Marilyn E. Scott

Parasitism: A cryptic determinant of animal community structure

Traditionally, the major biotic determinants of animal community structure were assumed to be competition and predation. Recent theoretical, experimental and field studies indicate that parasitism is also important. The spectrum of effects that parasites have on host individuals is as broad as the range of parasite-host interactions. This review considers evidence for direct effects of parasites on fecundity and survival that influence host demography, and for indirect effects on host physiology and behavior that interface with competition and predation.

Regulation of mouse colony abundance by Heligmosomoides polygyrus

Despite the ubiquitous presence of parasites, parasitism has not been considered among the list of regulatory factors in animal populations until recently. A detailed long-term study on the impact of the direct life-cycle nematode Heligmosomoides polygyrus on a breeding population of laboratory mice provides a clear example of the ability of helminths to regulate host abundance. In the absence of the parasite, the mouse population equilibrated at a density of 320 mice/m2 as a result of density-dependent effects on recruitment. When the parasite was added and transmission was maintained at high levels, infected mouse populations equilibrated at densities of less than 18 mice/m2. Reduced rates of parasite transmission and elimination of the parasite from the system both resulted in an increase in mouse density. These results have implications for both ecology and parasitology as they demonstrate a potentially important but often ignored component of host populations that may well influence host abundance and community structure.

Zinc Deficiency Impairs Immune Responses against Parasitic Nematode Infections at Intestinal and Systemic Sites

Research on the complex interactions among host nutritional status, parasitic infection and immune responsiveness has focused on the detrimental consequences of parasitic infections on host nutritional status and on mechanisms by which malnutrition impairs immunocompetence. Curiously, relatively few studies have examined the effects of malnutrition on the immune response in the parasite-infected host, and even fewer have considered the events occurring at the intestinal level, where absorption of nutrients occurs, intestinal parasites reside, and the gastrointestinal-associated lymphoid tissues play a role in directing both the local and the more systemic immune responses. Our work using a zinc-deficient nematode-infected mouse model reveals that parasites are better able to survive in the zinc-deficient hosts than in well-nourished hosts; that the production of interleukin-4 in the spleen of zinc-deficient mice is depressed, leading to depressed levels of IgE, IgG(1) and eosinophils; and that the function of T cells and antigen-presenting cells is impaired by zinc deficiency as well as by energy restriction. Given the paramount role of the gastrointestinal-associated lymphoid tissues in inducing and regulating immune responses to intestinal parasites and in orchestrating responses in the spleen and peripheral circulation, we conclude that zinc deficiency (in association with energy restriction) exerts profound effects on the gut mucosal immune system, leading to changes in systemically disseminated immune responses and, importantly, to prolonged parasite survival.

Assays to Detect β-Tubulin Codon 200 Polymorphism in Trichuris trichiura and Ascaris lumbricoides

Background The soil-transmitted helminths (STH) Ascaris lumbricoides and Trichuris trichiura are gastrointestinal parasites causing many disabilities to humans, particularly children. The benzimidazole (BZ) drugs, albendazole (ALB) and mebendazole (MBZ), are commonly used for mass treatment for STH. Unfortunately, there is concern that increased use of anthelmintics could select for resistant populations of these human parasites. In veterinary parasites, and lately in filarial nematodes, a single amino acid substitution from phenylalanine to tyrosine, known to be associated with benzimidazole resistance, has been found in parasite β-tubulin at position 200. We have developed pyrosequencer assays for codon 200 (TTC or TAC) in A. lumbricoides and T. trichiura to screen for this single nucleotide polymorphism (SNP). Method and Findings Pyrosequencing assays were developed and evaluated for detecting the TTC or TAC SNP at codon 200 in β-tubulin in A. lumbricoides and T. trichiura. Genomic DNA from individual worms, eggs isolated from individual adult worms or from fecal samples with known treatment history and origin, were sequenced at β-tubulin by pyrosequencing, and genotypes were confirmed by conventional sequencing. The assays were applied to adult worms from a benzimidazole-naïve population in Kenya. Following this, these assays were applied to individual worms and pooled eggs from people in East Africa (Uganda and Zanzibar) and Central America (Panama) where mass anthelmintic drug programs had been implemented. All A. lumbricoides samples were TTC. However, we found 0.4% homozygous TAC/TAC in T. trichiura worms from non-treated people in Kenya, and 63% of T. trichiura egg pools from treated people in Panama contained only TAC. Conclusion Although the codon 200 TAC SNP was not found in any of the A. lumbricoides samples analyzed, a rapid genotyping assay has been developed that can be used to examine larger populations of this parasite and to monitor for possible benzimidazole resistance development. The TAC SNP at codon 200, associated with benzimidazole resistance in other nematodes, does occur in T. trichiura, and a rapid assay has been developed to allow populations of this parasite to be monitored for the frequency of this SNP. Sample sizes were small, anthelmintic efficacy was not assessed, and treated and non-treated samples were from different locations, so these frequencies cannot be extrapolated to other populations of T. trichiura or to a conclusion about resistance to treatment. The occurrence of the TAC SNP at codon 200 of β-tubulin in T. trichiura may explain why benzimidazole anthelmintics are not always highly effective against this species of STH. These assays will be useful in assessing appropriate treatment in areas of high T. trichiura prevalence and in monitoring for possible resistance development in these STH.

Female mice mate preferentially with non-parasitized males

If parasitic infection is a driving force in female mate choice, then females should preferentially select parasite-free males. The role of infection on female mate choice in mammals was assessed using a 3-chambered apparatus. A female CD-1 mouse was allowed to choose between 2 tethered male mice, 1 uninfected and 1 infected with 200 larvae (L3) of the intestinal nematode Heligmosomoides polygyrus. Both uninfected and infected males were equally receptive to the oestrous females, and females did not differ in the number of visits and time spent exploring the 2 males. Female time preference was not a useful predictor of ultimate mate choice, whereas first mount preference of the female was a reliable indicator. Results indicate that female mice preferentially mated with uninfected males as evidenced by first ejaculation choice, but that male infection status did not significantly affect female reproductive success. Interestingly, litters sired by infected males contained a significantly higher percentage of females suggesting that parasite-induced hormonal changes may alter the sex ratio of the offspring. This study provides the first direct evidence of the impact of parasitic infection on mammalian mate choice.

Relationship of childhood protein-energy malnutrition and parasite infections in an urban African setting

Summary A clear understanding of protein‐energy malnutrition (PEM), parasite infection and their interactions is essential in formulating health and development policies. We studied the prevalence of PEM indicators and the prevalence and/or intensity of infection in 558 Zairian children aged 4 months to 10 years. Multivariate analyses were used to estimate relationships between PEM indicators and parasitic infection. Stunting was found in 40.3% of children, wasting in 4.9% and kwashiorkor in 5.1%. The risk of stunting was significantly higher in children with Ascaris lumbricoides. The risk of wasting was higher in children with A. lumbricoides or Trichuris trichiura, whereas the risk of kwashiorkor was high with T. trichiura but very reduced in those with A. lumbricoides. Plasmodium infection was not related to nutritional indicators. These relationships highlight important interactions, both synergistic and antagonistic, between nutrition and parasites in central Africa.

Multiple infection with Plasmodium and helminths in communities of low and relatively high socio-economic status.

A study was conducted in the city of Lubumbashi, Zaire: (1) to survey parasitic infections and clinical conditions in the local children and their mothers; (2) to identify combinations of parasites and clinical conditions that commonly occurred together in individuals; and (3) to determine whether single- and/or multiple-species infections were risk determinants of the observed clinical conditions. Overall, 1100 children and mothers from three subdivisions, two of low socio-economic status (LSES) and one of relatively high socio-economic status (HSES), provided stool and blood samples and were clinically examined. Plasmodium prevalence was higher in the two LSES subdivisions than in the HSES subdivision. Prevalence and intensity of Ascaris lumbricoides infection were low in the HSES subdivision and one of the two LSES subdivisions. In contrast, prevalence and intensity of Trichuris trichiura and of hookworms were similar in all subdivisions. Plasmodium and A. lumbricoides were the most frequently found single-species infections. The combination of A. lumbricoides and Plasmodium was the most frequent double-species infection and that of A. lumbricoides, Plasmodium and T. trichiura was the most frequent triple-species infection. Significant positive associations between parasite species were detected in the HSES subdivision, and in one of the two LSES subdivisions. Because the relationships were not consistently detected, it is hypothesized that the associations are determined by environmental conditions rather than synergy between the parasites in the host. The most commonly observed clinical conditions were abdominal pain, diarrhoea, fever, and low packed-cell volume (PCV). The occurrence of each was significantly lower in the HSES subdivision than in at least one of the two LSES subdivisions. Abdominal pain and low PCV were most common in individuals presenting with only a single clinical condition, and the combination of this symptom and sign was the most commonly observed pair of conditions. Abdominal pain, low PCV and diarrhoea was the most common combination in individuals with three clinical conditions. Logistic regression revealed that hookworm infection, T. trichiura infection, young age and residence in an LSES subdivision were determinants of diarrhoea. Trichuris trichiura infection, young age and living in an LSES subdivision were risk factors for abdominal pain. Plasmodium infection and young age were risk factors for fever. LSES was the only predictor of low PCV. Infection with A. lumbricoides did not enter any of the models. No significant interactions were detected among parasites, indicating that there was no synergism or antagonism among parasites in the induced disease.

Predisposition of individuals and families in Mexico to heavy infection with Ascaris lumbricoides and Trichuris trichiura.

Stool specimens from 90% of the population of 2 marginal communities in Coatzacoalcos, Mexico, were examined by the Kato-Katz technique for the estimation of eggs per gram of faeces (epg). Average epg values for Ascaris lumbricoides and Trichuris trichiura were calculated for each family in the community, based on the arithmetic mean epg of family members for each parasite. 39 families were classified as either lightly or heavily infected with A. lumbricoides and infected members were treated with mebendazole. A. lumbricoides expelled following treatment were collected from the children 2-10 years of age, and reinfection was monitored monthly for 6 months. At the end of the reinfection interval, a final stool specimen was collected from all family members and those infected were treated. Again, A. lumbricoides expelled by the children 2-10 years of age were collected. There was a significant correlation between the number of A. lumbricoides expelled by the children at the first and second treatments as well as in epg values before the first and second treatments for both A. lumbricoides and T. trichiura in children (2-10 years) and adults (greater than 19 years). At the family level, there were significant correlations between the values of family mean epg before each treatment for both A. lumbricoides and T. trichiura.

The distribution of intestinal helminth infections in a rural village in Guatemala

Fecal egg count scores were used to investigate the distribution and abundance of intestinal helminths in the population of a rural village. Prevalences of the major helminths were 41% with Ascaris lumbricoides, 60% with Trichuris trichiura and 50% with Necator americanus. All three parasites showed a highly aggregated distribution among hosts. Age/prevalence and age/intensity profiles were typical for both A. lumbricoides and T. trichiura with the highest worm burdens in the 5-10 year old children. For hookworm both prevalence and intensity curves were convex in shape with maximum infection levels in the 30-40 year old age class. Infected females had higher burdens of T. trichiura than infected males in all age classes of the population; there were no other effects of host gender. Analysis of associations between parasites within hosts revealed strong correlations between A. lumbricoides and T. trichiura. Individuals with heavy infections of A. lumbricoides and T. trichiura showed highly significant aggregation within households. Associations between a variety of household features and heavy infections with A. lumbricoides and T. trichiura are described.

Association between Response to Albendazole Treatment and β-Tubulin Genotype Frequencies in Soil-transmitted Helminths

Background Albendazole (ABZ), a benzimidazole (BZ) anthelmintic (AH), is commonly used for treatment of soil-transmitted helminths (STHs). Its regular use increases the possibility that BZ resistance may develop, which, in veterinary nematodes is caused by single nucleotide polymorphisms (SNPs) in the β-tubulin gene at positions 200, 167 or 198. The relative importance of these SNPs varies among the different parasitic nematodes of animals studied to date, and it is currently unknown whether any of these are influencing BZ efficacy against STHs in humans. We assessed ABZ efficacy and SNP frequencies before and after treatment of Ascaris lumbricoides, Trichuris trichiura and hookworm infections. Methods Studies were performed in Haiti, Kenya, and Panama. Stool samples were examined prior to ABZ treatment and two weeks (Haiti), one week (Kenya) and three weeks (Panama) after treatment to determine egg reduction rate (ERR). Eggs were genotyped and frequencies of each SNP assessed. Findings In T. trichiura, polymorphism was detected at codon 200. Following treatment, there was a significant increase, from 3.1% to 55.3%, of homozygous resistance-type in Haiti, and from 51.3% to 67.8% in Kenya (ERRs were 49.7% and 10.1%, respectively). In A. lumbricoides, a SNP at position 167 was identified at high frequency, both before and after treatment, but ABZ efficacy remained high. In hookworms from Kenya we identified the resistance-associated SNP at position 200 at low frequency before and after treatment while ERR values indicated good drug efficacy. Conclusion Albendazole was effective for A. lumbricoides and hookworms. However, ABZ exerts a selection pressure on the β-tubulin gene at position 200 in T. trichiura, possibly explaining only moderate ABZ efficacy against this parasite. In A. lumbricoides, the codon 167 polymorphism seemed not to affect drug efficacy whilst the polymorphism at codon 200 in hookworms was at such low frequency that conclusions cannot be drawn.