Identification of tungiasis infection hotspots with a low-cost, high-throughput method for extracting Tunga penetrans (Siphonaptera) off-host stages from soil samples-An observational study.

BACKGROUND: The sand flea, Tunga penetrans, is the cause of a severely neglected parasitic skin disease (tungiasis) in the tropics and has received little attention from entomologists to understand its transmission ecology. Like all fleas, T. penetrans has environmental off-host stages presenting a constant source of reinfection. We adapted the Berlese-Tullgren funnel method using heat from light bulbs to extract off-host stages from soil samples to identify the major development sites within rural households in Kenya and Uganda. METHODS AND FINDINGS: Simple, low-cost units of multiple funnels were designed to allow the extraction of >60 soil samples in parallel. We calibrated the method by investigating the impact of different bulb wattage and extraction time on resulting abundance and quality of off-host stages. A cross-sectional field survey was conducted in 49 tungiasis affected households. A total of 238 soil samples from indoor and outdoor living spaces were collected and extracted. Associations between environmental factors, household member infection status and the presence and abundance of off-host stages in the soil samples were explored using generalized models. The impact of heat (bulb wattage) and time (hours) on the efficiency of extraction was demonstrated and, through a stepwise approach, standard operating conditions defined that consistently resulted in the recovery of 75% (95% CI 63-85%) of all present off-host stages from any given soil sample. To extract off-host stages alive, potentially for consecutive laboratory bioassays, a low wattage (15-25 W) and short extraction time (4 h) will be required. The odds of finding off-host stages in indoor samples were 3.7-fold higher than in outdoor samples (95% CI 1.8-7.7). For every one larva outdoors, four (95% CI 1.3-12.7) larvae were found indoors. We collected 67% of all off-host specimen from indoor sleeping locations and the presence of off-host stages in these locations was strongly associated with an infected person sleeping in the room (OR 10.5 95% CI 3.6-28.4). CONCLUSION: The indoor sleeping areas are the transmission hotspots for tungiasis in rural homes in Kenya and Uganda and can be targeted for disease control and prevention measures. The soil extraction methods can be used as a simple tool for monitoring direct impact of such interventions.

Myth or truth: investigation of the jumping ability of Tunga penetrans (Siphonaptera: Tungidae).

Female sand fleas (Tunga penetrans Linnaeus, 1758, Siphonaptera: Tungidae) cause a severe parasitic skin disease known as tungiasis. T. penetrans is a small flea, measuring less than 1 mm in length. The females of this species burrow into the skin of human and animal hosts and mostly affect the feet. This has led to the anecdotal assumption that T. penetrans, unlike its relatives in the Siphonaptera family, would have a limited jumping ability potentially not reaching higher body parts. However, there is no data supporting this. This study evaluated the jumping capabilities of T. penetrans for height and distance using sticky tapes. The vertical jump of the female T. penetrans ranged from 4.5 to 100 mm with a mean of 40 mm whereas the vertical jump of the male T. penetrans ranged from 1.2 to 138 mm with a mean of 46 mm. The horizontal jump of the female T. penetrans ranged from 18 to 138 mm with a mean of 64 mm and that of the male ranged from 9 to 251 mm with a mean of 80 mm. Based on the literature, fleas of various species have been described as jumping vertically 50-100 times their size and horizontally 5-100 times their size. In this respect, sand fleas appear to have equal expert jumping abilities to their relatives. Their aggregation on people's feet is not likely a result of their poor jumping ability but might be an adaptation to the host's behavior which would require further investigations.

Environmental and Household-Based Spatial Risks for Tungiasis in an Endemic Area of Coastal Kenya.

Tungiasis is a cutaneous parasitosis caused by an embedded female sand flea. The distribution of cases can be spatially heterogeneous even in areas with similar risk profiles. This study assesses household and remotely sensed environmental factors that contribute to the geographic distribution of tungiasis cases in a rural area along the Southern Kenyan Coast. Data on household tungiasis case status, demographic and socioeconomic information, and geographic locations were recorded during regular survey activities of the Health and Demographic Surveillance System, mainly during 2011. Data were joined with other spatial data sources using latitude/longitude coordinates. Generalized additive models were used to predict and visualize spatial risks for tungiasis. The household-level prevalence of tungiasis was 3.4% (272/7925). There was a 1.1% (461/41,135) prevalence of infection among all participants. A significant spatial variability was observed in the unadjusted model (p-value < 0.001). The number of children per household, earthen floor, organic roof, elevation, aluminum content in the soil, and distance to the nearest animal reserve attenuated the odds ratios and partially explained the spatial variation of tungiasis. Spatial heterogeneity in tungiasis risk remained even after a factor adjustment. This suggests that there are possible unmeasured factors associated with the complex ecology of sand fleas that may contribute to the disease's uneven distribution.

Molecular identification and characterization of Sarcocystis spp. in horsemeat and beef marketed in Japan.

Human Sarcocystis infections are known to be caused by the ingestion of raw or undercooked beef or pork containing mature sarcocysts of Sarcocystis hominis or S. suihominis, respectively. In addition, several cases of parasitic food poisoning in Japan have recently been reported after consumption of raw horsemeat containing sarcocysts of S. fayeri. In this study, the presence of sarcocysts in 28 horsemeat and 121 beef samples collected in Tokyo was investigated. Sarcocysts of S. fayeri were found in 16 horsemeat samples. Sarcocysts of S. hominis were not detected in beef samples, while sarcocysts of S. cruzi were detected in 60 beef samples. In addition, S. hirsuta and S. bovini were isolated only from New Zealand beef samples. Bradyzoites in sarcocysts collected from 62/73 sarcocyst-positive refrigerated horsemeat and beef samples were determined to be viable. Molecular analysis of S. fayeri 18S rRNA gene sequences revealed that intraspecific variation among eight individual bradyzoites from a single sarcocyst was as high as 9.8%. In contrast, mitochondrial cytochrome c oxidase subunit 1 (mtDNA cox1) gene sequences from the six fragments of a single sarcocyst were 100% identical. Sarcocysts of S. bovini isolated from beef also exhibited intraspecific variation in 18S rRNA gene sequences and had to be cloned before sequencing, while mtDNA cox1 gene sequences were obtained by direct sequencing. Therefore, we conclude that molecular analysis of the mtDNA cox1 gene is the most useful for identification of Sarcocystis species. This study provides the first published partial sequence of the S. fayeri mtDNA cox1 gene.

A survey of gastrointestinal parasites of alpacas (Vicugna pacos) raised in Japan.

This study aimed to determine the prevalence of gastrointestinal parasites in alpacas raised in Japan. From December 2010 to October 2011, 53 alpacas (Vicugna pacos) raised at a farm in the Kanto region, Japan, were examined for gastrointestinal parasites by 3 fecal tests: direct smear, centrifuged flotation and formalin-ether sedimentation. Eggs of Nematodirus sp. were found in 13.2%, Trichuris sp. in 11.3%, Capillaria spp. in 5.7%, strongyle-type in 50.9% and Moniezia sp. in 1.9%. Oocysts of Eimeria punoensis and/or E. alpacae were found in 69.8%, E. lamae in 1.9% and E. macusaniensis in 7.5%. We found that alpacas raised in Japan have gastrointestinal parasitic fauna similar to those in other countries.

Feline demodicosis caused by an unnamed species.

A case of feline demodicosis is described in this report. A 13-year-old spayed female domestic short hair cat weighing 4.5 kg was being treated with cefovecin and alternately with prednisone or methylprednisolone. On further physical examination, the cat showed mild erythema and hair loss on the bridge of the nose, around the eyes, on the chin, on the side part of the breast and on the abdomen. A large number of Demodex mites were found in deep skin scrapings from the affected areas. The cat was then treated with ivermectin at 600 µg/kg administered SC daily. After 4 weeks of treatment, the cat was clinically normal with no mites detected in the skin scrapings from the face or breast areas. The mite responsible may represent a previously seen but as yet unnamed new species. This is third report that describes a case of feline demodicosis caused by a different, unnamed mite species that has different morphological characteristics to those of known Demodex mites and may represent a previously seen but as yet unnamed species.