Genetic variation in the immune system and malaria susceptibility in infants: a nested case-control study in Nanoro, Burkina Faso.

BACKGROUND: Genetic polymorphisms in the human immune system modulate susceptibility to malaria. However, there is a paucity of data on the contribution of immunogenetic variants to malaria susceptibility in infants, who present differential biological features related to the immaturity of their adaptive immune system, the protective effect of maternal antibodies and fetal haemoglobin. This study investigated the association between genetic variation in innate immune response genes and malaria susceptibility during the first year of life in 656 infants from a birth cohort survey performed in Nanoro, Burkina Faso. METHODS: Seventeen single nucleotide polymorphisms (SNPs) in 11 genes of the immune system previously associated with different malaria phenotypes were genotyped using TaqMan allelic hybridization assays in a Fluidigm platform. Plasmodium falciparum infection and clinical disease were documented by active and passive case detection. Case-control association analyses for both alleles and genotypes were carried out using univariate and multivariate logistic regression. For cytokines showing significant SNP associations in multivariate analyses, cord blood supernatant concentrations were measured by quantitative suspension array technology (Luminex). RESULTS: Genetic variants in IL-1ß (rs1143634) and FcγRIIA/CD32 (rs1801274)-both in allelic, dominant and co-dominant models-were significantly associated with protection from both P. falciparum infection and clinical malaria. Furthermore, heterozygote individuals with rs1801274 SNP in FcγRIIA/CD32 showed higher IL-1RA levels compared to wild-type homozygotes (P = 0.024), a cytokine whose production is promoted by the binding of IgG immune complexes to Fcγ receptors on effector immune cells. CONCLUSIONS: These findings indicate that genetic polymorphisms in genes driving innate immune responses are associated to malaria susceptibility during the first year of life, possibly by modulating production of inflammatory mediators.

Preliminary assessment of the computer-based Taenia solium educational program 'The Vicious Worm' on knowledge uptake in primary school students in rural areas in eastern Zambia.

OBJECTIVE: The zoonotic helminth Taenia solium is endemic in Zambia, causing human (taeniasis and (neuro)cysticercosis) and pig (porcine cysticercosis) diseases with high health, social and economic burdens. We aimed to evaluate the impact of a health educational program intended to lead to powerful and cumulative improvements in knowledge, attitudes and practices that decrease parasite transmission and disease occurrence. METHODS: Half-day health education workshops were conducted in three primary schools in the highly endemic Eastern Province of Zambia, using the computer-based T. solium educational program 'The Vicious Worm'. Questionnaires were administered before and after the educational component to determine the program's impact on knowledge uptake in primary school students. RESULTS: In total, 99 students participated: 38 males and 61 females, with a median age of 14 years (range 10-18 years). Baseline general knowledge of T. solium, including awareness of the different human and pig disease states, and disease diagnosis, treatment and prevention, was quite high (average score 62%) and consistent across all three study areas. Participants' knowledge had significantly increased after the educational component, particularly regarding parasite transmission and disease prevention. CONCLUSION: Preliminary assessment of 'The Vicious Worm' indicates it is an effective tool for the short-term T. solium education of primary school students in Zambia. Follow-up studies are planned to assess the longer term impact of the program on knowledge uptake in the study neighbourhoods. Inclusion of tailored 'The Vicious Worm' educational workshops should be considered in integrated cysticercosis control programs in endemic areas of sub-Saharan Africa.

Community-based tsetse fly control minimizes the effect of trypanosomosis on livestock in Metekel zone, Ethiopia.

Tsetse-related trypanosomosis threatens integrated crop-livestock production system in Ethiopia. Metekel zone is among the tsetse-infested areas in Ethiopia. Community-based tsetse fly and trypanosomosis control using targets was conducted from June 2011 to May 2012 in the zone. In coordination with the control project, a questionnaire survey was administered to assess the economic importance of the tsetse fly and trypanosomosis control conducted in the Zone. On average, there were 3 livestock deaths/household before control and this was decreased to 0.07 after control. This death was contributed from all possible livestock diseases. A household loss 4380 (95% CI = 4223.98-4541.79) EB (1 US$ = 17.9 EB) only due to the deaths of his/her livestock was reduced to 90.78 (95% CI = 89.15-92.42) EB after control. Morbidity of oxen in the study areas was significantly decreased by 70.7%. As a result, the monetary loss to rent for healthy draught oxen was decreased by 96.4%. When a household with one cattle harvest 10.9 quintals (1090 kg), his neighbor's household with ten cattle maximize his/her yield to about 50.5 quintals (5050 kg) of crops indicating that trypanosomosis has great economic impact in crop-livestock production system. As the community-based tsetse fly control approach is simple, cost effective, and appropriate for riverine tsetse species, we conclude that farmers can minimize the negative impact of tsetse flies and trypanosomosis in the tsetse-infested areas of African countries.

Bovine and Caprine Brucellosis in Bangladesh: Bayesian evaluation of four serological tests, true prevalence, and associated risk factors in household animals.

A cross-sectional study was carried out to estimate the true prevalence of Brucella spp. and identify allied risk factors/indicators associated with brucellosis in the Dinajpur and Mymensingh districts of Bangladesh. A total 320 stratified random blood samples were collected and tested in parallel for Brucella antibodies using Rose Bengal (RBT), slow agglutination (SAT), and indirect and competitive ELISA. In addition, a structured questionnaire was administered to each household herd owner to gather information regarding potential risk factors. Both univariate and multivariate logistic regression analyses were used to identify potential risk factors or indicators at animal level. A Bayesian approach was used to estimate the true prevalence of brucellosis along with the test performances (Se and Sp). The estimated animal level true prevalence in cattle was 9.70 % (95 % CPI 5.0-16 %) and in goat 6.3 % (95 % CPI 2.8-11.0 %). The highest sensitivity was achieved by SAT ranges from 69.6 to 78.9 %, and iELISA was found to be more specific (97.4 to 98.8 %) in comparison with other tests. On the other hand, a significant level of (P < 0.05) Brucella seropositivity was found in cattle that breed naturally compared with those that undergo artificial insemination. In goats, exotic breeds were significantly associated (P < 0.05) with Brucella seroprevalence compared with indigenous breeds. Goats with a previous records of abortion and/or retained placenta were also found to have significant levels (P < 0.05). Cows with previous abortion records showed higher odds (18 times) of being seropositive. None of the evaluated tests can be recommended to apply alone for the diagnosis of bovine and caprine brucellosis.

The unexpected discovery of Brucella abortus Buck 19 vaccine in goats from Ecuador underlines the importance of biosecurity measures.

Very few, mostly old, and only preliminary serological studies of brucellosis in goats exist in Ecuador. In order to assess the current epidemiological situation, we performed a cross-sectional serological study in the goat populations of Carchi (n = 160 animals), Pichincha (n = 224 animals), and Loja provinces (n = 2024 animals). Only two positive serological results (RB negative and SAT-EDTA ≥400 IU/ml) were obtained in lactating goats from the same farm in Quito (Pichincha province). Additionally, milk was sampled from 220 animals in Pichincha province. The present study indicates a low apparent prevalence in Pichincha province and absence in Carchi and Loja provinces. A total of 25 positive milk ring tests (MRT) were obtained in Pichincha province yielding a prevalence of MRT of 11.16%. Subsequent culture was performed on the positive MRT samples. All results were negative, apart from a single sample, obtained from a serologically positive goat in Quito, that was positive for Brucella abortus strain 19 (B19). Several hypotheses are forwarded concerning this unexpected result. The most likely hypothesis is the possible accidental use of a needle, previously used for vaccination of cattle with the said vaccine, for the administration of drug treatment to the goat. This hypothesis underlines the necessity of biosecurity measures to prevent this type of accidents.

Community-based tsetse fly control significantly reduces fly density and trypanosomosis prevalence in Metekel Zone, Northwest, Ethiopia.

African animal trypanosomosis is a great obstacle to livestock production where tsetse flies play a major role. Metekel zone is among the tsetse-infested areas. Community-based tsetse fly and trypanosomosis control using targets was conducted from June 2011 to May 2012 in Metekel zone, Ethiopia, to decrease trypanosomosis and tsetse fly. Cloth screen targets were developed, impregnated with 0.1 % deltamethrin, and deployed alongside rivers by the research team together with the community animal health workers. Monthly parasitological and entomological data were collected, processed, and compared with similar data collected before control. Overall average tsetse fly (Glossina tachinoides) density decreased from 1.13 to 0.18 fly/trap/day after control. The density was decreased in all sites with no significant difference among the sites. However, higher decrements were observed in the dry and late dry seasons by more than 12 and 6 times, respectively. The reduction in overall apparent prevalence of trypanosomosis caused by Trypanosoma congolense, Trypanosoma brucei, and Trypanosoma vivax from 12.14 % before to 3.61 % after control coincides with the tsetse fly reduction. In all the study sites, significant reduction was observed before and after control. The highest decrement was observed in the late dry season when the apparent prevalence was reduced from 7.89 to 1.17 % before and after control, respectively. As this approach is simple, cost-effective, and appropriate for riverine tsetse species, we recommend to be scaled up to other similar places.

Assessment of Risk Factors for a High Within-Batch Prevalence of Yersinia enterocolitica in Pigs Based on Microbiological Analysis at Slaughter.

The purpose of the current study was to find farm-level factors influencing the bacteriological prevalence of Yersinia enterocolitica in pigs at time of slaughter. On 100 farms, data concerning a broad range of farm aspects (e.g., management and housing system, biosecurity, and hygiene measurements) were collected using a face-to-face questionnaire. At the slaughterhouse, tonsils of on average 70 slaughter pigs per batch were sampled to determine the infection status of pigs. After univariable mixed-effect logistic regressions, variables that were related to the Yersinia prevalence (p<0.05) were included in a multivariable model. In this model, the factors remaining positively associated with a higher Y. enterocolitica carriage in the tonsils (p<0.1) were an increasing number of piglet suppliers, a high density of pig farms in the area, and the use of semislatted floors in the fattening pig unit. The proper use of a disinfection bath before entering the stables and a poor biosecurity level were protective factors, although a higher prevalence was associated with a significant positive interaction between the presence of pets in the stables and a poor biosecurity level. Reducing the number of piglet suppliers, using a disinfection bath properly, and prohibiting pets inside the stables could be easily implemented by pig farmers to lower the prevalence of Y. enterocolitica in pigs at slaughter.

Prevalence of bovine theileriosis in North Central region of Algeria by real-time polymerase chain reaction with a note on its distribution.

To determine the presence and distribution of bovine theileriosis in the North Central region of Algeria, 358 DNA samples and 359 blood smears were analyzed from nine provinces. Theileria DNA extracted from cattle blood was amplified by fluorescence resonance energy transfer polymerase chain reaction (FRET-PCR). Blood smears were examined for Theileria piroplasms by microscopical examination (ME) of Giemsa-stained slides. While microscopical identification revealed only 42 animals being infected with Theileria piroplasms, PCR-positive amplification using Theileria genus-specific primers was obtained from 132 Theileria spp. (P < 0.0001). Among the 132 positives, 108 animals (81.8 %) were found positive of Theileria annulata, while 24 (18.2 %) were found positive for Theileria sp. (P < 0.0001). However, melting curve analysis of these latter samples revealed the presence of two different peaks, 51.5 ± 0.5 °C corresponding to Theileria sp1 and 52.5 ± 0.5 °C for Theileria sp2. Cloning and sequencing of Theileria sp1 and Theileria sp2 using the Cox primers indicated that these species are very closely related to Theileria buffeli. There is a highly significant difference in the distribution of theileriosis between different provinces (P < 0.0001). This disparity between provinces is probably due to differences in tick contact, influenced by the subhumid bioclimatic gradient and differences in agricultural land use.

Relation between serology of meat juice and bacteriology of tonsils and feces for the detection of enteropathogenic Yersinia spp. in pigs at slaughter.

The association between positive serology and culture detection of Yersinia spp. in individual pigs was determined. Pieces of diaphragm from 370 pig carcasses were collected for serological analysis, and tonsils and feces of the same carcass were collected for bacteriological analysis. Detection of anti-Yersinia antibodies in meat juice samples was done using an indirect enzyme-linked immunosorbent assay (ELISA) based on Yops (Yersinia outer proteins). Tonsils and feces were tested for the presence of enteropathogenic Yersinia spp. by direct plating on cefsulodin-irgasan-novobiocin agar plates. Of the 370 meat juice samples, 241 (65.1%) gave a positive serological reaction using a cutoff value of 20%. Enteropathogenic Yersinia spp. (Yersinia enterocolitica serotype O:3 and Yersinia pseudotuberculosis) were found in tonsils of 161 pigs and feces of 30 pigs. Recovery of enteropathogenic Yersinia from the tonsils was highly correlated with positive serotiters, whereas no correlation was found between serology and fecal excretion. Results demonstrated that serology has an acceptable sensitivity, but a relatively low specificity for the rapid detection of enteropathogenic Yersinia spp. in tonsils of pigs at slaughter.

Evaluation of the ISO 10273:2003 method for the isolation of human pathogenic Yersinia enterocolitica from pig carcasses and minced meat.

Pig carcass swabs (n = 254) and minced meat samples (n = 82) were examined for pathogenic Yersinia enterocolitica using different routinely used enrichment protocols. All samples were obtained in the context of the official Yersinia monitoring program in Belgium. In total, 28 carcasses (11.0%) were contaminated with Y. enterocolitica bioserotype 4/O:3 and one (0.4%) with bioserotype 2/O:9. Four minced meat samples (4.9%) tested positive for Y. enterocolitica bioserotype 4/O:3. Using the ISO 10273:2003 method, eight out of the 29 Yersinia-positive carcasses (27.6%) and none of the contaminated minced meat samples (0.0%) were detected. Reducing the enrichment time in PSB from 5 to 2 days increased the number of positive samples. Overall, enrichment in PSB at 25 °C recovered more positive carcasses and minced meat samples than selective enrichment and cold enrichment. As the exclusive use of the ISO 10273:2003 method results in a strong underestimation of Y. enterocolitica positive carcasses and minced meats, efforts are needed to optimize the current version of the ISO method. In addition, isolation of pathogenic Y. enterocolitica requires experience and the use of a stereomicroscope to avoid false negative results.

Predictors of condom use and refusal among the population of Free State province in South Africa.

BACKGROUND: This study investigated the extent and predictors of condom use and condom refusal in the Free State province in South Africa. METHODS: Through a household survey conducted in the Free Sate province of South Africa, 5,837 adults were interviewed. Univariate and multivariate survey logistic regressions and classification trees (CT) were used for analysing two response variables 'ever used condom' and 'ever refused condom'. RESULTS: Eighty-three per cent of the respondents had ever used condoms, of which 38% always used them; 61% used them during the last sexual intercourse and 9% had ever refused to use them. The univariate logistic regression models and CT analysis indicated that a strong predictor of condom use was its perceived need. In the CT analysis, this variable was followed in importance by 'knowledge of correct use of condom', condom availability, young age, being single and higher education. 'Perceived need' for condoms did not remain significant in the multivariate analysis after controlling for other variables. The strongest predictor of condom refusal, as shown by the CT, was shame associated with condoms followed by the presence of sexual risk behaviour, knowing one's HIV status, older age and lacking knowledge of condoms (i.e., ability to prevent sexually transmitted diseases and pregnancy, availability, correct and consistent use and existence of female condoms). In the multivariate logistic regression, age was not significant for condom refusal while affordability and perceived need were additional significant variables. CONCLUSIONS: The use of complementary modelling techniques such as CT in addition to logistic regressions adds to a better understanding of condom use and refusal. Further improvement in correct and consistent use of condoms will require targeted interventions. In addition to existing social marketing campaigns, tailored approaches should focus on establishing the perceived need for condom-use and improving skills for correct use. They should also incorporate interventions to reduce the shame associated with condoms and individual counselling of those likely to refuse condoms.

Campylobacter contamination in broiler carcasses and correlation with slaughterhouses operational hygiene inspection.

This study investigates factors associated with Campylobacter contamination of broiler carcasses, using survey data collected from nine Belgian slaughterhouses in 2008 in accordance with a European Union baseline study. Campylobacter were detected in 51.9% (202/389) (95% confidence interval, 46.8%-56.9%) of broiler carcasses. Campylobacter concentration was <10 CFU/g in 49.6% of carcasses, while 20.6% were contaminated with ≥ 1000 CFU/g. The mean Campylobacter concentration, as calculated by maximum likelihood estimation for left-censored data, was 1.8 log(10) CFU/g, with a standard deviation of 1.9 log(10) CFU/g. There was statistically significant variation among slaughterhouses in prevalence and concentrations of Campylobacter in their sampled carcasses. Campylobacter prevalence (but not concentrations) was positively associated with increase in broilers age. Both Campylobacter prevalence and concentration were significantly higher in carcasses sampled during June and September (but not in July and August) than carcasses sampled in January. We also investigated the correlation (Spearman's rank correlation test) between the scores of official control inspections and Campylobacter prevalence for eight out of the nine slaughterhouses. The control inspections were routinely performed by the Belgian Federal Agency for the Safety of the Food Chain, and the concluded inspection scores were used as a general numerical indicator for the status of operational hygiene and quality of management in the slaughterhouses. Ranking of slaughterhouses based on their inspection scores was statistically correlated (Spearman's correlation coefficient = 0.857) with their ranking based on prevalence of Campylobacter. In the present study we demonstrate how the outcomes from a routine baseline survey could be coupled with other readily available data from national control authorities in order to enable a better insight over Campylobacter contamination status in broiler slaughterhouses. Findings from this work call for subsequent in-depth investigations on technical and hygiene management factors that could impact Campylobacter contamination across broiler slaughterhouses.

Effect of sampling and short isolation methodologies on the recovery of human pathogenic Yersinia enterocolitica from pig tonsils.

The objective of this study was to determine the effect of sampling (swab samples compared to destructive samples) on isolation rates of human pathogenic Yersinia enterocolitica from pig tonsils. Moreover, the relative efficiency of different rapid, routinely applicable isolation methods was evaluated. Therefore, swab and destructive samples from tonsils of 120 pigs at slaughter were analyzed in parallel using direct plating and different enrichment methods. Salmonella-Shigella-desoxycholate-calcium chloride (SSDC) agar, cefsulodin-irgasan-novobiocin (CIN) agar, and Yersinia enterocolitica chromogenic medium (YeCM) were used as selective agar media. For enrichment, irgasan-ticarcillin-potassium chlorate (ITC) broth and peptone-sorbitol-bile (PSB) broth were incubated at 25°C for 48 h. Overall, 55 tonsils (45.8%) were positive for Y. enterocolitica bioserotype 4/O:3. Recovery was significantly higher using the destructive method compared to the swabbing method. Direct plating resulted in 47 and 28 Y. enterocolitica-positive destructive and swab samples, respectively. Alkali treatment of PSB and ITC enrichment broths significantly increased recovery of pathogenic Y. enterocolitica from destructive tonsil samples. The performance of YeCM for qualitative and quantitative isolation of pathogenic Y. enterocolitica from pig tonsils was equal to SSDC and CIN. In conclusion, direct plating and ISO 10273: 2003 with minor modifications are suitable and rapid methods for isolation of pathogenic Y. enterocolitica from destructive tonsil samples.

Age-stratified Bayesian analysis to estimate sensitivity and specificity of four diagnostic tests for detection of Cryptosporidium oocysts in neonatal calves.

There is no gold standard diagnostic test for the detection of bovine cryptosporidiosis. Infection is usually highest in 2-week-old calves, and these calves also excrete high numbers of oocysts. These factors may give rise to variations in the sensitivity and specificity of the various diagnostic tests used to detect infection in calves of various ages. An age-stratified Bayesian analysis was carried out to determine the optimum diagnostic test to identify asymptomatic and clinical Cryptosporidium sp. infection in neonatal calves. Fecal samples collected from 82 calves at 1 week, 2 weeks, 3 weeks, and 4 weeks of age were subjected to the following tests: microscopic examination of smears stained with either phenol-auramine O or fluorescein isothiocyanate (FITC)-conjugated anti-Cryptosporidium monoclonal antibody, nested-PCR, and quantitative real-time PCR. The results confirmed a high prevalence of Cryptosporidium sp. infection, as well as a high level of oocyst excretion, in 2-week-old calves. The sensitivities of all the tests varied with the age of the calves. Quantitative real-time PCR proved to be the most sensitive and specific test for detecting infection irrespective of the age of the calf. The microscopic techniques were the least sensitive and exhibited only moderate efficiency with 2-week-old calves excreting large numbers of oocysts, the majority of which were diarrheic. It was concluded that, when interpreting the results of routine tests for bovine cryptosporidiosis, cognizance should be taken of the sensitivity of the tests in relation to the age of the calves and stage of infection.

Bayesian Estimation of the Prevalence and Test Characteristics (Sensitivity and Specificity) of Two Serological Tests (RB and SAT-EDTA) for the Diagnosis of Bovine Brucellosis in Small and Medium Cattle Holders in Ecuador.

In Ecuador, a national program for bovine brucellosis control has been in implementation since 2008. Given the costs, small- and medium-sized livestock holders are not completely committed to it. The objective of this study was to determine true prevalence (TP) of bovine brucellosis in small- and medium-sized herd populations, as well as the diagnostic sensitivity and specificity of the Rose Bengal (RB) test and the sero-agglutination test (SAT)-EDTA using a Bayesian approach. Between 2011 and 2016, 2733 cattle herds were visited, and 22,592 animal blood samples were taken in nineteen provinces on mainland Ecuador. Bayes-p and deviance information criterion (DIC) statistics were used to select models. Additionally, risk-factor analysis was used for herds according to their brucellosis test status. True prevalence (TP) in herds was estimated by pool testing. National seroprevalence of farms was 7.9% (95% CI: 6.79-9.03), and TP was 12.2% (95% CI: 7.8-17.9). Apparent prevalence (AP) in animals was 2.2% (95% CI: 1.82-2.67), and TP was 1.6% (95% CrI: 1.0-2.4). Similarly, the sensitivity of the RB was estimated at 64.6% (95% CrI: 42.6-85.3) and specificity at 98.9% (95% CrI: 98.6-99.0); for the SAT-EDTA test, sensitivity was 62.3% (95% CrI: 40.0-84.8) and 98.9% (95% CrI: 98.6-99.1) for specificity. Results of the two tests were highly correlated in infected and uninfected animals. Likewise, high spatial variation was observed, with the Coastal Region being the zone with the highest TP at 2.5%. (95% CrI: 1.3-3.8%) in individual animals and 28.2% (95% CI: 15.7-39.8) in herds. Risk factors include herd size, type of production (milk, beef, and mixed), abortions recorded, and vaccination. The results of this study serve to guide authorities to make decisions based on parallel testing at the beginning of a bovine brucellosis program for small livestock holders to increase sensitivity level of the screening tests in Ecuador.

Domestic animals and epidemiology of visceral leishmaniasis, Nepal.

On the Indian subcontinent, visceral leishmaniasis (VL) is considered an anthroponosis. To determine possible reasons for its persistence during interepidemic periods, we mapped Leishmania infections among healthy persons and animals in an area of active VL transmission in Nepal. During 4 months (September 2007-February 2008), blood was collected from persons, goats, cows, and buffaloes in 1 village. Leishmania infections were determined by using PCR. We found infections among persons (6.1%), cows (5%), buffaloes (4%), and goats (16%). Data were georeferenced and entered into a geographic information system. The bivariate K-function results indicated spatial clustering of Leishmania spp.-positive persons and domestic animals. Classification tree analysis determined that among several possible risk factors for Leishmania infection among persons, proximity of Leishmania spp.-positive goats ranked first. Although our data do not necessarily mean that goats constitute a reservoir host of L. donovani, these observations indicate the need for further investigation of goats' possible role in VL transmission.

Methodology for assessing determinants of manure use in urban areas of Africa.

The objectives of this study were to identify the major determinants (e.g. zootechnical and socio-economic parameters) of manure management practised by urban livestock keepers. Data were collected from four single-visit, multi-object surveys in three cities of Africa. The livestock keepers were classified either as 'good user' or 'bad user' of manure depending on how they use the manure. The results of this study indicate that the habit of manure utilization shows geographical differences. In general, livestock keepers engaged in agricultural activities are good manure users. Furthermore, manual workers, traders or the self-employed are better manure users than other professional groups. The manure utilization varies also in function of the livestock numbers. In Brazzaville, the habit of selling animals influenced manure management positively. Experience of keeping livestock before the civil war in Brazzaville was also a determining factor of manure use in the post-war urban livestock keeping in 2001. When designing policies to reduce the environmental effects of urban livestock keeping, the highlighted factors can assist city planners to understand how urban livestock keepers use manure. Using the non-parametric classification tree method CART, two approaches were used to select an optimal classification of 'bad' manure users: the 'Cook and Goldman rule' and the 'Classification with different misclassification cost ratios' method. The optimal tree was selected by using receiver operating characteristics curve information. This method can be useful in assessing other aspects of waste management.

Molecular screening of cattle ticks, tick-borne pathogens and amitraz resistance in ticks of Santo Domingo de los Tsáchilas province in Ecuador.

The province of Santo Domingo de los Tsáchilas in Ecuador is a strategic place for cattle production and trade. The lack of knowledge about tick species, tick control and tick-borne diseases in Ecuador prompted this study with the goal of identifying the cattle-tick species and tick-borne agents present in the area and molecularly determining the potential acaricide resistance to amitraz of the major cattle tick species. Eighty-four cattle farms were visited and in 88 % of them, cattle were infested with ticks. Additionally, 24 historical samples from other surrounding Ecuadorian provinces, were screened as well. Besides morphological keys, PCR-RFLP MspI was used to confirm the presence of the Rhipicephalus ticks. The tick samples were also screened for tick-borne agents using PCR-RFLP BseDI and Hhal tests to identify circulating Babesia sp. and Anaplasma spp. Furthermore, the PCR-RFLP EciI technique was used to identify the amitraz resistance gene in populations of Rhipicephalus microplus in the province. Pooled testing was used to determine prevalence at individual-tick level. The presence of R. microplus and Amblyomma cajennense sensu lato (s.l.) ticks was found in 83 % and 21 % of the cattle farms respectively, showing R. microplus is widespread in the province of Santo Domingo de los Tsáchilas. Regarding tick-borne agents, only Anaplasma marginale was observed in 50 % of the visited farms of the province, while about 27 % of the ticks tested positive according to estimations from the data of the tick pools. The presence of Babesia bigemina was only confirmed in samples collected outside the province. The amitraz resistance allele in R. microplus was found in 62 % of the farms, but the percentage of farms with cattle ticks completely resistant to this acaricide was low (2%). The findings of this study should prompt cattle producers and animal health authorities to monitor control strategies, which address the management of resistant tick populations and the epidemiologically-unstable areas of tick-borne diseases.

Impact of insecticide-treated nets on wild pyrethroid resistant Anopheles epiroticus population from southern Vietnam tested in experimental huts.

BACKGROUND: In this study, the efficacy of insecticide-treated nets was evaluated in terms of deterrence, blood-feeding inhibition, induced exophily and mortality on a wild resistant population of Anopheles epiroticus in southern Vietnam, in order to gain insight into the operational consequences of the insecticide resistance observed in this malaria vector in the Mekong delta. METHOD: An experimental station, based on the model of West Africa and adapted to the behaviour of the target species, was built in southern Vietnam. The study design was adapted from the WHO phase 2 guidelines. The study arms included a conventionally treated polyester net (CTN) with deltamethrin washed just before exhaustion, the WHO recommended long-lasting insecticidal net (LLIN) PermaNet 2.0 unwashed and 20 times washed and PermaNet 3.0, designed for the control of pyrethroid resistant vectors, unwashed and 20 times washed. RESULTS: The nets still provided personal protection against the resistant An. epiroticus population. The personal protection ranged from 67% for deltamethrin CTN to 85% for unwashed PermaNet 3.0. Insecticide resistance in the An. epiroticus mosquitoes did not seem to alter the deterrent effect of pyrethroids. A significant higher mortality was still observed among the treatment arms despite the fact that the An. epiroticus population is resistant against the tested insecticides. CONCLUSION: This study shows that CTN and LLINs still protect individuals against a pyrethroid resistant malaria vector from the Mekong region, where insecticide resistance is caused by a metabolic mechanism. In the light of a possible elimination of malaria from the Mekong region these insights in operational consequences of the insecticide resistance on control tools is of upmost importance.