Persistence of Trypanosoma cruzi vector-borne transmission among school-age children in the Bolivian Chaco documented by 24-month longitudinal serosurveillance.

BACKGROUND: Chagas disease represents a major public health concern in several Latin American countries, including Bolivia. METHODS: We present a longitudinal serosurvey for Trypanosoma cruzi antibodies among a cohort of 120 school-age children from rural communities in the Bolivian Chaco at three time points between 2017 and 2019. Serum samples extracted from dry blood spots collected on filter paper were tested for T. cruzi antibodies by enzyme-linked immunosorbent assay and rapid diagnostic test. RESULTS: T. cruzi antibodies were detected in 7/120 (5.8%), 8/120 (6.7%) and 11/120 (9.2%) samples in 2017, 2018 and 2019, respectively. An average incidence of 1.76 per 100 person-years was observed. CONCLUSIONS: Our findings support the persistence of vector-borne T. cruzi transmission in this area, highlighting the need for strengthening multidisciplinary efforts against Chagas disease.

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.

Relevant increase of CTX-M-producing Escherichia coli carriage in school-aged children from rural areas of the Bolivian Chaco in a three-year period.

OBJECTIVES: The aim of this study was to perform two cross-sectional surveys on the fecal carriage of CTX-M-producing Enterobacterales in school-aged children from rural areas of the Bolivian Chaco (2016 vs 2019). METHODS: A total of 757 fecal samples were collected from school-aged children living in nine indigenous communities (n=337, 2016; n=420, 2019). After a first passage onto MacConkey agar (MCA), samples were plated onto MCA plus cefotaxime 2 µg/mL (MCA-CTX), and a loopful of the bacterial growth was used as a template for the detection of group 1, 2, 8/25, and 9 blaCTX-M variants by multiplex reverse transcriptase polymerase chain reaction . Positive samples were tested again for detecting, identifying, and characterizing CTX-M-positive isolates. RESULTS: Growth onto MCA-CTX was obtained with 208 samples (27.5%; 62/337, 2016; 146/420, 2019), of which 201 (96.6%) were positive for blaCTX-M genes. Overall, a relevant increase of fecal carriage of CTX-M-producing Enterobacterales was observed in the study period: 17,5% (59/337) in 2016 compared with 33,8% (142/420) in 2019, p<0.01. Nonetheless, the relative group distribution of CTX-M groups remained stable, with group 1 being the prevalent, followed by group 9 and group 8/25. Group 2 was not detected. CONCLUSIONS: The present study demonstrated an alarming spread of CTX-M enzymes in rural areas of the Bolivian Chaco, where antibiotics consumption is limited. Further studies are encouraged to better understand the dissemination dynamics of such relevant resistance determinants.

Feasibility of a Combined Mobile-Health Electrocardiographic and Rapid Diagnostic Test Screening for Chagas-Related Cardiac Alterations.

BACKGROUND: Chronic Chagas cardiomyopathy (CChC) is the most common cause of death related to Chagas disease (CD). The aim of this study was to assess the feasibility of a combined rapid diagnostic test (RDT) and electrocardiographic (ECG) screening in a remote rural village of the Bolivian Chaco, with a high prevalence of CChC. METHODS: Consecutive healthy volunteers > 15 years were enrolled in the community of Palmarito (municipality of Gutierrez, Santa Cruz Department, Bolivia) in February 2019. All patients performed an RDT with Chagas Stat-Pak® (CSP, Chembio Diagnostic System, Medford, NY, USA) and an ECG by D-Heart® technology, a low-cost, user-friendly smartphone-based 8-lead Bluetooth ECG. RDTs were read locally while ECGs were sent to a cardiology clinic which transmitted reports within 24 h from recording. RESULTS: Among 140 people (54 men, median age 38(interquartile range 23-54) years), 98 (70%) were positive for Trypanosoma cruzi infection, with a linear, age-dependent, increasing trend (p < 0.001). Twenty-five (18%) individuals showed ECG abnormalities compatible with CD. Prevalence of ECG abnormalities was higher in infected individuals and was associated with higher systolic blood pressure and smoking. Following screening, 22 (16%) individuals underwent clinical evaluation and chest X-ray and two were referred for further evaluation. At multivariate analysis, positive CSP results (OR = 4.75, 95%CI 1.08-20.96, p = 0.039) and smoking (OR = 4.20, 95%CI 1.18-14.92, p = 0.027) were independent predictors of ECG abnormalities. Overall cost for screening implementation was <10 $. CONCLUSIONS: Combined mobile-Health and RDTs was a reliable and effective low-cost strategy to identify patients at high risk of disease needing cardiologic assessment suggesting potential future applications.

Cross-sectional study on intestinal parasite infections in different ecological zones of the Department of La Paz, Bolivia.

We performed a cross-sectional parasitological survey to assess the prevalence of protozoa and helminth infection among 275 school-age children (SAC) living in rural and peri-urban areas located in different ecological zones of the Department of La Paz, Bolivia. Public health activities for the control of STH, based on the biannual administration of mebendazole to preschool and school children are implemented at national level since 1986. We found an overall prevalence of 82.2% for intestinal parasites, including protozoa (80%) and helminths (23.3%). Blastocystis and Entamoeba coli were the most prevalent protozoa (found in 44% and 20.7% of the SAC enrolled); as for helminths, Ascaris lumbricoides and Hymenolepis nana were diagnosed in 14.5% and 3.3% of the children, respectively, followed by Trichuris trichiura 1.4%, Enterobius vermicularis 1.4%, Strongyloides stercoralis 0.7% and hookworms 0.7%. Molecular characterization of Blastocystis positive samples evidenced three different subtypes (ST1, ST2, ST3) highlighting the risk of transmission also from animal reservoir. We found a significant difference in the distribution of intestinal parasitic infection (IPIs) by ecological zone (44/74. 59% in Andean highlands, 94/170, 88% in tropical lowlands and 88/94, 94% in the Yungas, p < 0.001). Access to potable water (OR 0.1 95%CI 0.02-0.5, p = 0.004) and the habit of boiling drinking water (OR 0.3, 95% CI 0.2-0.7, p = 0.004) showed an independent association with a lower risk of all IPIs and STHs, respectively. The very low prevalence of STH infections of moderate heavy intensity demonstrate that periodical deworming has been successful in reducing the morbidity due to these parasites, however the high prevalence of protozoa demonstrate that sanitation is still problematic and there is a relevant contamination of the environment with human faeces. Significant efforts are still needed to reduce IPIs transmission and to improve health and sanitation in this area.

Skeletogenesis by transfated secondary mesenchyme cells is dependent on extracellular matrix-ectoderm interactions in Paracentrotus lividus sea urchin embryos.

In the sea urchin embryo, primary mesenchyme cells (PMCs) are committed early in development to direct skeletogenesis, provided that a permissive signal is conveyed from adjacent ectoderm cells. We showed that inhibition of extracellular matrix (ECM)-ectoderm cells interaction, by monoclonal antibodies (mAb) to Pl-nectin, causes an impairment of skeletogenesis and reduced expression of Pl-SM30, a spicule-specific matrix protein. When PMCs are experimentally removed, some secondary mesenchyme cells (SMCs) switch to skeletogenic fate. Here, for the first time we studied SMC transfating in PMC-less embryos of Paracentrotus lividus. We observed the appearance of skeletogenic cells within 10 h of PMCs removal, as shown by binding of wheat germ agglutinin (WGA) to cell surface molecules unique to PMCs. Interestingly, the number of WGA-positive cells, expressing also msp130, another PMC-specific marker, doubled with respect to that of PMCs present in normal embryos, though the number of SM30-expressing cells remained constant. In addition, we investigated the ability of SMCs to direct skeletogenesis in embryos exposed to mAbs to Pl-nectin after removal of PMCs. We found that, although phenotypic SMC transfating occurred, spicule development, as well as Pl-SM30-expression was strongly inhibited. These results demonstrate that ectoderm inductive signals are necessary for transfated SMCs to express genes needed for skeletogenesis.

UVB radiation prevents skeleton growth and stimulates the expression of stress markers in sea urchin embryos.

Ozone depletion results in an increased flux of biologically damaging radiations reaching the earth. Although ultraviolet (UV) penetration is attenuated by the seawater, harmful effects can be still observed at low depths where sea urchin embryos are living. We have used Paracentrotus lividus embryos to study the impacts of UV radiation on their development. Blastula cultures were exposed to different doses of UVB (312 nm) radiations and the resulting endpoint effects were evaluated in terms of embryonic morphological abnormalities, variations in specific gene expression, and changes in the levels of stress proteins. We found that embryos were moderately sensitive to 50 J/m2 UVB radiation; an increase in the number of developmentally delayed and malformed embryos was detected when increasing doses, up to 1000 J/m2, were used. Major developmental defects, observed 24 and 48 h after exposure, consisted in the failure of skeleton elongation and patterning. Accordingly, we found a reduction in the number of primary mesenchyme cells that expressed Pl-SM30, a gene coding for one of the specific matrix proteins of the skeleton. The morphological effects observed 1, 24, and 48 h after exposure were correlated with a dose-dependent increase in the level and in the activation of two recognized stress markers, namely hsp70 and p38 MAPk, respectively, consistent with their role in mediating cellular response to stress and suggesting a function in embryo survival.

Expression of univin, a TGF-beta growth factor, requires ectoderm-ECM interaction and promotes skeletal growth in the sea urchin embryo.

Pl-nectin is an ECM protein located on the apical surface of ectoderm cells of Paracentrotus lividus sea urchin embryo. Inhibition of ECM-ectoderm cell interaction by the addition of McAb to Pl-nectin to the culture causes a dramatic impairment of skeletogenesis, offering a good model for the study of factor(s) involved in skeleton elongation and patterning. We showed that skeleton deficiency was not due to a reduction in the number of PMCs ingressing the blastocoel, but it was correlated with a reduction in the number of Pl-SM30-expressing PMCs. Here, we provide evidence on the involvement of growth factor(s) in skeleton morphogenesis. Skeleton-defective embryos showed a strong reduction in the levels of expression of Pl-univin, a growth factor of the TGF-beta superfamily, which was correlated with an equivalent strong reduction in the levels of Pl-SM30. In contrast, expression levels of Pl-BMP5-7 remained low and constant in both skeleton-defective and normal embryos. Microinjection of horse serum in the blastocoelic cavity of embryos cultured in the presence of the antibody rescued skeleton development. Finally, we found that misexpression of univin is also sufficient to rescue defects in skeleton elongation and SM30 expression caused by McAb to Pl-nectin, suggesting a key role for univin or closely related factor in sea urchin skeleton morphogenesis.