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The contribution of hookworm and other parasitic infections to haemoglobin and iron status among children and adults in western Kenya.

A cross-sectional study of 729 children and adults in western Kenya investigated the impact of infection with hookworm, Ascaris lumbricoides, Trichuris trichiura, Schistosoma mansoni and malaria on iron status. In bivariate analyses, hookworm intensities as low as 300 eggs/g of faeces were negatively related to levels of haemoglobin (Hb) and serum ferritin (SF). Malaria parasitaemia was negatively related to Hb and positively related to SF, while S. mansoni intensities were negatively related to SF. Multivariate regression analysis was done to identify predictors of Hb and SF levels. In children, age (in years) was the only predictor for Hb (B = 1.7 g/L) and only malaria parasitaemia (negative, light, moderate, heavy) was retained in the model for log10 SF (B = 0.097 microgram/L). In adults, hookworm infection and malaria parasitaemia together with age, sex, pregnancy, SF levels < 12 micrograms/L and elevated body temperature were significant predictors of low Hb. The regression coefficient for hookworm egg count (for increments of 100 eggs/g) was -1.3 g/L. Significant interactions between sex and age and between sex and malaria parasitaemia were revealed. Age and malaria parasitaemia were significant predictors only among females, with a regression coefficient for malaria parasitaemia of -6.9 g/L. The regression coefficient for hookworm did not change when SF < 12 micrograms/L was taken out of the model, indicating that the effect of hookworm cannot be explained by low iron stores alone. Using SF as the dependent variable, hookworm and S. mansoni intensities together with age and sex were retained in the model. The regression coefficients for hookworm egg count (increments of 100 eggs/g) and S. mansoni egg count (increments of 10 eggs/g) were -0.011 microgram/L and -0.012 microgram/L, respectively. Iron deficiency was a problem in this population and hookworm infections contributed significantly to this situation.

Adolescent↗

Hookworm infection: new developments and prospects for control.

PURPOSE OF REVIEW: Hookworm infection remains a major health burden in developing countries. Successful control will likely be achieved through continued advances in our understanding of the epidemiology, molecular biology and immunopathogenesis of hookworm infection. This review summarizes recent advances in each of these fields, and discusses ongoing efforts to develop vaccines against hookworm anemia and growth delay. RECENT DEVELOPMENTS: Revised estimates indicate that hookworms afflict over 700 million persons in tropical and subtropical regions. Prevalence and intensity often vary considerably at both the regional and local levels, and may be influenced by climate, soil composition, education, and socioeconomic status. Immunoepidemiological studies suggest that hookworm infection likely induces a complex mixture of host-protective and pathological immune responses. There has been substantial progress in elucidating the molecular pathogenesis of hookworm disease, primarily through the identification of a number of parasite virulence factors. Mass chemotherapy remains a mainstay of hookworm control strategies although continued use of benzimidazole anthelminthics is perhaps contributing to the development of anthelminthic resistance. Consequently, there remains a need for innovative approaches, including the development of vaccines and new chemotherapeutic agents, in order to provide effective global control of hookworm disease. SUMMARY: Hookworm infection and disease is a significant threat to global health. Recent advances, particularly those at the molecular level, have provided a wealth of opportunities to better understand pathogenesis. This will likely allow for the development of novel measures such as vaccines to complement existing control methods.

Ancylostoma↗

Hemoquant determination of hookworm-related blood loss and its role in iron deficiency in African children.

Iron deficiency remains the most prevalent form of human malnutrition, and current interventions to control it have not decreased the global prevalence. Hookworm control activities are becoming more widely implemented, but the importance of these efforts to prevent anemia in populations is not well-defined. We studied the relationships among hookworm infection, intestinal blood loss, and iron status of 203 Zanzibari school children. Helminth infection intensity was quantified by fecal egg counts, and iron deficiency anemia was defined by low hemoglobin and serum ferritin concentrations. Intestinal blood loss was quantified by measuring fecal heme and heme breakdown products as porphyrin, a noninvasive method that has not been used previously to assess hookworm blood loss. Intestinal blood loss was strongly and linearly related to hookworm egg counts. The degree of degradation of fecal heme indicated that blood loss occurred in the upper gastrointestinal tract, compatible with the behavior of hookworms. Trichuris trichiura and Ascaris lumbricoides infections were also common, but did not contribute significantly to intestinal blood loss in this population. The prevalence of iron deficiency anemia increased steadily as hookworm infection intensity and intestinal blood loss increased. In the context of a poor diet, as exists in Zanzibar and many tropical countries, hookworm-related blood loss contributes dramatically to anemia. In such contexts, hookworm control is a feasible and essential component of anemia control. Determination of fecal heme is relatively simple and noninvasive and may be a useful tool for measuring the impact of hookworm control activities.

Anemia, Iron-Deficiency↗

Hookworm vaccines: past, present, and future.

Hookworms are gastrointestinal nematodes that infect almost 1 billion people in developing countries. The main clinical symptom of human hookworm infections is iron-deficiency anaemia, a direct consequence of the intestinal blood loss resulting from the parasite's feeding behaviour. Although treatment is available and currently used for the periodic removal of adult hookworms from patients, this approach has not effectively controlled hookworm in areas of rural poverty. Furthermore, treated individuals remain susceptible to reinfection following exposure to third-stage infective hookworm larvae in the soil as early as 4-12 months after drug treatment. Therefore, a prophylactic vaccine against hookworm infection would provide an attractive additional tool for the public-health control of this disease. The feasibility of developing a vaccine is based on the previous success of an attenuated larval vaccine against canine hookworm. Several laboratory and field studies have explored the development of a human anti-hookworm vaccine, describing potential protective mechanisms and identifying candidate antigens, one of which is now in clinical trials. The current roadmap that investigators have conceived has been influenced by vaccine development for blood-feeding nematodes of livestock and companion animals; however, recombinant vaccines have yet to be developed for nematodes that parasitise animals or human beings. The roadmap also addresses the obstacles facing development of a vaccine for developing countries, where there is no commercial market.

Global Health↗

Cloning and characterisation of an aspartyl protease inhibitor (API-1) from Ancylostoma hookworms.

Hookworm infection persists as a public health problem in developing nations. Vaccine-based strategies offer the best chance of long-term control. Aspartyl protease inhibitors from parasitic nematodes are highly immunogenic, and have been suggested as potential vaccine antigens. An aspartyl protease inhibitor, API-1, was cloned and characterised from the hookworms Ancylostoma caninum and Ancylostoma ceylanicum. Using sequence from the hookworm expressed sequence tag project, specific primers were designed and used to amplify Ac-api-1 from A. caninum infective L3 cDNA by PCR. Amplicons from the 5' and 3' ends were cloned, sequenced, and combined to create an 874-bp full-length composite sequence of the Ac-api-1 gene. The A. ceylanicum api-1 cDNA of 878 bp was cloned from L3 cDNA using the A. caninum primers. The amino acid sequences of hookworm orthologues were nearly identical, and database searching indicated they belonged to the aspin family, a group of nematode specific aspartyl protease inhibitors that includes the Ascaris pepsin inhibitor PI-3. Ac-api-1 mRNA was detected by reverse transcriptase PCR in eggs, L1, L3 and adult life cycle stages. A polyclonal antiserum against Escherichia coli expressed recombinant Ac-API-1 detected the protein in adult A. caninum excretory/secretory products, but not in those from activated infective larvae. Immunolocalisation experiments using the antiserum indicated that Ac-API-1 is present primarily in the pseudocoelomic fluid in adult hookworms. Soluble, yeast-expressed Ac-API-1 failed to inhibit pepsin or a hookworm gut aspartyl protease in vitro, but inhibited approximately 30% of the proteolytic activity of adult excretory/secretory products. The pseudocoleomic location, presence in all life cycle stages, lack of inhibitory activity against pepsin, and inhibitory activity against excretory/secretory products suggest that Ac-API-1 inhibits an unidentified, putative aspartyl protease secreted by adult hookworms, and may be released as an enzyme-inhibitor complex. The highly immunogenic properties of nematode aspins suggest that Ac-API-1 represents a promising target for a recombinant hookworm vaccine.

Amino Acid Sequence↗

An investigation of hookworm infection and reinfection following mass anthelmintic treatment in the south Indian fishing community of Vairavankuppam.

Hookworm infections, as assessed by counting worms expelled following anthelmintic treatment and by egg output, were found to be of low prevalence and intensity in a South Indian fishing community. The initial overall prevalence of infection in the community was 43%, and the average burden was estimated at 2.2 hookworms per person. The age profiles of prevalence and intensity differed between males and females, with the latter harbouring significantly higher levels of infection. Children of both sexes under 10 years of age rarely harboured hookworms. Treatment with pyrantel pamoate was estimated to be 91% effective in clearing hookworm infections. Egg counts made on stools collected during an 11-month period of reinfection indicated that female patients became reinfected soon after treatment, while little hookworm egg excretion was observed in males during the observation period following treatment. Females acquired a significantly higher number of worms during the reinfection period compared with males, although the average burden in females reached only 28% of the initial, pre-treatment level. The hookworm population consisted of predominantly Necator americanus, and less than 10% of Ancylostoma duodenale. The parasites were highly aggregated within the host population with 10% of the community harbouring over 65% of the total hookworms. Low values of the negative binomial aggregation parameter, k, (indicating extreme over-dispersion) were recorded in groups stratified by age and sex. Highly significant positive correlations were observed between the initial (pre-treatment) and reinfection worm burdens of female (but not of male) patients. It is suggested that occupational practices related to walking through areas contaminated with hookworm larvae play an important role in generating the observed patterns of infection within this community.

Adult↗

Effects of climatic, socio-economic and behavioural factors on the transmission of hookworm (Necator americanus) on two low-country plantations in Sri Lanka.

The climatic, socio-economic and behavioural factors influencing hookworm (Necator americanus) infection in Sri Lanka were explored between February 2000 and June 2001. In February 2000, a single stool sample was collected from each of the 477 subjects investigated, who were aged 2-74 years (median = 13 years) and lived on the 'lowcountry' Maliboda and Ayr plantations. The 'baseline' prevalence (28.5%) and intensity of hookworm infection (0- 4828.5 eggs/g faeces, with a mean of 128.4 eggs/g) were then determined by examining these initial samples, as Kato-Katz smears. Subsequently, each participant was treated with a single, 500-mg dose of mebendazole and then followed-up, at monthly intervals, for the next 15 months. Whenever a subject was found smear-positive for hookworm eggs at one of the monthly follow-ups, he or she was treated again with mebendazole. This approach allowed the monthly incidence of hookworm infection to be determined for each subject, assuming that subjects became smear-positive approximately 6 weeks post-infection. During the study period, rainfall and mean temperature were recorded daily and then converted to monthly values so that the relationship between the incidence of infection over each month and the rainfall and mean temperature over the same period could be explored. In addition, potentially relevant data on the socio-economic status and behaviour of each subject were collected, in questionnaire-based interviews with the adult subjects and the caregivers of the children investigated. Odds ratios (OR) and their 95% confidence intervals (CI) were then calculated for each factor that might increase the risk of hookworm infection. The monthly incidence of hookworm infection showed three peaks -- in September 2000 (21.3%), January 2001 (20.8%) and May 2001 (17.5%) -- at Maliboda, and two peaks -- in September 2000 (25.0%) and February 2001 (29.2%) -- at Ayr. With the data for all subjects combined, incidence showed a statistically significant correlation with mean temperature (r = -0.468; P = 0.018). The results of a multivariate analysis also revealed that mean temperature was significant (beta = -5.296; P = 0.01) in hookworm incidence. Bathing and washing with water from rock-pools formed by waterfalls (OR = 1.33; CI = 1.35-4.01), the use of wells (OR = 2.35; CI = 1.29-4.30), and a lack of toilets (OR = 1.60; CI = 1.01-2.53) each appeared to increase the risk of hookworm infection significantly. Those living on the two study plantations, and perhaps many other similar plantations in Sri Lanka, would clearly benefit from improved access to the public water supply (especially to pipe-borne water) and toilets.

Adolescent↗

[Epidemiological Study On The Hookworm Infections In Korea]

This study was undertaken to evaluate the present status of hookworm infections in Korea from October 1976 through September 1978. The stool specimens of 5,632 persons(male 3,689: female 1,943) collected from primary and middle schools, foundling asylums, and the general populations of various parts in Korea were examined The methods employed were brine floatation technique for the prevalence rate of hookworm infections, Kato's cellophane thick smear technique for the other helminthic infections, and Stoll's dilution egg counting technique for the intensity of hookworm infections. Ancylostoma duodenale and Necator americanus infections were classified by polyethylene tube coproculture method and by the evacuated adult worms after administration of anthelmintics against the patients infected with hookworms. The following results were obtained in this study. 1. The prevalence rate of hookworm infections in rural area was higher than in urban area in the ratio of 2.4:1. And in the age group of over 20, the prevalence rate in significantly higher than that in urban area in the ratio of 5.8:1. 2. In the urban area, difference in the prevalence rate of hookworm infections between the age group of over 20 and that of under 19 was not observed. But in the rural area, the ratio of the prevalence rate between the age group of over 20 to that of under 19 was 8:1. 3. The prevalence rate of hookworm infections in female was higher than that in male in all age group except the age group of under 9 and 20~29. The ratio of the prevalence rate between female to male was 1.5:1 in total population. 4. The area which showed the mean E.P.G. of over 1,000 was not found and almost all of the patients who were infected with hookworms gave the mean E.P.G. between l~999. 5. Necator americanus was newly found in Dangjin Gun, Boseong Gun, Habcheon Gun and Weonseong Gun. In these 4 newly found areas, Dangjin Gun and Boseong Gun showed the composition rate of 25.5% and 83.4% respectively.

Journal Article↗

T-lymphocyte subsets in patients with hookworm infection in Zaria, Nigeria.

Peripheral lymphocyte subsets CD3, CD4, CD8 were studied using monoclonal antibodies to determine the mechanism of immunosuppression observed in an earlier study with total T-cells using the sheep erythrocyte rosetting technique. The study was carried out in 37 Nigerian patients with hookworm infection (Necator americanus and Ancylostoma duodenale and 17 healthy Nigerians as controls. The leucocyte migration inhibition (LMI) test was also carried out to assess the functional integrity of the lymphocytes, while the hookworm status of the patients was assessed by the Stoll technique for measuring egg counts. The results of the T-cell studied showed that CD3 and CD4 cell percentages were significantly depressed in hookworm patients compared to controls (P<0.05). The difference between mean CD8 percentages and absolutes in hookworms patients and controls were not significant. Absolute CD3 and CD4 numbers were not significantly lowered compared to controls in the study and CD4/CD8 ratios were insignificantly reduced. Patients with polyparasitism including hookworm infection showed significant reduction in CD3 and CD4 cells when compared with controls and with patients with hookworm infection alone. The leucocyte migration inhibition response to purified protein derivative of M. tuberculosis (PPD) was significantly decreased in hookworm patients compared to controls, confirming that functionally cell mediated immunity is depressed in hookworm infection.

Adolescent↗

Evidence for an association between hookworm infection and cognitive function in Indonesian school children.

The association between helminth infection and cognitive and motor function was investigated in school-age children in Java, Indonesia. 432 children from 42 primary schools participated in the study. Children were stratified by age and sex into two age groups, 8-9 years and 11-13 years. Children infected with hookworm performed significantly worse than children without hookworm infection in 6 of the 14 cognitive or motor tests. After controlling for school (as a random effect) plus age, socio-economic status and parental education, sex, stunting (height-for-age < - 2sd), body mass index, haemoglobin concentration and the presence of A. lumbricoides and T. trichiura infections, infection with hookworm explained significantly lower scores on tests of Fluency (P < 0.01), Digit-Span Forwards (P < 0.01), Number Choice (P < 0.01), Picture Search (P < 0.03), Stroop Colour Word (P < 0.02) and Mazes (P < 0.001). In 4 of the 6-tests (Fluency, Number Choice, Picture Search and Mazes), there was a significant interaction between hookworm infection and age (P < 0.03), indicating that the association between hookworm and lower test scores increased with age. No associations were observed between hookworm infection and scores in tests of Digit-Span Backwards, Corsi-Block, Stroop Colour, Stroop Interference, Free Recall, Verbal Analogies, Bead Threading or the Pegboard (P > 0.05). Tests associated with helminths represented various functions of working memory. No significant associations between helminth infection and motor function were observed that could not be explained by chance. The results suggest that hookworm infection can have a significant adverse effect on children's working memory which may have consequences for a child's reasoning ability and reading comprehension. Although the results are only associational, the fact that differences in cognition were observed at baseline imply that preventing infection with helminths in school-age children could be of benefit.

Adolescent↗

Impact of repeated mass treatment on human Oesophagostomum and hookworm infections in northern Ghana.

Oesophagostomum bifurcum is a common parasite of humans causing disease in parts of northern Ghana and northern Togo. The impact of repeated mass treatment with albendazole on infection with O. bifurcum and hookworm is analysed and the results compared with those in a control area where no treatment was given. At baseline, O. bifurcum and hookworm prevalences were 53.0% and 86.9%, respectively (n=1011). After 12 months, following two rounds of albendazole treatment, prevalences decreased significantly to 5.4% for O. bifurcum and 36.8% for hookworm (n=535). Twenty-four months after the baseline survey and following a total of four rounds of treatment, prevalences were further reduced to 0.8% and 23.4% for O. bifurcum and hookworm, respectively (n=478). Overall, there was a significant decrease in the larval counts, measured as geometric mean larval count per 4 g of stool of O. bifurcum from 3.0 to 0.1 and of hookworm from 47.2 to 1.8. The fourth mass treatment was carried out in April 2003 by the Lymphatic Filariasis Elimination Programme. Overall, compliance to treatment varied from 70% to 80%. In the control area, Oesophagostomum prevalence increased from 18.5% to 37.0% and the intensity from 0.4 to 1.4. For hookworm, both prevalence (86.1-91.3%) and intensity (54.8-74.3) increased but not to a significant level. The prospects of eliminating human oesophagostomiasis from the intervention area, while simultaneously achieving an important reduction of hookworm prevalences by albendazole mass treatment, are discussed.

Adolescent↗

Epidemiology of hookworm infection in Soong nern district, Korat province, Thailand.

Stool surveys in Soong Nern district, Korat Province, were done by the MF concentration technique. The intensity of hookworm infection, egg counts, results of treatment by anthelminthic drugs, observations on new infections and reinfections following treatment throughout the year and the density of nematode larvae in soil in the study area were investigated. Hookworm infection rates were high in Na-Glang village, both in adult villagers and school-aged children. The intensity of hookworm infections was considered mild and after treatment the numbers of eggs in the stools diminished or disappeared. New cases and reinfection increased during the rainy month, due to socio-environmental factors in the village that favoured the development of hookworm larvai in the soil. Although in this study it was impossible to distinguish hookworm larvae among the nematode larvae collected form soil it was highly probable that hookworm larvae were present. Control of hookworm infection in the study area was proposed by improvement of environmental sanitation and health education.

Adolescent↗

['Intervening is representing': medical campaigns that helped identify the prevalence of hookworm].

This paper compares anti-hookworm campaigns conducted in the early twentieth century in France, Germany, Brazil and West India. The populations that suffer from hookworm are not identical in the North and in the South. In tropical and semi-tropical regions hookworm is mainly found among poor peasants and is related to lack of hygiene, while in temperate climates hookworm was a professional disease of miners, a highly organized professional segment. Nevertheless, major disparities in the pattern of hookworm control did not reflect the North-South divide, but a difference between campaigns. These aimed at the eradication of hookworm infection (Germany and West-India) and at alleviating the effects of this infection on populations (France and Brazil). Maps that represented the prevalence of hookworm mirrored the aims of the sanitary campaign in which they were used: eradication of parasitic worms versus the reduction of the handicap induced by these worms. In public health as well, representing is intervening. And vice versa: patterns of intervention shape representations.

Brazil↗

Hookworm anemia in the adult population of Jagapati village, Bali, Indonesia.

A cross sectional study was conducted in Jagapati Village, Bali to assess some characteristics of hookworm anemia among the adult population. Hookworm anemia was defined as an iron deficiency anemia in heavily infected individuals (EPG > 2,000). WHO criteria for anemia and criteria of Hercberg for iron deficiency were used. In this study, 15 cases of hookworm anemia were found among 454 total samples (3.3%), or among 123 cases of iron deficiency anemia (12.2%). The age varied between 16-69 years with male to female ratio of 1:2.8. Twelve cases were found with symptoms and signs of anemia, 1 case with full blown hookworm anemia, and 2 cases were asymptomatic. The hemoglobin level was found to be 4.5-12.9 g/dl, with 12 cases (80.0%) being classified as mild anemia, 2 cases (13.3%) as moderate anemia, and 1 case as severe anemia. The mean serum iron level was 39.6 mg/dl, mean transferrin saturation was 11.1%, and mean serum ferritin level was 9.6 ng/dl. Hypoalbuminemia was found in 9 cases (60.0%), eosinophilia in 8 cases (53.3%), and low serum folic acid level in 5 cases (38.5%). High total serum IgE level was found in all but one case, with mean total serum IgE level of 3,739 U/ml. The intensity of hookworm infection was moderate in 11 cases (73.3%) and severe in 4 cases (26.7%). It could be concluded that hookworm anemia was characterized by iron deficiency anemia with eosinophilia, high serum total IgE level, hypoalbuminemia and moderate to severe hookworm infection.

Adolescent↗

Quantitative genetic analysis of susceptibility to hookworm infection in a population from rural Zimbabwe.

Overdispersion is a common feature of population distribution patterns for hookworm infection in humans. Genetic factors may be partially responsible for this observed increased susceptibility in a fraction of the exposed population. However, the hypothesis that there are genetic components to susceptibility to this infectious disease has not been tested explicitly. The purpose of this study is to quantify the influence of genetic factors on patterns of hookworm infection in a rural population in Zimbabwe. A quantitative measure of hookworm load, number of hookworm eggs per gram of feces, as determined by the Kato thick smear technique, was available for 289 individuals. Of these, 279 individuals were members of 62 nuclear families and 10 were independent individuals. We analyzed the hookworm data in combination with the pedigree structure of the sampled individuals using quantitative genetic analysis techniques. Using this variance decomposition approach, we estimated the heritability of hook worm load to be 0.37 +/- 0.09 (p < 0.0001). This significant heritability indicates that 37% of the variation (after correcting for the effects of covariates) in hookworm eggs per gram observed in this population is attributable to genetic factors. The results suggest that further investigation and characterization of the genetic components influencing susceptibility to hookworm infection are warranted.

Adolescent↗

The prevalence of Trichuris, Ascaris and hookworm infection in Orang Asli children.

A community study on the age and sex related prevalence, intensity infection and frequency distribution of Ascaris, Trichuris and hookworm was carried out in 205 Orang Asli (Aborigines) children (95 boys, 110 girls) aged 1-13 years. The overall prevalence of Ascaris, Trichuris and hookworm was 62.9%, 91.7% and 28.8%, respectively. Almost two-thirds of the children were infected with moderate and severe intensity infection of Trichuris, 46.3% had moderate to severe intensity infection of Ascaris. However only 1.5% had moderate intensity of hookworm infection. The prevalence and mean intensity infection (measured by eggs per g) of Ascaris was age-dependent; lower in age group 1-4 years reached peak and stable at age group 5 years and above. The prevalence of Trichuris was high in all age groups and it fluctuated with age; the mean intensity of infection of Trichuris (measured by eggs per g) was age-dependent. Hookworm infection also rose with age and reached peak at 5-6 years, following that the prevalence declined. The frequency distributions of Ascaris, and hookworm were overdispersed. A strong positive correlation (p < 0.001) were observed between Ascaris and Trichuris and between hookworm and Trichuris. A positive correlation (p < 0.01) were also observed between Ascaris and hookworm.

Adolescent↗

Hookworm infection and disease: advances for control.

Advances in epidemiology and in control strategies of hookworm infection and associated disease were reviewed. Recent estimates indicate that hookworms infect approximately 1.3 billion people worldwide, and 96 million suffer from associated morbidity, including also insidious effects on nutritional status and on physical and intellectual development. Recent research studies on iron loss and iron-deficiency anaemia associated to hookworm infection were irondeficiency anemia associated to hookworm infection were reviewed. Recommendations for planning strategies for the control of hookworm infection were addressed and epidemiology, targets, chemotherapy, health education, sanitation, monitoring and evaluation in helminth control programmes were discussed. Special relevance was given to chemotherapy with new, single dose broad spectrum, safe anthelminthic drugs as the mainstay of control strategy to reduce intensity of infection, iron-deficiency anaemia and other morbidity indicators associated with hookworm infections. For the control of transmission of hookworm infection, periodic chemotherapy should be implemented in the context of ongoing improvement of sanitation and promotion of health education. These elements should be integrated into the prevailing system of primary health care and must be based on multisectoral collaboration to ensure sustainability of control programmes.

Ancylostoma↗

Characterization and localization of cathepsin B proteinases expressed by adult Ancylostoma caninum hookworms.

The hookworm Ancylostoma caninum induces human eosinophilic enteritis (EE), probably via allergic responses to its secretions. Cysteine and metallo-proteinases may be the components of these secretions that elicit hypersensitivity reactions. In order to characterize genes encoding cysteine proteinases (CP) secreted by A. caninum, an adult hookworm cDNA library was constructed and screened with a cloned fragment of a hookworm CP gene. This fragment was obtained using consensus oligonucleotide, CP-gene-specific primers in the polymerase chain reaction. cDNAs encoding two CPs were obtained from the library and sequenced. The first gene, AcCP-1, encoded a cathepsin B-like zymogen CP of 343 amino acids (aa), predicted to be processed in vivo into a mature CP of 255 aa. Closest nucleotide identities were to Haemonchus contortus cysteine protease (61%) and to human cathepsin B (60%). The second gene, AcCP-2, encoded a mature CP of 254 aa, that showed 86% identity to AcCP-1, and 58% and 47% identity to bovine cathepsin B and human cathepsin B, respectively. Rabbit antisera raised against recombinant AcCP-1 reacted with esophageal, amphidial and excretory glands in formalin-fixed, paraffin embedded sections of both male and female adult hookworms, and with an antigen of approx. 40 kDa in Western blot analysis of excretory/secretory products from adult hookworms. Together, these immuno-hybridization results strongly suggest that the CP encoded by the AcCP-1 gene is secreted by hookworms. These are the first reported CP genes from hookworms. Proteinases encoded by these genes may be responsible for the CP activity that we have shown previously to be secreted by adult A. caninum.

Amino Acid Sequence↗