1.1.1 Datos demográficos y clínicos de la población a estudio

Villages were first visited in May 2012 (van Kesteren et al., 2013), by a team of researchers from the University of Salford, the University of Zurich, the University of Franche-Comté and Bishkek Veterinary Institute. In Sary-Mogol (SM) and Taldu-Suu (TS), researchers travelled from house to house, georeferenced each occupied house using a Garmin® GPS60 unit, and administered an oral questionnaire in Kyrgyz or Russian with the person who answered the door (or the head of the household, if this was preferred). Details of the questionnaires administered are given in the appendix (A3). The “household questionnaire” contained information regarding household demographics, behaviour and dog and livestock. If dogs were owned and not based permanently in mountain pasture (“Jailoo”), a copy of the “dog questionnaire” was administered for each dog. This contained questions about dog demographics, dog management and recent praziquantel treatment. An attempt was also made to collect a faecal sample from each dog registered. If dogs were present, an attempt was made to collect faeces per rectum (by experienced veterinarians). However, if dogs were not present, could not be restrained, or had recently defaecated, samples were collected from the floor (with attempts made to ascertain faeces ‘ownership’ with the owner wherever possible). Fresh samples were obtained wherever possible due to the risk of DNA degradation in older samples. Samples were divided upon collection, with some stored in 35ml universal tubes containing 0.3% PBS Tween (Fisher Scientific, Loughborough, UK) buffer with 10% formalin (sourced locally) for coproantigen ELISA; and some stored in bijoux tubes or 15ml polypropylene tubes containing 70% ethanol (sourced locally) for PCR testing. Finally, a random sample of 40 households from Sary-Mogol and Kashka’Suu and 20 households from Taldu-Suu were selected to receive an additional questionnaire relating to healthcare and economic issues.

If dogs were present at the time of visit, they were dosed with praziquantel at a dosage of 5mg/kg. If dogs were not present (either in an unknown location or temporarily at summer pasture), an estimate of their weight was given by the owner and a suitable number of tablets were left with the owner for dosing upon return.

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The same process was repeated in Kashka’Suu (KS) and Kara-Kabak (KK), but without praziquantel dosing (with the exception of a number of dogs in KK selected to undergo arecholine purgation). In KS, households were selected in groups of six due to proximity to a number of randomly generated points within the village boundaries (as estimated visually from recent ’SPOT’ satellite imagery taken from Google Earth). A broad estimate of the required sample size for each of these two villages were estimated using the equation: 𝑛 = 𝑃(1−𝑃)_𝑑2 . Using an expected coproantigen prevalence (𝑃) of 20%, and 95% confidence intervals (1.96 × 𝑑) of ±10%, this was estimated at around 60-70 dogs. As population estimates for the villages were not available prior to the visit, no adjustment for finite population size was made, but approximately 25% of the households in KS were ultimately visited, and due to its small size all households in KK were visited. Maps of all sampled dogs in each of the four villages are shown in figures 2.2 and 2.3.

In TS and KK, a total of 33 dogs (convenience sampled with the assistance of the local veterinarian) underwent arecoline purgation. Owners restrained the dogs whilst they were administered a 0.4% solution of arecoline hydrobromide in water orally (7mg arecoline/kg body weight), which was repeated if there was no purge within 30 minutes. The initial faecal void was collected and stored as described above, and the purge was filtered and closely inspected for adult worms by an experienced veterinarian (Iskender Ziadinov). Due to logistical difficulties, it was not possible to match the purges collected from dogs in TS to individual households, although this was possible in KK. As a public health precaution, all purged dogs (including those in Kara-Kabak) were given praziquantel.

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Figure 2.2. Locations of dogs sampled from Sary-Mogol (west) and Taldu-Suu (east) in May 2012. Imagery from Google Earth (satellite image taken 20th Jan, 2012 from SPOT 5 satellite)

Figure 2.3. Locations of dogs sampled from Kashka’Suu (west) and Kara-Kabak (east) in May 2012. Imagery from Google Earth (satellite image taken 20th Jan, 2012 from SPOT 5 satellite)

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A second visit to the field site was made in October 2012 by two researchers from the University of Salford and one veterinarian from the Bishkek Veterinary Institute. Each household in SM, TS and KK which reported owning dogs during the previous visit was revisited again (including any household which had not been visited previously), and all dogs present were identified. Those in SM or TS were dosed with praziquantel as before (either supervised dosing, or tablets left with the owner). In KK, any dogs which had previously undergone arecholine purgation were also given praziquantel. A shortened version of the previous questionnaire was also administered (see appendix), and faecal samples were collected from all available dogs. Due to the difficulties previously experienced with rectal sampling, a decision was made to collect most faecal samples from the floor during this visit. Due to a large culling campaign in the village, fewer dogs were present at this visit, and therefore all available dogs were able to be registered and sampled in these three villages.

A random sample of households in KS were visited in order to sample any dogs present and conduct a questionnaire for each dog. Households were selected randomly from a sampling frame of houses generated from imagery collected by the ‘SPOT5’ satellite in 2010 (images taken from Google Earth), and households were sequentially visited until around 60 dogs had been sampled. Households without dogs were recorded but did not undergo a questionnaire.

Following the October visit, a World Bank -funded praziquantel dosing scheme was commenced in the area. Raion capitals provided praziquantel and a consent form for canine dosing (detailing the effect of the tablet and required faeces management measures following dosing), although local veterinarians and paraveterinarians needed to collect the tablets and make photocopies of the consent forms themselves. All signed consent forms were to be returned to the raion capital for monitoring. Therefore, on the subsequent visits to the field site in April and September 2013 and April and September 2014, a decision was made to not interfere with the ongoing dosing campaign (which was also providing funding to local vets). As such, only faeces were collected (from the ground) and questionnaires completed, with no overall census conducted and no praziquantel administered.

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