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PubMed2006

The detection of anthelmintic resistance in nematodes of veterinary importance.

Before revised World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines on the detection of anthelmintic resistance can be produced, validation of modified and new methods is required in laboratories in different parts of the world. There is a great need for improved methods of detection of anthelmintic resistance particularly for the detection of macrocyclic lactone resistance and for the detection of resistant nematodes in cattle. Therefore, revised and new methods are provided here for the detection of anthelmintic resistance in nematodes of ruminants, horses and pigs as a basis for discussion and with the purpose that they are evaluated internationally to establish whether they could in the future be recommended by the WAAVP. The interpretation of the faecal egg count reduction test has been modified and suggestions given on its use with persistent anthelmintics and continuous release devices. An egg hatch test for benzimidazole (BZ) resistance is described. A microagar larval development test for the detection of benzimidazole and levamisole resistance provides third stage larvae for the identification of resistant worms. The sensitivity of these two tests can be increased by using discriminating doses rather than LD(50) values. Details are given of a PCR based test for the analysis of benzimidazole resistance in strongyles of sheep and goats, horses and cattle. Although promising for ruminant trichostrongyles, quantitative determination of gene frequency using real time PCR requires further development before PCR tests will be used in the field. Apart from faecal egg count reduction tests there are currently no satisfactory tests for macrocylic lactone resistance despite the great importance of this subject. Except for treatment and slaughter trials there are no validated tests for fasciolicide resistance or for the detection of resistance in cestodes.

Animals↗
PubMed2006

Helminths of red foxes (Vulpes vulpes) in Denmark.

An epidemiological study of helminths in 1040 red foxes collected from various localities in Denmark during 1997-2002, revealed 21 helminth species at autopsy, including nine nematode species: Capillaria plica (prevalence 80.5%), Capillaria aerophila (74.1%), Crenosoma vulpis (17.4%), Angiostrongylus vasorum (48.6% from Northern Zealand (endemic area)), Toxocara canis (59.4%), Toxascaris leonina (0.6%), Uncinaria stenocephala (68.6%), Ancylostoma caninum (0.6%), and Trichuris vulpis (0.5%); seven cestodes: Mesocestoides sp. (35.6%), a number of Taeniid species (Taenia pisiformis, T. hydatigena, T. taeniaeformis, T. crassiceps, and unidentified Taenia spp.) (22.8%), and Echinococcus multilocularis (0.3%); four trematodes: Alaria alata (15.4%), Cryptocotyle lingua (23.8%), Pseudamphystomum truncatum (3.6% from Northern Zealand), and Echinochasmus perfoliatus (2.4% from Northern Zealand); one acanthocephalan: Polymorphus sp. (1.2%). Significant difference in prevalence was found for T. canis and A. vasorum according to host sex, and for T. canis, U. stenocephala, Mesocestoides sp., Taenia spp., A. alata, A. vasorum, and Capillaria spp. according to age groups (adult, young or cub). Prevalence and average worm intensity for each helminth species varied considerably according to geographical locality, season, and year. Aggregated distribution was found for several helminth species. The two species E. multilocularis and E. perfoliatus are first records for Denmark.

Age Factors↗
PubMed2006

Survey of helminths in adult sheep in Heilongjiang Province, People's Republic of China.

The prevalence of helminths in adult sheep was investigated in Heilongjiang Province, People's Republic of China between January 1999 and September 2003. A total of 326 adult sheep representing local breeds (Xingjiang Fine Wool Sheep, Dongbei Fine Wool Sheep) as well as introduced breeds (Merino and Charollais) from representative geographical locations in Heilongjiang Province were slaughtered and examined for the presence of helminths. The worms were examined, counted and identified to species according to existing keys and descriptions. A total of 26 helminth species were found representing 2 phyla, 3 classes, 13 families and 20 genera. All sheep were infected by more than one helminth species. Oesophagostomum columbianum, Haemonchus contortus and Trichostrongylus colubriformis were the most common nematode species, and Paramphistomum cervi, Orientobilharzia turkestanica and Fasciola hepatica were the most common trematode species, whereas the infection of adult sheep with cestodes was uncommon. The results of the present investigation provide relevant "base-line" data for Heilongjiang Province, China, for assessing the effectiveness of future control strategies against helminth infections in sheep.

Animals↗
PubMed2006

Control of the taeniosis/cysticercosis complex: future developments.

Cysticercosis is due to the establishment of the larval stage of the zoonotic cestode parasite Taenia solium. The infection causes substantial human morbidity and mortality, particularly in several Latin American countries and parts of Africa and Asia, as well as economic losses in pig husban dry due to condemnation of infected pork meat. The life cycle of T. solium includes human beings as definitive hosts and pigs as intermediate hosts. Cysticercosis is acquired by the ingestion of eggs released by human tapeworm carriers, who become infected after ingesting pork meat contaminated with cysticerci. Taenia solium transmission has been associated with poverty, lack of sanitary services and practices of rearing backyard pigs with free access to the areas that villagers use as toilets, as well as cultural behaviour. Nonetheless, due to the recent increase of migration and tourism, industrial countries are also reporting cases of human cysticercosis. There are many epidemiological studies that have been conducted mainly in Latin American countries that have evaluated intervention measures for control of cysticercosis including the development and testing of vaccines. Furthermore, the involvement of international agencies and institutions, such as the World Health Organization, the Food and Agriculture Organization and the International Livestock Research Institute, as well as the commitment of policymakers, scientists and field workers, are key means for the sustainable control and, hopefully, eradication of T. solium infections.

Animal Husbandry↗
PubMed2006

Dog echinococcosis in northern Spain: comparison of coproantigen and serum antibody assays with coprological exam.

A large sheep-dog population from the province of Alava (northern Spain) has been investigated in order to determine the prevalence of the cestode parasite Echinococcus granulosus. Worms were detected in 14.0% of 721 dog faecal supernatants by coproantigen ELISA, and in 9.1% of 754 dog serum samples by serum antibody ELISA. A weak but statistically significant correlation (Spearman's rho=0.103, 95% CI: 0.023-0.178) between the two immunoassay results was found. In addition, eggs of the family Taeniidae were detected in 10.3% of 726 faecal samples examined by coproparasitological (flotation and sedimentation) tests. The overall E. granulosus infection rate, based on a Bayesian latent class model that accounts for the imperfect sensitivities and specificities of all diagnostic tests used, was estimated to be 8.0% (95% credible interval: 5.4-11.4%), corroborating that sheep-dog is the dog class most vulnerable to acquiring the infection. Dog sex did not influence the prevalence of E. granulosus, independently of the diagnostic test used or the dog region of origin. No significant linear correlation was found between the coproantigen ELISA OD values and the dog age (Spearman's rho=-0.049, 95% CI: -0.234 to 0.135), suggesting that there were no differences in prevalence of E. granulosus between old and young dogs. The obtained results highlight the importance of initiating a control program based on regular treatment of the sheep-dogs with praziquantel in the province of Alava.

Age Factors↗
PubMed2006

Species diversity and acquisition of gastrointestinal parasites in calves aged 0-13 months in periurban livestock production in Mali.

Post-mortem inspections on 51 calves were conducted to investigate acquisition and spectrum of gastrointestinal parasites in young calves in periurban livestock production in Mali. Generalized linear models with herd as a repeated effect were used to test the influence of management, age and season of birth on parasite numbers and diversity. Parasite number and burden increased with age. In the age class 4-13 months animals carried already up to eight different gastrointestinal parasite species. Parasite spectrum found included 11 nematodes, 3 cestodes and 1 protozoan parasite. The most frequent parasite species found were Haemonchus placei (age class 0-1 month: 7%, 1-4 months: 38%, 4-13 months: 69%), Cooperia pectinata (0%, 33% and 44%) and C. punctata (0%, 33% and 38%). Calves born during the rainy season had higher parasite burden and species diversity than calves born during the dry season. Calves kept under modernised management excreted more Strongyloides papillosus eggs than calves kept under traditional management. Calves acquired most of the parasites occurring in adult bovines in West Africa within their first year of life.

Age Factors↗
PubMed2006

Ovine echinococcosis I. Immunological diagnosis by enzyme immunoassay.

Immunodiagnosis in sheep presents problems of sensitivity and specificity, limiting its applicability in surveillance systems. The objective of this study was to develop a sensitive, specific and accessible technique for diagnosing cystic echinococcosis in naturally infected sheep and to evaluate the validity of necropsy as a reference test. A total of 247 sheep were studied at slaughterhouses, confirming the parasitological diagnosis with histology. Serum was processed with enzyme immunoassay (EIA) using three antigen preparations: total hydatid liquid (LHT), purified fraction of LHT (S2B) and purified lipoprotein (B). Western Blot (WB) was used as a control. EIA proved effective for differentiating Echinococcus granulosus from larval stage of Taenia hydatigena and intestinal cestodes in all three antigen preparations. Serums from macroscopically negative sheep were reactive to EIA and positive with WB. In the whole flock, sensitivity was 89.2% for LHT, 80.0% for S2B and 86.4% for B. Sensitivity in lambs was 78.6% for LHT, 75.0% for S2B and 64.3% for B. Macroscopic diagnosis at the time of slaughter was found to have limitations as a reference test for immunodiagnosis of cystic equinococcosis in sheep, so it was necessary to include histology and WB as reference tests. LHT was the antigen preparation of greatest value and EIA proved to be a sensitive and specific technique, adequate for surveillance systems and for evaluating control programmes.

Abattoirs↗
PubMed2006

The complete mitochondrial genomes of Schistosoma haematobium and Schistosoma spindale and the evolutionary history of mitochondrial genome changes among parasitic flatworms.

Complete mitochondrial genome sequences for the schistosomes Schistosoma haematobium and Schistosoma. spindale have been characterized. S. haematobium is the causative agent of urinary schistosomiasis in humans and S. spindale uses ruminants as its definitive host; both are transmitted by freshwater snail intermediate hosts. Results confirm a major gene order rearrangement among schistosomes in all traditional Schistosoma species groups other than Schistosoma japonicum; i.e., species groups S. mansoni, S. haematobium, and S. indicum. These data lend support to the 'out of Asia' (East and Southeast Asia) hypothesis for Schistosoma. The gene order change involves translocation of atp6-nad2-trnA and a rearrangement of nad3-nad1 relative to other parasitic flatworm mt genomes so far sequenced. Gene order and tRNA secondary structure changes (loss and acquisition of the DHU and/or TPsiC arms of trnC, trnF, and trnR) between mitochondrial genomes of these and other (digenean and cestode) flatworms were inferred by character mapping onto a phylogeny estimated from nuclear small subunit rRNA gene sequences of these same species, in order to find additional rare genomic changes suitable as synapomorphies. Denser and wider taxon sampling of mt genomes across the Platyhelminthes will validate these putative characters.

Animals↗
PubMed2003

Predictive values and quality control of techniques for the diagnosis of Echinococcus multilocularis in definitive hosts.

By the end of the 1980s the only reliable technique for diagnosing the intestinal Echinococcus multilocularis infection of definitive hosts was parasite detection at necropsy. Currently, several techniques for the post mortem and in vivo diagnosis are available, including classical and modern methods. The sedimentation and counting technique (SCT) is used for the exact determination of the worm burden in the intestine after necropsy. The SCT has high sensitivity and specificity values (both close to 100%) and can be regarded as 'gold standard'. The principle of the intestinal scraping technique (IST) is the stereomicroscopic examination of at least 15 intestinal thick smears. This technique has a sensitivity of 78% and a specificity approximating 100%. In recent years, the IST has been successfully used in large post mortem surveys of foxes for E. multilocularis. The newer techniques for detecting coproantigens by ELISA (CA-ELISA) exhibit rather high sensitivities between 84 and 95%, combined with very high specificities of >96%, the latter regarding non-Echinococcus cestodes and other parasites. However, cross-reactivity may occur with E. granulosus. Copro-DNA detection by PCR is also highly sensitive (89-94%) and specific (100%). With the SCT, IST and Copro-PCR one person can only examine about 10-20 animals per day, whereas the CA-ELISA allows the examination of 200 samples. Therefore, the latter test is suited for mass-screening of definitive host populations. Both the CA-ELISA and the Copro-PCR allow the examination of materials from dead and living animals, including faecal samples collected in the field. Quite often diagnostic techniques have been used without adequate quality control and proper definition of their performance characteristics, including diagnostic sensitivity, specificity, predictive values and some other parameters. Examples are presented with the aim to demonstrate the need and the value of calculating the predictive values for assays used to diagnose the E. multilocularis infection in individual animals and in definitive host populations.

Animals↗
PubMed2003

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.

Animals↗
PubMed2003

Immunodiagnostic tools for taeniasis.

Most diagnostic work conducted on the Taenia species zoonoses has been carried out on the larval stage of Taenia solium in man, reflecting the relative severity of the pathology caused by this stage of that organism. This review will, however, concentrate on the immunodiagnosis of the adult intestinal stages of these parasites in humans. Diagnosis of T. solium will be examined in most detail because of the relative importance of this parasite but relevant work from other cestodes of man and animals will also be discussed. In addition both classical and molecular approaches to diagnosis will be briefly covered. There have been a number of advances in immunodiagnosis of taeniasis over recent years that have improved both diagnostic sensitivity and specificity. Techniques for the detection of Taenia specific coproantigens in human taeniasis infections have been shown to more than double the numbers of T. solium cases accurately diagnosed in epidemiological studies. More recently, work on the serological diagnosis of T. solium have led to the development of a sensitive and specific enzyme linked immuno-transfer blot for the detection of species and stage specific circulating antibodies to adult worm excretory-secretory antigens. Work is ongoing to further improve these assays.

Animals↗
PubMed1997

Polycystic echinococcosis in tropical America: Echinococcus vogeli and E. oligarthrus.

The paper reports available data on 72 known human cases of polycystic echinococcosis (PE) caused by Echinococcus vogeli (Ev) or E. oligarthrus (Eo). The patients' ages ranged from 6 to 78 years (median 44); one third were younger than 22 years and had severe lesions, demonstrating the aggressiveness of PE. There were no differences in occurrence between sexes. In 80% of the cases the lesions were in the liver alone or in combination with other organs; the rest were located in the lung or other single sites. The diagnosis of PE was based on the demonstration of polycystic larval cestode lesions by radiological imaging (X-ray, US, CT scan) in patients born in tropical sylvatic areas of America where wild carnivores (canids, felids) and rodents (pacas and other species) were present. Serologic tests often, but not always, supported the diagnosis. Species identification of Ev or Eo were based on morphological characteristics of rostellar hooks from protoscoleces. The most common clinical presentation was abdominal; hard, round masses in or connected with the liver, hepatomegaly, increased abdominal size, pain, marked weight-loss and fever. Signs of portal hypertension were also present in 25% of cases, all of whom died of the disease or surgical complications following biliary drainage or partial hepatectomy. Ten percent were in asymptomatic persons. Albendazole treatment resulted in clinical improvement and disappearance or reduction of the size of lesions in some patients observed up to 24 months. The author believes that albendazole treatment should be tried before any other treatment is implemented. Human PE was reported from 11 countries, from Nicaragua to Argentina, 31 case were due to E. vogeli, three to E. oligarthrus (two orbital from Venezuela and Surinam and one cardiac from Brazil) and 38 were due to Echinococcus sp. (no rostellar hooks were available). PE reported from Nicaragua, Costa Rica, Chile, Argentina and Uruguay, countries outside the geographical range of the bush dog (only natural host of Ev) may have been due to Eo rather than to Ev. Several felids are known to serve as final host of Eo, and they are present in those areas. Epidemiological field studies of autochthonous human populations in tropical areas may contribute to early detection and make possible greater success in treatment of polycystic echinococcosis.

Adult↗
PubMed2001

A comparison of the internal transcribed spacer of the ribosomal DNA for Eubothrium crassum and Eubothrium salvelini (Cestoda: Pseudophyllidea), parasites of salmonid fish.

The internal transcribed spacers (ITS-1 and ITS-2) and 5.8S subunit of the ribosomal RNA gene of Eubothrium crassum and Eubothrium salvelini, cestode parasites of salmonid fish, were sequenced and compared. The lengths and GC contents of the regions sequenced were 1119 bp and 50.1% for E. crassum, and 1117 bp and 50.5% for E. salvelini, respectively. Sequence alignment and comparison of both taxa showed 97.9% similarity. Within the ITS-1 and ITS-2 of both species, the restriction enzymes MspI and Bsh1236I possessed different cleavage sites and could be employed for rapid differentiation of both species of Eubothrium by PCR-RFLP.

Amino Acid Sequence↗
PubMed2001

Biochemical characterisation of a hydrophobic ligand binding protein from the tapeworm Hymenolepis diminuta.

The cestode Hymenolepis diminuta contains an abundant, cytoplasmic, hydrophobic ligand, binding protein (H-HLBP). Studies with polarity sensitive probes suggest a single hydrophobic binding site, the results also indicate that the single tryptophan in the molecule (Trp41) is involved in ligand binding. Of the possible physiological ligands tested, only haematin and retinoids (retinol and retinoic acid) show appreciable binding in addition to fatty acids. H-HLBP also binds a range of anthelmintics, again with K(D) values in the nM range. The interaction of anthelmintics with hydrophobic binding proteins may be important in determining drug specificity and site of action and could have a role in the development of drug resistance.

Animals↗
PubMed2001

Characterisation of a beta-tubulin gene from the liver fluke, Fasciola hepatica.

This study represents the first beta-tubulin sequence from a trematode parasite, namely, the liver fluke, Fasciola hepatica. PCR of genomic DNA showed that at least one beta-tubulin gene from F. hepatica contains no introns. A number of amino acids in the primary sequence of fluke tubulin are different from those described previously in various nematode species and the cestode, Echinococcus multilocularis. beta-Tubulin is an important target for benzimidazole anthelmintics, although (with the exception of triclabendazole) they show limited activity against F. hepatica. The amino acid differences in fluke beta-tubulin are discussed in relation to the selective toxicity of benzimidazoles against helminths and the mechanism of drug resistance.

Amino Acid Sequence↗
PubMed2001

Immunisation with Salmonella typhimurium-delivered glyceraldehyde-3-phosphate dehydrogenase protects mice against challenge infection with Echinococcus multilocularis eggs.

Recombinant glyceraldehyde-3-phosphate dehydrogenase of the cestode parasite Echinococcus multilocularis was expressed in Escherichia coli and in Salmonella typhimurium. The potential of different forms of the recombinant antigen to protect BALB/c mice against oral challenge infections with E. multilocularis eggs was evaluated. Oral or intraperitoneal immunisation with live attenuated S. typhimurium as a carrier for recombinant glyceraldehyde-3-phosphate dehydrogenase of the E. multilocularis resulted in significant protection, reducing the number of developing metacestodes up to 79.8%. The sera of protected animals did not contain detectable amounts of antibody against glyceraldehyde-3-phosphate dehydrogenase of E. multilocularis. By contrast, although anti-glyceraldehyde-3-phosphate dehydrogenase of E. multilocularis antibodies were detectable in the sera, immunisation with E. coli-expressed recombinant glutathione-S-transferase-fusion protein or with glyceraldehyde-3-phosphate dehydrogenase of E. multilocularis fused to a 6HIS-tag failed to protect the animals against oral challenge infections. These data emphasise that antigen delivery systems play a critical role in vaccination and the induction of protective immunity against helminth parasites.

Amino Acid Sequence↗
PubMed2002

Diversity in the Monogenea and Digenea: does lifestyle matter?

If the cestodes are excluded, then the parasitic platyhelminths of fishes divide neatly into the external and monoxenous Monogenea and the internal and heteroxenous Digenea. Both groups have apparently had long associations of coevolution, host switching and adaptation with fishes and have become highly successful in their respective habitats. Current estimates of species richness for the two groups suggest that they may be remarkably similar. Here we consider the nature of the diversity of the Monogenea and Digenea of fishes in terms of richness of species and higher taxa to determine what processes may be responsible for observed differences. The Monogenea includes at least two super-genera (Dactylogyrus and Gyrodactylus) each of which has hundreds of species; no comparable genera are found in the Digenea. Possible reasons for this difference include the higher host specificity of monogeneans and their shorter generation time. If allowance is made for the vagaries of taxonomic 'lumping' and 'splitting', then there are probably comparable numbers of families of monogeneans and digeneans in fishes. However, the nature of the families differ profoundly. Richness in higher taxa (families) in the Digenea is explicable in terms of processes that appear to have been unimportant in the Monogenea. Readily identifiable sources of diversity in the Digenea are: recolonisation of fishes by taxa that arose in association with tetrapods; adoption of new sites within hosts; adoption of new diets and feeding mechanisms; adaptations relating to the exploitation of ecologically similar groups of fishes and second intermediate hosts; and adaptations relating to the exploitation of phylogenetic lineages of molluscs. In contrast, most higher- level monogenean diversity (other than that associated with the subclasses) relates principally to morphological specialisation for attachment by the haptor.

Animals↗