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Molecular detection of predation by soil micro-arthropods on nematodes.

The relative importance of the factors driving change in the population dynamics of nematodes in the soil is almost completely unknown. Top-down control by micro-arthropod predators may have a significant impact on nematode population dynamics. We report experiments showing that mites and Collembola were capable of reducing nematode numbers in the laboratory and were feeding on a targeted nematode species in the field. A PCR-based approach was developed for the detection of predation on three species of slug- and insect-pathogenic nematodes: Phasmarhabditis hermaphrodita, Heterorhabditis megidis and Steinernema feltiae. The collembolan Folsomia candida and the mesostigmatid mite Stratiolaelaps miles were employed as model predators to calibrate post-ingestion prey DNA detection times. Fragments of cytochrome oxidase I (COI) mtDNA were sequenced and species-specific primers were designed, amplifying 154-, 154- and 203-bp fragments for each of the nematode species. Detection times for nematode DNA within the guts of Collembola were longer than in mites, with half-lives (50% of samples testing positive) of 08.75 h and 05.03 h, respectively. F. candida significantly reduced numbers of the nematode H. megidis, with rates of predation of approximately 0.4 nematode infective juveniles per collembolan per hour over 10 h. Four taxa of field-caught micro-arthropod that had been exposed to the nematode P. hermaphrodita for a period of 12 h were analysed and significant numbers of three taxa tested positive. This is the first application of PCR techniques for the study of nematophagy and the first time these techniques have been used to measure predation on nematodes in the field.

Animals↗

Relationships between the larval growth inhibition of Caenorhabditis elegans by apigenin derivatives and their structures.

Due to consumer reluctance to take synthetic drugs for nematode infections and the appearance of resistance to anthelminthic drugs, new drugs from natural products must be developed. Caenorhabditis elegans is one of the freely living nematodes and serves as a good model system for screening anthelminthic drugs. In this study, thirteen flavonoid derivatives were tested for anthelminthic activity and the relationships between their activities and structures were investigated. The structural information combined with the data for the larval growth inhibition of C. elegans provided meaningful structural insights in the search for new anthelminthic drugs.

Animals↗

Treatment of Baylisascaris procyonis infections in dogs with milbemycin oxime.

An examination was made as to the ability of Sentinel Flavor Tabs (milbemycin oxime/lufenuron) to treat Baylisascaris procyonis infections in dogs. The study was designed as a critical trial and included five naturally infected dogs and two dogs that were experimentally infected. Another dog from a prior clinical trial that was treated with Sentinel Flavor Tabs as part of the original FDA submission package for intestinal nematode infections was also included with the treated dogs. Of the five naturally infected dogs treated as part of the critical trial, three were cleared of their infections. These five dogs passed a total of 52 worms after treatment; one dog retained 23 worms and the other retained 1 worm at necropsy 7 days after treatment. Two of five experimentally infected Beagle dogs that had been given mice that had been fed 200 infectious eggs, developed patent infections with the parasite. These dogs were treated, and one of the dogs passed one worm and the other passed two worms after treatment with no worms being detected at necropsy 7 days after treatment. The one dog that was treated with milbemycin oxime as part of the FDA submission was clear of worms at necropsy. Overall, the mean efficacy of Sentinel Flavor Tabs was found to be 91.0%. Of the eight dogs that were treated, six were totally cleared of their infections, a cure rate of 75%. The two dogs that did not clear their infections had very large numbers of adult B. procyonis within their intestinal tracts at the time of treatment, one dog had 40 worms (23 remaining) and the other had 26 worms (1 remaining). It is suggested that the treatment of dogs with monthly Sentinel Flavor Tabs could markedly reduce the chance of infected dogs contaminating the environment. Also, additional monthly treatments are highly likely to clear dogs of any worms not killed with the initial treatment.

Animals↗

Current status of human parasitic infections in Taiwan.

The eradication of the 2 mosquito-borne parasitic diseases, malaria and lymphatic filariasis, is one of the greatest achievements of the parasite control campaigns in Taiwan. Most of the soil-transmitted nematode infections, with the exception of pinworm infection, are currently well controlled and limited to some aboriginal areas. Food-borne parasitic zoonosis such as infections with Angiostrongylus cantonensis, Clonorchis sinensis, and Taenia saginata asiatica are not rare, but the former is seasonal and the latter 2 are ethnically and geographically associated. Intestinal protozoal infections with Giardia lamblia and Cryptosporidium parvum are at low levels but may be widely distributed. Opportunistic protozoal infections among patients with acquired immunodeficiency syndrome, which included amebic colitis, Pneumocystis carinii pneumonia, and cerebral toxoplasmosis, are becoming increasingly important. The rapid increase in international travel and the introduction of large numbers of foreign workers from other countries in Southeast Asia may change the epidemiological patterns of parasitic infections in Taiwan.

AIDS-Related Opportunistic Infections↗

Helicobacter pylori vaccines and mechanisms of effective immunity: is mucus the key?

In this theoretical article, the hypothesis is proposed that immunization against gastric helicobacter infection is mediated by CD4+ T-cell induced changes in mucus production. Vaccine development for the gastric pathogen Helicobacter pylori has encountered several problems. Resolving these problems is impeded by our lack of understanding of the mechanisms by which the immune response influences bacterial colonization. Protective immunity requires CD4+ T cells, but the majority of helicobacters are located in the mucus of the gastric lumen, away from the epithelial surface. Evidence suggests that this mechanism functions independently of antibodies, so how this is achieved is unknown. Clues to this mechanism may be provided by immune clearance of nematode infection. Similar to H. pylori, expulsion of the intestinal nematode, Nippostrongylus brasiliensis, in rodents is mediated by CD4+ T-cell changes in the numbers of goblet cells and the type of mucins secreted into the gut. Immune-mediated changes in secretion of gastric mucins could similarly be responsible for the reductions in helicobacter colonization seen in immunized animals. Helicobacter pylori are highly motile bacteria that have evolved to inhabit their specialized niche. Alterations in their mucus environment could influence their motility, such that the bacteria cannot remain efficiently within the mucus and are flushed away.

Animals↗

High risk of hookworm infection among wastewater farmers in Pakistan.

The health risks of wastewater use in agriculture were investigated in the city of Faisalabad, Pakistan, by means of a cross-sectional study. The study showed an increased risk of intestinal nematode infection and hookworm infection, in particular, in wastewater farmers (OR = 31.4, 95% CI 4.1-243) and their children (OR = 5.7, 95% CI 2.1-16) when compared with farming households using regular (non-wastewater) irrigation water. Textile labourers living in the same village as the wastewater farmers showed a lower risk of hookworm infection than wastewater farmers but an increased risk compared with farming households using regular irrigation water. Many urban and peri-urban farmers make a living by using untreated wastewater in the production of fresh produce for the urban market. Banning the use of untreated wastewater would deprive these farmers of their livelihood and affect food supply for the urban population. If treatment of wastewater is not a feasible option, the promotion of footwear and improved hygiene, the construction of toilets, in combination with regular anthelminthic treatment, would be suitable alternatives to safeguard the health of wastewater farmers and their children.

Adolescent↗

A multicenter evaluation of the effectiveness of Quest Gel (2% moxidectin) against parasites infecting equids.

Controlled trials with a common protocol were conducted in Idaho, Illinois and Tennessee to evaluate anthelmintic effectiveness of Quest Gel (QG; 2% moxidectin) against lumenal parasites in horses. Candidate horses were required to have naturally acquired nematode infections, as confirmed by presence of strongylid eggs in feces. At each site, 24 equids were blocked on the basis of pretreatment strongyle fecal egg counts (EPG) and randomly assigned to treatments within blocks. Within each block of two animals, one received QG on Day 0 at a dosage of 0.4 mg moxidectin/kg b.w. and one was an untreated control. Body weights measured the day before treatment served as the basis for calculating treatment doses. Horses assigned to treatment with QG received the prescribed dose administered orally with the commercially packaged Sure Dial syringe. Horses were necropsied 12-14 days after treatment, and lumenal parasites and digesta were harvested separately from each of five organs, including the stomach, small intestine, cecum, ventral colon and dorsal colon. Parasites from stomachs and small intestines were identified to genus, species and stage. Micro- (i.e., < 1.5 cm) and macroparasites (i.e., > 1.5 cm) in aliquots from the cecum, ventral colon and dorsal colon were examined in aliquots of approximately 200 parasites until at least 600 parasites had been identified to genus, species and stage or until all parasites in the 5% aliquot were examined, whichever occurred first. Data were combined across sites and analyzed by mixed model analysis of variance to assess the fixed effect of treatment and random effects of site and block within site. Because QG does not contain a cestocide, efficacy of QG against tapeworms was not significant (P > 0.05). Based on geometric means, however, efficacy of QG was greater than 90% (P < 0.05) against 38 species and developmental stages of cyathostomes, strongyles, bots, larval pinworms and ascarids encountered in at least 6 of 36 control horses in the combined data set. None of the horses treated with moxidectin exhibited evidence of adverse effects. Study results demonstrate QG, administered to horses with naturally acquired endoparasite infections at a dosage of 0.4 mg moxidectin/kg b.w., was highly effective against a broad range of equine parasitic infections.

Analysis of Variance↗

Modulation of calf immune responses by Ostertagia ostertagi: the effect of diet during trickle infection.

The effects of Ostertagia ostertagi infection and diet on antibody responses to O. ostertagi third stage larval (L3) antigen and to an unrelated antigen, Keyhole Limpet Haemocyanin (KLH) were determined in calves experimentally infected with 3000 L3 on alternate days for 6 weeks. Calves were given one of two diets, and were either infected or not infected with O. ostertagi L3. The diets were either high (H) or low (L) in protein/energy and were within the range of normal husbandry practice in the UK. Both IgG1 and IgG2, but not IgA, responses to L3 antigen were increased in the L-diet compared with the H-diet. IgA responses to L3 antigen were not affected by dietary treatment. The effects of diet and infection on anti-KLH IgG1 were independent of each other; IgG1 anti-KLH responses were decreased by infection and by the L-diet compared with the H-diet. The data suggest that there is a strong interrelationship between diet and immunity during nematode infections.

Animals↗

Measuring the local burden of disease. A study of years of life lost in sub-Saharan Africa.

BACKGROUND: An effective health policy necessitates a reliable characterization of the burden of disease (BOD) by cause. The Global Burden of Disease Study (GBDS) aims to deliver this information. For sub-Saharan Africa (SSA) in particular, the GBDS relies on extrapolations and expert guesses. Its results lack validation by locally measured epidemiological data. METHODS: This study presents locally measured BOD data for a health district in Burkina Faso and compares them to the results of the GBDS for SSA. As BOD indicator, standard years of life lost (age-weighted YLL, discounted with a discount rate of 3%) are used as proposed by the GBDS. To investigate the influence of different age and time preference weights on our results, the BOD pattern is again estimated using, first, YLL with no discounting and no age-weighting, and, second, mortality figures. RESULTS: Our data exhibit the same qualitative BOD pattern as the GBDS results regarding age and gender. We estimated that 53.9% of the BOD is carried by men, whereas the GBDS reported this share to be 53.2%. The ranking of diseases by BOD share, though, differs substantially. Malaria, diarrhoeal diseases and lower respiratory infections occupy the first three ranks in our study and in the GBDS, only differing in their respective order. Protein-energy malnutrition, bacterial meningitis and intestinal nematode infections occupy ranks 5, 6 and 7 in Nouna but ranks 15, 27 and 38 in the GBDS. The results are not sensitive to the different age and time preference weights used. Specifically, the choice of parameters matters less than the choice of indicator. CONCLUSIONS: Local health policy should rather be based on local BOD measurement instead of relying on extrapolations that might not represent the true BOD structure by cause.

Africa South of the Sahara↗

Genomic tools to improve parasite resistance.

The natural genetic variability of the ruminant immune system provides a feasible means to control gastrointestinal (GI) parasite infection without anthelmintics. However, the paradigm of traditional selection has not been effectively applied to the moderately heritable traits of parasite resistance (h approximately equal to 0.3) due to the difficulty and expense of gathering accurate phenotypes in a commercial production setting. These characteristics make host traits related to GI nematode infection ideal candidates for genomics-based research. To initiate explanation of important allelic differences, economic trait loci (ETL) are being identified and mapped using a resource population of Angus cattle segregating for GI nematode resistance and susceptibility to the two most common nematode parasites of US cattle, Ostertagia ostertagi and Cooperia oncophora. The population is composed of five generations of half-sib progeny with complete phenotypic records produced from controlled infections. To detect the genomic locations of the three distinct phenotypic traits being expressed (innately immune, acquired immune, and immunologically non-responsive), genotypes have been generated for DNA markers (N=199) spaced at regular intervals (approximately 20cm intervals) throughout the entire genome (3000cm). Although initial ETL detection may be limited by half-sib family size, the unique structure of this population provides additional statistical power for refining map position of potential ETL. After allele frequency and contribution to phenotype are determined in this population, marker tests associated with ETL most beneficial for controlling parasite infection can be accurately used for selection. Comparative map and functional genomic information from humans and other species of biomedical importance will be utilized in further investigations to elucidate the genes underlying ETL.

Animals↗

Demonstration of a range of inflammatory mediators released in trichostrongylosis of sheep.

The levels of inflammatory mediators in the intestinal contents of sheep immunized with Trichostrongylus colubriformis larvae increased in the first 6 days after challenge. These mediators were histamine, leukotriene C4, 6-keto-prostaglandin F1 alpha (from prostacyclin) and thromboxane B2. Leukotriene C4 was released in the greatest quantities. Leukotriene B4 was present but its concentration remained unchanged after challenge. The presence of these particular mediators in the intestinal contents after challenge is consistent with antigen-induced mediator release from the mucosal mast cells found in immune sheep undergoing challenge infection. This is the first sequential analysis of mediator release in sheep that also demonstrates the release of prostacyclin and thromboxane into the intestine during expulsion of a nematode infection.

6-Ketoprostaglandin F1 alpha↗

The influence of host immunity on the epidemiology of trichostrongyle infections in cattle.

The acquisition of immunity to the important trichostrongyles of cattle is reviewed and the subsequent effect on the epidemiology of trichostrongylosis is discussed. Examples of factors which can delay or impair the development of immunity to these nematode infections are presented. These include the interactions with anthelmintic prophylactic programmes, nutrition, intercurrent infections, pregnancy and lactation.

Animal Nutritional Physiological Phenomena↗

Interleukin-3 and c-Kit/stem cell factor are required for normal eosinophil responses in mice infected with Strongyloides venezuelensis.

To evaluate the potential roles of Interleukin-3 (IL-3) and c-Kit, the tyrosine kinase receptor for stem cell factor (SCF), in eosinophil responses in vivo, we examined eosinophil numbers in uninfected or nematode-infected wild-type mice, IL-3-/- mice, and IL-3-/- mice that also have a marked reduction in SCF/c-Kit signaling (ie, Kit(W)/Kit(W-v), IL-3-/- mice). We found no significant differences in the numbers of eosinophils in the blood, bone marrow or various tissues of IL-3-/- vs IL-3+/+ mice, either at baseline or after the induction of bone marrow, blood or tissue eosinophilia in response to infection with Strongyloides venezuelensis (S.v.) or Nippostrongylus brasiliensis (N.b.). However, in mice with markedly impaired SCF/c-Kit signaling, IL-3 contributed significantly to the increased numbers of eosinophils that were observed in multiple tissues during S.v. infection, but not during infection with N.b.

Animals↗

[Application of a mermithid nematode for the control of mosquito larvae in natural conditions].

Field tests were carried out with the Romanomermis culicivorax nematode (Ross and Smith, 1976) for the control of 3 mosquito species--Anopheles albimanus (Wiedeman, 1821): Culex nigripalpus (Theobald, 1901); and Uranotaenia saphirina (Oster-Sacken, 1868)--in 10 natural reservoirs. The infective nematodes were disseminated in doses of 1 x 10(3) preparasites/m2 of surface area. Increased infestation indices were observed with values ranging from 1.2 to 3.4; mortality percentages fluctuated between 70 and 97% depending on the mosquito species found in the reservoirs. In those reservoirs containing specimens of the 3 species, anophelines showed more susceptibility to parasite infection; culicines showed a lower susceptibility in general. Mosquito larva populations were significantly reduced in the treated reservoirs.

Animals↗

Evidence of glutathione transferase complexing and signaling in the model nematode Caenorhabditis elegans using a pull-down proteomic assay.

Phage display techniques using random peptide interactions have supported the role of mammalian glutathione transferase (GST) as part of a signalling pathway for both oxidative stress and an apoptosis pathway. Little is known about the interaction of nonmammalian GST with other proteins. GSTs have been implicated in the development of chronic nematode infections by neutralising cytotoxic products arising from host immune initiated reactive oxygen species (ROS) assault. In this study we attached one of the key GSTs expressed in the model nematode Caenorhabditis elegans to an affinity support matrix and directly identified major interacting proteins by two-dimensional electrophoresis and peptide mass fingerprinting before and following oxidative stress. Nematode GST does not appear to be a stand-alone enzyme and interacts with many types of proteins in both normal and ROS stress conditions. Pull-down proteomic presents a flexible, label free, rapid and economical assay without specialised ligand fishing equipment to identify protein binding partners.

Amino Acid Sequence↗

Protection studies with a globin-enriched protein fraction of Ostertagia ostertagi.

The protective capacity of an adult stage Ostertagia ostertagi globin antigen was tested in four vaccination experiments in cattle. In a preliminary experiment, calves were vaccinated three times intraperitoneally with 250 microg globin in Freund's adjuvant and challenged with a trickled infection of 25,000 infective larvae. In three subsequent field studies, calves were vaccinated twice or three times intramuscularly with 80-100 microg globin in Quil A and challenged with a natural gastrointestinal nematode infection on pasture. Higher globin-specific antibody levels were detected in the vaccinated calves than in the control animals in all vaccine trials. In the preliminary experiment, geometric mean cumulative egg counts in the globin group were reduced by 52% and total worm burdens were reduced by 28%, compared to the controls. In the first field trial cumulative faecal egg counts were reduced by 63% in the vaccinated calves. However, the reduction in faecal egg output in these two experiments was not statistically significant and no reduction in faecal egg counts was observed in the vaccinated animals in the two last field trials. In conclusion, vaccination of calves with O. ostertagi globin resulted in highly variable protection levels after challenge infection.

Adjuvants, Immunologic↗

Parasitic infection of the seahorse (Hippocampus erectus)--a case report.

This report details the development of a microsporidan infection in a colony of seahorses (Hippocampus erectus) caused by Glugea heraldi. Of 76 animals imported from Florida Bay (USA), two survived. A myxosporidan (Sphaeromyxa sp.) and an unidentified nematode infection were also diagnosed in the same colony, but these infections were not considered important, causative factors in the mortalities.

Animals↗