Natural Definitive Host Research Articles

Canine Alveolar Echinococcosis: Morphology and Inflammatory Response

Echinococcus multilocularis is a tapeworm with zoonotic potential that is common in the northern hemisphere. Its natural definitive hosts are foxes and other canids and its intermediate hosts are rodents. Other mammalian species, including man, can become infected and develop infiltrative, tumour-like lesions that particularly affect the liver. The present study characterizes the relatively rare lesions of alveolar echinococcosis in 10dogs. Metacestodes in these dogs frequently contained no protoscolices and were thus sterile. Metacestodes were surrounded by fibrous tissue intermixed with inflammatory cells comprising variable numbers of macrophages, T and B lymphocytes, and plasma cells, as well as neutrophils and/or eosinophils. These lesions resemble those observed in man and are in contrast to those observed in the natural intermediate hosts, which have less fibrosis and more protoscolices.

Intrahepatic growth and maturation of Gnathostoma turgidum in the natural definitive opossum host, Didelphis virginiana

Gnathostoma turgidum is a gastric nematode parasite of opossums found in the Americas. We recently found that G. turgidum juveniles appear in the liver of the opossums where they become mature adults and almost synchronously move to the stomach during certain months of the year, suggesting the importance of the liver for the growth and maturation of this species in the final hosts. In this study we attempted to detect G. turgidum larvae in the liver of opossums, Didelphis virginiana that are the natural final hosts. The results show that tiny (< 3 mm in length) third stage larvae (L3) appeared in the liver of opossums around November and December. Also in the liver, we found large L3 of up to about 10 mm in length together with juveniles and mature adults from February to March. In spite of their length, large L3 have 4 rows of hooklets, and their gonads remained undeveloped. Morphological features of the small and large L3 of G. turgidum are described including scanning electron microscope images. The seasonal switching of the several growth stages of G. turgidum from small L3 to adult worms in the liver and eventual migration to the stomach in opossums suggests the unique feature of G. turgidum utilizing the liver as the maturation site.

Frequency distribution of Echinococcus multilocularis and other helminths of foxes in Kyrgyzstan

Echinococcosis is a major emerging zoonosis in central Asia. A study of the helminth fauna of foxes from Naryn Oblast in central Kyrgyzstan was undertaken to investigate the abundance of Echinococcus multilocularis in a district where a high prevalence of this parasite had previously been detected in dogs. A total of 151 foxes (Vulpes vulpes) were investigated in a necropsy study. Of these 96 (64%) were infected with E. multilocularis with a mean abundance of 8669 parasites per fox. This indicates that red foxes are a major definitive host of E. multilocularis in this country. This also demonstrates that the abundance and prevalence of E. multilocularis in the natural definitive host are likely to be high in geographical regions where there is a concomitant high prevalence in alternative definitive hosts such as dogs. In addition Mesocestoides spp., Dipylidium caninum, Taenia spp., Toxocara canis, Toxascaris leonina, Capillaria and Acanthocephala spp. were found in 99 (66%), 50 (33%), 48 (32%), 46 (30%), 9 (6%), 34 (23%) and 2 (1%) of foxes, respectively. The prevalence but not the abundance of E. multilocularis decreased with age. The abundance of D. caninum also decreased with age. The frequency distribution of E. multilocularis and Mesocestoides spp. followed a zero-inflated negative binomial distribution, whilst all other helminths had a negative binomial distribution. This demonstrates that the frequency distribution of positive counts and not just the frequency of zeros in the data set can determine if a zero-inflated or non-zero-inflated model is more appropriate. This is because the prevalences of E. multolocularis and Mesocestoides spp. were the highest (and hence had fewest zero counts) yet the parasite distribution nevertheless gave a better fit to the zero-inflated models.

Echinostome Infections in the Striped-Field Mouse, Apodemus agrarius, and the Ussuri White-Toothed Shrew, Crocidura lasiura, Caught Near the Demilitarized Zone, Gyeonggi-do (Province), Republic of Korea

A total of 1,498 small mammals (rodents and insectivores), including Apodemus agrarius (n = 1,366), Crocidura lasiura (54), Mus musculus (32), Micronytus fortis (28), Eothenomys regulus (9), Micronys minutes (6), and Cricetulus triton (3), were live-trapped in Gyeonggi-do (Province) (Paju-si, Pocheon-gun, and Yeoncheon-gun) near the demilitarized zone (DMZ) from December 2004 to September 2005. A. agrarius was found to be infected with 3 species of echinostomes (Echinostoma hortense, Echinostoma cinetorchis, and Euparyphium murinum), while C. lasiura was infected with 1 species (Echinochasmus japonicas) of echinostome. Other mammals were free from echinostome infections. Total 16 E. hortense were detected in 7 (0.5%) mice, 9 E. cinetorchis from 5 (0.4%), and 3 E. murinum from 2 (0.1%) out of 1.366 A. agrarius examined. E. japonicus was found only in 1 (1.9%; total 3 specimens) C. lasiura. These results demonstrate that A. agrarius and C. lasiura, inhabiting near the DMZ of Gyeonggi-do serve as the natural definitive hosts for several species of echinostomes, although their infection rates are low. This is the first record of natural infections of A. agrarius with E. cinetorchis and C. lasiura with E. japonicus in the Republic of Korea.

Intestinal Helminth Infections in Feral Cats and a Raccoon Dog on Aphaedo Island, Shinan-gun, with a Special Note on Gymnophalloides seoi Infection in Cats

Four feral cats and a raccoon dog purchased from a local collector on Aphaedo Island, Shinan-gun, where human Gymnophalloides seoi infections are known to be prevalent, were examined for their intestinal helminth parasites. From 2 of 4 cats, a total of 310 adult G. seoi specimens were recovered. Other helminths detected in cats included Heterophyes nocens (1,527 specimens), Pygidiopsis summa (131), Stictodora fuscata (4), Acanthotrema felis (2), Spirometra erinacei (15), toxocarids (4), and a hookworm (1). A raccoon dog was found to be infected with a species of echinostome (55), hookworms (7), toxocarids (3), P. summa (3), and S. erinacei (1). No G. seoi was found in the raccoon dog. The results indicate that feral cats and raccoon dogs on Aphaedo are natural definitive hosts for intestinal trematodes and cestodes, including G. seoi, H. nocens, and S. erinacei. It has been first confirmed that cats, a mammalian species other than humans, play the role of a natural definitive host for G. seoi on Aphaedo Island.

The Ruddy Turnstone, Arenaria interpres interpres, a New Definitive Host for Gynaecotyla squatarolae (Digenea: Microphallidae)

The ruddy turnstone, Arenaria interpres interpres, a migratory Korean bird, was proved to be a natural definitive host for Gynaecotyla squatarolae (Digenea: Microphallidae). The ruddy turnstone was found dead at the seashore of Okgueup, Gunsan-si, Jeollabuk-do. The intestinal tract was examined, and 98 unknown flukes were recovered. The worms were 600 284 micrometer in size, and had 2 ventral suckers. The seminal vesicle was large, the genital atrium was prominent, and the average egg size was 20 12.5 micrometer. Based on these results, the worms were identified as G. squatarolae. This is the first report on the ruddy turnstone as a natural definitive host of G. squatarolae in the Republic of Korea.

RENEWED HOPE FOR A VACCINE AGAINST THE INTESTINAL ADULT TAENIA SOLIUM

Review of experimental and observational evidence about various cestode infections of mammalian hosts revives hope for the development of an effective vaccine against adult intestinal tapeworms, the central protagonists in their transmission dynamics. As for Taenia solium, there are abundant immunological data regarding cysticercosis in humans and pigs, but information about human taeniasis is scarce. A single publication reporting protection against T. solium taeniasis by experimental primo infection and by vaccination of an experimental foster host, the immunocompetent female hamster, kindles the hope of a vaccine against the tapeworm to be used in humans, its only natural definitive host.

Apodemus agrarius as a new definitive host for Neodiplostomum seoulense

A total of 1,496 rodents and insectivores were live-trapped at Yeoncheon-gun (n = 351), Paju-shi (804), and Pocheon-gun (343), Gyeonggi-do (Province), and examined for intestinal helminths, including Neodiplostomum seoulense, seasonally from December 2004 to September 2005. Six species of rodents, including Apodemus agrarius (1,366), Mus musculus (32), Micronytus fortis (28), Eothenomys regulus (9), Micronys minutus (6), and Cricetulus triton (3), and 1 species of insectivores Crocidura lasiura (54) were collected. A total of 321 adult N. seoulense were collected from 19 (1.4%) A. agrarius. The worm burden ranged from 1 to 101 per A. agrarius (mean; 16.9). No N. seoulense was observed in other rodent or insectivore species examined. The infection rate during autumn (4.5%) was higher than those during spring (0.8%), summer (0.8%), and winter (0.5%). The average number of N. seoulense in infected A. agrarius was the highest in spring (66.0 specimens), followed by autumn (15.2), winter (4.5), and summer (3.3). This study first confirms that A. agrarius is a natural definitive host for N. seoulense, and demonstrates that the infection rates and intensities vary seasonally and geographically.

Plagiorchis muris infection in Apodemus agrarius from northern Gyeonggi-do (Province) near the demilitarized zone

The small intestines of 6 species of rodents and 1 species of insectivore were examined seasonally for Plagiorchis muris infection in 3 different localities in northern Gyeonggi-do (Province), near the demilitarized zone (DMZ). A total of 1,496 animals, including 1,366 Apodemus agrarius, 54 Crocidura lasiura (insectivore), 32 Mus musculus, 28 Micronytus fortis, 9 Eothenomys regulus, 6 Micronys minutus, and 3 Cricetulus triton, were live-trapped at Yeoncheon-gun (n = 351), Paju-shi (804) and Pocheon-gun (343) at 3-mo intervals from December 2004 to September 2005. A total of 1,647 P. muris were collected from 72 (5.3%) A. agrarius. The infection rate was the highest in Pocheon-gun (8.2%), followed by Yeoncheon-gun (5.0%) and Paju-shi (4.2%). A higher infection rate was observed in A. agrarius captured during September (19.4%) than those captured during December (3.0%), June (2.6%), or April (0%). However, the worm burden was the highest in June (av. 32.1/animal), followed by September (24.7), December (4.0), and April (0). None of the other animal species were found infected with P. muris. The results reveal that A. agrarius is a natural definitive host for P. muris, and infection rates and worm burdens vary seasonally and geographically.

Cranial helminths of Mustela vison Schreber, 1777 in Spain

A survey was carried out to investigate the presence of cranial helminths in 337 American minks ( Mustela vison) from Spain. This information was obtained partly in order to evaluate potential conservation problems and sanitary risks to the congeneric European mink ( Mustela lutreola), one of the most endangered carnivores in the world. Skulls and rectal faeces of each specimen were simultaneously analysed. Troglotrema acutum and Skrjabingylus nasicola were found in 5.6% of the M. vison analysed. No cranial lesions were seen in any of the examined skulls. The finding of both helminths in Spanish free-living M. vison specimens enlarges their natural definitive host spectrum in Western Europe. One relatively important focus of T. acutum in M. vison was detected (30.4%) in the Spanish Álava province while S. nasicola was found to be very infrequent. The suitability of both analytical methods was assessed in order to know to what degree coprological analysis reflects the real prevalence of cranial helminths in this host. It is possible to conclude that coprological analysis can be used instead of necropsies to analyse the possible incidence of pathogenic cranial helminths in mustelids. This aspect is very important and useful when trying to analyse the helminthological status of endangered species such as the native mink ( M. lutreola) particularly in areas where both congeneric species are present and strict competition occurs.

Development of Taenia saginata asiatica metacestodes in SCID mice and its infectivity in human and alternative definitive hosts

Development of Taenia saginata asiatica metacestodes in SCID mice, and its infectivity in humans, golden hamsters, and Mongolian gerbils as alternative definitive hosts, were investigated. Cysticerci were recovered from SCID mice that were subcutaneously injected with hatched oncospheres of T. s. asiatica. The morphological changes of metacestodes were observed. The recovered cysticerci were fed to gerbils, hamsters and humans, to check for their infectivity. Tapeworms were recovered from gerbils and hamsters fed with 20 to 45 week-old cysticerci, and proglottids excretions were observed in human volunteers fed with 45 week-old cysticerci. However, no tapeworms were recovered from gerbils fed with 10 week-old cysticerci. Our results suggest that T. s. asiatica oncospheres needed more than 20 weeks to develop to maturity in SCID mice to be infective to both their natural and alternative definitive hosts.

High incidence of the gullet worm, Gongylonema pulchrum, in a squirrel monkey colony in a zoological garden in Japan

Histological examination revealed the gullet worm ( Gongylonema pulchrum) embedded in the lingual mucosa of two of four dead Bolivian squirrel monkeys ( Saimiri boliviensis) from a zoological garden in Kyushu, Japan. The zoo had kept the monkeys as a colony of approximately 80–100 individuals in a moated, open ground since 1981. To assess the incidence of G. pulchrum infection in this colony, lingual scraping with disposable plastic sticks was conducted in February during 2 consecutive years (2003 and 2004). The oval, thick-shelled egg containing a larva was found in 15 of 27 arbitrarily-chosen adult monkeys (55.5%) in 2003, and 27 of 106 monkeys (25.5%) in 2004. Infection of other zoo-kept mammals with G. pulchrum was not assessed. Since the gullet worm infects a variety of mammals including primates as natural definitive hosts, and dung beetles and cockroaches as intermediate hosts, the zoological garden may provide an ideal environment for the parasite. Zoo veterinarians should be aware of this disease in kept mammals, and should consider in the case of primates, monthly or bimonthly prophylactic anthelmintic treatment.

Light and scanning electron microscopy of the miracidium of Echinostoma paraensei (Trematoda, Echinostomatidae)

Echinostoma paraensei was described 1967 by Lie and Basch. Recently its natural definitive host, the aquatic rodent Nectomys squamipes , endemic in Brazil, was identified. As most of the echinostomatids, this species presents a zoonotic potential. The morphology and topography of the E. paraensei miracidium obtained from adult worms collected from the natural definitive host N. squamipes is described by light and electron microscopy. The arrangement and the dimensions of the epidermal plates are given. The eyespots are composed by two pairs of lenses measuring 6.03 μm. SEM observations shows that the miracidium body is covered by cilia except at the terebratorium region and the presence of α-tubulin in the cilia of the larvae is first recorded by immunelabeling. Nineteen papilla-like structures arranged in three axes and four groups were observed at the terebratoriun, this structure is retractable, presenting folds and a corrugated surface with profiles of cytoplasmic expansions, forming network of anatomizing folds.

Light and scanning electron microscopy of the miracidium of Echinostoma paraensei (Trematoda, Echinostomatidae)

Echinostoma paraensei was described 1967 by Lie and Basch. Recently its natural definitive host, the aquatic rodent Nectomys squamipes, endemic in Brazil, was identified. As most of the echinostomatids, this species presents a zoonotic potential. The morphology and topography of the E. paraensei miracidium obtained from adult worms collected from the natural definitive host N. squamipes is described by light and electron microscopy. The arrangement and the dimensions of the epidermal plates are given. The eyespots are composed by two pairs of lenses measuring 6.03 μm. SEM observations shows that the miracidium body is covered by cilia except at the terebratorium region and the presence of α-tubulin in the cilia of the larvae is first recorded by immunelabeling. Nineteen papilla-like structures arranged in three axes and four groups were observed at the terebratoriun, this structure is retractable, presenting folds and a corrugated surface with profiles of cytoplasmic expansions, forming network of anatomizing folds.

Relationship between intermediate host taxon and infection by nematodes of the genus Rhabdochona

We describe the intermediate and definitive hosts of the fish nematodes Rhabdochona coronacauda and R. denudata honshuensis and discuss the relationships between parasitism and the feeding habitats of their intermediate hosts. We found that the principal intermediate hosts of the two nematodes were filter-feeding mayflies of the genera Ephemera, Photamanthus and Isonychia. Ephemera strigata seemed to be the most important intermediate host of these nematodes. Adult R. coronacauda were found mainly in Hemibarbus longirostris and Rhinogobius flumineus, which are benthic fishes that feed on benthic aquatic insects, including E. strigata. For R. coronacauda, therefore, the feeding habits of the definitive hosts facilitate host alternation by this species. However, adult R. denudata honshuensis were found in cyprinids. In particular, Zacco temmincki was the principal natural definitive host in our study area. Since Z. temmincki is a swimming predator, E. strigata nymphs that burrow in the substrate are not the main prey of this species. This indicates that the transmission of R. denudata honshuensis hardly occurs from E. strigata nymph to Z. temmincki, suggesting another, unknown transmission route.

Red foxes ( Vulpes vulpes) are a natural intermediate host of Neospora caninum

The present study was undertaken to determine if red foxes are natural intermediate and/or definitive host for Neospora caninum and to study the importance of infection of N. caninum in this species in North-eastern Spain. Faecal samples and brain tissues were obtained from 122 foxes from 21 rural areas of Catalonia. Faeces collected were examined for parasite eggs and coccidian oocysts using sucrose flotation. For PCR-based diagnosis of N. caninum in brain tissues, the specific genomic Nc5 region was selected as the target sequence for DNA amplification. To control for PCR failure and facilitate identification of truly negative samples, the competitor pNc5C molecule was added to all negative samples in a second round of PCR reactions. Of the 122 foxes analysed, 13 (10.7%) were positive by PCR for N. caninum. Signal intensities of all positive samples were relatively weak with the exception of one sample from a 3-month male animal, that also showed the highest repeatability. No differences were observed by sex, age or area of sampling analysis. Detection of stages of N. caninum in brain from naturally infected red foxes demonstrated that red foxes are a natural intermediate host for N. caninum. Faecal samples were analysed for the presence of N. caninum oocysts, however, no oocysts compatible with N. caninum were found. A widespread latent infection of red foxes in North-eastern Spain found in the present study indicates that red foxes could have a very important role in the epidemiology of neosporosis in our area.

Sex-specific genetic structure in Schistosoma mansoni: evolutionary and epidemiological implications.

We studied the population genetic structure of 360 and 1247 adult Schistosoma mansoni using seven microsatellite and seven random amplified polymorphic DNA (RAPD) markers, respectively. Parasites were collected from their natural definitive host Rattus rattus in Guadeloupe (West Indies). We found a sex-specific genetic structure, a pattern never before reported in a parasitic organism. Male genotypes were more randomly distributed among rats than female genotypes. This interpretation was consistent with a lower differentiation between hosts for males relative to females, the higher genetic similarity between females in the same host and the observed local (i.e. within-individual-host) differences in allele frequencies between the two sexes. We discuss our results using ecological and immunological perspectives on host-parasite relationships. These results change our view on the epidemiology of schistosomiasis, a serious disease affecting humans in African and American intertropical zones.

Life cycle of Brachylaima mascomai n. sp. (Trematoda: Brachylaimidae), a parasite of rats in the Llobregat delta (Spain).

The terrestrial triheteroxenous life cycle of Brachylaima mascomai n. sp. (Trematoda: Brachylaimidae) is elucidated. Operculated, assymetric, embryonated eggs (25.4 x 12.7 microm) are passed with feces of the natural (Rattus norvegicus and R. rattus) and experimental (albino and wild mice, albino rats, Apodemus sylvaticus, Mus spretus [Muridae] and the golden gerbil) definitive hosts and ingested by the helicid gastropod Pseudotachea splendida, the only natural and experimental first intermediate host. Microcaudate cercariae harbored in branched sporocysts in the digestive gland emerge from this snail and contact P. splendida, Otala punctata, Theba pisana, and Helix (C.) aspersa snails developing into unencysted infective metacercariae in the kidney. Definitive hosts are infected by ingestion of infected snails; the adult parasites inhabit the small intestine. Chaetotaxic cercarial pattern specific at acetabular (S(II) 8-10 papillae) and cephalic (C(III) 13-15 papillae, H 16 papillae) levels. Three types of cercarial papillae are observed by scanning electron microscopy (SEM) and their arrangement is correlated with chaetotaxy for the first time in trematodes: argentophilic papillae with fingerlike process (cephalic, body, and acetabular levels), argentophilic papillae with opening (2 papillae in the M body level), and nonargentophilic dome-shaped papillae (alternated with argentophilic S(II) papillae on the ventral sucker). SEM detected interlacing network of ridges covering the metacercarial body. Adults with multidigitate tegumentary spines were observed by SEM. Subequal suckers; the acetabulum located in the posterior part of anterior fifth of body. Vitellaria extend from between middle level and anterior margin of anterior testis to between middle level and posterior margin of acetabulum. Uterus almost reaches the intestinal bifurcation.

Description of the adult worms of a new Brazilian isolate of Echinostoma paraensei (Platyhelminthes:Digenea) from its natural vertebrate host Nectomys squamipes by light and scanning electron microscopy and molecular analysis.

Echinostoma paraensei Lie and Basch, 1967 (Echinostomatidae:Platyhelminthes), a 37 collar spine echinostome of the "revolutum group", has been used extensively as a model organism to study the interactions of digenetic trematodes with both their snail and vertebrate hosts. This worm was first isolated from the snail Biomphalaria glabrata from Belo Horizonte (BH isolate), Minas Gerais State, Brazil, by Lie and Basch [J Parasitol (1967) 53:1192-1199]. The natural definitive host for the BH isolate was never determined, and it has been maintained in the laboratory since 1967 in B. glabrata and hamsters. In this study, using light and scanning electron microscopy and molecular analysis, we describe an echinostome recently obtained from its natural vertebrate host, the wild rodent Nectomys squamipes (Rodentia: Sigmodontinae) from Sumidouro, Rio de Janeiro State, Brazil (RJ isolate). This echinostome was also compared to the laboratory-maintained BH isolate of E. paraensei. We observed that adult worms of both BH and RJ isolates could be differentiated from other echinostome species by the relatively small size of the dorsal collar spines relative to lateral and corner collar spines. SEM confirmed the similarity of this morphological character between the two isolates. As additional diagnostic features, the tegumentary spines are scale-like and the region between the genital pore and the acetabulum lacks scales. There is a folded protuberance with a pore just posterior to the genital pore. The tegument of the acetabulum is unspined and radially wrinkled, and there are numerous randomly distributed small, domed, ciliated papillae. The sequences of the internal transcribed spacers of the nuclear rDNA complex of the RJ and BH isolates are identical. Together these shared features provide strong evidence that both isolates are the same and can be referred to as E. paraensei. In conclusion, we have identified, for the first time, one of the natural definitive hosts for E. paraensei, the rodent N. squamipes, and have extended the known geographical distribution of this species to include Sumidouro in Rio de Janeiro State, Brazil.

A review of Gymnophalloides seoi (Digenea: Gymnophallidae) and human infections in the Republic of Korea.

Studies on Gymnophalloides seoi (Digenea: Gymnophallidae) and human infections are briefly reviewed. This minute intestinal fluke was first discovered from a Korean woman suffering from acute pancreatitis and gastrointestinal troubles. It was described as a new species by Lee, Chai and Hong in 1993. The southwestern coastal village where the patient resided was found to be a highly endemic area, and additional endemic areas have been identified. The parasite is very small, 0.33-0.50 mm long and 0.23-0.33 mm wide, and characterized by the presence of a ventral pit. The first intermediate host remains unknown, but the second intermediate host has been found to be the oyster Crassostrea gigas. Man and the Palearctic oystercatcher Haematopus ostralegus have been shown to be natural definitive hosts, and wading birds including the Kentish plover Charadrius alexandrinus are highly susceptible to experimental infection. Gerbils, hamsters, cats, and several strains of mice were also susceptible laboratory hosts. In experimentally infected mice, the parasites inhabit the small intestine, pinching and sucking the root of villi with their large oral suckers, but they did not invade beyond the mucosa in immunocompetent mice. However, they were found to invade the submucosa in immunosuppressed mice. Human G. seoi infections have been found in at least 25 localities; 23 islands on the Yellow Sea or the South Sea, and 2 western coastal villages. The highest prevalence was found in a village on Aphaedo, Shinan-gun (49% egg positive rate); other areas showed 0.8-25.3% prevalence. Infected people complained of variable degrees of gastrointestinal troubles and indigestion. The infection can be diagnosed by recovery of eggs in the feces; however, an expert is needed to identify the eggs. Praziquantel, 10 mg/kg in single dose, is effective for treatment of human infections. Eating raw oysters in endemic areas should be avoided.