Long-Standing International Cooperation in Parasitology Research: A Summary of 35 Years of Activities in the Bolivian Chaco.

The Bolivian Chaco is a semiarid region with a low population density, situated in the southeast part of the Plurinational State of Bolivia. Here, despite the improvements of the last 15 years, poverty remains high in rural areas, where social vulnerability is widespread. The Guaraní ethnic group often lives in isolated communities with a low standard of hygiene and sanitation. This epidemiological scenario favors the spread of transmissible diseases, including several parasitic infections belonging to the neglected tropical diseases (NTDs) group. In this area, a long-standing research activity, built upon the synergism between local and foreign institutions, has been established since the late 1980s and helps to fill in the knowledge gap about the epidemiology dynamics of soil-transmitted helminths, vector-borne parasites, and other parasitic diseases. A 35-year history of cooperation programs in parasitology research has contributed to informing local health authorities of the NTD burden in the Bolivian Chaco and, ultimately, supports local healthcare providers in the management of parasitic diseases.

Trypanosoma cruzi infection in the human population of the Bolivian Chaco: four serosurveys over a 26-year period (1987-2013).

INTRODUCTION: Chagas disease (CD) remains a public health concern in several Latin American countries. At global level, Bolivia has the highest CD burden and the Chaco region, in the southeast of the country, is the most affected area. We report the results of four serosurveys for Trypanosoma cruzi antibodies, carried out approximately ten years apart from each other, during the lapse 1987-2013, in different localities of the Bolivian Chaco. METHODOLOGY: Four cross-sectional surveys were conducted in various localities, mostly rural, of the Bolivian Chaco, during the period 1987-2013. RESULTS: Although a reliable analysis of CD epidemiological trend is challenging, a partial reduction of anti-T. cruzi seroprevalence over the past four decades in the Bolivian Chaco may be assumed. In particular, in 1987 the exposure to T. cruzi in rural setting was universal since the first years of life, while it resulted gradually lower and age-dependent thereafter. Moreover, T. cruzi seroprevalence among women of reproductive age (15-45 years) has been persistently high in rural areas. CONCLUSIONS: T. cruzi transmission is still active and CD remains a concern throughout the Bolivian Chaco. More efforts are needed in order to achieve a sustainable interruption of vector-borne CD transmission in this area.

Decline in Total Serum IgE and Soluble CD30 in the Context of Soil-Transmitted Helminth Decline in Bolivia.

In the Bolivian Chaco, recent surveys documented a dramatic decrease in the prevalence of soil-transmitted helminth (STH) infections as compared with the 1980s after thirty years of preventive chemotherapy (PC). Concomitant immunological rearrangements are expected. Because nematode infections are associated with increased levels of circulating IgE and glycoprotein CD30 soluble form (sCD30), this study aims to evaluate changes in serological markers of T helper (Th)2-cells activity between 1987 (high STH prevalence) and 2013 (low STH prevalence) in rural communities in the Bolivian Chaco area. We collected 151 sera during two different surveys in 1987 (n = 65) and 2013 (n = 86) and measured the concentration of total IgE and sCD30 by immunoassays. We found a statistically significant age-independent decrease in the total IgE (P < 0.0001) and sCD30 (P < 0.0001) from 1987 to 2013. The significant decrease in serological Th2 markers (IgE and sCD30) between 1987 and 2013 is consistent with the drop in STH prevalence in this geographical area during the same period of time. Further studies might elucidate the clinical and epidemiological impact of these serological rearrangements.

Canine Trypanosoma cruzi infection in the Bolivian Chaco.

A cross-sectional study on Trypanosoma cruzi was carried out in 2013 to evaluate the role of dogs as possible source of infection for humans in two rural communities of the highly endemic Bolivian Chaco (Bartolo, Chuquisaca Department, n = 57 dogs; and Ivamirapinta, Santa Cruz Department, n = 48 dogs). Giemsa-stained thick and thin smears, rapid immunochromatographic test (ICT) (Chagas Quick test, Cypress Diagnostic, Belgium) and polymerase chain reaction for T. cruzi on dried blood spots were performed. All smears proved negative by microscopic examination, whereas 23/103 (22%) were positive by ICT and 5/105 (5%) blood samples contained T. cruzi DNA, evidencing the potential role of dogs in the domestic transmission of the parasite.

Hymenolepis nana-An Emerging Intestinal Parasite Associated with Anemia in School Children from the Bolivian Chaco.

Tropical anemia can have multiple causes, whether socioeconomic, dietary, or infectious. In the Bolivian Chaco, soil-transmitted helminthiases (STH), malaria, and Chagas disease are potential infectious causes of anemia among school-aged children (SAC). Following years of preventive chemotherapy with mebendazole, the prevalence of STH among SAC living in that area is now negligible, whereas protozoan infections are still highly prevalent (81%); Hymenolepis nana is the most frequent intestinal helminth (∼13%). We present results of hemoglobin (Hb) assessment and the association between parasitic infections and Hb levels of that SAC population. Overall, 511 SAC (girls:boys ratio 1:1, mean age 9.4 years [95% confidence interval {CI}: 9.3-9.5]) had Hb levels measured by using a point of care testing (HemoCue® Hb 301 System; HemoCue, Angelhome, Sweden). The prevalence of anemia was 23% (117/511), with mean and median Hb level = 12.2 g/dL (95% CI: 12.1-12.3; range 9.2-15.4 g/dL). By multivariate analysis, H. nana infection was associated with an increased risk of anemia (odds ratio 2.9, 95% CI: 1.5-5.7, P = 0.002). Two samples (0.5%) were positive for Trypanosoma cruzi and none for Plasmodium spp. by polymerase chain reaction of the 439 children tested. Anemia is still a concern among SAC living in the Bolivian Chaco. Our findings call for a greater attention to fecal-oral emerging pathogens, such as H. nana, and highlight the importance of water, sanitation, and hygiene improvements for disadvantaged population such as those living in the Bolivian Chaco.

Scaling down of a deworming programme among school-age children after a thirty-year successful intervention in the Bolivian Chaco.

OBJECTIVE: Preventive chemotherapy is the WHO-recommended control method for soil-transmitted helminthiases. In the Bolivian Chaco, 6-monthly single-dose mebendazole delivery to school-age children achieved a dramatic decrease in soil-transmitted helminthiases prevalence between 1987 and 2013. Consequently, in September 2016, preventive chemotherapy delivery was interrupted in nine rural communities. In compliance with WHO recommendations, we intensified surveillance to monitor soil-transmitted helminthiases prevalence and detect potential changes that would require interventions. METHODS: We conducted two cross-sectional parasitology surveys 12 months apart (September 2016-2017) among school-age children living in the communities where preventive chemotherapy delivery had been halted. Study design, methods of sampling and sample analysis technique (direct microscopy, Kato-Katz technique) followed WHO recommendations, aiming to obtain data representative of the Bolivian Chaco ecological zone. RESULTS: We collected 426 samples in 2016 and 520 in 2017. Soil-transmitted helminthiasis prevalence was unremarkable: 0.7% (95% CI 0-1.5%) in 2016 and 0.8% (0-1.5%) in 2017. Conversely, the prevalence of tapeworms (13% in 2016, 12% in 2017) and intestinal protozoan infections (81% in 2016 and 75% in 2017) continued to be high. CONCLUSIONS: Our findings support the role of preventive chemotherapy in reducing soil-transmitted helminthiases transmission, as otherwise poor hygienic and health conditions persist in the Bolivian Chaco. A national survey, involving areas from all the ecological zones of Bolivia, is now warranted.

Seroepidemiological trend of strongyloidiasis in the Bolivian Chaco (1987-2013) in the absence of disease-specific control measures.

OBJECTIVES: The prevalence of Strongyloides stercoralis infections is grossly underestimated because infections go mostly undetected, although they can persist for a lifetime due to the auto-infective cycle. In the Bolivian Chaco, the prevalence of soil-transmitted nematodes dropped dramatically in the past 25 years, but the mebendazole used for preventive chemotherapy has no effect on S. stercoralis. Meanwhile, the prevalence of intestinal protozoan infections remains unchanged. We compared S. stercoralis seroprevalence in rural communities of the Bolivian Chaco from 1987 to 2013. METHODS: Sera collected during two previous serosurveys, conducted in the Chaco region in 1987 and 2013, were tested for S. stercoralis using a commercial assay (Bordier-ELISA, Bordier Affinity Products, Switzerland). RESULTS: Overall, 355 sera were analysed, 122 from the 1987 survey and 233 from the 2013 survey. Seropositivity for S. stercoralis was significantly more prevalent in 1987 (19/122, 16% in 1987 vs. 15/233, 6% in 2013, P = 0.006), accounted for by a drop from 17% to 3% in people under 26 years of age. Multivariate analysis showed a significant association between seropositivity for S. stercoralis and age in the 2013 population (OR 1.03 for each one-year increase, 95%CI 1.00-1.05, P = 0.04), but none in 1987. CONCLUSIONS: The significant reduction in S. stercoralis seroprevalence in Bolivian Chaco cannot be explained by preventive chemotherapy or improved social-sanitary conditions. As the drop is seen in younger generations, it is consistent with little transmission occurring. However, the risk of transmission still exists, as prevalence is persistently high in older individuals, who present a potential reservoir due to the lifelong nature of S. stercoralis infections.

Intestinal parasitic infections and associated epidemiological drivers in two rural communities of the Bolivian Chaco.

INTRODUCTION: In 2013 a coproparasitological survey was carried out in two rural communities of the Bolivian Chaco to determine the prevalence of intestinal parasitic infections (IPIs) and to investigate on possible infection drivers through a questionnaire interview. METHODOLOGY: Faecal samples were examined by microscopy. Samples positive for Entamoeba histolytica complex and Blastocystis were molecularly examined to identify the species/subtypes involved. RESULTS: The overall infection rate was 86%, identical in both communities and mostly due to protozoa. Soil-transmitted helminths were detected in <3% of children and adults. DISCUSSION: The protozoa detected, including Blastocystis subtypes, indicate faecal contamination of the environment by both humans (as confirmed by the presence of Hymenolepis nana) and animals. Nested-PCR identified E. histolytica, thus signalling the possible occurrence of invasive amoebosis. Lack of safe water, environmental contamination, poor sanitation and hygiene, shared by both communities, are the main drivers of IPIs. In addition, unlike gender and socioeconomic factors, childhood (only for some species), crowding and cohabitation with animals proved to be further significant protozoon infection risk factors. CONCLUSIONS: These results highlight the need for the promotion of access to clean water, improved sanitation and better hygiene, thus reducing the frequency of preventive chemotherapy for STHs while continuing to monitor the population for possible recrudescence.

Human Babesiosis, Bolivia, 2013.

To investigate human babesiosis in the Bolivian Chaco, in 2013 we tested blood samples from 271 healthy persons living in 2 rural communities in this region. Microscopy and PCR indicated that 3.3% of persons were positive for Babesia microti parasites (US lineage); seroprevalence was 45.7%. Appropriate screening should mitigate the risk for transfusion-associated babesiosis.

Dramatic decrease in prevalence of soil-transmitted helminths and new insights into intestinal protozoa in children living in the Chaco region, Bolivia.

We assessed the prevalence of intestinal parasites among 268 2-12-year-old children living in rural areas, small villages, and semi-urban areas of the Chaco region, south-eastern Bolivia. The overall parasitism was 69%. Only protozoa, helminths, or co-infections were observed in 89.2%, 5.9%, or 4.9% of the positive children, respectively. A significant progressive increase in overall parasite prevalence was found when passing from rural areas to small villages and semi-urban areas. The most commonly found species were Entamoeba coli (38.4%), Giardia intestinalis (37.7%), and Blastocystis spp. (16%). Hymenolepis nana was the most prevalent helminth (5.6%), followed by Ascaris lumbricoides and hookworms (1.5% and 0.4%) evidenced only in rural areas and in villages. Molecular diagnostics identified Blastocystis subtypes 9 and 2, and 5 infections by Entamoeba histolytica and 4 by Entamoeba dispar. The dramatic decrease in prevalence of soil-transmitted helminths with respect to that observed about 20 years ago (> 40%) evidences the success of the preventive chemotherapy intervention implemented in 1986. Health education and improved sanitation should be intensified to control protozoan infections.

Evaluation of immunodiagnostics for toxocarosis in experimental porcine cysticercosis.

We assessed whether immunodiagnostic tests for cysticercosis can cross-react with the currently available immunodiagnostic tests for Toxocara canis in an established animal model for cysticercosis infection in pigs, known host for Toxocara. We examined by TES-enzyme-linked immunosorbent test and immunoblot assay for toxocarosis and cysticercosis the baseline and final follow-up sera of 10 pigs, before and after (3 months) infection with Taenia solium. After successful cysticercosis infection, the nine evaluable pigs became seropositive to T. solium (enzyme-linked immunoelectrotransfer blot assay), but did remain seronegative for Toxocara in both assays, documenting the lack of cross-reactivity with anti-T. solium antibodies in both T. canis assays. These findings should help clinicians better interpret serology for toxocariosis and cysticercosis in endemic areas for both helminth infections.

Anticuerpos contra Trichinella spiralis en la población rural de la provincia Cordillera, Bolivia / Seroprevalence of antibodies to Trichinella spiralis among rural residents of Cordillera province, Bolivia

Se realizó un estudio seroepidemiológico para determinar la prevalencia de anticuerpos contra Trichinella spiralis en el área rural de la provincia Cordillera del Departamento de Santa Cruz en Bolivia. Se examinaron 234 muestras de suero mediante el ensayo de inmunoabsorción enzimática (ELISA) y se detectaron los anticuerpos en siete muestras (3 por cien). Los resultados documentan por primera vez la presencia de infestación humana por Trichinella en Bolivia y sugieren la necesidad de fortalecer la vigilancia sanitaria de la triquinosis en los mataderos municipales, impedir la faena clandestina de animales y, sobre todo, lograr que los productores y pobladores tomen conciencia de los peligros de esta zoonosis

A seroepidemiological study was conducted to determine the prevalence of antibodies to Trichinella spiralis among rural residents of Cordillera province, Santa Cruz Department, Bolivia. Using the enzyme-linked immunosorbent assay (ELISA), 234 serum samples were examined, and antibodies were detected in seven of the samples (3%). The results document for the first time the presence of human infestation with Trichinella in Bolivia and suggest the need to strengthen trichinelosis surveillance in the municipal slaughterhouses, to prevent the clandestine slaughter of animals, and particularly to ensure that residents and meat producers in the area become aware of the dangers of this zoonosis