Molecular identification of trematode parasites infecting the freshwater snail Bithynia siamensis goniomphalos in Thailand.

Digenetic trematodes are important parasites of humans and animals. They have complex life cycles and typically infect a gastropod as the first intermediate host. Bithynia siamensis goniomphalos, the first intermediate host of the liver fluke, Opisthorchis viverrini, harbours a wide variety of other trematode species. Morphological details of cercariae of 20 trematode taxa from B. s. goniomphalos, collected mainly in Thailand from 2009 to 2014, were provided in an earlier paper. Correct identification to the species or genus level based on morphology of these cercariae is generally not possible. Therefore, we used molecular data to improve identification and to investigate the diversity of the species of trematodes infecting B. s. goniomphalos. We were successful in extracting, amplifying and sequencing portions of the 28S ribosomal RNA (rRNA) gene for 19 of these 20 types of cercaria, and the internal transcribed spacer 2 region for 18 types. BLAST searches in GenBank and phylogenetic trees inferred from the 28S rRNA sequences identified members of at least nine superfamilies and 12 families. Only a few cercariae could be assigned confidently to genus or species on the basis of the sequence data. Matching sequence data from named adult trematodes will be required for definitive identification. There is clearly a great diversity of trematode species utilizing B. s. goniomphalos in Thailand.

Untangling the Complexity of Liver Fluke Infection and Cholangiocarcinoma in NE Thailand Through Transdisciplinary Learning.

This study demonstrates how a transdisciplinary learning approach provided new insights for explaining persistent Opisthorchis viverrini infection in northern Thailand, as well as elucidating problems of focusing solely on the parasite as a means of addressing high prevalence of cholangiocarcinoma. Researchers from diverse backgrounds collaborated to design an investigative homestay program for 72 Singaporean and Thai university students in five northeast Thai villages. The students explored how liver fluke infection and potential cholangiocarcinoma development are influenced by local landscape dynamics, aquatic ecology, livelihoods, food culture and health education. Qualitative fieldwork was guided daily by the researchers in a collaborative, co-learning process that led to viewing this health issue as a complex system, influenced by interlinked multidimensional factors. Our transdisciplinary experience has led us to believe that an incomplete understanding of these linkages may reduce the efficacy of interventions. Further, viewing liver fluke infection and cholangiocarcinoma as the same issue is inadvisable. Although O. viverrini infection is an established risk factor for the development of cholangiocarcinoma, multiple factors are known to influence the likelihood of acquiring either. Understanding the importance of the current livelihood transition, landscape modification and the resulting mismatch between local cultures and new socio-ecological settings on cholangiocarcinoma initiation and liver fluke transmission is of critical importance as it may help readjust our view of the respective role of O. viverrini and other socioeconomic risk factors in cholangiocarcinoma etiology and refine intervention strategies. As demonstrated in this study, transdisciplinary approaches have the potential to yield more nuanced perspectives to complex diseases than research that focuses on specific aspects of their epidemiology. They may therefore be valuable when designing effective solutions to context-sensitive diseases such as liver fluke infection and cholangiocarcinoma.

Trematode diversity in the freshwater snail Bithynia siamensis goniomphalos sensu lato from Thailand and Lao PDR.

In order to obtain a comprehensive understanding of trematode diversity in Bithynia siamensis goniomphalos sensu lato, the first intermediate host of the liver fluke Opisthorchis viverrini s.l., the prevalence of larval trematode species was investigated in different localities in Thailand and Lao People's Democratic Republic (Lao PDR). In Thailand, snail samples were collected from 29 localities in the nine provinces: Buri Ram, Surin, Chaiya Phum, Maha Sarakham, Khon Kaen, Kalasin, Mukdahan, Sakon Nakhon and Nakhon Phanom. In Lao PDR, snail samples were collected from 21 localities in Vientiane Province and six localities in Savannakhet Province. Snails were identified by standard morphological criteria and then examined for trematode infection using the cercarial shedding method. Twenty different types of cercariae were detected and identified, based on morphological criteria. Virgulate type 1 emerged as the most common cercaria, with an average prevalence of 10.90% (range 0.26-54.22%) in Thailand and 6.58% (range 1.15-89.77%) in Lao PDR. Opisthorchis viverrini s.l. cercariae were the fourth most common in Thailand, with an average prevalence of 1.59% (0.15-6.93), while in Lao PDR their prevalence was 0.96% (0.08-8.37). The high diversity of trematode cercariae observed in this study indicates that B. s. goniomphalos s.l. is highly susceptible to infection with a variety of trematode species. However, the role of non-opisthorchiid trematodes as fish-borne parasites in human health is not fully known and further molecular identification is required.

The influence of human settlements on the parasite community in two species of Peruvian tamarin.

Although there is a growing recognition that the transfer of diseases between humans and non-human primates can be of great significance for conservation biology, there have been only a few studies focusing on parasites. In this study, saddleback (Saguinus fuscicollis) and moustached tamarin (Saguinus mystax) from the rainforest of the Peruvian lowlands were used as models to determine helminth parasite associations between canopy-dwelling primate species and a nearby human settlement. The human population showed high prevalences of infestation with a number of nematodes, including Ascaris lumbricoides (88.9%), Trichuris trichiura (37%) and hookworms (55.6%). However, the ova of these geohelminths were not detectable in tamarin faeces. Thus, no direct parasite transfer from humans to non-human primates could be documented. However, tamarin groups with more frequent contact to humans and their facilities had significantly higher prevalences and egg output of Prosthenorchis elegans, an important primate pathogen, than a forest group. In contrast, a cestode was significantly more common with more egg output in sylvatic than in human-associated groups. Human alteration of the habitat is likely to play a major role in determining the occurrence, prevalence and intensity of helminth infestation of wild non-human primates.

Impact of temporal changes and host factors on the genetic structure of a population of Opisthorchis viverrini sensu lato in Khon Kaen Province (Thailand).

The population genetics of 317 individual Opisthorchis viverrini from Khon Kaen Province Thailand, from 4 different years and 4 cyprinid fish species was examined using multilocus enzyme electrophoresis of enolase (Enol), phosphoglucomutase (Pgm) and triose phosphate isomerase (Tpi). Allele and genotype frequencies for Enol and Pgm were consistent irrespective of year or host species. No heterozygote deficiency was detected for Enol. Significant heterozygote deficiencies were detected in 3 of 4 years for Pgm. For Tpi, allele frequencies of the most common allele and genotype frequency varied between years and among individuals from different host species. Heterozygote deficiencies for Tpi were detected in 2 years. No significant heterozygous deficiencies were detected among O. virerrini from different fish species in 2005, except at Pgm and Tpi from Puntioplites protozsron. There was no statistical significance in pairwise FST values between O. viverrini from Cyclocheilichthys armatus in different years or different host species in 2005. Significant departures from Hardy-Weinberg expectations and a high rate of gene flow in a population of O. viverrini are discussed in terms of self- and cross-fertilisation, natural selection, non-random mating, the Wahlund effect, presence of null alleles, intensity of infection, biology and ecology of their intermediate cyprinid hosts.

Mitochondrial DNA sequence variation among geographical isolates of Opisthorchis viverrini in Thailand and Lao PDR, and phylogenetic relationships with other trematodes.

The present study compared the genetic variation among 14 different geographical isolates of Opisthorchis viverrini sensu lato from Thailand and Lao PDR using sequence data for 2 mitochondrial DNA genes, the subunit 1 of NADH dehydrogenase gene (nad1) and cytochrome c oxidase gene (cox1). Four different nad1 haplotypes were detected among isolates, all of which were identical at the amino acid sequence level. Nucleotide sequence variation among 14 isolates ranged from 0 to 0.3% for nad1. Two different cox1 haplotypes were detected among isolates. These two haplotypes differed at 2 nucleotide positions, one of which resulted in a change in the amino acid sequence. Nucleotide sequence variation among isolates for cox1 ranged from 0 to 0.5%. Comparison of cox1 sequences of O. viverrini to those of other trematodes revealed nucleotide differences of 13-31%. A phylogenetic analysis of the cox1 sequence data revealed strong statistical support for a clade containing O. viverrini and 2 other species of opisthorchid trematodes; O. felineus and Clonorchis sinsensis.

Argas persicus sensu stricto does occur in Australia.

The status of Argas persicus in Australia has been a matter of controversy for over 25 years. Although early records of A. persicus sensu stricto are common, a major revision of the genus indicated that these records refer to A. robertsi, first described in 1968 from northern Australia, and to an "undescribed" member of the complex occurring in the south. Here, we show that A. persicus sensu stricto does occur in southern Australia and is the only species of Argas in the area recorded from poultry. Another undescribed species belonging to the A. persicus complex, from crows' nests near Lake Eyre in South Australia, was also discovered. This information is of considerable epidemiological significance, as A. persicus sensu stricto is a major vector for a number of highly pathogenic diseases of poultry, not all of which have yet been recorded from Australia.

Striped mice, Rhabdomys pumilio, and other murid rodents as hosts for immature ixodid ticks as in the Eastern Cape Province.

Striped mice, Rhabdomys pumilio, were trapped over a period of 17 months in the Thomas Baines Nature Reserve, and placed in cages, over water, until all the ticks they harboured had detached. The mice were then returned to the reserve. Four ixodid tick species were recovered from the mice of which the larvae and nymphs of Rhipicephalus follis and Rhipicephalus simus were the most numerous. Most larvae of R. follis detached from mice trapped from March to July, and most nymphs in March and from June to September. Most larvae of R. simus detached from mice trapped from December to March, and most nymphs from January to March and during May and June. Seven ixodid tick species were collected from striped mice, house rats, Rattus rattus, vlei rats, Otomys spp. and Praomys sp. captured in the vicinity of human dwellings or animal holding facilities in the Grahamstown district. The striped mice captured in the Thomas Baines Reserve harboured considerably larger numbers of ticks than any of the rodent species in the more urbanized localities.

Environmental, cultural and social changes and their influence on parasite infections.

The changes taking place within the societies, cultures and the environments in which we live are massive and complex. By referring to simple epidemiological models it is possible to build an objective framework with which to look at these changes in terms of their likely impact on the epidemiology of parasitic diseases within human communities. These parameters are listed for hosts and both micro- and macroparasites, as are epidemiologically significant cultural, social and environmental variables. Changes in these variables may be either detrimental or beneficial to human health and may, in addition, interact in complex ways. Examples of the complexity of changes which can influence epidemiology are provided for a cultural template of the population living in the north-east of Thailand.

A simplified method of cleaning ixodid ticks for microscopy.

The cleaning of ixodid ticks for microscopy can be achieved quickly and efficiently using a combination of a wax solvent and an ultrasonic cleaner. The technique involves minimum handling of specimens, produces no detectable damage and is suitable for cleaning many specimens at the same time.

Transmission risk of Borrelia burgdorferi sensu lato from Ixodes ricinus ticks to humans in southwest Germany.

The risk of Borrelia burgdorferi infection and the value of antibiotic prophylaxis after tick bite are controversial. In this study, performed in two areas of southwestern Germany, ticks were collected from 730 patients and examined by the polymerase chain reaction (PCR) for B. burgdorferi. To assess whether transmission of B. burgdorferi occurred, the patients were clinically and serologically examined after tick removal and during follow-up examinations. Data from all tick bites gave a total transmission rate of 2.6% (19 patients). Eighty-four ticks (11.3%) were PCR positive. Transmission occurred to 16 (26.7%) of 60 patients who were initially seronegative and could be followed up after the bite of an infected tick. These results indicate that the transmission rate from infected ticks in Europe is higher than previously assumed. Examination of ticks and antibiotic prophylaxis in the case of positivity appears to be indicated.

Multiparasite communities in animals and humans: frequency, structure and pathogenic significance.

Individual humans and animals are subject to infection by a variety of parasites (broadly defined to include viruses, bacteria and other non-protozoan microparasites) at any one time. Multiple parasite infections occur frequently in populations of wild animals as well as in humans from developing countries. In some species and regions, hosts with multiple infections are more common than hosts with either no infection or a single infection. Studies, predominantly on animals, show that a wide variety of environmental and host-dependent factors can influence the structure and dynamics of the communities of parasites that make up these multiple infections. In addition, synergistic and competitive interactions can occur between parasite species, which can influence the likelihood of their successful transmission to other hosts and increase or decrease their overall pathogenic impact. This review summarises aspects of our current knowledge on the frequency of multiparasite infections, the factors which influence them, and their pathogenic significance.

Ecological implications of control strategies: arthropods of domestic and production animals.

The control of arthropods of veterinary importance represents a disturbance for the ecosystem and its animal community. This disturbance can influence the densities of target and non-target organisms and their associated indirect interactions in the food web, leading to reductions in the species richness and diversities in the communities involved. Finely tuned control affecting the target organism alone can have unexpected consequences, depending on the relationships between the target and other organisms within the food web. Broad-spectrum control methods will have more wide ranging and less predictable consequences. Modifications within the community caused by arthropod control can take place over generations and may not be immediately apparent. Most modern techniques of control are designed to minimize environmental impact by concentrating specifically on the target organism. The effects of control of arthropod pests of domestic and production stock must be seen against a background of more general effects caused by rural development which acts to modify the environment continuously over a long time span.

Tick infestation patterns and prevalence of Borrelia burgdorferi in ticks collected at a veterinary clinic in Germany.

A total of 4803 domestic and wild animals which were presented for examination at a veterinary clinic in north Baden, Germany over a period of 1 year were examined for tick infestation. A total of 434 nymphal and adult ticks were collected from 175 hosts. Ticks found belonged to the species Ixodes ricinus (385), Ixodes hexagonus (48), and Ixodes ventalloi (one). The polymerase chain reaction was used to examine 132 I. ricinus and 21 I. hexagonus for the presence of Borrelia burgdorferi. Twenty-two per cent of adult I. ricinus were infected as were one female and one larval I. hexagonus.

Comparison of urinary bladder and ear biopsy samples for determining prevalence of Borrelia burgdorferi in rodents in central Europe.

PCR was used to compare urinary bladder and ear biopsy samples from four European species of wild rodents for the presence of Borrelia burgdorferi sensu lato. From 60 paired comparisons of bladder and ear biopsy samples, the PCR results were concordantly positive or negative in 43 samples (71.7%). Of the 17 which differed, 14 bladder samples were positive and ear samples were negative while the converse occurred for three samples. Thus ear biopsy samples led to a significantly lower estimate of infection than bladder biopsy samples. This suggests that the use of ear biopsy samples in epidemiological studies of B. burgdorferi in Central European rodents is likely to lead to underestimates of the prevalence.

[Natural epidemiology of Lyme borreliosis with reference to clustered incidence of illnesses in the suburbs of a North Baden community]. / Untersuchungen zur natürlichen Epidemiologie der Lyme-Borreliose anlässlich des gehäuften Auftretens von Erkrankungen in einem Vorort einer nordbadischen Gemeinde.

Human infection with Borrelia burgdorferi, the causative agent of Lyme disease, is unusually common in the "Scheelkopf" area of the town Bruchsal in north Baden (Germany), a situation which has led to considerable publicity and public concern. This study was carried out in order to clarify this situation by determining the prevalence of B. burgdorferi in both the free-living tick populations (Ixodes ricinus) and the rodent population from the "Scheelkopf" as well as from surrounding control areas. The polymerase chain reaction (PCR) was used to determine the presence of infection in whole tick preparations and in mouse bladders. The prevalence of B. burgdorferi in freeland ticks ranged from 19% to 44% and in mice from 6% to 29% depending on the area studied. The "Scheelkopf", with prevalences for ticks and mice of 33% and 10% respectively, was not significantly different from the control areas. Our results indicate that there is a high risk of human infection throughout the study area. This is probably related to the intensive use of the area for gardens and the related recreational behaviour of the human population in conjunction with the high rate of infection prevailing in I. ricinus.

Small and medium sized mammals as predators of ticks (Ixodoidea) in South Africa.

A total of 1,640 stomach contents from 56 South African species of small and medium sized mammals were examined for ticks. Only six ticks were found in three specimens of the caracal Felis caracal. Although identification of the tick species was not possible owing to fragmentation and partial digestion, the genus Haemaphysalis was represented in four of these cases. We conclude that predation by small and medium sized mammals on ticks is uncommon in South Africa.