Survival of Ancylostoma caninum on bluegrass pasture.

The survival of infective larvae of Ancylostoma caninum on outdoor grass plots was studied in 40 experiments over 1 year. Weather data were collected over the period. Mean larval survival from August to early November was 24 days (range 1 to 49), from December through February was 0 days, and from March to mid-August was 6.6 days (range 0 to 21). Moderate to high temperatures and substantial rainfall favored larval survival; low temperatures and rainfall favored larval destruction.     

NADH oxidase and fumarate reductase of Ancylostoma ceylanicum.

Ancylostoma ceylanicum, the hookworm parasite of cat, dog and man, was found to contain NADH and/or NADPH oxidase as well as fumarate reductase activities. Both the enzyme systems were predominantly located in the membranes of mitochondrial-rich preparations. The membranes also exhibited the presence of a reduced pyridine nucleotide transhydrogenase activity which transferred hydrogen from NADPH to NAD. Amongst respiratory inhibitors, rotenone (Site I inhibitor) markedly depressed both NADH oxidase and fumarate reductase while others, namely antimycin-A, KCN and azide, had a lesser effect.     

Mechanism of skin penetration by Ancylostoma tubaeforme larvae.

Skin penetration by infective Ancylostoma tubaeforme larvae has been investigated cinematographically and using in vitro techniques. The dermal tissue appears to cause little hinderance to larval migration but complete penetration through the skin from the dermal direction did not occur, although total penetration from the epidermal surface was frequently accomplished. No evidence could be found for enzymic secretions emanating from the worms under conditions that gave positive results from Necator americanus and Strongyloides fülleborni infective larvae. The results indicated that A. tubaeforme was able to penetrate without the use of enzymic secretions and an alternative, mechanical mechanism for penetration is advanced.     

Ancylostoma duodenale: maintenance through six generations in helminth-native pups.

An Indian strain of Ancylostoma duodenale with a known propensity for arrested development in man was passaged serially in 2-month-old, helminth-naive, male beagles. The sixth passage was initiated about 500 days after laboratory maintenance began. Experimental animals were infected by stomach tube with 1500 larvae each. These infections became patent in 22–31 days. In a line passaged in dogs without the use of a corticosteroid (prednisolone), adaptation, as judged by fecal egg count, increased through the first three generations and then declined sharply. This line appears destined for extinction. In another line passaged in prednisolone-treated pups, the strain's original viability is being maintained. Infections were long lived, frequently surviving for 90 days or more. Maximum egg output in terms of eggs per gram of feces (EPG) was highly variable between generations and between individuals within generations, but commonly exceeded 1500 EPG. In pups necropsied between 44 and 55 days, harboring fifth- and sixth-generation infections, adult worm burdens ranged from 47–146 and 34–128 in the nonprednisolone and prednisolone lines, respectively. Arrested larvae have been found in the same pups and apparently contribute to a turnover in the population of adult worms. Although A. duodenale has yet to be fully adapted to dogs in the sense that indefinite survival without prednisolone is assured, the present system yields a variety of life history stages in quantities suitable for many experimental purposes. Thus it provides a utilitarian laboratory model for the investigation of one of man's major hookworms.     

Determinants of the longevity of third-stage infective larvae of Ancylostoma tubaeforme.

The effects of some extrinsic factors on the lipid (energy) reserves and longevity of third-stage larvae of the cat hookworm Ancylostoma tubaeforme, were investigated under controlled laboratory conditions. In nonstressful microenvironmental conditions, larval longevity was directly related to the rate of utilisation of the lipid reserves. The effects of the various environmental stresses on longevity could also be explained largely on the basis of their deleterious effects on the lipid metabolism of the larvae.     

Biological control of infective larvae of Ancylostoma spp. in beach sand

BackgroundGeohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission.AimsThe aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L₃) of Ancylostoma spp. in beach sand under laboratory conditions.MethodsIn the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L₃ in pots containing 30 g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L₃ and distilled water in pots with 30 g of sand.ResultsEvidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L₃: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p < 0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9 cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L₃ and 5 g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L₃. At the end of 7 days the non-predation L₃ of Petri dishes using the method of Baermann were recovered. Difference (p < 0.01) between groups on reducing the average number of Ancylostoma spp. L₃ (percent reduction of 84%) was observed.ConclusionsThe results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae.     

Effects of alcohol on natural resistance to Ancylostoma caninum in mice.

Experiments were performed to study the effect of alcohol on natural immunity of Swiss albino mice during Ancylostoma caninum infection. Natural resistance is lowered in alcoholic mice; 40% alcohol by mouth produced significant effect after 20 daily injections. The effect of the drug on natural immunity depended upon the concentration and the period of consumption. Animals retained maximum number of larvae during alcohol treatment.     

Phosphoinositide-3-OH-kinase inhibitor LY294002 prevents activation of Ancylostoma caninum and Ancylostoma ceylanicum third-stage infective larvae

The developmentally arrested hookworm infective larva resumes development only after encountering specific host-mediated cues during invasion. These cues activate a signaling pathway that culminates in the resumption of development. In Ancylostoma caninum, activation is characterised by the resumption of feeding and the release of excretory/secretory products required for infection. The dauer stage of the free-living nematode Caenorhabditis elegans is a developmentally arrested stage analogous to the hookworm infective larva. Dauer larvae exit developmental arrest in response to environmental cues that indicate favorable conditions for reproduction and growth. Because of the similarity between dauer recovery and activation, exit from dauer provides a model for hookworm larval activation. An insulin-signaling pathway has been implicated in controlling exit from developmental arrest in both C. elegans dauers and A. caninum larvae. To further investigate the role of insulin signaling in hookworm larval activation, the phosphatidylinositol-3-OH kinase inhibitor LY294002 was tested for its effect on in vitro activation using the resumption of feeding as a marker for activation. LY294002 prevented feeding in A. caninum infective larvae stimulated with host serum filtrate and a glutathione-analogue, the muscarinic agonist arecoline, or the cell permeable cGMP-analogue 8-bromo-cGMP. Similar results were seen with the congeneric hookworm Ancylostoma ceylanicum. These data suggest that an insulin-signaling pathway mediates activation in hookworm larvae, as in C. elegans, and that the phosphatidylinositol-3-OH kinase inhibitor acts downstream of the cGMP and muscarinic signaling steps in the pathway. In A. caninum, LY294002 had no effect on the release of excretory/secretory products associated with activation, suggesting that the secretory pathway diverges from the activation pathway upstream of the phosphatidylinositol-3-OH kinase step. These results provide additional support for the insulin-signaling pathway as the primarily pathway for activation to parasitism in hookworm larvae.     

Ancylostoma ceylanicum infections in humans in Vietnam

Ancylostoma ceylanicum is recognized as the only zoonotic hookworm species that is able to mature into adult stage in the human intestine. While human infections caused by this hookworm species have been reported from neighboring countries and this hookworm is prevalent in dogs in Vietnam, human infection has never been reported in Vietnam. The present study, therefore, aimed to identify human infections with A. ceylanicum in Vietnam. A total of 526 fecal samples from the residents in Long An Province were collected and the presence of hookworm eggs was detected by the Kato-Katz method. The results indicated that the overall prevalence of human hookworm infection was 85/526 (16.2%). After filter paper culture, 3rd stage larvae were successfully obtained from 48 egg-positive samples. The larvae were identified for their species using semi-nested PCR-RLFP on the cox1 gene. As a result, two hookworm species were confirmed; single species infections with Necator americanus or A. ceylanicum, and mixed infections with both species were found in 47.9%, 31.3%, and 20.8% of the samples, respectively.     

Enzymes of the tricarboxylic acid cycle in Ancylostoma ceylanicum and Nippostrongylus brasiliensis.

Enzymes of the tricarboxylic acid (TCA) cycle and glyoxylate pathway were investigated in adults and infective larvae of Ancylostoma ceylanicum and Nippostrongylus brasiliensis, and their activities were compared with those obtained in rat liver. A complete sequence of enzymes of the TCA cycle, with most of them showing activities quite similar to those in the rat liver homogenate, was detected in adults of both species. All the enzymes except fumarase and malate dehydrogenase were located predominantly in mitochondria where they showed a variable distribution of activities between the soluble and the membranes fractions. Malate dehydrogenase and fumarase were found in both the mitochondria and the 9,000-g supernatant fraction. Succinyl CoA synthetase, which was present in minimum activity, appeared rate limiting. Enzymes of the glyoxylate pathway, particularly isocitrate lyase, seemed to aid the functioning of the Krebs cycle by allowing the formation of succinate from isocitrate. The infective larvae of both species also were found equipped with all the enzymes of the Krebs cycle. Nonetheless, only isocitrate lyase of the glyoxylate pathway could be detected in these parasites.