Genetic characterization of nodular worm infections in Asian Apes.

Parasitic nematodes of Oesophagostomum spp., commonly known, as 'nodular worms' are emerging as the most widely distributed and prevalent zoonotic nematodes. Oesophagostomum infections are well documented in African non-human primates; however, the taxonomy, distribution and transmission of Oesophagostomum in Asian non-human primates are not adequately studied. To better understand which Oesophagostomum species infect Asian non-human primates and determine their phylogeny we analysed 55 faecal samples from 50 orangutan and 5 gibbon individuals from Borneo and Sumatra. Both microscopy and molecular results revealed that semi-wild animals had higher Oesophagostomum infection prevalence than free ranging animals. Based on sequence genotyping analysis targeting the Internal transcribed spacer 2 of rDNA, we report for the first time the presence of O. aculeatum in Sumatran apes. Population genetic analysis shows that there is significant genetic differentiation between Bornean and Sumatran O. aculeatum populations. Our results clearly reveal that O. aculeatum in free-ranging animals have a higher genetic variation than those in semi-wild animals, demonstrating that O. aculeatum is circulating naturally in wildlife and zoonotic transmission is possible. Further studies should be conducted to better understand the epidemiology and dynamics of Oesophagostomum transmission between humans, non-human primates and other wild species and livestock in Southeast Asia.

Oesophagostomiasis in non-human primates of Gombe National Park, Tanzania.

Oesophagostomum sp. is a parasitic nematode that frequently infects wild chimpanzees. Although nodular lesions are commonly associated with infection, some wild chimpanzee populations seem to tolerate Oesophagostomum nodular lesions while those at Gombe and other sites suffer from associated morbidity and mortality. From August 2004 to December 2013, we examined demographic (i.e., age, sex) and individual correlates (i.e., fecal consistency, Oesophagostomum egg production) to Oesophagostomum-associated pathology in 14 individually recognized chimpanzees at Gombe Stream National Park, Tanzania. In addition, we characterized Oesophagostomum-associated pathology in 14 individual sympatric primates including baboons, colobus, and cercopithecid monkeys. In five chimpanzees, there was no evidence of any significant underlying disease aside from oesophagostomiasis to explain the thin condition or diarrhea. All five of these chimpanzees had moderate to numerous parasitic nodules. In general, nodules were more numerous in older chimpanzees. Three of four chimpanzees with the highest average Oesophagostomum egg counts in feces collected during the year prior to their death had numerous parasitic nodules at necropsy. In contrast, the four chimpanzees with the lowest egg counts had only moderate numbers of nodules. No association (P = 0.74) was noted between frequency of diarrhea in the year prior to death and the number of nodules noted at necropsy. Nodules were also present in all baboons examined documenting pathology associated with Oesophagostomum infection in wild baboons. In contrast, no lesions were noted in colobus or cercopithecid monkeys, although it is uncertain if they are infected as no fecal studies have been completed in these species to date at Gombe. Sequence of DNA isolated from nodules in chimpanzees matched (99%) Oesophagostomum stephanostomum. Further research is needed to identify the types of Oesophagostomum causing lesions in baboons and to determine if baboons suffer from these infections. Am. J. Primatol. 80:e22572, 2018. © 2016 Wiley Periodicals, Inc.

Seasonal dynamics of endoparasitic infections at an organic goat farm and the impact of detected infections on milk production.

This study evaluated patterns and species composition of parasitic infections detected over a 1-year period at an organic goat farm. As a result of coprological examination, the overall prevalence of observed strongylids (99%), coccidia of the genus Eimeria (98%), and Muellerius capillaris lungworms (93%) was calculated. The most prevalent strongylids recovered from incubated fecal samples were Haemonchus contortus (42%), genera Trichostrongylus (23%), Oesophagostomum columbianum (13%), and Teladorsagia circumcincta (11%). A maximum intensity of coccidia infection 5150 oocysts per gram, strongylids infection 9900 eggs per gram and lungworm infection 867.26 larvae per gram were detected. The various effects (including environment, host, and parasites) on milk yield, lactose, protein, and fat were evaluated using generalized linear mixed models. Milk yield (P < 0.0001), milk fat (P < 0.01), and lactose (P < 0.0001) were affected by month, i.e., these parameters were influenced by the month of the year, regardless of the individual goat. With the intensity of infection detected in our study, only protein content was affected (P < 0.01) by parasitic infection (exclusively caused by strongylids). Correlation between measurements from one individual revealed that the goat itself can substantially decrease protein content but has much less of an effect on fat, milk yield, and lactose. Based on our results, we can conclude that a low intensity of parasitic infections does not significantly affect milk yield and the qualitative parameters of milk.

Prevalence of three Oesophagostomum spp. from Tibetan Pigs analyzed by Genetic Markers of nad1, cox3 and ITS1.

Parasitic nematodes of Oesophagostomum spp., commonly known as 'nodular worms' are one of the most widely distributed and prevalent emerging zoonotic nematodes. However, little is known about the prevalence and gene characteristics of those parasites in Tibetan pigs. Therefore, a study was carried out to investigate the prevalence, isolation and identification of Oesophagostomum spp from Tibetan pigs by genetic markers of nad1,cox3 and ITS1 for the first time. The results revealed that the infection rate of O. dentatum and O. quadrispinulatum by genetic markers of nad1 was 81.13%; 35 (66.04%); the O. dentatum and O. quadrispinulatum by genetic markers cox3 was 66.04%, and O. dentatum and O. stephanostomum by genetic markers ITS1 was found to be 77.36%. Interestingly, the O. stephanostomum specie was identified and isolated from 90.48% stomach and 69.23% colon samples by genetic markers of ITS1. The present study, for the first time has described the presence and genetic characterization of Oesophagostomun spp of O. dentatum, O. quadrispinulatum and especially O. stephanostomum in Tibetan pigs from the high and remote Tibetan plateau. A public concern should be raised in terms of economical losses and severe public health problem.

Molecular identification of the strongyloid nematode Oesophagostomum aculeatum in the Asian wild elephant Elephas maximus.

The transmission of zoonoses by wildlife, including elephants, is a growing global concern. In this study, we screened for helminth infections among Asian wild elephants (Elephas maximus) of the Salakpra Wildlife Sanctuary, Kanchanaburi, Thailand. Elephant faecal samples (45) were collected from the sanctuary grounds during January through November 2013 and assayed individually using the tetranucleotide microsatellite technique. Microscopic examination indicated a high prevalence of strongylids (93.0%) and low prevalences of trichurids (2.3%) and ascarids (2.3%). To identify the strongylid species, small subunit (SSU) rDNA sequences were amplified from copro-DNA and compared with sequences in GenBank. The generated SSU-rDNA sequences comprised five distinct haplotypes that were closely related to Oesophagostomum aculeatum. A phylogenetic analysis that incorporated related nematodes yielded a tree separated into two main clades, one containing our samples and human and domestic animal hookworms and the other consisting of Strongyloides. The present results indicate that O. aculeatum in local elephants is a potential source of helminthiasis in human and domestic animals in this wild-elephant irrupted area.

Nodular Worm Infections in Wild Non-human Primates and Humans Living in the Sebitoli Area (Kibale National Park, Uganda): Do High Spatial Proximity Favor Zoonotic Transmission?

BACKGROUND: Nodular Oesophagostomum genus nematodes are a major public health concern in some African regions because they can be lethal to humans. Their relatively high prevalence in people has been described in Uganda recently. While non-human primates also harbor Oesophagostomum spp., the epidemiology of this oesophagostomosis and the role of these animals as reservoirs of the infection in Eastern Africa are not yet well documented. METHODOLOGY/PRINCIPAL FINDINGS: The present study aimed to investigate Oesophagostomum infection in terms of parasite species diversity, prevalence and load in three non-human primates (Pan troglodytes, Papio anubis, Colobus guereza) and humans living in close proximity in a forested area of Sebitoli, Kibale National Park (KNP), Uganda. The molecular phylogenetic analyses provided the first evidence that humans living in the Sebitoli area harbored O. stephanostomum, a common species in free-ranging chimpanzees. Chimpanzees were also infected by O. bifurcum, a common species described in human populations throughout Africa. The recently described Oesophagostomum sp. found in colobine monkeys and humans and which was absent from baboons in the neighboring site of Kanyawara in KNP (10 km from Sebitoli), was only found in baboons. Microscopic analyses revealed that the infection prevalence and parasite load in chimpanzees were significantly lower in Kanyawara than in Sebitoli, an area more impacted by human activities at its borders. CONCLUSIONS/SIGNIFICANCE: Three different Oesophagostomum species circulate in humans and non-human primates in the Sebitoli area and our results confirm the presence of a new genotype of Oesophagostomum recently described in Uganda. The high spatiotemporal overlap between humans and chimpanzees in the studied area coupled with the high infection prevalence among chimpanzees represent factors that could increase the risk of transmission for O. stephanostomum between the two primate species. Finally, the importance of local-scale research for zoonosis risk management is important because environmental disturbance and species contact can differ, leading to different parasitological profiles between sites that are close together within the same forest patches.

Nodule worm infection in humans and wild primates in Uganda: cryptic species in a newly identified region of human transmission.

INTRODUCTION: Soil-transmitted helminths (STHs) are a major health concern in tropical and sub-tropical countries. Oesophagostomum infection is considered endemic to West Africa but has also been identified in Uganda, East Africa, among primates (including humans). However, the taxonomy and ecology of Oesophagostomum in Uganda have not been studied, except for in chimpanzees (Pan troglodytes), which are infected by both O. bifurcum and O. stephanostomum. METHODS AND FINDINGS: We studied Oesophagostomum in Uganda in a community of non-human primates that live in close proximity to humans. Prevalence estimates based on microscopy were lower than those based on polymerase chain reaction (PCR), indicating greater sensitivity of PCR. Prevalence varied among host species, with humans and red colobus (Procolobus rufomitratus) infected at lowest prevalence (25% and 41% by PCR, respectively), and chimpanzees, olive baboons (Papio anubis), and l'hoest monkeys (Cercopithecus lhoesti) infected at highest prevalence (100% by PCR in all three species). Phylogenetic regression showed that primates travelling further and in smaller groups are at greatest risk of infection. Molecular phylogenetic analyses revealed three cryptic clades of Oesophagostomum that were not distinguishable based on morphological characteristics of their eggs. Of these, the clade with the greatest host range had not previously been described genetically. This novel clade infects humans, as well as five other species of primates. CONCLUSIONS: Multiple cryptic forms of Oesophagostomum circulate in the people and primates of western Uganda, and parasite clades differ in host range and cross-species transmission potential. Our results expand knowledge about human Oesophagostomum infection beyond the West African countries of Togo and Ghana, where the parasite is a known public health concern. Oesophagostomum infection in humans may be common throughout Sub-Saharan Africa, and the transmission of this neglected STH among primates, including zoonotic transmission, may vary among host communities depending on their location and ecology.

Pathologic lesions in chimpanzees (Pan trogylodytes schweinfurthii) from Gombe National Park, Tanzania, 2004-2010.

During a population decline or disease outbreak, the true risk of specific diseases to a wild population is often difficult to determine because of a lack of baseline disease information. To better understand the risk of disease in an endangered and scientifically important population of chimpanzees (Pan trogylodytes schweinfurthii), a health monitoring program was initiated in Gombe National Park, Tanzania. As part of this health monitoring program, comprehensive necropsies with histopathology were conducted on chimpanzees (n = 11; 5 male, 6 female), ranging in age from fetal to 44 yr, that were found dead between August 2004 and January 2010. In contrast to previous reports, respiratory disease was not noted as a cause of morbidity or mortality. Trauma was the most common cause of death in these 11 chimpanzees. All of the chimpanzees greater than 1 yr of age had intestinal and mesenteric parasitic granulomas associated with true strongyles consistent with Oesophagostomum spp. The relative numbers of granulomas increased with age and, in some cases, may have been a cause of weight loss and diarrhea. Simian immunodeficiency virus (SIV)cpz infection was documented in four deceased apes, all of whom exhibited varying amounts of lymphoid depletion including two females with marked CD4+ T cell loss consistent with endstage SIVmac or human immunodeficiency virus infections. Myocardial megalokaryosis was common in chimpanzees greater than 1 mo of age; yet myocardial interstitial fibrosis, a common lesion in captive chimpanzees, was uncommon and only noted in two aged chimpanzees. These findings provide important information on causes of morbidity and mortality in wild chimpanzees, information that can be used to interpret findings during population declines and lead to better management of this population in the context of disease risk.

[Nematodes of the genus Oesophagostomum: an emerging risk for humans and apes in Africa?]. / Les nématodes du genre Oesophagostomum. Un risque émergent pour l'homme et les grands singes en Afrique?

Nematodes of the genus Oesophagostomum are common intestinal parasites found in cattle, pigs and primates. They can cause severe illness, resulting from the formation of granulomas, caseous lesions and abscesses in the intestinal wall. Human oesophagostomosis is endemic in northern Ghana and Togo. In these regions, epidemiological investigations have been conducted to determine the biological characteristics, transmission dynamics and optimal management of clinical cases. Nodular oesophagostomosis has also been described in free-ranging chimpanzees and gorillas. Clinical signs associated with nodules have been observed in great apes raised in sanctuaries, while the health status of their wild counterparts does not seem to be significantly affected It has been suggested that some nonhuman primates may act as reservoirs for human oesophagostomosis. In Ghana, identification of genetic differences among Oesophagostomum nematodes infecting different primate hosts suggests that oesophagostomosis is a rare zoonosis. In Uganda, where the situation is diferent, cross-infection is probably more frequent.

Nodular worm infection in wild chimpanzees in Western Uganda: a risk for human health?

This study focused on Oeosophagostomum sp., and more especially on O. bifurcum, as a parasite that can be lethal to humans and is widespread among humans and monkeys in endemic regions, but has not yet been documented in apes. Its epidemiology and the role played by non-human primates in its transmission are still poorly understood. O. stephanostomum was the only species diagnosed so far in chimpanzees. Until recently, O. bifurcum was assumed to have a high zoonotic potential, but recent findings tend to demonstrate that O. bifurcum of non-human primates and humans might be genetically distinct. As the closest relative to human beings, and a species living in spatial proximity to humans in the field site studied, Pan troglodytes is thus an interesting host to investigate. Recently, a role for chimpanzees in the emergence of HIV and malaria in humans has been documented. In the framework of our long-term health monitoring of wild chimpanzees from Kibale National Park in Western Uganda, we analysed 311 samples of faeces. Coproscopy revealed that high-ranking males are more infected than other individuals. These chimpanzees are also the more frequent crop-raiders. Results from PCR assays conducted on larvae and dried faeces also revealed that O. stephanostomum as well as O. bifurcum are infecting chimpanzees, both species co-existing in the same individuals. Because contacts between humans and great apes are increasing with ecotourism and forest fragmentation in areas of high population density, this paper emphasizes that the presence of potential zoonotic parasites should be viewed as a major concern for public health. Investigations of the parasite status of people living around the park or working inside as well as sympatric non-human primates should be planned, and further research might reveal this as a promising aspect of efforts to reinforce measures against crop-raiding.

Faecal egg output and herbage contamination with infective larvae of species of Ostertagia and Oesophagostomum from naturally infected farmed sika deer Cervus nippon in Lithuania.

Faecal egg outputs and subsequent herbage larval contamination with third stage larvae (L3) of Ostertagia spp. and Oesophagostomum spp. from a herd of naturally infected sika deer Cervus nippon were examined in the same pasture in 2001/2002 in Lithuania. Sika deer were infected with Ostertagia circumcincta, O. kolchida, O. spiculoptera, Oesophagostomum radiatum, O. columbianum and O. venulosum. Faecal egg output in adult deer peaked in the spring during the periparturient period and also in late August, compared with a peak in egg output in calves during September to November. Herbage contamination with L3 of Ostertagia spp. peaked in June but larvae were not present on pastures from the end of September. Hence the highest risk of infection was in early born calves grazed on pastures in July. Infective larvae of Oesophagostomum spp. did not survive during the winter, but the nematodes were reintroduced onto the pastures by adult deer in the spring.

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.

Annual mass treatment with albendazole might eliminate human oesophagostomiasis from the endemic focus in northern Ghana.

As a follow-up to the study by Ziem et al., in this issue, efforts to control human oesophagostomiasis and hookworm infections in northern Ghana were pursued, and the results evaluated in collaboration with the Lymphatic Filariasis Elimination Programme. This phase of evaluation of the impact of mass treatment was no longer limited to a small-scale research setting: it was done both in the context of an operationally viable national control programme and as a continuation of the Oesophagostomum Intervention Research Project (OIRP). The methods of evaluation included classical stool examination with Kato thick smears, stool culture and ultrasound examination of the colon wall. The results showed that yearly population-based albendazole-ivermectin treatment in 11 villages scattered over north-eastern Ghana, with a treatment coverage of 70-75%, resulted in a reduction of Oesophagostomum prevalence from about 20% pre-intervention to less than 1% after 2 years of mass treatment. Simultaneously, hookworm prevalence went down from 70% to approximately 15%. The data, however, cannot be readily compared with those of Ziem et al. because of the relatively crude diagnostic (single stool cultures) screening system that had to be used for the evaluation of the large-scale control programme. In the research area of the OIRP, interruption of mass treatment resulted in a rising hookworm prevalence. The Oesophagostomum prevalence, on the other hand, continued to go down. Transmission of human oesophagostomiasis appears interruptible and small numbers of persistent cases of Oesophagostomum infection were shown insufficient to serve as a nucleus of renewed spread of the infection. The data suggest that both the infection with and the pathology due to human oesophagostomiasis can be eliminated and that elimination is likely to be achieved through operationally feasible albendazole-ivermectin treatment as used by the Global Alliance for the Elimination of Lymphatic Filariasis.

Oesophagostomum dentatum and Trichuris suis infections in pigs born and raised on contaminated paddocks.

Transmission of Oesophagostomum dentatum and Trichuris suis was studied in outdoor reared pigs. Six farrowing paddocks were naturally contaminated in May to mid-June 2001 by experimentally infected seeder pigs. In early July 1 sow farrowed on each paddock and starting at week 3 post-partum (p.p.) the offspring was slaughtered serially every 2 weeks for parasite recovery. Faeces were collected regularly for parasite egg counts and acid-insoluble ash (AIA) content as an indicator of geophagy. Weaning took place at week 7 p.p. by removing the sow. Paddock infection levels were estimated in mid-June (O. dentatum) and late November (O. dentatum and T. suis) using helminth-naïve tracer pigs. Soil and vegetation samples were collected regularly. Despite a high initial contamination by the seeder pigs, O. dentatum paddock infectivity was negligible to low throughout the raising of the experimental piglets, which had a slow accumulation of nodular worms ending with a mean of 422 worms/pig at week 19 p.p. As only few eggs developed to infectivity overall T. suis transmission was minimal. The first T. suis were recovered at week 11 p.p. and the highest mean burden of 21 worms/pig was recorded at week 19 p.p. The experimental pigs initially had a high faecal level of AIA although it decreased over time. The results are discussed in relation to the biological characteristics of the 2 parasites and their occurrence in organic pig production.

Mass treatment with albendazole reduces the prevalence and severity of Oesophagostomum-induced nodular pathology in northern Ghana.

Previous surveys conducted in northern Ghana where Oesophagostomum bifurcum is endemic showed that O. bifurcum-induced nodular pathology could be detected in up to 50% of the inhabitants. The impact of albendazole-based mass treatment to control both infection and morbidity is assessed and compared with the situation in a control area where no mass treatment has taken place. A significant reduction in the prevalence of infection based on stool cultures was achieved following two rounds of mass treatment in one year: from 52.6% (361/686) pre treatment to 5.2% (22/421) 1 year later (chi(1)(2)=210.1; P<0.001). At the same time, the morbidity marker of ultrasound-detectable nodules declined from 38.2% to 6.2% (chi(1)(2)=138.1; P<0.001). There was a shift from multinodular pathology, often seen in heavy infections, to uninodular lesions. In the control villages where no treatment took place, O. bifurcum infection increased from 17.8% (43/242) to 32.2% (39/121) (chi(1)(2)=9.6; P<0.001). Nodular pathology decreased slightly from 21.5% to 19.0%, but a higher proportion of these subjects developed multinodular pathology compared with baseline (chi(1)(2)=5.5; P=0.019). It is concluded that repeated albendazole treatment significantly reduces O. bifurcum-induced morbidity.

Distribution and clustering of Oesophagostomum bifurcum and hookworm infections in northern Ghana.

Human Oesophagostomum infections are locally common in northern Ghana. The present study describes the results of a cross-sectional survey involving 1011 subjects, selected by a compound-based random sampling method from 1227 compounds in 24 villages. Selected persons were examined by both Kato and coproculture methods. Hookworm-like eggs, representing ova of Oesophagostomum bifurcum and hookworm were detected in 87.5% of the Kato smears. The geometric mean egg count of the infected subjects was 1018. Upon coproculture, third-stage larvae of O. bifurcum and hookworm were detected in 53.0% and 86.9% of subjects respectively. Oesophagostomum infections were clustered but no clear explanation for aggregation of infections could be found as yet. Subjects infected with hookworm had a 5-fold higher risk of being infected with O. bifurcum. Infection rates in adult women were higher than in adult men. No association was found with family size, level of hygiene or with the presence of animals in the compounds. Representatives of the Bimoba-tribe were significantly more infected than those of the other tribes. It appears, however, that this tribal association is a geographical phenomenon: Bimoba are mostly living in villages with the highest infection rates.

Insights into the epidemiology and genetic make-up of Oesophagostomum bifurcum from human and non-human primates using molecular tools.

The nodule worm Oesophagostomum bifurcum (Nematoda: Strongylida) is a parasite of major human health importance predominantly in northern Togo and Ghana. Currently, it is estimated that 0.25 million people are infected with this nematode, and at least 1 million people are at risk of infection. Infection with this parasite causes significant disease as a consequence of encysted larvae in the wall of the large intestine. In spite of the health problems caused by O. bifurcum, there have been significant gaps in the knowledge of the biology, transmission and population genetics of the parasite. This review provides an account of some recent insights into the epidemiology and genetics of the parasite from human and non-human primate hosts in specific regions of Africa using molecular tools. Recent research findings are discussed mainly in relation to non-human primates being reservoirs of infection, and the consequences for the prevention and control of oesophagostomiasis in humans are briefly discussed.

Oesophagostomum bifurcum in non-human primates is not a potential reservoir for human infection in Ghana.

In northern Togo and Ghana, human infection with the parasitic nematode Oesophagostomum bifurcum is of major health importance. Elsewhere, oesophagostomiasis is considered a zoonotic infection, non-human primates being the natural host. We examined 349 faecal samples of the olive baboon, mona monkey and black and white colobus monkey from two geographically distinct areas in Ghana, outside the region endemic for O. bifurcum in humans. Using both microscopy and species-specific PCR, we found a high prevalence of O. bifurcum (75-99%) in olive baboons and mona monkeys. The majority of the test-positive faecal samples contained large numbers of larvae after copro-culture (>100). No O. bifurcum was detected in the faeces of the black and white colobus monkeys. Observational studies on the behaviour of the non-human primates, focusing on defecation, food consumption and the sharing of habitat with the local human population, indicated favourable conditions for zoonotic transmission. Given that no human infection with O. bifurcum has been reported from either study area, the present findings support the hypothesis that O. bifurcum from humans in the north of Ghana, and O. bifurcum from olive baboons and/or mona monkeys are distinct.

Definition of genetic markers in nuclear ribosomal DNA for a neglected parasite of primates, Ternidens deminutus (Nematoda: Strongylida)--diagnostic and epidemiological implications.

Ternidens deminutus (Strongylida) is a parasitic nematode infecting non-human and human primates in parts of Africa, Asia and the Pacific islands. The present study genetically characterized T. deminutus and defined genetic markers in nuclear ribosomal DNA (rDNA) as a basis for developing molecular-diagnostic tools. The sequences of the second internal transcribed spacer (ITS-2) of rDNA were determined for adult specimens of T. deminutus (Nematoda: Strongylida: Oesophagostominae) from the Olive baboon and the Mona monkey. The length and G+C content of the ITS-2 sequences was 216 bp and approximately 43%, respectively. While there was no sequence variation among individual T. deminutus specimens from the baboon, 6 (2.8%) nucleotide differences were detected in the ITS-2 between the parasite from baboon and that of the Mona monkey, which is similar to the difference (3.2%) between 2 other species of Oesophagostominae (Oesophagostomum bifurcum and O. stephanostomum) from non-human primates, suggesting significant population variation or the existence of cryptic (i.e. hidden) species within T. deminutus . Pairwise comparisons of the ITS-2 sequences of the 2 operational taxonomic units of T. deminutus with previously published ITS-2 sequences for selected members of the subfamilies Oesophagostominae and Chabertiinae indicated that species from primates (including those representing the subgenera Conoweberia and Ihleia) are closely related, in accordance with previous morphological studies. The sequence differences (27-48.3%) in the ITS-2 between the 2 taxonomic units of T. deminutus and hookworms (superfamily Ancylostomatoidea) enabled their identification and delineation by polymerase chain reaction (PCR)-based mutation scanning. The genetic markers in the ITS-2 provide a foundation for improved, PCR-based diagnosis of T. deminutus infections and for investigating the life-cycle, transmission patterns and ecology of this parasite.