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Soil-transmitted helminths (STHs) are a group of parasites that are globally distributed and are the most prevalent neglected disease (NTD) in Latin America and the Caribbean (LAC); their presence is associated with health and development problems. In Argentina, their distribution is heterogenous, and there are highly endemic areas in the north of the country. The World Health Organization (WHO) recommends the mass deworming of children as a first-line strategy for the prevention and control of STHs and recently also encourage the development of more sensitive diagnostic tests. The aim of this study was to determine the prevalence of STHs in Tartagal (Salta, Argentina) after four years of deworming interruption with albendazole and ivermectin. A total of 437 fecal samples were analyzed using standard coprological techniques, a subset of which were selected to molecularly typify protozoan parasites; 257 blood samples were analyzed for the presence of specific antibodies to the STH Strongyloides stercoralis. The most prevalent protozoan species were G. intestinalis (19.6–49.2%) and B. hominis (19.1–38.5%). Molecular characterization allowed us to evidence possible zoonotic or human-to-human transmission pathways for Giardia intestinalis or Blastocystis spp., while serology for S. stercoralis proved to be a useful screening tool for monitoring this parasite after treatment. In general, a decrease in the prevalence of STHs was observed in the area, from 60% to 2.9–20% for hookworms and from 51% to 1–9.3% for S. stercoralis four years after treatment, demonstrating the effectiveness and duration of anthelmintic treatment with these two drugs. [ABSTRACT FROM AUTHOR]

Background: In resource-limited countries, stool microscopy is the diagnostic test of choice for intestinal parasites (soil-transmitted helminths and/or intestinal protozoa). However, sensitivity and specificity is low. Improved diagnosis of intestinal parasites is especially important for accurate measurements of prevalence and intensity of infections in endemic areas. Methods: The study was carried out in Orán, Argentina. A total of 99 stool samples from a local surveillance campaign were analyzed by concentration microscopy and McMaster egg counting technique compared to the analysis by multi-parallel quantitative real-time polymerase chain reaction (qPCR). This study compared the performance of qPCR assay and stool microscopy for 8 common intestinal parasites that infect humans including the helminths Ascaris lumbricoides, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Trichuris trichiura, and the protozoa Giardia lamblia, Cryptosporidium parvum/hominis, and Entamoeba histolytica, and investigated the prevalence of polyparasitism in an endemic area. Results: qPCR showed higher detection rates for all parasites as compared to stool microscopy except T. trichiura. Species-specific primers and probes were able to distinguish between A. duodenale (19.1 %) and N. americanus (36.4 %) infections. There were 48.6 % of subjects co-infected with both hookworms, and a significant increase in hookworm DNA for A. duodenale versus N. americanus (119.6 fg/μL: 0.63 fg/μL, P < 0.001) respectively. qPCR outperformed microscopy by the largest margin in G. lamblia infections (63.6 % versus 8.1 %, P < 0.05). Polyparasitism was detected more often by qPCR compared to microscopy (64.7 % versus 24.2 %, P < 0.05). Conclusions: Multi-parallel qPCR is a quantitative molecular diagnostic method for common intestinal parasites in an endemic area that has improved diagnostic accuracy compared to stool microscopy. This first time use of multi-parallel qPCR in Argentina has demonstrated the high prevalence of intestinal parasites in a peri-urban area. These results will contribute to more accurate epidemiological survey, refined treatment strategies on a public scale, and better health outcomes in endemic settings. [ABSTRACT FROM AUTHOR]

Background: In resource-limited countries, stool microscopy is the diagnostic test of choice for intestinal parasites (soil-transmitted helminths and/or intestinal protozoa). However, sensitivity and specificity is low. Improved diagnosis of intestinal parasites is especially important for accurate measurements of prevalence and intensity of infections in endemic areas. Methods: The study was carried out in Orán, Argentina. A total of 99 stool samples from a local surveillance campaign were analyzed by concentration microscopy and McMaster egg counting technique compared to the analysis by multi-parallel quantitative real-time polymerase chain reaction (qPCR). This study compared the performance of qPCR assay and stool microscopy for 8 common intestinal parasites that infect humans including the helminths Ascaris lumbricoides, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Trichuris trichiura, and the protozoa Giardia lamblia, Cryptosporidium parvum/hominis, and Entamoeba histolytica, and investigated the prevalence of polyparasitism in an endemic area. Results: qPCR showed higher detection rates for all parasites as compared to stool microscopy except T. trichiura. Species-specific primers and probes were able to distinguish between A. duodenale (19.1 %) and N. americanus (36.4 %) infections. There were 48.6 % of subjects co-infected with both hookworms, and a significant increase in hookworm DNA for A. duodenale versus N. americanus (119.6 fg/μL: 0.63 fg/μL, P < 0.001) respectively. qPCR outperformed microscopy by the largest margin in G. lamblia infections (63.6 % versus 8.1 %, P < 0.05). Polyparasitism was detected more often by qPCR compared to microscopy (64.7 % versus 24.2 %, P < 0.05). Conclusions: Multi-parallel qPCR is a quantitative molecular diagnostic method for common intestinal parasites in an endemic area that has improved diagnostic accuracy compared to stool microscopy. This first time use of multi-parallel qPCR in Argentina has demonstrated the high prevalence of intestinal parasites in a peri-urban area. These results will contribute to more accurate epidemiological survey, refined treatment strategies on a public scale, and better health outcomes in endemic settings. [ABSTRACT FROM AUTHOR]

Objective To determine the ability of recombinant antigens to detect cases of infection with Trypanosoma cruzi among cases of infection with Leishmania spp. by serological methods. Methods Sera from 41 patients infected with Leishmania spp. were evaluated with ELISA using single ( FRA, CP1 and TSSAVI) or pooled (commercial Rec- ELISA) recombinant proteins or homogenate antigens (commercial H- ELISA). As there is no gold standard antigen to discriminate Chagas disease from leishmaniasis, the correlation of results between defined antigens and the homogenate was made with Kappa Index ( KI), the level of correlation considered being used as a criterion of specificity. Results Single recombinant antigens and Rec- ELISA showed good correlation ( KI > 0.8). A low correlation ( KI < 0.66) was observed between the results from single recombinant antigens or the commercial recombinant kit and H- ELISA. Conclusions The highly correlated results between T. cruzi single or pooled recombinant proteins are indicative of the usefulness of recombinant antigens for Chagas diagnosis. Our results also indicate that in the city of Oran in Argentina, between 12% and 17% of patients with leishmaniasis are also infected with Chagas disease. The high KI values between TSSAVI and the other recombinant proteins suggest that in these patients, the infection may be caused by T. cruzi II and/or V and/or VI lineages. [ABSTRACT FROM AUTHOR]

Background: The WHO has established a control strategy for Strongyloides stercoralis in school-aged children as well as targets and to maintain control programs for Ascaris lumbricoides, Trichuris trichiura and hookworms. For an efficient development of control programs, it is necessary to know the target countries around the world, as well as the areas within each country where efforts should be focused. Therefore, maps that provide information on the areas at risk for soil-transmitted helminth (STH) infections on a national and sub-national scale would allow for a better allocation of resources. Methods: We used the ecological niche models MaxEnt and Kuenm R library to estimate the global distribution of S. stercoralis and hookworms. We used occurrence points of both species extracted from surveys of two literature reviews and from the Global Atlas of Helminth Infection database, together with 14 raster maps of environmental variables. Results: We obtained two raster maps with the presence probability of S. stercoralis and hookworm infections at a global level and then estimated the global population at risk to be 2.6 and 3.4 billion, respectively. The population at risk was also estimated at the country level using estimations for areas as small as 25 km2. A relationship was found between the probability of the presence of S. stercoralis and its prevalence, and a raster map was generated. Annual precipitation, annual temperature, soil carbon content and land cover were the main associated environmental variables. The ecological niches of Strongyloides stercoralis and hookworms had an overlap of 68%. Conclusions: Here we provide information that can be used for developing more efficient and integrated control strategies for S. stercoralis and hookworm infections. This information can be annexed to the study of other risk factors or even other diseases to assess the health status of a community. Graphical Abstarct: [ABSTRACT FROM AUTHOR]

Background: Strongyloidiasis and Chagas disease are endemic in northern Argentina. In this study we evaluate the association between S. stercoralis and T. cruzi infections in villages with diverse prevalence levels for these parasites. Further understanding in the relationship between these Neglected Tropical Diseases of South America is relevant for the design of integrated control measures as well as exploring potential biologic interactions. Methodology: Community based cross-sectional studies were carried in different villages of the Chaco and Yungas regions in Argentina. Individuals were diagnosed by serology for S. stercoralis and T. cruzi. The association between S. stercoralis and T. cruzi, and between anemia and the two parasites was evaluated using two approaches: marginal (Ma) and multilevel regression (Mu). Results: A total of 706 individuals from six villages of northern Argentina were included. A total of 37% were positive for S. stercoralis, 14% were positive for T. cruzi and 5% were positive for both. No association was found between infection with S. stercoralis and T. cruzi in any of the models, but we found a negative correlation between the prevalence of these species in the different villages (r = -0.91). Adults (> 15 years) presented association with S. stercoralis (Ma OR = 2.72; Mu OR = 2.84) and T. cruzi (Ma OR = 5.12; Mu OR = 5.48). Also, 12% and 2% of the variance of infection with S. stercoralis and T. cruzi, respectively, could be explained by differences among villages. On the other hand, anemia was associated with infection with S. stercoralis (Ma OR = 1.73; Mu OR = 1.78) and was more prevalent in adults (Ma OR = 2.59; Mu OR = 2.69). Conclusion: We found that coinfection between S. stercoralis and T. cruzi is not more frequent than chance in endemic areas. However, the high prevalence for both parasites, raises the need for an integrated strategy for the control of STH and Chagas disease. Author summary: Infections with S. stercoralis and T. cruzi belong to the group of neglected tropical diseases (NTDs). NTDs are influenced by climate and the presence of their vectors and host reservoirs in areas with structural poverty. S. stercoralis and T. cruzi have high prevalence in northern Argentina. Due to the high prevalence and chronicity of infections by these parasites, coinfection with both is possible. We conducted a cross sectional study in six villages in the north of Argentina. These villages have different prevalence for the two parasites and are located in different ecoregions (Chaco, Yungas and the border between these two). We found that coinfection between S. stercoralis and T. cruzi occurs only by chance; and the most affected groups for S. stercoralis and T. cruzi infection are adults. We also found that adults or individual with infection with S. stercoralis were more likely to have anemia. In addition, each village represents a different setting for infection with S. stercoralis and T. cruzi, and individuals in different villages presented differences in the probability of infection. [ABSTRACT FROM AUTHOR]

Background The efficacy of currently available anthelminthics against Trichuris trichiura infections is significatively lower than for other soil-transmitted helminths. The combination of ivermectin (IVM) and albendazole (ALB) has shown significant improvements in efficacy. Methods Safety and efficacy randomized controlled clinical trial comparing 3 experimental regimens against ALB monotherapy for the treatment of T. trichiura infections in northern Honduras. Infected children were randomized to 4 treatment arms: arm 1, single-dose ALB (400 mg); arm 2, single-dose ALB (400 mg) plus IVM (600 μg/kg); arm 3, ALB (400 mg) for 3 consecutive days; or arm 4, ALB (400 mg) plus IVM (600 μg/kg) for 3 consecutive days. Efficacy was measured based on the egg reduction and cure rates, both assessed 14–21 days after treatment, using the Kato-Katz method. Safety was evaluated by analyzing the frequency and severity of adverse events. Results Of 176 children randomized to 1 of the 4 treatment arms, 117 completed treatment and follow-up. The egg reduction rates for arms 1, 2, 3, and 4 were 47.7%, 96.7%, 72.1%, and 100%, respectively; with P values <.001 for comparisons between IVM groups and ALB-only arms. The cure rates were 4.2%, 88.6%, 33.3%, and 100%, respectively. A total of 48 adverse events (85.4% mild) were reported in 36 children. Conclusions The combined use of ALB and high-dose IVM is a highly effective and well tolerated treatment for the treatment of T. trichiura infections, offering significantly improved treatment for the control of this infection. Clinical Trials Registration NCT04041453. [ABSTRACT FROM AUTHOR]

Introduction The objective of this cross-sectional study was to describe the main symptoms associated with COVID-19, and their diagnostic characteristics, to aid in the clinical diagnosis. Methods An analysis of all patients diagnosed by RT-PCR for SARS-CoV-2 between April and May 2020 in Argentina was conducted. The data includes clinical and demographic information from all subjects at the time of presentation (n=67318, where 12% were positive for SARS-CoV-2). The study population was divided into four age groups: pediatric (0-17 years), young adults (18-44 years), adults (45- 64 years), and elderly (65-103 years). Multivariate logistic regression was used to measure the association of all symptoms and to create a diagnostic model based on symptoms. Results Symptoms associated with COVID-19 were anosmia, dysgeusia, headache, low-grade fever, odynophagia, and malaise. However, the presentation of these symptoms was different between the different age groups. In turn, at the time of presentation, the symptoms associated with respiratory problems (chest pain, abdominal pain, and dyspnea) had a negative association with COVID-19 or did not present statistical relevance. On the other hand, the model based on 16 symptoms, age and sex, presented a sensitivity of 80% and a specificity of 46%. Conclusions There were significant differences between the different age groups. Additionally, there were interactions between different symptoms that were highly associated with COVID-19. Finally, our findings showed that a regression model based on multiple factors (age, sex, interaction between symptoms) can be used as an accessory diagnostic method or a rapid screening of suspected COVID-19 cases. [ABSTRACT FROM AUTHOR]

Background: Approximately 30% of children worldwide are infected with gastrointestinal parasites. Depending on the species, parasites can disrupt intestinal bacterial microbiota affecting essential vitamin biosynthesis. Methods: Stool samples were collected from 37 asymptomatic children from a previous cross-sectional Argentinian study. A multi-parallel real-time quantitative PCR was implemented for Ascaris lumbricoides, Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Trichuris trichiura, Cryptosporidium spp., Entamoeba histolytica and Giardia duodenalis. In addition, whole-genome sequencing analysis was conducted for bacterial microbiota on all samples and analyzed using Livermore Metagenomic Analysis Toolkit and DIAMOND software. Separate analyses were carried out for uninfected, Giardia-only, Giardia + helminth co-infections, and helminth-only groups. Results: For Giardia-only infected children compared to uninfected children, DNA sequencing data showed a decrease in microbiota biodiversity that correlated with increasing Giardia burden and was statistically significant using Shannonʼs alpha diversity (Giardia-only > 1 fg/µl 2.346; non-infected group 3.253, P = 0.0317). An increase in diversity was observed for helminth-only infections with a decrease in diversity for Giardia + helminth co-infections (P = 0.00178). In Giardia-only infections, microbiome taxonomy changed from Firmicutes towards increasing proportions of Prevotella, with the degree of change related to the intensity of infection compared to uninfected (P = 0.0317). The abundance of Prevotella bacteria was decreased in the helminths-only group but increased for Giardia + helminth co-infections (P = 0.0262). Metagenomic analysis determined cobalamin synthesis was decreased in the Giardia > 1 fg/µl group compared to both the Giardia < 1 fg/µl and the uninfected group (P = 0.0369). Giardia + helminth group also had a decrease in cobalamin CbiM genes from helminth-only infections (P = 0.000754). Conclusion: The study results may provide evidence for an effect of parasitic infections enabling the permissive growth of anaerobic bacteria such as Prevotella, suggesting an altered capacity of vitamin B12 (cobalamin) biosynthesis and potential impact on growth and development in children. [ABSTRACT FROM AUTHOR]

Background: Soil Transmitted Helminth (STH) infections negatively impact physical and mental development in human populations. Current WHO guidelines recommend morbidity control of these infections through mass drug administration (MDA) using albendazole (ABZ) or mebendazole. Despite major reductions in STH associated morbidity globally, not all programs have demonstrated the expected impact on prevalence of parasite infections. These therapeutic failures may be related to poor programmatic coverage, suboptimal adherence or the exposure of parasites to sub-therapeutic drug concentrations. As part of the DeWorm3 project, we sought to characterize the serum disposition kinetics and pattern of urinary excretion of ABZ and its main metabolites ABZ sulphoxide (ABZSO) and ABZ sulphone (ABZSO2) in humans, and the assessment of the duration and optimal time point where ABZ and/or its metabolites can be measured in urine as an indirect assessment of an individual’s adherence to treatment. Methodology/Principal findings: Consecutive venous blood and urine samples were collected from eight (8) human volunteers up to 72 h post-ABZ oral administration. ABZ/metabolites were quantified by HPLC. The ABZSO metabolite was the main analyte recovered both in serum and urine. ABZSO Cmax in serum was 1.20 ± 0.44 μg/mL, reached at 4.75 h post-treatment. In urine, ABZSO Cmax was 3.24 ± 1.51 μg/mL reached at 6.50 h post-ABZ administration. Conclusion/Significance: Pharmacokinetic data obtained for ABZ metabolites in serum and urine, including the recovery of the ABZ sulphoxide derivative up to 72 h in both matrixes and the recovery of the amino-ABZ sulphone metabolite in urine samples, are suggesting the possibility of developing a urine based method to assess compliance to ABZ treatment. Such an assay may be useful to optimize ABZ use in human patients. Trial registration: ClinicalTrials.gov . [ABSTRACT FROM AUTHOR]

Background: Previous studies showed that a naturally attenuated strain from Trypanosoma cruzi triggers an immune response mainly related to a Th2-type profile. Albeit this, a strong protection against virulent challenge was obtained after priming mice with this attenuated strain. However, this protection is not enough to completely clear parasites from the host. In T. cruzi infection, early Interferon-gamma (IFN-γ) is critical to lead type 1 responses able to control intracellular parasites. Therefore we evaluated whether the co-administration of a plasmid encoding murine IFN-γ could modify the immune response induced by infection with attenuated parasites and improve protection against further infections.Methods: C57BL/6J mice were infected intraperitoneally with three doses of live attenuated parasites in combination with plasmid pVXVR-mIFN-γ. Before each infection dose, sera samples were collected for parasite specific antibodies determination and cytokine quantification. To evaluate the recall response to T. cruzi, mice were challenged with virulent parasites 30 days after the last dose and parasite load in peripheral blood and heart was evaluated.Results: As determined by ELISA, significantly increase in T. cruzi specific antibodies response was detected in the group in which pVXVR-mIFN-γ was incorporated, with a higher predominance of IgG2a subtype in comparison to the group of mice only inoculated with attenuated parasites. At our limit of detection, serum levels of IFN-γ were not detected, however a slight decrease in IL-10 concentrations was observed in groups in which pVXVR-mIFN-γ was supplemented. To analyze if the administration of pVXVR-mIFN-γ has any beneficial effect in protection against subsequent infections, all experimental groups were submitted to a lethal challenge with virulent bloodstream trypomastigotes. Similar levels of challenge parasites were detected in peripheral blood and heart of mice primed with attenuated parasites alone or combined with plasmid DNA. Expansion of IgG antibodies was not significant in TCC+ pVXVR-mIFN-γ; however, the overall tendency to sustain a Th2 profile was maintained.Conclusions: Overall, these results suggest that administration of plasmid pVXVR-mIFN-γ could have beneficial effects on host specific antibody production in response to T. cruzi attenuated infection; however, this outcome is not reflected in an improved protection against further virulent infections. [ABSTRACT FROM AUTHOR]

Background: Molecular-based surveys have indicated that Ancylostoma ceylanicum, a zoonotic hookworm, is likely the second most prevalent hookworm species infecting humans in Asia. Most current PCR-based diagnostic options for the detection of Ancylostoma species target the Internal Transcribed Spacer (ITS) regions of the ribosomal gene cluster. These regions possess a considerable degree of conservation among the species of this genus and this conservation can lead to the misidentification of infecting species or require additional labor for accurate species-level determination. We have developed a novel, real-time PCR-based assay for the sensitive and species-specific detection of A. ceylanicum that targets a non-coding, highly repetitive genomic DNA element. Comparative testing of this PCR assay with an assay that targets ITS sequences was conducted on field-collected samples from Argentina and Timor-Leste to provide further evidence of the sensitivity and species-specificity of this assay. Methods/Principal findings: A previously described platform for the design of primers/probe targeting non-coding highly repetitive regions was used for the development of this novel assay. The assay’s limits of detection (sensitivity) and cross-reactivity with other soil-transmitted helminth species (specificity) were assessed with real-time PCR experiments. The assay was successfully used to identify infections caused by A. ceylanicum that were previously only identified to the genus level as Ancylostoma spp. when analyzed using other published primer-probe pairings. Further proof of sensitive, species-specific detection was provided using a published, semi-nested restriction fragment length polymorphism-PCR assay that differentiates between Ancylostoma species. Conclusions/Significance: Due to the close proximity of people and domestic/wild animals in many regions of the world, the potential for zoonotic infections is substantial. Sensitive tools enabling the screening for different soil-transmitted helminth infections are essential to the success of mass deworming efforts and facilitate the appropriate interpretation of data. This study describes a novel, species-specific, real-time PCR-based assay for the detection of A. ceylanicum that will help to address the need for such tools in integrated STH deworming programs. Trial registration: ClinicalTrials.gov [ABSTRACT FROM AUTHOR]

Background. Endemic areas of tegumentary leishmaniasis (TL) in Salta, Argentina, present some overlap zones with the geographical distribution of Chagas disease, with mixed infection cases being often detected. Objectives. The purpose of this study was to determine the magnitude of Leishmania sp. infection and potential associated risk factors, the serologic prevalence of T. cruzi, and the presence of T. cruzi-Leishmania sp. mixed infection in a region of the northwest of Argentina. Methods. Cross-sectional studies were conducted to detect TL prevalence and T. cruzi seroprevalence. A case-control study was conducted to examine leishmaniasis risk factors. Results. Prevalence of TL was 0.17%, seroprevalence of T. cruzi infection was 9.73%, and mixed infection proportion—within the leishmaniasic patients group—was 16.67%. The risk factors associated with TL transmission were sex, age, exposure to bites at work, staying outdoors more than 10 hours/day, bathing in the river, and living with people who had lesions or were infected during the study. Discussion. The endemic pattern of TL seems to involve exposure of patients to vectors in wild as well as peridomestic environment. Cases of T. cruzi infection are apparently due to migration. Therefore, a careful epidemiological surveillance is necessary due to the contraindication of antimonial administration to chagasic patients. [ABSTRACT FROM AUTHOR]

Many infectious diseases arise from co-infections or re-infections with more than one genotype of the same pathogen. These mixed infections could alter host fitness, the severity of symptoms, success in pathogen transmission and the epidemiology of the disease. Trypanosoma cruzi, the etiological agent of Chagas disease, exhibits a high biological variability often correlated with its genetic diversity. Here, we developed an experimental approach in order to evaluate biological interaction between three T. cruzi isolates belonging to different Discrete Typing Units (DTUs TcIII, TcV and TcVI). These isolates were obtained from a restricted geographical area in the Chaco Region. Different mixed infections involving combinations of two isolates (TcIII + TcV, TcIII + TcVI and TcV + TcVI) were studied in a mouse model. The parameters evaluated were number of parasites circulating in peripheral blood, histopathology and genetic characterization of each DTU in different tissues by DNA hybridization probes. We found a predominance of TcVI isolate in blood and tissues respect to TcIII and TcV; and a decrease of the inflammatory response in heart when the damage of mice infected with TcVI and TcIII + TcVI mixture were compared. In addition, simultaneous presence of two isolates in the same tissue was not detected. Our results show that biological interactions between isolates with different biological behaviors lead to changes in their biological properties. The occurrence of interactions among different genotypes of T. cruzi observed in our mouse model suggests that these phenomena could also occur in natural cycles in the Chaco Region. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi calreticulin (TcCRT) is a virulence factor that binds complement C1, thus inhibiting the activation of the classical complement pathway and generating pro-phagocytic signals that increase parasite infectivity. In a previous work, we characterized a clonal cell line lacking one TcCRT allele (TcCRT+/−) and another overexpressing it (TcCRT+), both derived from the attenuated TCC T. cruzi strain. The TcCRT+/− mutant was highly susceptible to killing by the complement machinery and presented a remarkable reduced propagation and differentiation rate both in vitro and in vivo. In this report, we have extended these studies to assess, in a mouse model of disease, the virulence, immunogenicity and safety of the mutant as an experimental vaccine. Balb/c mice were inoculated with TcCRT+/− parasites and followed-up during a 6-month period. Mutant parasites were not detected by sensitive techniques, even after mice immune suppression. Total anti-T. cruzi IgG levels were undetectable in TcCRT+/− inoculated mice and the genetic alteration was stable after long-term infection and it did not revert back to wild type form. Most importantly, immunization with TcCRT+/− parasites induces a highly protective response after challenge with a virulent T. cruzi strain, as evidenced by lower parasite density, mortality, spleen index and tissue inflammatory response. TcCRT+/− clones are restricted in two important properties conferred by TcCRT and indirectly by C1q: their ability to evade the host immune response and their virulence. Therefore, deletion of one copy of the TcCRT gene in the attenuated TCC strain generated a safe and irreversibly gene-deleted live attenuated parasite with high immunoprotective properties. Our results also contribute to endorse the important role of TcCRT as a T. cruzi virulence factor. Author Summary: Trypanosoma cruzi is a protozoan parasite which infects 9 million people in Latin America. Currently there is no vaccine to prevent this disease. Therefore, different approaches or alternatives are urgently needed to identify new protective immunogens. Live vaccines are likely to be most effective in inducing protection; however, safety issues associated with their use have been raised. Hence, we genetically manipulated an attenuated strain of T. cruzi as a safety device to rule out the possibility of reversion to the virulent phenotype. The genetically modified parasites were highly susceptible to killing by the complement machinery and presented a reduced propagation and differentiation rate. We have extended these studies to assess, the virulence, immunogenicity and safety of the mutant as an experimental vaccine. Accordingly, we show that genetically modified parasites present attenuated virulence in mice. The genetic alteration was stable and, after long term infection, it did not revert back to wild type form. Furthermore, after challenge with a virulent T. cruzi strain, mutant immunization induces a highly protective response evidenced by significantly lowered parasite density, mortality, spleen weight index and tissue inflammatory response. Our study provides new insights into the host-pathogen interactions and into the use and evaluation of irreversibly gene-deleted live attenuated parasites to protect against Chagas disease. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi calreticulin (TcCRT) is a virulence factor that binds complement C1, thus inhibiting the activation of the classical complement pathway and generating pro-phagocytic signals that increase parasite infectivity. In a previous work, we characterized a clonal cell line lacking one TcCRT allele (TcCRT+/−) and another overexpressing it (TcCRT+), both derived from the attenuated TCC T. cruzi strain. The TcCRT+/− mutant was highly susceptible to killing by the complement machinery and presented a remarkable reduced propagation and differentiation rate both in vitro and in vivo. In this report, we have extended these studies to assess, in a mouse model of disease, the virulence, immunogenicity and safety of the mutant as an experimental vaccine. Balb/c mice were inoculated with TcCRT+/− parasites and followed-up during a 6-month period. Mutant parasites were not detected by sensitive techniques, even after mice immune suppression. Total anti-T. cruzi IgG levels were undetectable in TcCRT+/− inoculated mice and the genetic alteration was stable after long-term infection and it did not revert back to wild type form. Most importantly, immunization with TcCRT+/− parasites induces a highly protective response after challenge with a virulent T. cruzi strain, as evidenced by lower parasite density, mortality, spleen index and tissue inflammatory response. TcCRT+/− clones are restricted in two important properties conferred by TcCRT and indirectly by C1q: their ability to evade the host immune response and their virulence. Therefore, deletion of one copy of the TcCRT gene in the attenuated TCC strain generated a safe and irreversibly gene-deleted live attenuated parasite with high immunoprotective properties. Our results also contribute to endorse the important role of TcCRT as a T. cruzi virulence factor. [ABSTRACT FROM AUTHOR]

Simple Summary: Scours is the most common disease in dairy calves, and it is a multifactorial syndrome complex. Cryptosporidium sp., rotavirus group A, and bovine coronavirus are the three main pathogens associated with scours. The objective of this study was to identify potential factors associated with scours and these three pathogens in preweaned dairy calves. The results of this study indicated that scours is a prevalent disease in farms of Salta, Argentina, and that rotavirus and Cryptosporidium sp. infections, along with specific farm management practices, might be important contributing factors that could increase the chance of scours in dairy farms. Scours is the most common disease in dairy calves, and it is a multifactorial syndrome complex. Cryptosporidium sp. (C. sp.), rotavirus group A (RVA), and bovine coronavirus (BCoV) are the three main pathogens associated with scours. The objective of this study was to identify potential factors associated with scours, C. sp., RVA, and BCoV infections in preweaned dairy calves from Lerma Valley in Salta Province, Argentina. A total of 488 preweaned calves from 19 dairy farms located in the Lerma Valley were enrolled in this observational study. One fecal sample was collected from each calf between one week and two months of age for assessment of C. sp., RVA, and BCoV infection status. Cryptosporidium sp. oocysts and RVA and BCoV antigens in fecal samples were assessed using microscopic observation and indirect enzyme-linked immune sorbent assay (iELISA), respectively. A voluntary questionnaire was developed and used to collect data regarding management practices from the participants' farms. The data were analyzed using multivariable logistic regression models. Scours incidence was 35.4%, and a greater proportion of calves younger than 20 days were affected. Of the fecal samples, 18% and 9.5% tested were positives for C. sp. and RVA, respectively, while BCoV was detected only in two calves. Furthermore, 84.2% and 63.1% of the farms tested positive for Cryptosporidium sp. and RVA, respectively. In addition, the following variables were associated with higher odds of having scours: (1) herd size (>300 milking cows; OR = 1.7), (2) calf age (<20 days of age; OR = 2.2), (3) RVA and C. sp. test (positive test; RVA OR = 2.6; C. sp. OR = 3), calf feeding practices (feeding milk replacer; OR = 1.81), and newborn calf management practices (calf moved from maternity pen <6 h after calving; OR = 1.7). Concerning RVA infection, calves less than 20 days of age (OR = 2.6) had a higher chance of testing positive for RVA, while calves that remained in the calving pen for less than 6 h after calving had a lower chance (OR = 0.3). On the other hand, for C. sp. infection, large farm size (>300 milking cows; OR = 1.2) and young calf age (<20 days of age; OR = 4.4) indicated a higher chance of testing positive for C. sp., while calves belonging to farms that fed frozen colostrum (OR = 0.2) had a lower chance of becoming infected with C. sp. The result of this study indicated that scours is a prevalent disease in farms of the Lerma Valley, Salta, Argentina, and that RVA and C. sp. infections, along with specific farm management practices, might be important contributing factors that could increase the chance of NCS in dairy farms. [ABSTRACT FROM AUTHOR]

A set of 65 Trypanosoma cruzi stocks from dogs, opossums, insect vectors and humans was isolated in a geographically restricted endemic area for Chagas’ disease in Argentina and was analysed by multilocus enzyme electrophoresis for 15 loci. The results show that at least five multilocus genotypes (clonets) circulate in the study area, one belonging to T. cruzi IIe, one to T. cruzi IId and three clonets belonging to T. cruzi I; and they confirm the presence of these lineages in the country. The three clonets attributed to T. cruzi I were identical to each other for all loci except for Sod-2, where three different patterns were identified. These patterns suggest the presence of two homozygous genotypes and one heterozygous genotype. Our results also suggest association of clonet IIe with dogs, clonet IId with humans and the three T. cruzi I clonets with Didelphis albiventris. On the other hand, there was no significant association between Triatoma infestans and any particular clonet circulating in the area. These findings are consistent with the hypothesis of natural selection, from mixed populations of T. cruzi in vectors, toward more restricted populations in mammals. The epidemiological implications of the possible selection of different clonets by different mammal hosts and the significance of two homozygous genotypes and one heterozygous genotype for the Sod-2 locus are discussed. [Copyright &y& Elsevier]