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PubMed1990

Hepatic microsomal cytochrome P450 system during experimental hookworm infection.

Experimental infection of golden hamsters with the hookworm, Ancylostoma ceylanicum, caused a profound decline in the hepatic microsomal cytochrome P450 content. Concomitant decrease was also noticed in aminopyrine N-demethylase and benzo[a]pyrene hydroxylase activities. However, aniline hydroxylase activity was only marginally elevated during the infection. Microsomal markers, viz., cytochrome b5, NADH-cytochrome-c reductase, and glucose-6-phosphatase, were not significantly altered. Hepatic tissue exhibited an accumulation of lipids, especially phospholipids, triglycerides, and cholesterol, resulting in fatty necrosis around the central vein region. Isolated hepatic microsomes showed a decrease in phosphatidylcholine content. Impairment in hepatic mixed function oxidase (MFO) activities was further confirmed by prolongation in hexobarbital sleeping time and zoxazolamine-induced paralysis. The hepatic MFO system of A. ceylanicum-infected hamsters responded qualitatively and quantitatively in a manner similar to that of control hamsters, upon stimulation with selective chemical inducers like phenobarbitone and 3-methylcholanthrene. Kinetic and in vitro substrate binding studies revealed that for aminopyrine the substrate affinity and the maximum enzyme activity (Vmax) were decreased, while for aniline the binding affinity was decreased and the binding capacity was enhanced. Results indicate specific/selective impairment of the hepatic microsomal cytochrome P450 system during hookworm infection and may have many practical implications in toxicology and pharmacology.

Ancylostomiasis↗

PubMed1994

Apparent feeding behaviour of ensheathed third-stage infective larvae of human hookworms.

The third-stage ensheathed infective larvae (L3) of hookworms are reputed to be non-feeding stages, and the L2-derived sheath is considered to be a barrier to macromolecular uptake and secretion. During a study of the lectin-binding characteristics of en- and exsheathed L3 larvae of Ancylostoma duodenale, fluorescein isothiocyanate conjugated lectins (con A and Ulex europaeus agglutinin) were repeatedly seen in the oesophageal lumen, as well as on the surface of L3 larvae encased in the L2 sheaths. Repetition of these experiments using Necator americanus L3 larvae yielded similar results. These observations suggest that ensheathed hookworm larvae can obtain/utilise and secrete macromolecules, despite the presence of a sheath.

Ancylostoma↗

PubMed1987

Hookworm infection in a rural community in south India and its association with haemoglobin levels.

Faecal samples were obtained from 1113 persons living in a rural area in South India, and the hookworm ova load (Necator americanus) was determined using Kato's thick smear method. Evidence of hookworm infection was present in 92%, 77% having a count of under 100 epg, 11% a count of 1000 to 1999 epg, and 4% having counts between 2000 and 12,000 epg. Females had significantly higher ova counts than males on the average, but age did not appear to have any effect. Haemoglobin was also estimated: 80% of adult males, 87% of adult females and 90% of children were anaemic. There was a significant negative association between ova load and haemoglobin level, and the decrease in haemoglobin for a doubling of the ova load was estimated by regression analysis to be 0.18, 0.29 and 0.16 g/dl in adult males, adult females and children, respectively. There was nearly perfect agreement in the ranking of 10 clusters by mean ova count and mean haemoglobin level or percentage with anaemia.

Adolescent↗

PubMed1990

Human eosinophilic enteritis caused by dog hookworm Ancylostoma caninum.

An epidemic of eosinophilic enteritis (93 cases) has occurred in Townsville, northern Queensland, Australia. A hookworm was found attached to a resected, inflamed ileal segment from 1 patient but the species could not be identified. An adult hookworm of species Ancylostoma caninum was recovered at colonoscopy from the terminal ileum of a later patient. All of 38 patients interviewed in an epidemiological survey described behaviour which could have exposed them to infective larvae of this widespread dog parasite.

Adolescent↗

PubMed2006

The dingo non-long terminal repeat retrotransposons from the genome of the hookworm, Ancylostoma caninum.

Members of the retrotransposable element (RTE) clade of non-long terminal repeat (LTR) retrotransposon are widely distributed among eukaryote taxa, with representatives known from Caenorhabditis elegans, mammals, mosquitoes, schistosomes, and other taxa. An RTE retrotransposon has not, however, been characterized in detail from a parasitic nematode. Here, we characterize two discrete copies of an RTE-like non-LTR retrotransposon from the genome of the dog hookworm, Ancylostoma caninum. The elements were named dingo-1 and dingo-2. The full-length dingo-1 and dingo-2 elements were 3421 and 3171bp in length, respectively. They exhibited 54% nucleotide sequence identity to one another across their entire length and 40%/58% amino-acid sequence identity/similarity across their open reading frames. dingo-1 and dingo-2 exhibited hallmark structures and sequences of non-LTR retrotransposons of the RTE family including a single open reading frame encoding apurinic-apyrimidinic endonuclease (EN) and reverse transcriptase (RT), in that order. Phylogenetic analyses targeting the RT and the EN domains both confirmed that dingo-1 and dingo-2 were members of the RTE clade and that they were closely related to RTE-1 from C. elegans, to BDDF from Bos taurus and to SR2 from Schistosoma mansoni. Dot blot hybridization indicated that as many as 100-1000 copies of dingo-1 reside within the genome of A. caninum, while detection by RT-PCR of transcripts encoding dingo-like elements suggested that dingo-1 and -2 may be retrotranspositionally active within the genome of A. caninum. The dingo elements are the first retrotransposons to be characterized from a hookworm genome.

Ancylostoma↗

PubMed2006

A survey of the intestinal transcriptomes of the hookworms, Necator americanus and Ancylostoma caninum, using tissues isolated by laser microdissection microscopy.

The gastrointestinal tracts of multi-cellular blood-feeding parasites are targets for vaccines and drugs. Recently, recombinant vaccines that interrupt the digestion of blood in the hookworm gut have shown efficacy, so we explored the intestinal transcriptomes of the human and canine hookworms, Necator americanus and Ancylostoma caninum, respectively. We used Laser Microdissection Microscopy to dissect gut tissue from the parasites, extracted the RNA and generated cDNA libraries. A total of 480 expressed sequence tags were sequenced from each library and assembled into contigs, accounting for 268 N. americanus genes and 276 A. caninum genes. Only 17% of N. americanus and 36% of A. caninum contigs were assigned Gene Ontology classifications. Twenty-six (9.8%) N. americanus and 18 (6.5%) A. caninum contigs did not have homologues in any databases including dbEST-of these novel clones, seven N. americanus and three A. caninum contigs had Open Reading Frames with predicted secretory signal peptides. The most abundant transcripts corresponded to mRNAs encoding cholesterol-and fatty acid-binding proteins, C-type lectins, Activation-Associated Secretory Proteins, and proteases of different mechanistic classes, particularly astacin-like metallopeptidases. Expressed sequence tags corresponding to known and potential recombinant vaccines were identified and these included homologues of proteases, anti-clotting factors, defensins and integral membrane proteins involved in cell adhesion.

Amino Acid Sequence↗

PubMed2001

Giardia trophozoites in dog hookworm, Ancylostoma caninum: accident or hyperparasitism?

During the course of transmission electron microscopic studies of adult Ancylostoma caninum removed from a dog, several Giardia trophozoites were found in sections of the buccal cavity, oesophagus and intestine of several hookworms. Although the protozoa appeared viable, this unusual finding probably represents accidental uptake by, rather than an established infection of, the hookworm. It is feasible, however, that the trophozoites might have survived and even multiplied in this aberrant site.

Ancylostoma↗

PubMed1996

Human enteric infection with Ancylostoma caninum: hookworms reappraised in the light of a "new" zoonosis.

Recent studies in northeastern Australia indicate that enteric infection with Ancylostoma caninum is a leading cause of human eosinophilic enteritis. Much more frequent accompaniments of this infection are obscure abdominal pain with or without blood eosinophilia, while a large part of the population is probably infected asymptomatically. These conclusions are based on extensive serological investigations in patients and control subjects, as well as 15 cases in which single, adult hookworms were identified in situ in patients. In no case has more than one worm been identified, and none has been fully mature, so the infections have never been patent. Aphthous ulcers of the terminal ileum, caecum and colon have been seen in association with this infection and have also been observed in almost 5% of patients who are colonoscoped in north Queensland. Serodiagnosis has relied on an IgG and IgE ELISA using excretory-secretory antigens from adult A. caninum, but Western blot using these antigens to identify IgG4 antibodies to a protein of molecular weight 68 kDa (Ac68) promises to be more specific and sensitive. However, identical antigens appear to be secreted by the anthropophilic hookworms as well. The clinical, public health and biological significance of these findings are discussed in detail.

Adult↗

PubMed1998

Electron and light microscopy of peritoneal cellular immune responses in mice vaccinated and challenged with third-stage infective hookworm (Ancylostoma caninum) larvae.

The role of peritoneal macrophages in a murine model of immunity to living hookworm third-stage larvae (L3) was investigated. Mice immunized orally with 500 L3 once every 2 weeks for three times were challenged intraperitoneally with 2000 L3 1 week after the final immunization. The challenged larvae were collected from the peritoneal cavity at intervals between 2 and 72 h and then examined by inverted light microscopy, scanning electron microscopy and transmission electron microscopy. Peritoneal cellular responses in non-immunized mice served as negative controls. The numbers of peritoneal macrophages in immunized mice were 6-7-fold higher than in non-immunized mice. In the peritoneal cavity of immunized mice, host macrophages adhered to the challenged L3 within 2 h and created a cocoon-like encasing which surrounded the parasite. Extensive damage to the L3 was observed which included swelling, collapse and deformation of the larval cuticle. Lysis and vacuolization of the parasite's internal structures were also observed. In contrast, no significant cellular adherence and damage were observed in L3 recovered from non-immunized mice. L3-specific antibody levels were also elevated in the peritoneum of immunized mice relative to non-immunized controls. These studies implicate macrophages as important effector cells in hookworm larval vaccine immunity.

Ancylostoma↗

PubMedDate not supplied

Necator americanus: maintenance through one hundred generations in golden hamsters (Mesocricetus auratus). I. Host sex-associated differences in hookworm burden and fecundity.

The human hookworm Necator americanus was maintained through one hundred generations in the golden hamster (Mesocricetus auratus). The parasite strain employed here was ultimately adapted to hamsters without the requirement for exogenous steroids or other immunosuppressive agents. Moreover, there was no requirement to use neonatal hamsters--successful infections were obtained in 9- to 10-week-old hamsters infected subcutaneously with 250 hookworm larvae. This unique adaptation of N. americanus to hamsters permits its use for purposes of anthelminthic drug and vaccine development.

Animals↗

PubMed1999

Sequence differences in the internal transcribed spacers of DNA among four species of hookworm (Ancylostomatoidea: Ancylostoma).

The two ribosomal DNA internal transcribed spacers (1 and 2) of the hookworms Ancylostoma caninum, A. tubaeforme, A. ceylanicum and A. duodenale were sequenced. The sequence lengths were similar among the four species, except that A. ceylanicum had slightly longer (by 5-7 bp) internal transcribed spacer 1 and 2 sequences. The predicted secondary structure of the internal transcribed spacer 2 precursor rRNA was similar for all species, despite interspecific differences in primary sequence ranging from 0.9% to 13.2%. Interspecific differences in internal transcribed spacer 1 sequence ranged from 0.9% to 7.5%. A cladistic analysis of the sequence data, using the human hookworm Necator americanus as the outgroup, provided little resolution of the phylogenetic relationships, except that A. ceylanicum occurred on a branch external to the other three species. Nonetheless, internal transcribed spacers 1 and 2 may provide useful phylogenetic information at higher taxonomic levels within the superfamily Ancylostomatoidea.

Ancylostoma↗

PubMedDate not supplied

The use of human faeces for fertilizer is associated with increased intensity of hookworm infection in Vietnamese women.

To investigate different factors associated with hookworm infections we conducted 2 studies in a commune in northern Viet Nam. The first was part of a larger study on anaemia and covered 213 women (15-49 years of age) and their 92 children (6 months to 5 years of age) in one commune; 90% of the families reported using human faeces for fertilizer. Women who reported using fresh human faeces as fertilizer had significantly higher hookworm egg counts than women who either used treated human faeces or who did not use human faeces as fertilizer. The second study examined how human faeces were used for fertilizer in 30 selected families. Women participated in preparation and application of human faeces to crops in 81% of the families using human faeces for fertilizer. Two methods of preparing the faeces were described: 48% of the families mixed the faeces with ash before applying them to the field; 18% mixed the faeces with water; 33% used both methods.

Adolescent↗

PubMedDate not supplied

Comparison of the efficacy of mebendazole, albendazole and pyrantel in treatment of human hookworm infections in the southern region of Mali, West Africa.

A randomized, placebo-controlled trial of the efficacy of pyrantel (single dose 12.5 mg/kg bodyweight), mebendazole (single 500 mg dose) and albendazole (single 400 mg dose) in the treatment of hookworm infections (Necator americanus) was carried out in January 1998 in the Southern Region of Mali, West Africa, during the period of Ramadan (Islamic fast). Statistical analysis of the pre-intervention faecal egg counts showed that there was a significant pre-treatment chance bias, despite randomization of subjects into treatment groups, arising from the main effect of sex (heavier infections among males) and a sex x treatment interaction (the sex bias was not evident in the pyrantel-treatment group). The participants were re-examined 10 days after treatment, and after controlling for the drift in faecal egg counts in the placebo-treated subset, age, sex, fasting and intensity of infection, albendazole was clearly the most effective drug showing consistently efficacies in the range 92.1 to 99.7%, depending on the method of evaluation and the particular subset of the treatment group. Neither mebendazole nor pyrantel was as effective, with efficacies ranging from 60.9 to 89.8% and 4.8 to 89.7%, respectively. Fasting made no difference to drug efficacy. On the basis of our results the single 400 mg dose of albendazole is the treatment of choice for hookworm infections in this region of Mali. We emphasize the need for standardization of the methods used for trial designs, for calculation of summary data relating to drug efficacies and the accompanying statistical tests.

Adolescent↗

PubMed1999

Antibody responses in pregnancy-induced transmammary transmission of Ancylostoma caninum hookworm larvae.

Third stage larvae of the Ancylostoma caninum hookworm nematode have the capacity to infect a dog, abort the normal maturation pathway to become blood-feeding intestinal worms, and instead distribute throughout the body in a developmentally arrested state that is relatively resilient to most chemotherapeutic agents. During pregnancy, a percentage of the arrested larvae reactivate and transmit via the mammary glands to infect the nursing puppies with resulting iron-deficiency anemia and potential mortality. To determine if the suppression of parasite-specific antibody responses during pregnancy facilitates the reactivation and transmammary transfer of hookworm larvae, a murine model of A. caninum infection was used to compare the infected versus uninfected animals that were either bred or not bred. Initial comparisons of genetically divergent BALB/c versus C57BL/6 mice showed that both the strains mounted strong Th2 biased IgG1 and IgE antibody responses to A. caninum infection. Using the BALB/c strain for the breeding analyses, it was confirmed that larval transfer to the mouse pups only occurred during the post-partum lactational period. In the dams, levels of total and antigen-specific IgG1 and total IgE were highly correlated with parasite burden. During most phases of pregnancy and lactation, infected dams had lower total IgG1, IgG2a and IgE levels as compared to unbred mice at comparable times post-infection; this downward modulation of antibody responses supports the established dogma of a generalized immunosuppression associated with pregnancy. However, at parturition and post-partum lactation, antigen-specific IgG1 levels measured at 1:5000 serum dilutions were comparable between bred and unbred mice, and antigen-specific IgG2a levels at 1:100 serum dilutions were also not significantly different except for a marginal reduction in the bred mice at the lactational timepoint. The comparable anti-A. caninum IgG1 levels between bred and unbred mice, and low correlation between IgG2a levels and larval burden suggest that parasite-specific antibody responses do not play a major role in the pregnancy-associated transmammary transmission of A. caninum larvae. This conclusion does not rule out the possibility that underlying fluxes in the levels of specific cytokines associated with pregnancy and infection may be involved in the process of larval reactivation and transmission.

Ancylostoma↗

PubMed2002

Molecular cloning and characterization of Ac-mep-1, a developmentally regulated gut luminal metalloendopeptidase from adult Ancylostoma caninum hookworms.

A zinc metalloendopeptidase cDNA (Ac-mep-1) was cloned from Ancylostoma caninum adult hookworms. Ac-mep-1 is encoded by a 2.8 kb mRNA with a predicted open reading frame (ORF) of 870 amino acids (predicted pI=5.5, m.w.=98.7 kDa) that contains four potential N-linked glycosylation sites and predicted zinc-binding domains (HExxH and ENxADxGG). These domains represent signature sequences of the Neutral Endopeptidase 24.11 (neprilysin) family of enzymes. The ORF corresponding to Ac-MEP-1 exhibited strong similarity to metalloproteases from the trichostrongyle Haemonchus contortus as well as Caenorhabditis elegans. RT-PCR analysis of A. caninum eggs, L1, non-activated and activated L3 and adult cDNA identify transcription of Ac-MEP-1 only in the adult stage of the parasite. Mouse antibody raised to the expressed protein recognized proteins of approximately 90 and 100 kDa in adult hookworm extracts. Adult worm sections probed with these antisera localized Ac-mep-1 to the microvilli of the worm gastrointestinal tract suggesting a possible role for this enzyme in digestion of the parasite blood meal.

Amino Acid Sequence↗

PubMed2000

Purification of a diagnostic, secreted cysteine protease-like protein from the hookworm Ancylostoma caninum.

The enteric infection of humans with the canine hookworm Ancylostoma caninum varies in its clinical presentation, ranging from asymptomatic to eosinophilic gastroenteritis requiring surgical intervention. Infections are not patent, but can be diagnosed immunologically by detecting antibodies to an immunodominant secreted hookworm protein termed Ac68. To characterise Ac68, we purified the native protein from A. caninum excretory/secretory products using size exclusion followed by anion exchange chromatography. The epitopes in the purified protein recognised by human infection sera were shown to be proteins and not carbohydrates. The N-terminal amino acid sequence of the purified Ac68 was determined and six of the 11 residues obtained were shared with a previously characterised cysteine protease of A. caninum, AcCP1.

Amino Acid Sequence↗

PubMed1993

Observations on the feeding behaviour of parasitic third-stage hookworm larvae.

The feeding behaviour of parasitic 3rd-stage larvae (L3) of the hookworms Ancylostoma caninum, A. ceylanicum and Necator americanus was examined. Less than 11% of A. caninum L3 recovered from the small intestines of dogs infected orally were feeding at 4-48 h post-infection (p.i.), and none of the A. ceylanicum L3 recovered from the intestines of orally infected hamsters had resumed feeding. All L4 of both species recovered at 36 and 48 h p.i. had resumed feeding. On the other hand, approximately 16% of the A. ceylanicum L3 recovered from the skin of percutaneously infected hamsters at 18 h were feeding, and the percentage feeding increased to nearly 58% at 44 h p.i. Necator americanus L3 recovered from the skin of percutaneously infected neonatal hamsters resumed feeding at 6-12 h p.i. and reached 90-94% by 18 h. Feeding began to decline at 66 h, and reached 29% at 120 h p.i. This decrease was associated with the migration of larvae from the skin to the lungs. By 192 h p.i. over 95% of the larvae had reached the small intestine, and all had moulted to the L4. The results indicate that parasitic L3 resume feeding in the skin during percutaneous infections, and suggest that feeding by hookworm L3 correlates with the resumption of development.

Ancylostoma↗

PubMed2006

Saposin-like proteins from the intestine of the blood-feeding hookworm, Ancylostoma caninum.

Hookworms feed on blood, utilizing haemoglobin for nutrition, growth and reproduction. The haemoglobin digestion cascade has been partially elucidated, but the process immediately preceding this event, haemolysis, has received considerably less attention. We have cloned and expressed Ancylostoma caninum mRNAs encoding 2 proteins belonging to the saposin-like protein (SAPLIP) family, termed Ac-slp-1 and Ac-slp-2. The open reading frames of SLP-1 and SLP-2 were used to identify expressed sequence tags encoding SAPLIPs from the 4 major clades of animal parasitic nematodes. Both Ac-slp-1 and slp-2 mRNAs were shown to be expressed in all life stages assessed, with slp-1 predominantly being expressed in third-stage larvae (L3) before and after activation with dog serum. Recombinant SLP-1 and SLP-2 were expressed in insect cells and used to raise specific antisera in mice. These antisera were used as probes in fluorescence microscopy to localize the anatomic expression sites of both proteins to small, punctate organelles or vesicles within the intestinal cells of adult worms; weak staining was detected on the microvillar brush border of the intestine. Using transmission electron microscopy, both proteins were localized to similar vesicles in the intestinal cells of the L3. Recombinant proteins contained C-terminal purification tags that potentially precluded dimerization and possibly interfered with the subsequent detection of haemolytic activity. Their expression in the gut of the L3 and adult stages suggests a role for these hookworm SAPLIPs in the lysis of host cells during tissue migration and/or feeding.

Amino Acid Sequence↗