tereza. Fecal examination was carried out for counting of eggs per gram of faeces, identification and quantification of larvae, diagnosis of habronemosis and lungworms. Significant occurrence of large and small strongyles and Dictyocaulus sp. was observed but no Habronema spp. larvae was detected.

Taeniasis in Mongolia, 2002-2006.

Survey on secondary data of taeniasis/cysticercosis was carried out in Mongolia in 2002-2006. A total of 118 taeniid proglottids, a diphyllobothriid segment, and 59 serum samples from 118 taeniasis cases were collected at National Center for Communicable Diseases, Ulaanbaatar, Mongolia. In 2006, 14 serum samples were collected from local people who had histories of epileptic seizures in Selenge Province where pig husbandry was the main business. The 118 proglottids were confirmed to be Taenia saginata by mitochondrial DNA analysis using cytochrome c oxidase subunit 1 and cytochrome b genes. T. saginata taeniasis was widely distributed at least in 10 of 21 provinces. No variation in the nucleotide sequences of the two genes was observed among T. saginata isolates from Mongolia. There was no evidence of Taenia solium taeniasis/cysticercosis or Taenia asiatica taeniasis. A diphyllobothriid segment was confirmed to be Diphyllobothrium latum by mitochondrial DNA analysis.

Sympatric occurrence of Taenia solium, T. saginata, and T. asiatica, Thailand.

We confirmed sympatric occurrence of Taenia solium, T. saginata, and T. asiatica in western Thailand. DNA analysis of morphologically identified T. saginata, in a dual infection with T. solium, indicated it was T. asiatica. To our knowledge, this report is the first of T. asiatica and a dual Taenia infection from Thailand.

Taeniasis/cysticercosis in a Tibetan population in Sichuan Province, China.

The results of a preliminary survey of taeniasis/cysticercosis in Yajiang County, Ganze Tibetan Prefecture in southwest Sichuan Province, China, indicated a very high prevalence of taeniasis (22.5%), with Taenia saginata as the dominant species. There was also a significant occurrence of late-onset epilepsy (8.5% prevalence and 16.4% seropositive for Taenia solium antibodies) attributable in large part to probable neurocysticercosis caused by T. solium. The poor sanitation and hygiene in this Tibetan community likely contributed to a high risk of human cysticercosis despite a low level of T. solium taeniasis (actually no T. solium carriers were detected amongst the 21 proven Taenia carriers). In addition, three taeniasis cases were confirmed by DNA genotyping as Taenia asiatica, which is the first report of this tapeworm in Tibetans, the first report for Sichuan Province and only the third report for mainland China.

Usefulness of severe combined immunodeficiency (scid) and inbred mice for studies of cysticercosis and echinococcosis.

The topics in this review are the usefulness of immunodeficient and inbred mice for studies of developmental biology, drug efficacy and host specificity in cysticercosis and echinococcosis. In non-obese diabetic severe combined immunodeficiency (NOD/Shi-scid) mice of both sexes, in vitro hatched oncospheres of all three human taeniid species (Taenia solium, Taenia saginata and Taenia asiatica) developed into cysticerci comparable to or bigger than those developed in their known intermediate host animals, whereas only females were susceptible to these infections in other scid mice of BALB/c, C57BL or C.B-17 inbred strains. Detailed morphological observation from post-oncospheral to cysticercus developmental stages is expected to be easy when we use NOD/Shi-scid mice experimentally inoculated with in vitro hatched oncospheres. Metacestocidal effect of oxfendazole was evaluated in NOD/Shi-scid mice experimentally inoculated with oncospheres of T. solium. In Echinococcus multilocularis infection, larval tissue proliferated without induction of inflammatory host responses in scid mice, thus facilitating isolation of the larval vesicles and protoscoleces for biochemical and molecular biological studies. Trans portal inoculation of metacestode tissues resulted in proliferation of secondary echinococcal foci localized exclusively in the liver without metastasis to other tissues or organs. The advantages of a mouse model for Echinococcus granulosus are also described.

Neurocysticercosis: clinical manifestation, neuroimaging, serology and molecular confirmation of histopathologic specimens.

Diagnosis of neurocysticercosis (NCC) is usually based on neuroimaging and/or immunological analysis of cerebrospinal fluid (CSF) and/or serum samples for detection of specific antibodies against T. solium antigens. Additional confirmative diagnosis may be possible by morphological and molecular confirmation of resected histopathologic specimens. The majority of NCC cases do not always show typical neuroimaging figures with invaginated scolex. So, serology using highly specific antigens of T. solium, either semi-purified native or recombinant antigens, is essential for confirming NCC cases. There is some debate about the usefulness of CSF and serum for immunodiagnosis. When NCC cases with a solitary cyst or with calcified lesions are examined, serology is not always sensitive to differentiating such cases. Malignant brain tumor is most commonly suspected in Japan and is often treated surgically as an urgent task, if the clinicians have no experience of NCC cases. Only histopathological specimens are expected to show direct evidence of T. solium cysticercosis. Morphology is not always sufficient for identification of the Taenia species, even if the majority of cysticerci in the human brain are expected to be T. solium. Crucial confirmation is based on molecular identification. In this review, these four issues are briefly summarized.

Towards the international collaboration for detection, surveillance and control of taeniasis/ cysticercosis and echinococcosis in Asia and the Pacific.

Both cysticercosis and echinococcosis are potentially among the most serious helminth zoonoses threatening human health worldwide. However, due to the lack of reliable tools for confirmation or identification of patients or infected animals, epidemiological data are expected to be underestimated. Conversely, sometimes, such data are over estimated due to the lack of specificity. The most important issue for doing field surveys is that they use evidence based science. In this communication, advanced immunological and molecular tools for detection of individuals infected with either metacestodes or adult tapeworms are briefly overviewed, and the applications of such tools for epidemiological surveys in Indonesia, China and other countries are introduced. As immunological tools are based on antigen-antibody responses, there may exist some cross-reactions. Therefore, immunodiagnostic tools are expected to be useful for primary screening, and should be combined with confirmation of direct parasitological evidence (morphology or DNA), and imaging techniques for cysts. As a risk factor for human cysticercosis is the presence of tapeworm carriers, detection of taeniasis cases and differentiation of the three human Taenia species (Taenia solium, T. saginata and T. asiatica) in Asia and the Pacific requires consideration. Similarly, in northwest China, Echinococcus granulosus and E. multilocularis are coendemic and differentiation of these species is required in humans and definitive hosts. It is stressed that combination of several tools for identification of the parasite and for confirmation of diseases is important for obtaining highly reliable data before consideration of control of these zoonoses. Recent projects coordinated by Asahikawa Medical College have concentrated on immunological and molecular diagnostic techniques transferable to colleagues from endemic regions of Asia and the Pacific, and on organization of two international symposia to establish a platform for further collaboration in the future.

Multiple genotypes of Taenia solium--ramifications for diagnosis, treatment and control.

Mitochondrial DNA sequences of Taenia solium have fully been analyzed. Analysis of the full length of cytochrome c oxidase subunit 1 (1620 bp) and cytochrome b (1068 bp) genes of T. solium, isolated from Asia (China, Thailand, Indonesia and India), from Latin America (Mexico, Ecuador, Bolivia, Peru and Brazil) and from Africa (Tanzania, Mozambique and Cameroon), has revealed that the two phylogenies obtained were similar to each other regardless of the genes examined. The isolates from Asia formed a single cluster, whereas those from Latin America combined with those from Africa to form an additional cluster. It was estimated that these two genotypes emerged approximately 4-8 x 10(5) years ago. These results together with recent study of the ancient of human taeniid cestodes emerged several MYA in Africa, historical data on swine domestication, distribution of pigs and colonization patterns suggest that T. solium was introduced recently into Latin America and Africa from different regions of Europe during the colonial age, which started 500 years ago, and that T. solium of another origin independently spread in Asian countries, perhaps from China. Why did not T. solium of European origin invade or spread into Asia during the colonial age? Analysis of T. solium distribution must include other Taenia species, especially T. saginata and T. asiatica, which can not be differentiated from each other morphologically. BESS T-base analysis for differentiation of all human Taenia species including the two genotypes of T. solium, and T. saginata and T. asiatica has also been characterized. BESS T-base analysis differentiates African isolates from Latin American isolates as well but more samples should be analyzed for obtaining conclusive evidence for the latter. Serological analysis of cyst fluid of T. solium cysticerci obtained in China and Indonesia and from Mozambique and Ecuador indicates geographical differences in their banding patterns. These differences are discussed in the light of possible differences in pathology of T. solium worldwide. As it has been speculated that the ancient T. solium emerged several million years ago in Africa, it is necessary to analyze more isolates from Africa. Such working hypothesis may be evaluated combined with symptomatology and serology when we get additional DNA data from such areas, since there are some varieties of manifestation of neurocysticercosis with or without subcutaneous cysticercosis and of antigens of cyst fluid of T. solium from Asia and from Africa and/or America. Transfer of techniques of molecular identification and sero- and immuno-diagnoses between researchers and technicians from endemic countries using their own materials should be promoted with the aim of better international cooperation for the control of cysticercosis.

Echinococcosis and cysticercosis in Asia: evaluation of the modern technology for epidemiological study.

The recent emergence of zoonotic parasitic diseases of public health importance represents a growing global concern. Among zoonotic helminthic diseases, both echinococcosis and cysticercosis are the most serious diseases threatening human life. Neurocysticercosis (NCC) caused by ingestion of eggs of the pork tapeworm, Taenia solium, is spreading worldwide and not rare even in Muslim or Jewish communities. Alveolar echinococcosis (AE) caused by the proliferation of metacestodes of the fox tapeworm, Echinococcus multilocularis, is the most potentially lethal parasitic infection of the non-tropical northern hemisphere, whereas cystic echinococcosis (CE) caused by the proliferation of metacestodes of the dog tapeworm, E. granulosus, has rather a cosmopolitan distribution. As the life cycles of T. solium, E. multilocularis and E. granulosus are completed through predator-prey interactions, including humans, it is crucial to interrupt the cycle for control of these zoonotic cestodiases. Both NCC and CE are expected to be eradicable, since the principal life cycles of T. solium and E. granulosus are maintained between humans and pigs and between dogs and herbivorous domestic animals, respectively. In contrast, AE is perhaps not eradicable, since the life cycle of E. multilocularis is maintained between wild foxes and rodents. Modern technologies, including imaging, immunology and molecular biology, have been applied for epidemiological surveys. In the present review, we introduce such technologies applied in Japan, China and Indonesia, and point out the problems that need to be solved for control of these three zoonotic cestodiases.

Neurocysticercosis and epilepsy in Cameroon.

During January 2002 the frequency of Taenia solium cysticercosis was studied in a series of 504 epileptic patients from 3 rural localities in the West and North-West provinces of Cameroon using an enzyme-linked immunosorbent assay for both circulating antigen (Ag-ELISA) and antibody (Ag-ELISA) detection. Taenia solium antigens were detected in the sera of 1.2% of the patients whereas specific antibodies against the parasite were present in 44.6% of the patients. Significantly less seropositive results in Ab-ELISA were recorded in Batibo than in Bandjoun and Bamendjou, whereas a borderline significant difference was recorded with increasing age. Furthermore, 56.2% of patients with late-onset epilepsy showed antibodies against cysticercosis. Taenia solium cysticercosis appears to be an important cause of epilepsy in Cameroon.

Evaluation of an enzyme-linked immunosorbent assay (ELISA) with affinity-purified Em18 and an ELISA with recombinant Em18 for differential diagnosis of alveolar echinococcosis: results of a blind test.

Alveolar echinococcosis (AE) is the most potentially lethal parasitic zoonosis of the nontropical areas in the northern hemisphere, where cystic echinococcosis (CE) is also endemic. Both AE and CE are highly endemic in China, and both serologic detection of echinococcosis, either AE or CE, and differentiation of AE from CE are crucial problems. Evaluation of Western blot analysis (WB) and enzyme-linked immunosorbent assay (ELISA) for the Em18 antigen, using affinity-purified and recombinant Em18, was carried out "blindly" using 60 human sera from patients diagnosed in France. The results were compared with those obtained using a commercially available Echinococcus WB immunoglobulin G (IgG) kit developed in France. The Em18 WB and Echinococcus WB IgG showed very similar results for detection of AE. Both affinity-purified Em18 or a recombinant Em18 WB and Echinococcus WB IgG seem useful for identification of AE, and the latter seems appropriate for both AE and CE, whereas affinity-purified Em18 ELISA and the newly developed recombinant Em18 ELISA appear to be suitable for detection of AE, especially for epidemiological surveys.