Comparison of the Kato-Katz and FLOTAC techniques for the diagnosis of soil-transmitted helminth infections

Decisions on individual or community treatment and evaluation of chemotherapy based control programs depend on parasitological diagnostic techniques. The aim of this study was to compare the accuracy of a single Kato-Katz thick smear and a single FLOTAC for the determination of the prevalence and intensity of soil-transmitted helminth infections. A total of 271 faecal specimens were collected from schoolchildren in Ethiopia, and microscopically examined using the Kato-Katz method (41.7 mg stool per slide) and the FLOTAC technique. The combined results from the Kato-Katz and FLOTAC methods were used as diagnostic 'gold' standard for reference in the analysis. Agreement between the two methods showed kappa values of 0.74, 0.73 and 0.28 for Ascaris lumbricoides, Trichuris trichiura and hookworm, respectively. A single FLOTAC revealed significantly more infections than a single Kato-Katz for each of the three soil-transmitted helminths (p<0.01). The sensitivities of a single Kato-Katz for diagnosis of T. trichiura, A. lumbricoides and hookworm infections were 76.6%, 67.8% and 19.6%, respectively, while the sensitivity of FLOTAC was 100% for all the three soil-transmitted helminth species. A single Kato-Katz yielded considerably higher mean faecal egg counts (FECs) (729.1, 145.2 and 60.7 eggs per gram of stool (EPG) for A. lumbricoides, T. trichiura and hookworm, respectively) compared with a single FLOTAC (142.5, 54.5 and 14.6 EPG, respectively) (p<0.05). Our study confirms that a single FLOTAC is more sensitive than a single Kato-Katz for the diagnosis of soil-transmitted helminth infections, but results in lower FECs. Further standardization and validation are still required in different epidemiological settings with varying levels of intensity of infections before recommending FLOTAC for large-scale community diagnosis.     

Site selection by intestinal helminths of the Virginia opossum (Didelphis virginiana)

Digestive tract helminths of the Virginia opossum (Didelphis virginiana) were examined for site specificity and interspecific interactions. Thirty opossums were live-trapped at 2 localities in Georgia, killed, and their digestive tracts removed and divided into the following sections; esophagus, stomach, small intestine (20 equal sections), cecum, and large intestine (5 equal sections). Helminths present in each section were identified and counted. Three trematode, 1 cestode, 2 acanthocephala, and 6 nematode species were found. Two parasites (the acanthocephalan Centrorhynchus spinosus and the nematode Viannaia hamata) are new state records for Georgia and this is the first report of C. spinosus from an opossum. There was considerable overlap between the spatial distributions of many of the helminth species found in the small intestine though most species had different modal locations. Helminth species locations were unaffected by host sex or trapping locality. There was no evidence that the presence, absence, or intensity of any helminth species affected the location or intensity of other helminth species in the digestive tract.     

Malaria, schistosomiasis, and intestinal helminths in relation to microdams in Tigray, northern Ethiopia.

A survey was undertaken in Tigray, Northern Ethiopia, to assess the prevalence of malaria, schistosomiasis, and intestinal helminths in relation to microdams. The survey took place from March to June 1995, during the dry season, at 41 microdams. At each site the village nearest the dam (within thirty minutes walk) was selected, ten households were randomly chosen, and all family members were examined for malaria and intestinal parasites. The overall study sample was 2271 people, of all age groups. Plasmodium falciparum infection was documented in four villages (at 10% of microdams); prevalence was 1.2% (range 0-20% by village). Larvae of Anopheles gambiae s.l. were found at one microdam. Infection with intestinal schistosomiasis was documented in 20 villages (at 49% of microdams), and one third of those infected had moderate to heavy infections. Biomphalaria species, the intermediate host snails of Schistosoma mansoni, were found at 16 microdams (39%), and snails infected by mammalian cercariae were found in one locality. Infections with soil-transmitted nematodes were prevalent: hookworm was detected in more than two thirds of the villages, and Ascaris lumbricoides and Trichuris trichiura were present in almost half of the villages. Out of 2078 stool examinations, the prevalence of S. mansoni infection was 7.2% (range 0-48% by village), of A. lumbricoides 2.3% (range 0-31%), of T. trichiura 2.4% (range 0-21%), and of hookworm 8.9% (range 0-78%). The prevalence of malaria, S. mansoni and hookworm was higher at altitudes below 2000 metres above sea level. S. mansoni was more prevalent in microdams built more than 5 years before the survey, while T. trichiura was more prevalent at recently constructed microdams. The widespread distribution of schistosomiasis and intestinal helminths, and the presence of malaria infection during the dry season confirm that microdams create favourable conditions for the transmission of these parasitic diseases. Health safeguards must be incorporated into the planning, construction, and operation of microdams and irrigation systems in order to prevent or reduce these diseases. In areas with high prevalence, control activities should be intensified.     

Transport of helminths to Hawaii via the brown anole, Anolis sagrei (Polychrotidae)

Sixty-two brown anoles, Anolis sagrei, from Oahu, Hawaii were examined for helminths. Anolis sagrei was introduced to Hawaii, presumably from the Caribbean. Two species of trematodes, Mesocoelium monas and Platynosomum fastosum, 3 species of nematodes, Atractis scelopori, Physaloptera squamatae, and Physocephalus sp., 1 acanthocephalan, Acanthocephalus bufonis, and 1 pentastome, Raillietiellafrenatus, were found. Atractis scelopori and P. squamatae, previously unknown in Hawaii, are widely distributed in the Caribbean and were most likely transported to Hawaii with the introduced anoles. Mesocoelium monas, P. fastosum, Physocephalus sp., A. bufonis, and R. frenatus have been previously reported from Hawaiian herptiles; A. sagrei most likely acquired infections of these parasites from Hawaiian populations. This study indicates that helminths can be transported with their introduced hosts and become established in the colonized areas and that introduced lizards may quickly acquire species of previously established helminthes.     

Epidemiological study of the intestinal helminths of wild boar (Sus scrofa) and mouflon (Ovis gmelini musimon) in central Italy

Since 1995 the population of wild ungulates increased significantly in the "Parco provinciale dei Monti Livornesi" (Livorno, Tuscany, Central Italy). We studied the intestinal macroparasites of two hosts, the wild boar (Sus scrofa) and the mouflon (Ovis gmelini musimon). In the case of wild boars we found a dominant parasite species, Globocephalus urosubulatus. For this parasite the frequency distribution of the number of parasites per host agrees with a negative binomial distribution. There is not a significant correlation between the age of the animals and the parasitosis. Furthermore the mean parasite burden of male and female wild boars does not differ significantly. In the case of mouflons we found a dominant parasite species Nematodirus filicollis with Trichuris ovis as codominant species.     

Diet and intestinal helminths in a population of perch, Perca flavescens

In 1967 and 1968 1310 Perca flavescens (Mitchill) were examined from Lake Opeongo, Ontario for changes in five diet categories and in incidence of eight intestinal helminths in relation to seasons, host size and sex. Male fish were less common than females and were smaller. A high proportion of fish (57.7 %) had empty stomachs, but of the four types of foods taken microcrustacea were high in spring, late summer and again in early winter; insects increased in summer with another peak in autumn; crayfish had a small peak in spring and a larger one in autumn; and fish were lowest in summer. Males ate more smaller foods (microcrustacea) than females, but females ate more larger foods (crayfish and fish); no sex difference was seen with regard to insects or empty stomachs. Male fish ate very few larger items, but females showed an increase in food size with increasing size of fish. There was no seasonality in incidence of fish free of all parasites, but two trematodes Bunodera sacculata Van Cleave & Mueller and B. luciopercae (Müller), and one cestode Proteocephalus pearsei La Rue showed seasonality with populations lowest in fish in summer; and one trematode Crepidostomum cooperi Hopkins and one nematode Dacnitoides cotylophora Ward & Magath showed seasonality with populations highest in summer. In addition one cestode Bothriocephalus sp., one nematode Spinitectus gracilis Ward & Magath and one acanthocephalan Leptorhynchoides thecatus (Linton) showed no seasonality, perhaps on account of the low incidence in the case of Bothriocephalus sp. and L. thecatus. No parasites were more common in males, but two (P. pearsei and L. thecatus) were more common in females. In both males and females, with increasing size fish, (i) fish free of all parasites decreased, but (ii) incidence of C. cooperi increased. Incidence of B. luciopercae, Bothriocephalus sp., S. gracilis and L. thecatus all increased with increasing size of fish only in females. The relationship between diet and incidence of intestinal helminths is discussed in the light of what is known of their life cycles. The exercise illustrates our ignorance of population ecology of fish parasites and the interactions of parasites and their host populations.     

The potential role of habitat on intestinal helminths of mountain hares, Lepus timidus

Over the last century in the uplands of Scotland, the extent of heather moorland which supports high densities of mountain hares Lepus timidus has diminished and has gradually been replaced by large-scale commercial forestry plantations or expanding natural woodlands. The potential impact of such a change in land use on host-parasite interactions was investigated by comparing the intensity and prevalence of infection of hares by parasites in two separate habitats: a large hare-fenced young forestry plantation and the adjacent open moorland. Carcasses were collected in November 1990 from within both habitats and after the woodland had been enclosed for nine months. Age, sex, fatness (kidney fat index) and degree of infection of hares were noted. Two parasites were recorded: the nematode Trichostrongylus retortaeformis and the cestode Mosgovoyia pectinata. Clear differences in the intensity of infection of adults occupying the different habitats had occurred in the nine months since woodland enclosure. Adult mountain hares in the woodland had levels of infections approaching four times that observed in hares occupying the open moorland and although not significant, the prevalence of infection was greater in hosts inhabiting the woodland than the open moorland. It is suggested that the parasite-host relationship differs between the two habitats and as heather-dominated moorland landscapes become more fragmented with the increasing establishment of woodlands, the impact of parasites on the life history strategies of mountain hares needs to be reconsidered.     

A comparison of the sensitivity and fecal egg counts of the McMaster egg counting and Kato-Katz thick smear methods for soil-transmitted helminths

Background

The Kato-Katz thick smear (Kato-Katz) is the diagnostic method recommended for monitoring large-scale treatment programs implemented for the control of soil-transmitted helminths (STH) in public health, yet it is difficult to standardize. A promising alternative is the McMaster egg counting method (McMaster), commonly used in veterinary parasitology, but rarely so for the detection of STH in human stool.

Methodology/Principal Findings

The Kato-Katz and McMaster methods were compared for the detection of STH in 1,543 subjects resident in five countries across Africa, Asia and South America. The consistency of the performance of both methods in different trials, the validity of the fixed multiplication factor employed in the Kato-Katz method and the accuracy of these methods for estimating ‘true’ drug efficacies were assessed. The Kato-Katz method detected significantly more Ascaris lumbricoides infections (88.1% vs. 75.6%, p<0.001), whereas the difference in sensitivity between the two methods was non-significant for hookworm (78.3% vs. 72.4%) and Trichuris trichiura (82.6% vs. 80.3%). The sensitivity of the methods varied significantly across trials and magnitude of fecal egg counts (FEC). Quantitative comparison revealed a significant correlation (Rs >0.32) in FEC between both methods, and indicated no significant difference in FEC, except for A. lumbricoides, where the Kato-Katz resulted in significantly higher FEC (14,197 eggs per gram of stool (EPG) vs. 5,982 EPG). For the Kato-Katz, the fixed multiplication factor resulted in significantly higher FEC than the multiplication factor adjusted for mass of feces examined for A. lumbricoides (16,538 EPG vs. 15,396 EPG) and T. trichiura (1,490 EPG vs. 1,363 EPG), but not for hookworm. The McMaster provided more accurate efficacy results (absolute difference to ‘true’ drug efficacy: 1.7% vs. 4.5%).

Conclusions/Significance

The McMaster is an alternative method for monitoring large-scale treatment programs. It is a robust (accurate multiplication factor) and accurate (reliable efficacy results) method, which can be easily standardized.     

Clinical efficacy of antiparasite treatments against intestinal helminths and haematic protozoa in Gallotia caesaris (lizards)

Intestinal helminths and blood protozoa are common parasites of lizards. In captivity they can be a serious problem, but no information on treatments is available. In this study, several antiparasitic drugs were studied in Gallotia caesaris (Lacertida) from La Gomera, the Canary Islands, Spain. A lack of efficacy of chloroquine, and a decrease in haemogregarine (Apicomplexa) infection by atovaquone-proguanil was demonstrated. Furthermore, a positive effect of fenbendazole against intestinal nematode infection was observed. In the Canary Islands, Gallotia bravoana and Gallotia simonyi (Lacertida) are included in a captive breading recovery plan. The present results are useful in order to keep these animals in good condition and to control their parasites. Moreover, these results are important for reptiles kept as pets, due to the previous lack of knowledge of parasite management.     

Prevalence, incidence, intensity and associated morbidity of intestinal helminths in south Thailand

Intensive surveys for Ascaris lumbricoides, Trichuris trichiura and hookworm were carried out in two villages in Phang Nga Province, southern Thailand, in order to measure prevalence, estimate incidence and determine the relation between intensity of infection and morbidity before and after chemotherapy. The study populations were a small upland village community (Nai Tone) and a grade school in a small coastal village (Boh Saen). About half of the Nai Tone villagers were given a broad spectrum antihelminthic (albendazole), and the Boh Saen students were all treated successively with three drugs: piperazine citrate to treat for Ascaris, pyrantel pamoate to treat for hookworm, and mebendazole to treat for Trichuris. Stool examinations were made using the quick Kato smear technique, questionnaires were administered concerning a variety of possible symptoms, and anthropometric and blood biochemical parameters were measured both before and after treatment. The prevalence of Ascaris was 31.0 and 22.6%, hookworm was 89.1 and 88.0% and Trichuris was 59.7 and 77.8% in the Nai Tone and Boh Saen study populations, respectively. Average intensity of Ascaris was highest in the 0-9 year age class (greater than 32,000 epg) in Nai Tone Village. Hookworm intensity of infection was higher in males than in females in all age classes, and in Nai Tone Village at least 25% of males and 20% of females had 8000 or more epg of faeces. Trichuris intensity of infection was highest between 5 and 10 years of age in both populations. The only signs or symptoms showing a significant (P less than 0.05) difference between high and low classes of intensity of infection and a significant improvement (P less than 0.01) after drug treatment, were headache and flatulance in the case of hookworm infection in Boh Saen School. The presence of multiple infections made testing of hypotheses concerning particular parasite species difficult.     

Physalopteroides arnoensis n. sp. (Nematoda: Physalopteroidea) and other intestinal helminths of the mourning gecko, Lepidodactylus lugubris (Sauria: Gekkonidae), from Arno Atoll, Republic of the Marshall Islands, Oceania

Physalopteroides arnoensis n. sp. (Nematoda: Physalopteroidea) from the intestinal tract of the gecko Lepidodactylus lugubris is described and illustrated. Physalopteroides arnoensis is the 18th species to be assigned to the genus and is distinguished from other oriental species by the distribution pattern of caudal papillae of the male: 22 posterior subventral pedunculated papillae; 8 precloacal papillae in symmetrical pairs; 14 postcloacal papillae, 6 papillae immediately postcloacal in symmetrial pairs and remaining 8 papillae asymmetrically placed, 3 on right, 5 on left. Other helminths found include Pharyngodon lepidodactylus, Hedruris hanleyae, Maxvachonia chabaudi, and larvae of Ascarops sp. Our review of Physalopteroides removes all subspecies designations; Physalopteroides minor Caballero, 1969 n. comb is proposed.     

Sensitivity of double centrifugation sugar fecal flotation for detecting intestinal helminths in coyotes (Canis latrans)

Fecal analysis is commonly used to estimate prevalence and intensity of intestinal helminths in wild carnivores, but few studies have assessed the reliability of fecal flotation compared to analysis of intestinal tracts. We investigated sensitivity of the double centrifugation sugar fecal flotation and kappa agreement between fecal flotation and postmortem examination of intestines for helminths of coyotes (Canis latrans). We analyzed 57 coyote carcasses that were collected between October 2010 and March 2011 in the metropolitan area of Calgary and Edmonton, Alberta, Canada. Before analyses, intestines and feces were frozen at -80 C for 72 hr to inactivate Echinococcus eggs, protecting operators from potential exposure. Five species of helminths were found by postmortem examination, including Toxascaris leonina, Uncinaria stenocephala, Ancylostoma caninum, Taenia sp., and Echinococcus multilocularis. Sensitivity of fecal flotation was high (0.84) for detection of T. leonina but low for Taenia sp. (0.27), E. multilocularis (0.46), and U. stenocephala (0.00). Good kappa agreement between techniques was observed only for T. leonina (0.64), for which we detected also a significant correlation between adult female parasite intensity and fecal egg counts (R(s)=0.53, P=0.01). Differences in sensitivity may be related to parasite characteristics that affect recovery of eggs on flotation. Fecal parasitologic analyses are highly applicable to study the disease ecology of urban carnivores, and they often provide important information on environmental contamination and potential of zoonotic risks. However, fecal-based parasitologic surveys should first assess the sensitivity of the techniques to understand their biases and limitations.     

Associations and interactions among intestinal helminths of the brown trout, Salmo trutta, in northern Italy

Species co-occurrences and interspecific associations between intensity of infection were studied in helminth communities of three populations of brown trout, Salmo trutta, from northern Italy. Of the eight helminth species, only four were common enough to be included in the analyses: Pomphorhynchus laevis, Acanthocephalus anguillae, Echinorhynchus truttae and Cyathocephalus truncatus. The observed frequencies of co-occurrences of the different species, based on presence/absence data, did not differ from those predicted by a null model derived from prevalence data. However, the intensity of infection (number of individuals per fish) of different helminth species were generally, but not always significantly, negatively correlated in tests of pairwise associations. Variation in fish sizes and its effect on infection levels, and whether or not two helminth species used the same or different intermediate hosts, had no influence on these findings. Of the few significant negative associations found between pairs of helminth species, none was found in more than one fish population. This suggests that interspecific associations may be condition-dependent: even in apparently similar localities, the same combinations of helminth species show different associations. Without evidence of replicability, it is almost impossible to conclude to the consistent role of competition between any pair of helminth species in the field.