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Cryopreservation of the infective larvae of the common nematodes of ruminants.

Exsheathed infective larvae (L 3) of 19 species of nematodes were tested for infectivity in either sheep or cattle after they had been frozen in 0,9% NaCl solution, stored for a relatively short time in the gas phase of liquid nitrogen and subsequently thawed. In addition, 13 of these species were tested after similar storage for up to 18 months. In sheep, Haemonchus contortus, Ostertagia circumcincta, Trichostrongylus axei, Trichostrongylus colubriformis, Nematodirus spathiger and Oesophagostomum columbianum were viable after 2 years of cryopreservation, a mean of greater than 90% of the L 3 being alive when thawed after this period. Similar results were obtained with Chabertia ovina L 3 after 18 months and with Marshallagia marshalli, Trichostrongylus falculatus and Dictyocaulus filaria, after a short period of freezing. On the other hand, Gaigeria pachyscelis and Strongyloides papillosus survived freezing for up to 7 months but neither was viable at the end of this period, nor was exsheathed G. pachyscelis viable without freezing. Most of these infestations were established by inoculating the infective larvae into the abomasum and/or duodenum. M. marshalli, T. falculatus and C. ovina also proved infective after oral dosing. D. filaria, the only other species tested by this route, was not infective when dosed per os after thawing. The infective larvae of the bovine nematodes, Haemonchus placei, Ostertagia ostertagi, Nematodirus helvetianus, Oesophagostomum radiatum, Cooperia pectinata and Cooperia punctata survived freezing for a mean of 26 months, greater than 90% being alive on thawing, but infectivity was generally lower than with the same genera in sheep. Even when not frozen, exsheathed Bunostomum phlebotomum was non-infective. When Cooperia spp. after thawing were tested for infectivity by the oral route, more worms developed in one calf infested orally than in another infested by inoculation into the duodenum. Ova of H. contortus, M. marshalli, O. circumcincta, T. colubriformis, T. falculatus, N. spathiger, C. ovina, H. placei, O. ostertagi, Cooperia spp. and N. helvetianus were recovered from the faeces of animals infested with cryopreserved L 3. No ova of O. columbianum or O. radiatum were recovered from faeces, because differential larval counts were performed before they were patent. Nevertheless, gravid females were obtained post-mortem. Frozen L 3 of N. helvetianus were used to re-establish a pure strain in calves, 2,3 million ova being recovered from infestations with 10 670 L 3 frozen for 26 months. The infectivity of the progeny of frozen L 3 was tested with M. marshalli and C. ovina. In both instances infectivity was high and the worms which developed also produced ova, thus completing the cycle. This appears to be the first report of infective larvae of parasitic nematodes retaining their infectivity after being frozen in liquid nitrogen (gas phase) for longer than 2 years. This is also apparently the first time that M. marshalli T. colubriformis, T. falculatus, T. axei, N. spathiger, C...

Ancylostomatoidea↗

Antibiotic production in relation to bacterial growth and nematode development in Photorhabdus--Heterorhabditis infected Galleria mellonella larvae.

The population of Photorhabdus luminescens C9, bacterial symbiont of the entomopathogenic nematode, Heterorhabditis megidis 90, increased rapidly to 1.2-2.6x10(9) cells g(-1) wet Galleria mellonella larvae within 24 h of nematode infection of the larvae, and maintained a relatively constant level (1.2-2.0x10(10) cells g(-1)) through the entire 14-day period of nematode development. The antibiotic, 3, 5-dihydroxy-4-isopropylstilbene, was produced by P. luminescens C9 after 24 h of nematode infection, increased rapidly at 2-5 days postinfection and remained at a level of 3000-3600 microg g(-1) wet larvae until about 21 days, decreasing gradually thereafter. The early production and continued presence of a relatively large amount of 3,5-dihydroxy-4-isopropylstilbene in the infected insect supports the hypothesis that the antibiotics produced by the bacterial symbiont help minimize competition from other microorganisms and prevents the putrefaction of the nematode-infected insect cadaver.

Animals↗

Current status of soil-transmitted helminths in Vietnam.

This paper provides a review of surveys on soil-transmitted helminths that were done in Vietnam between 1990 and 2001. Prevalence estimates could be obtained for 29 of the 61 provinces. Extrapolating from this, it is estimated that 33.9 million people in Vietnam are infected with Ascaris (prevalence 44.4%), 17.6 million with Trichuris (prevalence 23.1%), and 21.8 million with hookworm (prevalence 28.6%). Prevalence of Ascaris and Trichuris showed a declining trend from the north to the south of the country. This is probably related to differences in climatic conditions. Hookworm prevalence is more evenly distributed throughout the country, but is concentrated in peri-urban and rural agricultural areas. Vegetable cultivation in which nightsoil is used as fertilizer is a risk factor for hookworm infection, especially among adult women. Helminth control programs should be targeted at school-age children in the northern provinces. Specific interventions are needed throughout the country for women of agricultural communities that are at risk for hookworm infection. There is clearly a need for more detailed analysis of risk factors to quantify the relative contribution of climatic, environmental, and human behavioral factors in the transmission of intestinal nematode infections in Vietnam.

Adolescent↗

Response to challenge infection with Haemonchus contortus and Trichostrongylus colubriformis in dairy goats. Differences between high and low-producers.

The objective of the study was to analyse the influence of the level of milk production in dairy goats on the ability to develop resistance to gastrointestinal nematode infection. Two groups of 50 goats were initially exposed (Group 'Previously Infected' PI) or not exposed (Group 'Not Previously Infected' NPI) to repeated infections with a mixture of Haemonchus contortus and Trichostrongylus colubriformis. After anthelmintic treatment, goats from both groups remained free of parasites for 2 months around kidding. One month after kidding, 24 lactating goats in each group were challenged with Haemonchus contortus and Trichostrongylus colubriformis. Parasitological, pathophysiological and milk production parameters were then examined fortnightly for 3 months following challenge. Within the PI and NPI group, the response to challenge was compared between the 25% goats exhibiting the lowest level of milk production and the 25% with the highest level of production. In the low producers, previous contact with the parasites was associated with a decrease in egg excretion which was not apparent in the high producers. Previous exposure to parasites was also associated with more severe pathophysiological disturbances in the high producers. Lastly, the response to challenge also induced more severe depression in milk production after previous infection in the animals with the highest productive potential. These results confirmed that the level of milk production in dairy goats could influence the resistance and/or resilience to parasitism with more severe consequences detected in the high producers. In addition, the results suggest that such differences could in fact be mainly related to differences in the host response to parasites.

Animals↗

Biochemical and genetic studies of two Heterodera avenae resistance genes transferred from Aegilops ventricosa to wheat.

Two Heterodera avenae resistance genes, Cre2 from Aegilops ventricosa AP-1 and Cre5 from Ae. ventricosa #10, were shown to confer a high level of resistance to the Spanish pathotype Ha71. No susceptible plants were found in the F(2) progeny from the cross between the two accessions of Ae. ventricosa, suggesting that their respective resistance factors were allelic. However, genes Cre2 and Cre5 apparently were transferred to a different chromosomal location in the wheat line H-93-8 and in the 6M(v)(6D) substitution, respectively, as proved by F(2) segregation of their cross progeny. The induction of several defence responses during early infection by the same H. avenae pathotype in resistant lines carrying Cre2 or Cre5 genes was studied. Isoelectrofocusing (IEF) isozyme analysis revealed that peroxidase, esterase and superoxide dismutase activity increased after nematode infection, in roots of resistant lines in comparison with their susceptible parents. Differential induced isoforms were also identified when IEF patterns of resistant lines were compared. A DNA marker, absent in Cre5-carrying genotypes, was found to be linked, thought not very tightly, to the Cre2 gene in the H-93-8 line. The differences observed between the Cre2 and Cre5 genes with respect to their chromosomal location in wheat introgression lines, de-toxificant enzyme induction and behaviour against different pathotypes, suggest they are different H. avenae resistance sources for wheat breeding.

Animals↗

The role of endosymbiotic Wolbachia bacteria in the pathogenesis of river blindness.

Parasitic filarial nematodes infect more than 200 million individuals worldwide, causing debilitating inflammatory diseases such as river blindness and lymphatic filariasis. Using a murine model for river blindness in which soluble extracts of filarial nematodes were injected into the corneal stroma, we demonstrated that the predominant inflammatory response in the cornea was due to species of endosymbiotic Wolbachia bacteria. In addition, the inflammatory response induced by these bacteria was dependent on expression of functional Toll-like receptor 4 (TLR4) on host cells.

Animals↗

Albendazole in the treatment of intestinal helminthiasis in children.

A single dose of 2% albendazole suspension (400 mg in 20 ml) was administered to 77 patients (42 males and 35 females), ages ranging between 2 to 12 years, with helminthic infections. Ascariasis was the most prevalent infection. Patients were followed-up for 3 weeks. The results showed that albendazole was highly effective against Ascaris lumbricoides, Ancylostoma duodenale and Enterobius vermicularis. Significant improvement was also observed in patients having infections due to Trichuris trichiura. Albendazole was well tolerated and did not produce any significant side-effects. Single dose albendazole appears to be appropriate for mass chemotherapy to control intestinal nematode infections in highly infected communities.

Albendazole↗

Enhancement of disease and pathology by synergy of Trichuris suis and Campylobacter jejuni in the colon of immunologically naive swine.

Campylobacter jejuni, a leading cause of bacterial gastroenteritis, has different age distribution and disease expression in developing and developed countries, which may be due to the endemnicity of infection and the age of acquisition of immunity. Differences in disease expression are not solely dependent on the C jejuni strain or virulence attributes. Another modulating factor in developing countries may be endemic nematode infections such as Trichuris, which drive type 2 cytokine responses and down-regulate type 1 immune responses. In this study, three-day-old germ-free pigs given dual infections with Trichuris suis and C jejuni had more frequent, more severe diarrhea and severe pathology than pigs given no pathogens, only T. suis, or only C jejuni. These pigs had significant hemorrhage and inflammatory cell infiltrates in the proximal colon where adult worms were found, and abscessed lymphoglandular complexes in the distal colon with intracellular C jejuni. Pigs given only C jejuni had mild clinical signs and pathology, and bacteria in feces or extracellular sites. Pigs given T. suis or no pathogens had no disease and minimal pathology. Thus, these agents synergized to produce significant disease and pathology, which was site specific.

Animals↗

Enhancement of disease and pathology by synergy of Trichuris suis and Campylobacter jejuni in the colon of immunologically naive swine.

Campylobacterjejuni, a leading cause of bacterial gastroenteritis, has different age distribution and disease expression in developing and developed countries, which may be due to the endemnicity of infection and the age of acquisition of immunity. Differences in disease expression are not solely dependent on the C. jejuni strain or virulence attributes. Another modulating factor in developing countries may be endemic nematode infections such as Trichuris, which drive type 2 cytokine responses and down-regulate type 1 immune responses. In this study, three-day-old germ-free pigs given dual infections with Trichuris suis and C. jejuni had more frequent, more severe diarrhea and severe pathology than pigs given no pathogens, only T. suis, or only C. jejuni. These pigs had significant hemorrhage and inflammatory cell infiltrates in the proximal colon where adult worms were found, and abscessed lymphoglandular complexes in the distal colon with intracellular C. jejuni. Pigs given only C. jejuni had mild clinical signs and pathology, and bacteria in feces or extracellular sites. Pigs given T. suis or no pathogens had no disease and minimal pathology. Thus, these agents synergized to produce significant disease and pathology, which was site specific.

Animals↗

Husbandry management, worm control practices and gastro-intestinal parasite infections of sows in pig-breeding farms in Münsterland, Germany.

A cross-sectional survey was performed on 144 farms randomly selected from 3 breeding organisations in Münsterland, Germany, to obtain representative data on husbandry management, worm control practices and their possible associations with gastro-intestinal parasite infections in sows. Straw bedding was provided in the majority of gestation units; lactating sows were frequently kept strawless. Pasturing of gilts and non-lactating sows were performed in 37% of the farms. Sows were treated with anthelmintics (mainly benzimidazoles, ivermectin or levamisole) in 98% of the farms. The proportions of farmers performing the 'targeted' (mainly prefarrowing) or 'non-targeted' treatment strategy (treatment of all animals several times per year) were similar. Piglets were anthelmintically dosed in only 19% of the farms. Patent infections of sows with Oesophagostomum spp., Ascaris suum, Trichuris suis and Eimeria spp. were detected in 79%, 7%, 8% and 29% of all the farms, respectively. Hyostrongylus rubidus infections were only found in farms with pasturing. The within-farm prevalences of both the strongylid and Eimeria infections were negatively correlated with the herd size but positively with the weaning age. Risk factors for strongylid infections were straw bedding in pens and pasturing. Pasturing was a risk factor for Eimeria infections too. There was a positive association between the time interval to last deworming of sows and strongylid egg counts. However, neither the treatment strategy nor the anthelmintic class used was associated with nematode infections. No relations were found between different cleaning methods in stables and endoparasite infections.

Animal Husbandry↗

Root-parasitic nematodes enhance soil microbial activities and nitrogen mineralization.

Obligate root-parasitic nematodes can affect soil microbes positively by enhancing C and nutrient leakage from roots but negatively by restricting total root growth. However, it is unclear how the resulting changes in C availability affect soil microbial activities and N cycling. In a microplot experiment, effects of root-parasitic reniform nematodes ( Rotylenchulus reniformis) on soil microbial biomass and activities were examined in six different soils planted with cotton. Rotylenchulus reniformis was introduced at 900 nematodes kg(-1) soil in May 2000 prior to seeding cotton. In 2001, soil samples were collected in May before cotton was seeded and in November at the final harvest. Extractable C and N were consistently higher in the R. reniformis treatments than in the non-nematode controls across the six different soils. Nematode inoculation significantly reduced microbial biomass C, but increased microbial biomass N, leading to marked decreases in microbial biomass C:N ratios. Soil microbial respiration and net N mineralization rates were also consistently higher in the nematode treatments than in the controls. However, soil types did not have a significant impact on the effects of nematodes on these microbial parameters. These findings indicate that nematode infection of plant roots may enhance microbial activities and the turnover of soil microbial biomass, facilitating soil N cycling. The present study provides the first evidence about the direct role of root-feeding nematodes in enhancing soil N mineralization.

Animals↗

Antiparasitic agents.

Several important developments have occurred in recent years in the chemotherapy for and prophylaxis of parasitic infections. Although mefloquine is clearly the most effective agent for prevention of chloroquine-resistant falciparum malaria, its use has been compromised by side effects, both real and imagined. Well-designed studies have shown that side effects occur no more frequently with low-dose mefloquine than with chloroquine. Use of mefloquine in pregnant women has not been associated with birth defects, but the incidence of stillbirths may be increased. Malarone is a new agent that combines atovaquone and proguanil, and it may be as effective as mefloquine; however, it is not yet available in the United States. Several newer agents have appeared in response to the development of multidrug resistant Plasmodium falciparum, especially in Southeast Asia. Halofantrine is available for the treatment of mild to moderate malaria due to P. falciparum and for P. vivax infections. Because of severe toxic effects, use of halofantrine should be restricted to only those unusual and rare situations in which other agents cannot be used. Artemisinin (an extract of the Chinese herbal remedy qinghaosu) and two derivatives, artesunate and artemether, are active against multidrug resistant P. falciparum and are widely used in Asia in oral, parenteral, and rectal forms. The antibacterial azithromycin in combination with atovaquone or quinine has now been reported to treat babesiosis effectively in experimental animals and in a few patients. Azithromycin in combination with paromomycin has also shown promise in the treatment of cryptosporidiosis (and toxoplasmosis when combined with pyrimethamine) in patients with the acquired immunodeficiency syndrome (AIDS). Albendazole is currently the only systemic agent available for treatment of microsporidiosis, an infection primarily of patients with AIDS. In addition, albendazole and ivermectin have emerged as effective broad-spectrum antihelminthics, with albendazole becoming the drug of choice for hydatid disease (echinococcosis), neurocysticercosis, and most intestinal nematode infections (except strongyloidiasis and trichuriasis). Liposomal amphotericin B is the first drug approved by the Food and Drug Administration for the treatment of visceral leishmaniasis.

Antiparasitic Agents↗

Ingestion of Cryptosporidium oocysts by Caenorhabditis elegans.

Cryptosporidium parvum has been associated with outbreaks of human illness by consumption of contaminated water, fresh fruits, and vegetables. Free-living nematodes may play a role in pathogen transmission in the environment. Caenorhabditis elegans is a free-living soil nematode that has been extensively studied and serves as a good model to study possible transmission of C. parvum oocysts that may come into contact with produce before harvest. The objective of this study was to determine whether C. elegans could serve as a potential mechanical vector for transport of infectious C. parvum and Cyclospora cayetanensis in agricultural settings and whether C. elegans could ingest, excrete, and protect oocysts from desiccation. Seventy to 85% of worms ingested between 0 and 500 oocysts after 1 and 2 hr incubation with oocysts. Most of the nematodes ingested between 101 and 200 oocysts after 2 hr. Intact oocysts and empty shells were excreted by nematodes. Infectivity was determined by the neonatal assay with different treatments of worms (intact or homogenized) or oocysts or both. Adult C. elegans containing C. parvum kept in water were infective for mice. In conclusion, C. elegans adults can ingest and excrete C. parvum oocysts. Caenorhabditis elegans containing C. parvum oocysts can infect mice but does not seem to protect oocysts from extreme desiccation at 23 C incubation of a day or longer. Cyclospora oocysts were not ingested by C. elegans. The role of free-living nematodes in produce contamination needs to be further examined.

Animals↗

A role for IgE in intestinal immunity. Expression of rapid expulsion of Trichinella spiralis in rats transfused with IgE and thoracic duct lymphocytes.

In these experiments we characterize the protective antibodies in immune serum that interact synergistically with immune thoracic duct lymphocytes (TDL) to induce rapid expulsion (RE) of Trichinella spiralis in adult rats. Antibodies with both reaginic and nonreaginic activity mediated RE upon passive transfer to adult rats that had been adoptively transfused with immune TDL 7 days earlier. In serum collected 28 days after a primary infection, the most important antibody was homocytotropic IgE. Native IgE produced by active infection was isolated from 28-day immune serum by salt precipitation and/or by sequential affinity chromatography. The murine mAb A2 and B5 (anti-rat IgE) were conjugated separately to Sepharose 4B affinity columns for affinity separations. IgE was shown to be pure by gel electrophoresis and Western blots and its m.w. was estimated at approximately 190,000. As little as 183 micrograms of purified IgE could induce RE after passive transfer to adult rats. The IgE was shown to be functional by PCA activity, Ag-binding on Western blots, and skin sensitization; the latter could be blocked by pretreatment with 1R162, a rat myeloma IgE. Monoclonal IgG of any isotype transferred in amounts up to 35 mg/rat could not transfer RE to rats previously transfused with TDL cells. Immune serum collected 3 mo after the primary infection contained insufficient IgE to transfer RE, but complex non-IgE fractions were protective. The data thus demonstrate that IgE is a functional Ig in the rat capable of mediating the rejection of challenge nematode infections of the gut in the absence of other specific Ig. Secondly, other Ig may also play a role, in particular, several weeks after the primary infection when specific IgE levels in serum have declined.

Animals↗

Safety of imidacloprid plus moxidectin topical solution applied to cats heavily infected with adult heartworms (Dirofilaria immitis).

A topically applied formulation containing 10% imidacloprid+1% moxidectin (Advocate/Advantage multi) has been developed for monthly application to cats for the prevention of feline heartworm (HW) disease caused by Dirofilaria immitis; and for the treatment and control of flea infestations, ear mite infestations, and intestinal nematode infections. A study model was designed to evaluate the safety of this product in cats harboring adult D. immitis infections. Eighty adult cats (40 males/40 females) were each inoculated with 60 third-stage D. immitis larvae on test day (TD) 1. On TD 243-245 echocardiographic imaging was performed on each cat to confirm and estimate the number of adult D. immitis residing in the cardiovascular system. A total of 35 cats were subsequently eligible for safety evaluation based on inclusion criteria. Four treatment groups were established and randomly selected for treatment: imidacloprid+moxidectin solution at the label dose (n=9) (group 1), imidacloprid+moxidectin solution at 5x the Iabel dose (n=9) (group 2), 6% selamectin topical solution (Revolution) at the label dose (positive control, n=8) (group 3), and topical treatment with placebo (negative control, n=9) (group 4). All cats were treated on TD 250. Treatments for groups 1, 3, and 4 were repeated on TDs 278 and 306. Group 2 cats were euthanized and examined for adult D. immitis on TD 288. All other cats were euthanized and examined for adult D. immitis on TD 334. No adverse events attributable to treatment with the test articles were observed during the study. The geometric mean numbers of adult D. immitis recovered at necropsy from treatment groups 1-4 were 2.9, 3.2., 4.0, and 2.7, respectively. There were no statistically significant differences in the comparison of adult D. immitis recovered at necropsy (ANOVA overall group effect P-value of 0.5356). The results of this study demonstrate that imidacloprid+moxidectin topical solution can be used safely in cats heavily infected with adult D. immitis.

Animals↗

Endogenous testosterone levels do not affect filarial worm burdens in mice.

It has been observed for several years that males of many mammalian species exhibit higher prevalence and intensity of parasitic infections than females. It has been theorized that this sexual dichotomy may be due to the immunosuppressive properties of testosterone. The administration of exogenous testosterone to laboratory animals often results in higher yields of several parasitic nematodes, lending credence to this hypothesis. We sought to determine if worm burdens in individual mice were related to serum testosterone concentrations. We found that in a cage of five male mice, one or two individuals exhibited high circulating testosterone levels, while the rest had low to undetectable levels. Serum testosterone did not correlate with worm recoveries. The implications of these findings are twofold. First, our data suggest that high endogenous testosterone does not necessarily decrease the ability of mice to resist nematode infection and second, that results obtained from administration of exogenous testosterone should be interpreted with caution.

Animals↗

Intestinal helminthiases and their control with albendazole among primary schoolchildren in riverine communities of Ondo State, Nigeria.

A study to establish the prevalence of intestinal helminthiases among schoolchildren of riverine communities in the Ilaje-Ese Odo Local Government Area of Ondo State, Nigeria was conducted. Ninety-four percent of the children studied were infected with intestinal helminths. Trichuris trichiura infection was the commonest (84%), followed by Ascaris lumbricoides (75.3 %) and hookworm (7.6 %). Dual helminthic infections were recorded, with Ascaris-Trichuris having the highest prevalence among the children. Poor environmental sanitation and personal hygiene combined with the absence of potable water and a lack of awareness of the effects of nematode infection were identified as the possible reasons for the high rate of infection. Treatment with albendazole (200 mg) brought about reductions in the level of Trichuris trichiura (to 41.7%), Ascaris lumbricoides (to 4.2%) and Hookworm (to 0.7%). The estimated rates of reduction were 94.4%, 49.7%, and 90.2% for Ascaris lumbricoides, Trichuris trichiura and hookworm respectively. Post-treatment helminthic reduction, as found in this study, is expected to enhance the mental and physical development of the children. Community mobilization with health education messages aimed at improving personal and community hygiene was initiated with an emphasis on creating a sustained reduction in the burden of helminthic infection.

Adolescent↗