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PubMed2003

Quantitative chromatographic determination of several benzimidazole anthelmintic molecules in parasite material.

A specific, precise and accurate high-performance liquid chromatographic (HPLC) analytical method has been developed for the quantitative determination of different benzimidazole (BZD) anthelmintics in parasite material (Moniezia benedeni). Mebendazole (MBZ), oxibendazole (OBZ), flubendazole (FLBZ), albendazole (ABZ) ricobendazole (RBZ), albendazole sulphone (ABZSO(2)), fenbendazole (FBZ), oxfendazole (OFZ) and fenbendazole sulphone (FBZSO(2)) were measured simultaneously in M. benedeni, a sheep and cattle cestode parasite used as a model of the biological matrix. The recovery, linearity, precision, accuracy, limits of detection and quantification of the method were determined. Drug extraction from the parasite's tissue homogenate was performed using methanol (liquid phase extraction), and after solvent evaporation, the residual material was cleaned up by solid phase extraction prior to analysis by reversed-phase HPLC. The resolution of all the BZD molecules assayed was obtained on a C(18) reversed-phase (5 microm, 250 mm x 4.6 mm) column using acetonitrile and ammonium acetate as the mobile phase and ultraviolet (UV) detection at 292 nm. Regression analyses for all the BZD compounds assayed were linear at concentrations ranging from 1.61 to 64.21 nmol/100mg protein (triplicate determinations) showing correlation coefficients greater than 0.9922. The developed method is efficient for the simultaneous determination of several benzimidazole anthelmintic molecules in parasite material and useful for the ex vivo and in vivo characterisation of the kinetics of drug uptake/diffusion in target parasites, which seems to be relevant to optimise parasite control both in human and veterinary medicine.

Animals↗
PubMed2004

Jejunal perforation caused by morphologically abnormal Taenia saginata saginata infection.

We report a case of jejunal perforation caused by infection with taeniid tapeworm possessing unusual morphology that has not been previously described. Among five cestodes recovered during operation, one exhibited a bifurcation at the posterior end and the other harbored a lateral branching of immature segments from the main strobila. By sequence analysis of the mitochondrial cytochrome c oxidase I gene and the 28S rRNA gene has confirmed them to be Taenia saginata saginata.

Adult↗
PubMed2006

Structure and putative function of dark- and light-adapted as well as UV-exposed eyes of the food store pest Psyllipsocus ramburi Sélys-longchamps (Insecta: Psocoptera: Psyllipsocidae).

The psocopteran Psyllipsocus ramburi Sélys-Longchamps can render food stuffs unpalatable and may serve as an intermediate host for cestodes. Its two circular compound eyes consist of about 26 facets, capped by strongly convexly curved corneae of 10-18 microm in diameter. Corneal nipples or interommatidial hairs are not developed. Beneath each corneal lens a cluster of four cone cells, enveloped by two primary pigment cells, separates an ommatidial group of eight retinula cells from the inner corneal surface. Membrane specializations of the retinula cells, known as the microvilli, measure 60 nm in diameter, and collectively make up the rhabdom, which is columnar in shape and has a distal diameter of 4 or 5 microm, depending on whether it is day- or night-adapted. Cone cell lengths measure 4.5 microm during the day and 8.5 microm at night and retinula cell screening pigments closely approach the edge of the rhabdom during the day. A 1-h exposure to UV-A (lambda(max)=351 nm) of ca. 1200 lx causes an almost total destruction of the photoreceptive membranes of the rhabdom and bleached all retinula cell screening pigments, but not the pigment grains of the primary pigment cells. Calculations, based on the anatomical data, suggest that the eyes are adapted to function under dim light levels, but cannot produce sharp images since their best possible acceptance angles are 22 degrees and 28 degrees in light- and dark-adapted states, respectively. Destruction of vision, likely affecting biorhythm and reproduction, by exposing the insects to UV-A may offer an alternative to the use of chemicals in controlling these insects.

Adaptation, Physiological↗
PubMed2006

Endosymbionts of Ctenocephalides felis felis (Siphonaptera: Pulicidae) obtained from dogs captured in Belo Horizonte, Minas Gerais, Brazil.

Specimens of fleas Ctenocephalides felis felis (1052 female symbol/448 male symbol), obtained from 150 dogs in Centro de Controle de Zoonoses de Belo Horizonte, Minas Gerais, Brazil, were dissected and examined for endosymbionts. Three protozoan, Nolleria pulicis, a gregarine (Actinocephalidae) and Leptomonas sp., together with one cestode, Dipylidium caninum were identified. Infections by N. pulicis and Leptomonas sp. occurred mainly in the warm-rainy period. The prevalence and distribution of these endosymbionts in fleas derived from Brazil and South America, and the their variation according to sex and season, are reported for the first time.

Animals↗
PubMed2006

EmsB, a tandem repeated multi-loci microsatellite, new tool to investigate the genetic diversity of Echinococcus multilocularis.

In order to explore the genetic diversity within Echinococcus multilocularis (E. multilocularis), the cestode responsible for the alveolar echinococcosis (AE) in humans, a microsatellite, composed of (CA) and (GA) repeats and designated EmsB, was isolated and characterized in view of its nature and potential field application. PCR-amplification with specific primers exhibited a high degree of size polymorphism between E. multilocularis and Echinococcus granulosus sheep (G1) and camel (G6) strains. Fluorescent-PCR was subsequently performed on a panel of E. multilocularis isolates to assess intra-species polymorphism level. EmsB provided a multi-peak profile, characterized by tandemly repeated microsatellite sequences in the E. multilocularis genome. This "repetition of repeats" feature provided to EmsB a high discriminatory power in that eight clusters, supported by bootstrap p-values larger than 95%, could be defined among the tested E. multilocularis samples. We were able to differentiate not only the Alaskan from the European samples, but also to detect different European isolate clusters. In total, 25 genotypes were defined within 37 E. multilocularis samples. Despite its complexity, this tandem repeated multi-loci microsatellite possesses the three important features for a molecular marker, i.e. sensitivity, repetitiveness and discriminatory power. It will permit assessing the genetic polymorphism of E. multilocularis and to investigate its spatial distribution in detail.

Animals↗
PubMed2005

Genetic variation in Taenia solium.

Neurocysticercosis is a major zoonotic larval cestode infection that has a worldwide distribution and is of significant public health importance. Knowledge of the genetic structure of Taenia solium can be applied to the epidemiology and transmission of this disease, since genetic variants may differ in infectivity and pathogenicity. Molecular epidemiological approaches can also enable detailed studies of transmission. On a global scale, mitochondrial markers have differentiated between T. solium isolates from Asia and isolates from Africa/Latin America. Intraspecific variation in T. solium has been detected to some extent, using RAPD markers to differentiate between T. solium populations from different regions within Mexico. Markers currently available for T. solium have not been used to analyse genetic variation at the community or local level. The development of highly polymorphic markers such as microsatellites in T. solium can provide the means to examine genetic heterogeneity of tapeworm infection at the household, community and regional level. Preliminary studies suggest it is possible to analyse population genetic variation in communities using a range of polymorphic markers.

Animals↗
PubMed2005

The genome project of Taenia solium.

We have constituted a consortium of key laboratories at the National Autonomous University of Mexico to carry out a genomic project for Taenia solium. This project will provide powerful resources for the study of taeniasis/cysticercosis, and, in conjunction with the Echinococcus granulosus and Echinococcus multilocularis genome project of expressed sequence tags (ESTs), will mark the advent of genomics for cestode parasites. Our project is planned in two consecutive stages. The first stage is being carried out to determine some basic parameters of the T. solium genome. Afterwards, we will evaluate the best strategy for the second stage, a full blown genome project. We have estimated the T. solium genome size by two different approaches: cytofluorometry on isolated cyton nuclei, as well as a probabilistic calculation based on approximately 2000 sequenced genomic clones, approximately 3000 ESTs, resulting in size estimates of 270 and 251 Mb, respectively. In terms of sequencing, our goal for the first stage is to characterize several thousand EST's (from adult worm and cysticerci cDNA libraries) and genomic clones. Results obtained so far from about 16,000 sequenced ESTs from the adult stage, show that only about 40% of the T. solium coding sequences have a previously sequenced homologue. Many of the best hits are found with mammalian genes, especially with humans. However, 1.5% of the hits lack homologues in humans, making these genes immediate candidates for investigation on pharmaco-therapy, diagnostics and vaccination. Most T. solium ESTs are related to gene regulation, and signal transduction. Other important functions are housekeeping, metabolism, cell division, cytoskeleton, proteases, vacuolar transport, hormone response, and extracellular matrix activities. Preliminary results also suggest that the genome of T. solium is not highly repetitive.

Animals↗
PubMed2005

Transmission dynamics of Taenia solium and potential for pig-to-pig transmission.

Taenia solium taeniasis/cysticercosis is one of few potentially eradicable infectious diseases and is the target of control programs in several countries. The larval stage of this zoonotic cestode invades the human brain and is responsible for most cases of adult-onset epilepsy in the world. Our current understanding of the life cycle implicates humans as the only definitive host and tapeworm carrier, and thus the sole source of infective eggs that are responsible for cysticercosis in both human and pigs through oral-faecal transmission. Here we review transmission dynamics of porcine cysticercosis including an alternative pig-to-pig route of transmission, previously not suspected to exist. Second-hand transmission of T. solium eggs could explain the overdispersed pattern of porcine cysticercosis, with few pigs harbouring heavy parasite burdens and many more harbouring small numbers of parasites.

Animals↗
PubMed2006

What is Asian Taenia?

Asian Taenia is a human tapeworm which was first recognized in Taiwan aborigines and subsequently from Asian countries: Korea, Indonesia, Vietnam, and China. It was originally described as T. asiatica Eom and Rim, 1993 based on the morphology in its adult and larval stage. A taxonomic disagreement on whether it is species or subspecies level is mainly due to the morphological similarity of this tapeworm with T. saginata, but a sympatric distribution of these two tapeworms is also known in China. The life cycle is quite distinct from T. saginata in using the pig as intermediate host and parasitizing visceral organs such as liver, lung and omentum. A long unresolved question in Asian countries concerns the inconsistency between worm ratio and the food preferences was clarified with this viscerotropic tapeworm. Molecular biological differentiation is possible with DNA techniques and a complete genome of mtDNA was sequenced recently which may provide a resource for comparative mitochondrial genomics and systematic studies of parasitic cestodes. This is a young parasite discovered most recently with many research questions yet to be clarified.

Animals↗
PubMed2005

Echinococcus shiquicus, a new species from the Qinghai-Tibet plateau region of China: discovery and epidemiological implications.

In Shiqu County of the Qinghai-Tibet plateau, many wild and domestic mammals are involved in the transmission cycles of Echinococcus spp. Echinococcus multilocularis and Echinococcus granulosus genotype G1 (sheep strain) are sympatrically distributed in the plateau. In 1995, we identified a unique strobilate stage of Echinococcus from the Tibetan fox, Vulpes ferrilata, but considered it to be a variant of E. multilocularis. Subsequent molecular genetic studies revealed that a hydatid cyst from the plateau pika, Ochotona curzoniae, had unique mitochondrial DNA sequences which are dissimilar to any published sequences of Echinococcus. The same sequences were subsequently found in adult worms from Tibetan foxes. Morphological, genetic and ecological features of the cestode led us to designate a new species Echinococcus shiquicus. E. shiquicus has been found at other areas surveyed on the plateau; however, no infections in humans caused by E. shiquicus have been yet identified.

Animals↗
PubMed2006

Progress in diagnosis, treatment and elimination of echinococcosis and cysticercosis.

Here I review the current status of geographical occurrence and public health significance of echinococcosis (Echinococcus spp. infections) and cysticercosis (Taenia solium infection) with special emphasis on the remarkable technologic progress achieved in recent decades that has led to greater understanding of the biology and epidemiology of these cestode infections. The greatest remaining challenges are to apply this knowledge and technology to improved medical management and prevention of these infections.

Animals↗
PubMed2005

The impact of natural helminth infections and supplementary protein on growth performance of free-range chickens on smallholder farms in El Sauce, Nicaragua.

Three on-farm studies were conducted in Nicaragua during three consecutive years (1999-2001) to assess the impact of natural helminth infections on growth performance of free-range chickens aged 3-4 months. On all participating farms, half of the chickens were treated regularly with anthelmintics (Trifen avicola - a combined formulation of piperazine, phenothiazine and dichlorophen - or albendazole) to express the growth potential of non-infected birds, whereas the other half served as non-treated controls. In 1999, treated chickens had a 39% higher weight gain compared to the control group 6 weeks after the first treatment on 15 farms. In 2000 and 2001, treated chickens had similar weight gain as the control group 10 weeks after the first treatment on 7 farms and 12 farms, respectively. The main reason for the very-different weight gain figures seems to be the weather conditions. In 1999, the study site experienced a rainy season with precipitation far above average, whereas in 2000 and 2001 the rainy seasons had precipitations far below average. Based on these findings, routine use of anthelmintics in the study area would only be recommended in wet years when production losses due to helminth infections seem to be pronounced. In 2001, the study set-up included an assessment of the effect of protein supplementation (soybean) on growth on six farms. Supplemented chickens (treated and non-treated with anthelmintics) had 17% higher weight gain than non-supplemented. Protein supplementation affected neither worm burdens nor faecal egg counts for any of the studied helminths. The post-mortem examinations showed that Trifen reduced burdens of Ascaridia galli, Heterakis gallinarum, and cestodes (efficacies of 100, 100 and 67%, respectively). Albendazole reduced burdens of H. gallinarum (efficacy of 100%). Efficacies against other helminths were difficult to assess due to low worm burdens. Chickens treated with albendazole had lower Ascaridia and Heterakis faecal egg counts than non-treated chickens.

Animals↗
PubMed2004

Factors contributing to the public health and economic importance of waterborne zoonotic parasites.

This is the first of a series of review articles in a Special Issue publication on waterborne zoonotic parasites. A brief historical overview of the occurrence and importance of waterborne parasites, dating from early civilization is presented. The article considers the diversity of parasites including protozoa, nematodes, cestodes and trematodes and the related zoonotic organism microsporidia. Many of the life cycle stages and their characteristics, which make parasites environmentally resistant and suitable for waterborne transmission are discussed. Surfaces of transmission stages consist of multiple layers of proteins, lipids, chitin or other substances capable of withstanding a variety of physical and chemical treatments. Delivery of waterborne parasites is facilitated by various mass distribution systems to consumers, and by transport and intermediate hosts such as fish and filter-feeding invertebrates which are consumed by humans. The article discusses the trends in global warming and climate change and potential for concurrent rise in waterborne disease outbreaks due to parasites. Impacts of technological modernization and globalization on the transmission of zoonotic waterborne zoonotic parasites are considered, including the effects of large-scale agricultural practices, rapid transportation of goods, and widespread movement of individuals and animals. Finally, transmission features and parasite attributes which contribute to concerns about accidental or orchestrated waterborne disease outbreaks are discussed.

Animals↗
PubMed2004

Field evaluation of an intravital diagnostic test of Echinococcus multilocularis infection in red foxes.

Echinococcus multilocularis parasitizes the small intestine of red foxes (Vulpes vulpes) and other carnivores, and has a wide distribution throughout the northern hemisphere. This cestode is the causative agent of human alveolar echinococcosis, a life-threatening helminth zoonosis. In 2000-2002, 2130 red foxes were examined for its presence in Slovakia, with a total prevalence of 30.7%. The data on occurrence were obtained by the combination of necropsy of small intestines from red foxes and coproantigen detection in faecal samples. The correlation between the number of detected specimens and the value of optical density of copro-ELISA test was found. When worm burdens were low (1-25 specimens) the sensitivity of the method was 31.3+/-8.64%, when worm burdens were >50 specimens, 81.8+/-0.66%, and with high worm burdens (>1000 specimens) the sensitivity reached 100+/-0.34%. E. multilocularis presence was detected using the nested PCR method from the eggs in the faecal samples with a 100% specificity. In epidemiological surveys of this zoonosis, it is of crucial importance to detect animals with a high level of infection, which are responsible for the bulk of environmental contamination. The advantage of copro-ELISA test lies in allowing the intravital diagnostics to be employed within the epidemiological survey of E. multilocularis occurrence in the protected and urban areas.

Animals↗
PubMed2004

Ecological analysis and description of the intestinal helminths present in dogs in Mexico City.

The objective of the present study was to determine the prevalence of parasitoses in stray dogs from Mexico City, as well as to describe the structure of the intestinal helminth communities present in said hosts. One hundred and twenty dog intestines were collected and dissected to evaluate their parasitic load; finding parasites in 102 dogs (85%). The cestodes collected were: Dipylidium caninum in 72 dogs (60%), Taenia hydatigena in three (2.5%), Taenia pisiformis in two (1.6%), Echinococcus granulosus, Mesocestoides vogae and Mesocestoides variabilis in only one animal each (0.83%), the latter two being collected for the first time ever in Mexico City. Nematodes collected were: Ancylostoma caninum in 75 (62.5%) animals, Toxocara canis in 16 (13.3%) and Toxascaris leonina in 5 (4.16%). By age, D. caninum and A. caninum were the most prevalent species in older animals, while T. canis was more prevalent in young animals (P<0.05). By season, T. canis was most common in the dry season (P<0.05). At the parasitic community level, the diversity value was 0.32, while that of equality was 0.1; the dominant species was M. vogae (Berger-Parker index of 0.95). At the infracommunity level, both diversity and equality were low (0.31 and 0.32, respectively) as compared to that observed in wild canines; the dominant species was A. caninum (Berger-Parker index of 0.37). Qualitative similarity was 33.8%, while quantitative similarity was 32.2%. Based upon the low values for mean diversity, equality and species richness, it is possible to deduce that helminth fauna in dogs from Mexico City is weakened, especially if one compares it to that of other wild canines.

Age Factors↗
PubMed2005

Observations on helminth infections of free-living and captive rheas (Rhea americana) in Brazil.

The present work describes helminth infection of eight free-living and 12 captive rheas (Rhea americana) from, respectively, Pantanal of Mato Grosso do Sul State, and Jaboticabal, Sao Paulo State, Brazil. Captive birds were young and had a high mortality rate, while free-living birds were adult and apparently healthy. Infections were evaluated by post-mortem examination of internal organs and recovery of helminths using standard parasitological procedures. Seven species of nematodes (Sicarius uncinipenis, Torquatoides crotophaga, Deletrocephalus dimidiatus, D. cesarpintoi, Paradeletrocephalus minor, Capillaria venteli and Dicheilonema rheae) and two species of cestodes (Houttuynia struthionis and Chapmania tauricolis) were identified. P. minor, which inhabits the large intestine, was the most common helminth in free-living birds (63.9%). In captive rheas, a mean parasitic load of 173 helminths per host was found. The gizzard of these birds was the most parasitized organ and S. uncinipenis was most common (92.5%). Parasitism of free-living and captive birds and associated pathology are discussed.

Animals↗
PubMed2005

Effect of Paecilomyces lilacinus on the viability of oncospheres of Taenia hydatigena.

Paecilomyces lilacinus (Thom) Samson is a saprophytic hyphomycete from the soil with biological activity on helminth eggs. We evaluated the influence in vitro of P. lilacinus on the viability of the oncospheres from Taenia hydatigena, a parasite cestode of dogs and sheep. The eggs were exposed to the fungus strain in sterile distilled water and observed by light microscopy at days 4, 7 and 14 post-inoculation, and the viability was evaluated. The viability found in the exposed P. lilacinus oncospheres was significantly different in all observations. P. lilacinus exercised a negative biological activity on T. hydatigena eggs in vitro.

Animals↗
PubMed2005

Vaccines against veterinary helminths.

The majority of attempts to develop commercial vaccines for veterinary helminths have focussed on identifying protein antigens, which could be formulated as protective vaccines. Notable successes have been achieved for some cestode parasites, where recombinant proteins have been developed into highly effective vaccines. Although effective protection can also be obtained using some nematode proteins in their native forms, it has not yet been possible to formulate commercially successful vaccines for other helminth parasites of veterinary significance. Increasing evidence suggests that parasite glycan moieties may provide an alternative source of vaccine antigens, and increased attention is now being given to this class of compounds. In addition to identifying candidate protective antigen(s), an increased research effort is needed to develop appropriate strategies for the formulation and delivery of helminth vaccines.

Animals↗