Uncinaria hamiltoni (Nematoda: Ancylostomatidae) in South American sea lions, Otaria flavescens, from northern Patagonia, Argentina

Thirty-one South American sea lion pups (Otaria flavescens) found dead in Punta León, Argentina, during the summer of 2002, were examined for hookworms (Uncinaria hamiltoni). Parasite parameters were analyzed in 2 locations of the rookery, i.e., a traditional, well-structured breeding area and an expanding area with juveniles and a lax social structure. Prevalence of hookworms was 50% in both localities, and no difference was observed in prevalence between pup sexes (P > 0.05). Hookworms were concentrated in the small intestine. Transmammary transmission is assumed because only adult hookworms were found in the pups. The mean intensity of hookworms per pup was 135; the mean intensity in females (92.78) was significantly different (P < 0.05) from that of males (230.25). No difference (P > 0.05) in intensity was found between the 2 breeding areas, although prevalence was higher in the traditional breeding area than in the other area. Location was the only factor affecting hookworm prevalence (P log-linear model: 0.9552; chi2: 1.5629). No apparent trend between body condition and intensity of hookworms was observed.     

Australian hookworms (Ancylostomatoidea): a review of the species present,their distributions and biogeographical origins

Ancylostomatoidea or hookworms recorded in Australia are reviewed and the attempt is made to provide the biogeographical background to their occurrence. The poor representation of this nematode superfamily is probably a reflection of the fact that they are primarily parasites of Carnivora, Artiodactyla, Insectivora, Rodentia, Edentata, Proboscoidea and primates, eutherian mammals which are either absent from the Australian fauna or which have only recently reached the continent. The principal genera of hookworms recorded to date from Australia are Ancylostoma, Bunostomum, Necator and Uncinaria. The majority of the ancylostomatoid fauna is represented by introduced species of man and domestic animals. Native or endemic species of hookworms are restricted to members of the genus Uncinaria with two species occurring in rodents and pinnipeds. Only a single endemic species of hookworm is known, U. hydromyidis, which is found in the small intestine of a rat. Significant problems remain in understanding the systematics, epidemiology and evolutionary relationships of the Australian ancylostomatoid fauna.     

Hookworms in the Americas: An alternative to trans-Pacific contact

Recent New World archaeoparasitological discoveries of hookworm in pre-European contexts have revived a long-standing debate about the origin of hookworms in the Americas. Historically, the climatic conditions of Beringia encountered by migrating people have been considered too harsh for tropical hookworms to survive, suggesting that hookworms must have been introduced into South America by 'storm-tossed' fisherman or explorers from Asia. Here, John Hawdon and Susan Johnston review the history of hookworms in the Americas, and propose an alternative to their trans-Pacific introduction based on the unique natural history of one of the human hookworms.     

Genes and genomes of Necator americanus and related hookworms

The human hookworms (Necator americanus and Ancylostoma duodenale) infect over one billion people. The phylogenetic relationships of the human hookworms suggest independent acquisition of the human host. The hookworms probably have a haploid chromosome number n = 6, and an XX-XO sex determination mechanism is likely to be used. Genetic and molecular research on hookworms is in its infancy, but several important genes and gene products have already been identified. Of note are cathepsin genes, a family of secreted proteins known as Ancylostoma activation-associated proteins and a family of anticoagulants. The inception of an expressed sequence tag program on the human hookworm, N. americanus, promises to yield many new genes with novel functions in the biology of these important parasites.     

Utility of capsule endoscopy for evaluating anthelmintic efficacy in fully conscious dogs

The current accepted standard for evaluating the efficacy of gastrointestinal anthelmintic drugs is necropsy of infected animals followed by a comparison of worm counts between treated and non-treated groups. In this study capsule endoscopy, a minimally invasive method of imaging the small intestine of humans, is evaluated as a possible alternative to necropsy for the purposes of worm quantification in dogs. Eighteen Beagle dogs were included in this study. These dogs were part of a separate trial intended to determine the efficacy of various candidate parasiticides against Ancylostoma caninum via the necropsy standard. Dogs were inoculated with A. caninum L3s 4 weeks prior to treatment with one of the candidate compounds; a control group (n=8) received no treatment. Capsule endoscopy was performed 6-14 days post-treatment, followed by necropsy the following day. Seventeen dogs had complete examinations, i.e. the capsule traversed the small intestine and reached the colon within the battery life of the capsule. A strong correlation (r(s)=0.87, P<0.0001) was observed between the worm counts acquired by capsule endoscopy and necropsy. There was no clear relationship between the ability of the capsule endoscope to detect hookworms and either visibility of the intestinal lumen or small intestinal transit time. Generation of a virtual spatial record of hookworm location from the capsule endoscopy data revealed a temporal trend, with the majority of worms present in the proximal small intestine in the morning versus the central to distal small intestine in the afternoon. Worm distribution as determined by capsule endoscopy closely resembled post-mortem findings. In conclusion, capsule endoscopy shows promise as an alternative to necropsy for the enumeration of A. caninum in the canine small intestine, although further work is required to improve completion rates and optimise intestinal examination.     

Molecular and morphometric evidence for separate species of Uncinaria (Nematoda: Ancylostomatidae) in California sea lions and northern fur seals: hypothesis testing supplants verification

California sea lions (Zalophus californianus) and northern fur seals (Callorhinus ursinus) are each believed to host distinct hookworm species (Uncinaria spp.). However, a recent morphometric analysis suggested that a single species parasitizes multiple pinniped hosts, and that the observed differences are host-induced. To explore the systematics of these hookworms and test these competing hypotheses, we obtained nucleotide sequences of nuclear ribosomal DNA (D2/D3 28S, D18/D19 28S, and internal transcribed spacer [ITS] regions) from 20 individual hookworms parasitizing California sea lion and northern fur seal pups where their breeding grounds are sympatric. Five individuals from an allopatric population of California sea lions were also sampled for ITS-1 and D18/D19 28S sequences. The 28S D2/D3 sequences showed no diagnostic differences among hookworms sampled from individual sea lions and fur seals, whereas the 28S D18/D19 sequences had one derived (apomorphic) character demarcating hookworms from northern fur seals. ITS sequences were variable for 7 characters, with 4 derived (apomorphic) states in ITS-1 demarcating hookworms from California sea lions. Multivariate analysis of morphometric data also revealed significant differences between nematodes representing these 2 host-associated lineages. These results indicate that these hookworms represent 2 species that are not distributed indiscriminately between these host species, but instead exhibit host fidelity, evolving independently with each respective host species. This evolutionary approach to analyzing sequence data for species delimitation is contrasted with similarity-based methods that have been applied to numerous diagnostic studies of nematode parasites.     

The mucosal cellular response to infection with Ancylostoma ceylanicum

Although hookworms are known to stimulate inflammatory responses in the intestinal mucosa of their hosts, there is little quantitative data on this aspect of infection. Here we report the results of experiments conducted in hamsters infected with Ancylostoma ceylanicum. Infection resulted in a marked increase in goblet cells in the intestinal mucosa, which was dependent on the number of adult worms present and was sustained as long as worms persisted (over 63 days) but returned to baseline levels within 7 days of the removal of worms by treatment with ivermectin. Increased mast cell responses were also recorded. Levels were again dependent on the intensity of worm burdens and lasted as long as 63 days after infection. When worms were eliminated, mast cell numbers took over 2 weeks to return to normal. Paneth cell numbers fell soon after infection, the degree of reduction being dependent on the worm burden. After clearance of worms, Paneth cell numbers returned to normal within a week, but then rebounded and numbers rose to higher levels than those in control na?ve animals. The time course of the response was similar whether animals experienced a chronic low-intensity infection without loss of worms or a higher intensity infection during the course of which worm burdens were gradually reduced. Clearly, A. ceylanicum was able to induce a marked inflammatory response in its host's intestine which was sustained for over 9 weeks after infection, and which hamsters appeared able to tolerate well. Our data draw attention to the resilience of hookworms which, unlike many other nematodes, are able to survive for many weeks in a highly inflamed intestinal tract.     

The survival strategies of hookworms

Carefully conducted field studies of human necatoriasis have failed to demonstrate the presence of strong immunity, indicating that hookworms have evolved survival strategies to counteract specific immune responses that undoubtedly occur in the host. David Pritchard focuses on these recent field data and on our current knowledge of the biochemical nature of molecules produced by hookworms which have the potential to neutralize host defences and ensure parasite survival.     

Effect of vaccinations with recombinant fusion proteins on Ancylostoma caninum habitat selection in the canine intestine

Laboratory dogs were vaccinated subcutaneously with 3 different recombinant fusion proteins, each precipitated with alum or calcium phosphate. The vaccinated dogs were then challenged orally with 400 third-stage infective larvae (L3) of the canine hookworm, Ancylostoma caninum. The 3 A. caninum antigens selected were Ac-TMP, an adult-specific secreted tissue inhibitor of metalloproteases; Ac-AP, an adult-specific secreted factor Xa serine protease inhibitor anticoagulant; and Ac-ARR-1, a cathepsin D-like aspartic protease. Each of the 3 groups comprised 6 male beagles (8 +/- 1 wk of age). A fourth group comprised control dogs injected with alum. All of the dogs vaccinated with Ac-TMP or Ac-APR-1 exhibited a vigorous antigen-specific antibody response, whereas only a single dog vaccinated with Ac-AP developed an antibody response. Dogs with circulating antibody responses exhibited 4.5-18% reduction in the numbers of adult hookworms recovered from the small intestines at necropsy, relative to alum-injected dogs. In contrast, there was a concomitant increase in the number of adult hookworms recovered from the colon. The increase in colonic hookworms was as high as 500%, relative to alum-injected dogs. Female adult hookworms were more likely to migrate into the colon than were males. Anti-enzyme and anti-enzyme inhibitor antibodies correlated with an alteration in adult hookworm habitat selection in the canine gastroinntestinal tract.     

Hookworm antigens: the potential for vaccination

Hookworms rank with Ascaris as the most prevalent soil-transmitted helminths of man. Up to 1000 million people may be infected. Not all suffer the life-threatening anaemia that reflects heavy burdens of hookworms, but even mild iron-deficiency anaemia due to hookworms can lead to intellectual and growth retardation - especially among infected children. Health education, mass chemotherapy and the sanitary disposal of faeces have been the traditional mainstays of hookworm control, but more recently chemotherapy targeted only to heavily infected individuals who appear predisposed to hookworm anaemia has emerged as a realistic alternative. Ultimately, however, knowledge of the molecular immunology of hookworm infection may provide the basis for rational vaccine development. Although the evidence for acquired immunity to hookworms in man is not strong, work is now underway to identify and to characterize antigens from third and fourth larval stages and adults of Necator americanus and species of Ancylostoma (Table 1). As this article shows, the work provides real promise for improved immunodiagnosis and possible vaccination.     

Intestinal helminthiases and schistosomiasis among school children in an urban center and some rural communities in southwest Nigeria

Intestinal helminths and schistosomiasis among school children were investigated in an urban and some rural communities of Ogun State, southwest Nigeria. Fecal samples of 1,059 subjects (524 males, 535 females) aged 3-18 years were examined using direct smear and brine concentration methods between June 2005 and November 2006. The pooled prevalence of infection was 66.2%. Ascaris lumbricoides showed the highest prevalence (53.4%) (P < 0.001) followed by hookworms (17.8%), Trichuris trichiura (10.4%), Taenia sp. (9.6%), Schistosoma mansoni (2.3%), Strongyloides stercoralis (0.7%), Schistosoma haematobium (0.6%), and Enterobius vermicularis (0.3%). The prevalences of A. lumbricoides, hookworms, Taenia sp., S. mansoni, and S. stercoralis in the urban centre were similar (P > 0.05) to those in the rural communities. The fertile and infertile egg ratios of A. lumbricoides in the urban centre and the rural communities were 13: 1 and 3.7: 1, respectively. Each helminth had similar prevalences among both genders (P > 0.05). The prevalence of A. lumbricoides increased significantly with age (P < 0.001). The commonest double infections were Ascaris and hookworms, while the commonest triple infections were Ascaris, hookworms, and Trichuris. The study demonstrates the need for urgent intervention programmes against intestinal helminthiases and schistosomiasis in the study area.     

DNA technological progress toward advanced diagnostic tools to support human hookworm control

Blood-feeding hookworms are parasitic nematodes of major human health importance. Currently, it is estimated that 740 million people are infected worldwide, and more than 80 million of them are severely affected clinically by hookworm disease. In spite of the health problems caused and the advances toward the development of vaccines against some hookworms, limited attention has been paid to the need for improved, practical methods of diagnosis. Accurate diagnosis and genetic characterization of hookworms is central to their effective control. While traditional diagnostic methods have considerable limitations, there has been some progress toward the development of molecular-diagnostic tools. The present article provides a brief background on hookworm disease of humans, reviews the main methods that have been used for diagnosis and describes progress in establishing polymerase chain reaction (PCR)-based methods for the specific diagnosis of hookworm infection and the genetic characterisation of the causative agents. This progress provides a foundation for the rapid development of practical, highly sensitive and specific diagnostic and analytical tools to be used in improved hookworm prevention and control programmes.     

Mechanistic and Single-Dose In Vivo Therapeutic Studies of Cry5B Anthelmintic Action against Hookworms

Background

Hookworm infections are one of the most important parasitic infections of humans worldwide, considered by some second only to malaria in associated disease burden. Single-dose mass drug administration for soil-transmitted helminths, including hookworms, relies primarily on albendazole, which has variable efficacy. New and better hookworm therapies are urgently needed. Bacillus thuringiensis crystal protein Cry5B has potential as a novel anthelmintic and has been extensively studied in the roundworm Caenorhabditis elegans. Here, we ask whether single-dose Cry5B can provide therapy against a hookworm infection and whether C. elegans mechanism-of-action studies are relevant to hookworms.

Methodology/Principal Findings

To test whether the C. elegans invertebrate-specific glycolipid receptor for Cry5B is relevant in hookworms, we fed Ancylostoma ceylanicum hookworm adults Cry5B with and without galactose, an inhibitor of Cry5B-C. elegans glycolipid interactions. As with C. elegans, galactose inhibits Cry5B toxicity in A. ceylanicum. Furthermore, p38 mitogen-activated protein kinase (MAPK), which controls one of the most important Cry5B signal transduction responses in C. elegans, is functionally operational in hookworms. A. ceylanicum hookworms treated with Cry5B up-regulate p38 MAPK and knock down of p38 MAPK activity in hookworms results in hypersensitivity of A. ceylanicum adults to Cry5B attack. Single-dose Cry5B is able to reduce by >90% A. ceylanicum hookworm burdens from infected hamsters, in the process eliminating hookworm egg shedding in feces and protecting infected hamsters from blood loss. Anthelmintic activity is increased about 3-fold, eliminating >97% of the parasites with a single 3 mg dose (∼30 mg/kg), by incorporating a simple formulation to help prevent digestion in the acidic stomach of the host mammal.

Conclusions/Significance

These studies advance the development of Cry5B protein as a potent, safe single-dose anthelmintic for hookworm therapy and make available the information of how Cry5B functions in C. elegans in order to study and improve Cry5B function against hookworms.     

Structural and functional characterization of a secreted hookworm Macrophage Migration Inhibitory Factor (MIF) that interacts with the human MIF receptor CD74

Hookworms, parasitic nematodes that infect nearly one billion people worldwide, are a major cause of anemia and malnutrition. We hypothesize that hookworms actively manipulate the host immune response through the production of specific molecules designed to facilitate infection by larval stages and adult worm survival within the intestine. A full-length cDNA encoding a secreted orthologue of the human cytokine, Macrophage Migration Inhibitory Factor (MIF) has been cloned from the hookworm Ancylostoma ceylanicum. Elucidation of the three-dimensional crystal structure of recombinant AceMIF (rAceMIF) revealed an overall structural homology with significant differences in the tautomerase sites of the human and hookworm proteins. The relative bioactivities of human and hookworm MIF proteins were compared using in vitro assays of tautomerase activity, macrophage migration, and binding to MIF receptor CD74. The activity of rAceMIF was not inhibited by the ligand ISO-1, which was previously determined to be an inhibitor of the catalytic site of human MIF. These data define unique immunological, structural, and functional characteristics of AceMIF, thereby establishing the potential for selectively inhibiting the hookworm cytokine as a means of reducing parasite survival and disease pathogenesis.     

Secretory cholinesterase of Ancylostoma ceylanicum: effect of tubulin binding agents and benzimidazole anthelmintics.

Ancylostoma ceylanicum, the human hookworm parasite, exhibited significant secretion of cholinesterase when maintained in vitro in RPMI-1640 medium. Secretion of the enzyme was linear up-to 4 hours of incubation. About 40 percent of the total cholinesterase activity was localized in the soluble fraction, while remaining activity was associated with the particulate fraction of the nematode. Exposure of the hookworms to colchicine in vitro caused significant inhibition in secretion of the enzyme by the parasite with concomitant accumulation of cholinesterase within the adult worms. Vinblastine did not show noticeable effect on the enzyme secretion as well as activity within the parasite. Incubation of hookworms with some benzimidazole anthelmintics viz., mebendazole or albendazole significantly reduced the capacity of the worms to secrete cholinesterase and increase in enzyme activity within the parasite. Adult worms recovered from mebendazole treated hamsters exhibited about 3 fold greater activity of cholinesterase as well as significantly lower capacity to secrete cholinesterase in vitro as compared to the worms recovered from untreated animals. These observations indicate role of microtubules in the secretion of cholinesterase by hookworms and as a target for the action of benzimidazole anthelmintics.     

四川动物体内发现的钩口线虫

报道寄生于四川17种动物体内的钩口线虫12种,它们隶属于钩口科的6个属,亦记述了部份宿主的感染情况。     

Iron deficiency in an area of Bolivia and high prevalence of intestinal helminthiasis

Between November and December 1987, 201 subjects from three communities of the Cordillera Province, Santa Cruz Department, South-East Bolivia (Camiri, Boyuibe, and Gutierrez) were examined for serum iron levels. Sixty-three (31.3%) were iron deficient. There were no significant differences between males and females (37.2% and 26.9%, respectively) and among the three localities studied. Iron deficiency was highest in the 3- to 9-year-old age group and lowest in the 30- to 39-year-old group. Considering the prevalence in the same localities of infections by hookworms, Trichuris trichiura, Strongyloides stercoralis, and Trichostrongylus, it is suggested that hookworms and possibly other intestinal parasites may be an important cause of the observed iron deficiency.     

Genetic structure of populations of the human hookworm, Necator americanus, in China

Twenty-one to 58 individual Necator americanus were sampled from each of four villages in south-western China. Each nematode was sequenced for 588 bp of the mitochondrial cytochrome oxidase I gene. Allelic and nucleotide diversity varied two-fold among villages. Overall FST among populations was approximately 0.28, but this large value resulted from one low-diversity population that had a large genetic distance to the other three populations (F(ST) = 0.10 without that population). There was no correlation between geographical and genetic distance among sites. Thus, the genetic structure of this species in China may be characterized by variable effective sizes and uneven movement among sites. We discuss the implications of this genetic structure for vaccine development and the spread of drug resistance in human hookworms, and compare the genetic structure of hookworms with that of other nematodes.