Assessment of crAssphage as a biological variable for SARS-CoV-2 data normalization in wastewater surveillance.

AIMS: This study aimed to assess the use of cross-assembled phage (crAssphage) as an endogenous control employing a multivariate normalization analysis and its application as a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) data normalizer. METHODS AND RESULTS: A total of 188 twelve-hour composite raw sewage samples were obtained from eight wastewater treatment plants (WWTP) during a 1-year monitoring period. Employing the N1 and N2 target regions, SARS-CoV-2 RNA was detected in 94% (177) and 90% (170) of the samples, respectively, with a global median of 5 log10 genomic copies per liter (GC l-1). CrAssphage was detected in 100% of the samples, ranging from 8.29 to 10.43 log10 GC l-1, with a median of 9.46 ± 0.40 log10 GC l-1, presenting both spatial and temporal variabilities. CONCLUSIONS: Although SARS-CoV-2 data normalization employing crAssphage revealed a correlation with clinical cases occurring during the study period, crAssphage normalization by the flow per capita per day of each WWTP increased this correlation, corroborating the importance of normalizing wastewater surveillance data in disease trend monitoring.

Gut microbiome signature and nasal lavage inflammatory markers in young people with asthma.

Background: Asthma is a complex disease and a severe global public health problem resulting from interactions between genetic background and environmental exposures. It has been suggested that gut microbiota may be related to asthma development; however, such relationships needs further investigation. Objective: This study aimed to characterize the gut microbiota as well as the nasal lavage cytokine profile of asthmatic and nonasthmatic individuals. Methods: Stool and nasal lavage samples were collected from 29 children and adolescents with type 2 asthma and 28 children without asthma in Brazil. Amplicon sequencing of the stool bacterial V4 region of the 16S rRNA gene was performed using Illumina MiSeq. Microbiota analysis was performed by QIIME 2 and PICRUSt2. Type 2 asthma phenotype was characterized by high sputum eosinophil counts and positive skin prick tests for house dust mite, cockroach, and/or cat or dog dander. The nasal immune marker profile was assessed using a customized multiplex panel. Results: Stool microbiota differed significantly between asthmatic and nonasthmatic participants (P = .001). Bacteroides was more abundant in participants with asthma (P < .05), while Prevotella was more abundant in nonasthmatic individuals (P < .05). In people with asthma, the relative abundance of Bacteroides correlated with IL-4 concentration in nasal lavage samples. Inference of microbiota functional capacity identified differential fatty acid biosynthesis in asthmatic compared to nonasthmatic subjects. Conclusion: The stool microbiota differed between asthmatic and nonasthmatic young people in Brazil. Asthma was associated with higher Bacteroides levels, which correlated with nasal IL-4 concentration.

Accuracy of Stool Antigen Test in the Diagnosis of Helicobacter pylori Infection in the Dominican Republic.

Introduction: Helicobacter pylori is a well-studied infectious agent due to its pathogenic potential for peptic ulcers and gastric cancer. It has a high prevalence worldwide and has several diagnostic methods, both invasive and non-invasive. It is important to address the diagnostic efficacy of these tests, as the data vary by location and the specific population in which they are used. Therefore, an effective testing method should be obtained, evaluating the possibility of substantially reducing invasive procedures and, therefore, associated costs. OBJECTIVE: This study proposes to define the diagnostic accuracy of the stool antigen test for H. pylori infection in the Dominican Republic. METHODS: An observational, retrospective, and cross-sectional study was conducted. The results of the stool antigen test for H. pylori infection were compared with the results of the gastric biopsy, as a gold standard test. Patients over 18 years of age with an indication for endoscopy due to suspicion of H. pylori infection, who attended the gastroenterology clinic in 2021, were included in the study. RESULTS: It was shown that the stool antigen test for H. pylori infection has a 61.54% sensitivity and 59.65% specificity. According to the study population, the positive predictive value (PPV) was 67.60% and the negative predictive value (NPV) was 53.13%. CONCLUSION: Low numbers of both sensitivity and specificity were determined, which is why it is pertinent to study alternative non-invasive methods. However, it is important to assess the antibiotic exposure of the study population, since the diagnostic accuracy of the stool test can be influenced by this factor.

Infectious SARS-CoV-2 Particles from Rectal Swab Samples from COVID-19 Patients in Brazil.

The main objective of this study was to investigate the dynamic of SARS-CoV-2 viral excretion in rectal swab (RS), saliva, and nasopharyngeal swab (NS) samples from symptomatic patients and asymptomatic contacts. In addition, in order to evaluate the replication potential of SARS-CoV-2 in the gastrointestinal (GI) tract and the excretion of infectious SARS-CoV-2 from feces, we investigated the presence of subgenomic nucleoprotein gene (N) mRNA (sgN) in RS samples and cytopathic effects in Vero cell culture. A prospective cohort study was performed to collect samples from symptomatic patients and contacts in Rio de Janeiro, Brazil, from May to October 2020. One hundred and seventy-six patients had samples collected at home visits and/or during the follow up, resulting in a total of 1633 RS, saliva, or NS samples. SARS-CoV-2 RNA was detected in 130 (73.9%) patients who had at least one sample that tested positive for SARS-CoV-2. The presence of replicating SARS-CoV-2 in RS samples, measured by the detection of sgN mRNA, was successfully achieved in 19.4% (6/31) of samples, whilst infectious SARS-CoV-2, measured by the generation of cytopathic effects in cell culture, was identified in only one RS sample. Although rare, our results demonstrated the replication capacity of SARS-CoV-2 in the GI tract, and infectious viruses in one RS sample. There is still a gap in the knowledge regarding SARS-CoV-2 fecal-oral transmission. Additional studies are warranted to investigate fecal or wastewater exposure as a risk factor for transmission in human populations.

Diagnostic Accuracy of Fecal Immunochemical Test (FIT) in Bleed-positive and Bleed-negative Colorectal Cancer (CRC) Among a Cohort of 5,090 Subjects who Participated in the Colorectal Neoplasia (CRN) Screening in Brazil.

BACKGROUND/AIM: This study assessed the diagnostic accuracy (DA) of fecal immunochemical test (FIT) ColonView (CV) and guaiac-based fecal occult blood test (HemoccultSENSA) among bleed-positive (history or signs of intestinal bleeding) and bleed-negative participants (no history or signs of intestinal bleeding) (n=5,090) in colorectal neoplasia (CRN) screening in Brazil. PATIENTS AND METHODS: The eligible patients for the study (n=506) collected three consecutive stool samples, to be analyzed by both assays (CV, SENSA). Finally, 421/5090 (8.3%) patients returned both samples, which were subjected to final analysis. Receiver operating characteristic (ROC) analysis with different cut-offs was performed to assess the DA. RESULTS: The area under curve (AUC) values for i) visually analyzed (VA) CV for bleed-positive CRC, ii) automatically analyzed (AA) CV for bleed-positive CRC, iii) VA CV for bleed-negative CRC, and iv) AA CV for bleed-negative CRC as endpoints were as follows: i) AUC=0.864, ii) AUC=0.933, iii) AUC=0.836, and iv) AUC=0.892. In roccomp analysis, the differences in AUC values were: between i) and ii) p=0.068; between i) and iii) p=0.497; between i) and iv) p=0.488; between ii) and iii) p=0.0058; between ii) and iv) p=0.229; and between iii) and iv) p=0.138. CONCLUSION: This is the first investigation where two modes of CV test, VA, and AA, for bleed-positive and bleed-negative CRC patients were used as the endpoint. The AA reading of the CV test showed higher DA in bleed-positive than in bleed-negative CRC patients.

Cost-Effectiveness Analysis of Universal Screening for Biliary Atresia in Japan.

OBJECTIVE: To evaluate the cost-effectiveness of universal newborn screening using stool color card or direct bilirubin (DB) testing when comparing with no screening for biliary atresia in Japanese setting. STUDY DESIGN: A decision analytic Markov microsimulation model was developed to evaluate the universal screening for biliary atresia. Our screening strategies included stool color card, DB, or no screening. The outcomes of all newborns undergoing 3 strategies were simulated to analyze event-free life-years defined as liver transplant-free survival, costs, and incremental cost-effectiveness ratio (ICER) over a 25-year period with an annual discount rate of 2% applied for both costs and outcomes. A 1-way sensitivity analysis was performed to assess the uncertainty. RESULTS: There were 941 000 newborn infants in our cohort and 114 cases of biliary atresia. The base case analysis showed that the stool color card strategy was $14 927 337 higher than no screening with an increase in 44 more event-free life-years gained, resulting in an ICER of $339 258 per event-free life-year gained. The DB screening strategy compared with stool color card was $138 994 060 higher with an increase in 271 more event-free life-years gained and an ICER of $512 893 per event-free life-year gained. The DB screening strategy compared with no screening resulted in an ICER of $488 639 per event-free life-year gained. The DB screening resulted in 16 fewer liver transplants than stool color card and stool color card had 2 fewer liver transplants than no screening. CONCLUSIONS: Universal screening for biliary atresia could be cost-effective depending on the willingness to pay thresholds for health benefits.

Genome of a husavirus from Southern Brazil

ABSTRACT New viruses of the Picornavirales order have been discovered with the increase in the number of sequences obtained by high-throughput sequencing, as well as human stool-associated RNA virus (husavirus [HuV]), found in human stool samples. However, there is much to be clarified about HuV. Its cellular host, evolutionary history, and other biological characteristics are still unknown. Therefore, samples collected from human beings and environmental samples in a watershed in Southern Brazil were processed for the metagenomic library. Upon metagenomic analysis, we identified a HuV (husavirus LMM_67754 OP019707) genome with 8,846 bp, which was reported for the first time in Southern Brazil. The new genome presents only 37% of nucleotide identity with Brazilian strains and more than 90% with genomes from China, Vietnam, Venezuela, and the Netherlands. The HuV phylogeny presents significant differences among genomes, probably because multiple introductions of the virus may have occurred. Many questions still need to be answered about HuV. Therefore, more sequences and studies on this virus are necessary to improve the comprehension of the unknown origin of Picornavirales.

¿Reconocemos adecuadamente la acolia/ hipocolia en los lactantes?

Objetivo: El objetivo de este estudio fue valorar el nivel de acierto del personal de salud en la identificación de hipocolia/acolia en un escenario simulado. Con respecto a esto afirmamos que la hipocolia/acolia son signos clínicos que indican obstrucción biliar, relacionándose con enfermedades colestásicas obstructivas, siendo la atresia de vías biliares una de las principales causas en los primeros meses de vida; en esta, el pronóstico del manejo quirúrgico depende del diagnóstico temprano. Métodos: Estudio descriptivo/ prospectivo basado en la realización de una prueba virtual al personal de salud de Medellín, dividiéndose según sitio de trabajo, edad, sexo y profesión. Fueron utilizadas cinco imágenes de la tabla de colores de materia fecal, utilizada en Taiwán, previa autorización de sus autores. Se consideró reconocimiento adecuado cuando el participante identificaba como anormales las tres imágenes que representaban la acolia/hipocolia. Resultados: Se obtuvo respuesta de 442 participantes (se excluyeron 53) y solo 253 (65 %) lograron reconocimiento adecuado. En el análisis por subgrupos se encontró que hubo un mayor acierto en los participantes del Hospital Pablo Tobón Uribe con respecto a otras instituciones (69 y 54.6 %, respectivamente - p. 0.005). Los participantes con menor porcentaje de acierto fueron los médicos generales en un (45 %). Conclusión: Durante la valoración simulada sobre la identificación adecuada de acolia/hipocolia se encontró que solo el 65 % de la población participante acertó en la identificación correcta de este signo clínico, lo que sugiere una mayor atención en cuanto al entrenamiento para la identificación de acolia/hipocolia.

Objetive: The objective of this study was to assess the level of success of healthcare worker in the identification of hypocholia / acholia in a simulated scenario. With respect to this, we affirm that hypocholia/acholia are clinical signs indicative of biliary obstruction, related to obstructive cholestatic diseases, being bile duct atresia one of the main causes in the first months of life. In this, the prognosis of surgical management depends on the early diagnosis. Methods: Descriptive and prospective study, based on the analysis of a virtual survey to Medellín's healthcare workers, subgroup analysis was done according to workplace, age, sex and profession. Five images were used from the Taiwan Stool Color Card, with the authorization from their authors. Adequate recognition was considered when the participant identified the three images representing acholia / hypocholia as abnormal. Results: a response was obtained from 442 participants (53 were excluded), and only 253 (65 %) achieved adequate recognition. In the analysis by subgroups, it was found that there was a greater success in the Pablo Tobón Uribe Hospital participants with respect to other institutions (69 % and 54.6 %, respectively - p. 0.005). The participants with the lowest percentage of correct answers were general practitioners (45%). Conclusion: During the simulated assessment on the adequate identification of acholia / hypocolia, it was found that only 65% of the participating population was correct in the right identification of this clinical sign, which suggests greater attention in training for the identification of acholia / hypocolia.

Evaluation of SmITS1-LAMP performance to diagnosis schistosomiasis in human stool samples from an endemic area in Brazil.

Schistosomiasis is a life-threatening infectious disease categorized by the World Health Organization as a public health issue. New molecular diagnostic alternatives for intestinal schistosomiasis caused by Schistosoma mansoni, such as the loop-mediated isothermal amplification (LAMP), a fast and simple amplification technique, have been proposed for control of this NTD in low-endemicity locations. A LAMP assay was performed to detect the internal transcribed spacer 1 ribosomal gene of S. mansoni (SmITS1-LAMP) in 322 DNA extracted from stool samples from schistosomiasis endemic area in Brazil. Kato-Katz analysis of human stool samples was used as the gold standard test, detecting 144 positive samples. SmITS1-LAMP detection limit achieved a maximum analytical sensitivity of 10 fg/µL using S. mansoni genomic DNA, subsequently detecting 17/144 (11.8%) positive samples. SmITS1-LAMP sensitivity and specificity were 12% (95%CI: 7%-18%) and 93% (95%CI: 89%-96%), respectively. Positive predictive value (PPV) and negative predictive value (NPV) were 59% (95%CI: 39% - 76%); and 57% (95%CI: 51% - 62%), respectively. Most cases involved men (61.8%), predominantly young adults (20-39 years old) in cases diagnosed by Kato-Katz and adults (40-59 years old) in cases diagnosed by LAMP. The low number of eggs per gram of stool (1-99 EPG) was the most frequently identified by both Kato-Katz and LAMP. Further studies are needed to evaluate the applicability of SmMIT-LAMP on Schistosoma mansoni diagnosis and surveillance of schistosome infections.

Neonatal Antibiotic Treatment Can Affect Stool Pattern and Oral Tolerance in Preterm Infants.

Preterm neonates are at high risk of infectious and inflammatory diseases which require antibiotic treatment. Antibiotics influence neonatal gut microbiome development, and intestinal dysbiosis has been associated with delayed gastrointestinal transit. Neonates who take less time to pass meconium have a better tolerance to enteral feeding. We analyzed the effect of neonatal antibiotic treatment on the stool pattern and oral tolerance in 106 preterm infants < 33 weeks gestational age. Neonates were classified in 3 groups according to neonatal antibiotic (ABT) treatment days: no antibiotics, 3−7 d ABT, and ≥8 d ABT. Preterm infants from the ≥8 d ABT group took longer to pass meconium and to start green and yellow stools, took longer to reach 100 and 150 mL/kg/day, and reached reduced volumes in enteral feeds at day of life 14 and 28 than infants from no ABT and 3−7 d ABT groups. Multiple linear regression models showed that neonatal antibiotic treatment, birth weight, invasive mechanical ventilation, surfactant, enteral feeding start day, neonatal parenteral nutrition, and neonatal fasting days are associated with the stool pattern and oral tolerance in preterm infants.

Only Low Effects of Water Filters on the Enteric Carriage of Gastrointestinal Pathogen DNA in Colombian Indigenous People.

Water filtration is a common strategy of water sanitation in resource-poor tropical settings. Here, we assessed the intermediate term effect of this preventive procedure including specific filter-related as well as general hygiene training on the molecular detection of enteric pathogens in stool samples from Colombian Indigenous people. From a total of 89 individuals from an Indigenous tribe called Wiwa, stool samples were assessed by real-time PCR for enteropathogenic microorganisms prior to the implementation of water filtration-based infection prevention. Three years after the onset of the preventive strategy, a follow-up assessment was performed. A significantly beneficial effect of water filtration could be shown for Ascaris spp. only (p = 0.035) and a tendency (p = 0.059) for Hymenolepis nana. No hints for effects on the gastrointestinal shedding of Giardia duodenalis, Entamoeba histolytica, Cryptosporidium spp., Campylobacter spp., Shigella spp./enteroinvasive Escherichia coli, Necator americanus, Strongyloides stercoralis, Trichuris trichiura, and Taenia spp. were seen. In conclusion, the study indicates that water filtration can only be an element of a multi-modal hygiene concept to reduce enteric pathogen carriage in inhabitants of resource-poor tropical settings in spite of tendencies of beneficial effects.

Effects of Bifidobacterium animalis subspecies lactis supplementation on gastrointestinal symptoms: systematic review with meta-analysis.

CONTEXT: The effects of probiotics on gastrointestinal (GI) symptoms have been increasingly investigated, particularly that of Bifidobacterium animalis. Clinical trials so far have shown differing evidence regarding these effects in healthy adults. OBJECTIVE: To synthesize the published evidence on the effects of B. animalis subspecies lactis on GI symptoms (GIS) in healthy adults. DATA SOURCE: A search of the Medline, Embase, Lilacs, Scopus, Web of Science, ProQuest, and Google Scholar databases was conducted for reports on randomized controlled trials published up to October 2021. DATA EXTRACTION: Population characteristics and data on colonic transit time (CTT), stool consistency, defecation frequency, abdominal pain, bloating, flatulence, volunteer compliance, and adverse events were extracted. A random-effects model was used to estimate the effect of probiotic treatment on these variables. DATA SYNTHESIS: In total, 1551 studies were identified, of which 14 were included in the qualitative synthesis and 13 in the meta-analysis. Overall, probiotic supplementation increased defecation frequency (standardized mean difference [SMD], 0.26; 95%CI, 0.13-0.39). Subgroup analysis revealed a decrease in CTT (SMD, -0.34; 95%CI, -0.62 to -0.07) in short-term treatment (≤14 d) and an improvement in stool consistency (SMD, 0.76; 95%CI, 0.44-1.08) in individuals without GIS. No improvement in abdominal pain and bloating was found. CONCLUSIONS: B. animalis subspecies lactis supplementation may increase defecation frequency and, in short-term treatment, may reduce CTT in healthy adults and improve stool consistency in individuals without GIS. More high-quality randomized controlled trials are needed to develop a clinical protocol for the use of this strain to improve these symptoms. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration no. CRD42020154060.

Escalas validadas para avaliação das fezes: revisão integrativa / Validated scales for stool assessment: integrative review

Introdução: A eliminação intestinal é uma necessidade fisiológica humana básica, que deve ser considerada e avaliada em toda a assistência prestada pelos profissionais de saúde. Objetivo: identificar as escalas validadas utilizadas na atualidade para avaliação das fezes em adultos e idosos. Método: revisão integrativa, protocolada na plataforma Open Science Framework, cujo registro está disponível sob o DOI https://doi.org/10.17605/OSF.IO/9CTMB, que incluiu estudos primários publicados na íntegra, que utilizaram escalas para avaliação de fezes, publicados nos idiomas inglês, português ou espanhol, no período de janeiro de 2001 a julho de 2021. As buscas foram realizadas nas bases de dados eletrônicas PubMed, LILACS, CINAHL e EMBASE em 16 de julho de 2021. Em seguida, as referências identificadas foram exportadas para o aplicativo Rayyan para remoção das duplicatas, seleção e avaliação dos estudos por dois revisores de forma independente e mascarada; os casos de divergência foram avaliados por um terceiro revisor. As evidências foram sintetizadas de forma descritiva. Resultados: a busca nas bases de dados resultou em 1.567 estudos. Após o processo de seleção e leitura na íntegra, 353 estudos foram considerados nesta revisão, dos quais 339 utilizaram escalas validadas (333 estudos utilizaram a Escala de Bristol e seis outras escalas) e 14 escalas não validadas. Também se verificou aumento na quantidade de publicações que avaliaram a consistência das fezes a partir do ano de 2015. Após a leitura dos 353 estudos, foram identificadas cinco escalas validadas: Escala de Forma das Fezes de Bristol, King's Stool Chart, Escala de consistência das fezes segundo Anastasi e Capili (2001) e Hansen (1981), Escala pictórica validada do Diarrhea Questionnaire e a ferramenta de avaliação fecal de Ohno. Conclusão: foram identificadas cinco escalas validadas e utilizadas nos últimos 20 anos para avaliação das fezes. Adicionalmente, os dados mostraram aumento mundial do uso das escalas e a necessidade de desenvolvimento de uma escala para avaliação das fezes com estudos de validação mais robustos

Introduction: Bowel elimination is a basic human physiological need, which must be considered and evaluated in all care provided by health professionals. Aim: to identify the validated scales used to assess stool in adults and the elderly. Method: an integrative review, registered on the Open Science Framework platform and available under DOI 10.17605/OSF.IO/9CTMB, which included primary studies published in full that used scales for stool assessment, published in English, Portuguese or Spanish, between January 2001 and July 2021. Searches of the PubMed, LILACS, CINAHL, and EMBASE electronic databases were performed on July 16, 2021. Afterward, the identified references were exported to the Rayyan application for removal of duplicates, selection, and independent and masked evaluation of studies by two reviewers; cases of divergence were evaluated by a third reviewer. The evidence was synthesized descriptively. Results: the database search resulted in 1,567 studies. After the selection process and reading in full, 353 studies were considered in this review. Of these 353, 339 used validated scales (333 studies used the Bristol Scale and six studies used other scales) and 14 studies used non-validated scales. There was also an increase in the number of publications that evaluated stool consistency from the year 2015. After reading the 353 studies, five validated scales were identified: Bristol Stool Form Scale, King's Stool Chart, Stool Consistency Scale according to Anastasi and Capili (2001) and Hansen (1981), the Diarrhea Questionnaire's validated pictorial scale, and Ohno's fecal assessment tool. Conclusion: Five validated scales were identified and used in the last 20 years for stool assessment. Additionally, the data showed a worldwide increase in the use of scales and the need to develop a scale to assess stool with more robust validation studies

The Effect of Infant Formulas With 4 or 8 g/L GOS/FOS on Growth, Gastrointestinal Symptoms, and Behavioral Patterns: A Prospective Cohort Study.

There is a lack of studies investigating the clinical benefits of prebiotic-supplemented infant formula. In this study, healthy infants that started, on medical recommendation, artificial feeding with one of 2 infant formulas containing 4 g/L of GOS/FOS (9:1) (IF4 group; n = 60) or 8 g/L of GOS/FOS (9:1) (IF8 group; n = 60) were followed for 30 days to the evaluation of growth and gastrointestinal symptoms. Exclusively breastfed infants (EBF; n = 60) were followed up as a reference. Both infant formulas supported adequate weight gain, however, IF4 formula promotes growth more similar to breastfed infants. There was no additional benefit of a dose higher than 4 g/L of GOS/FOS on gastrointestinal symptoms.

Methods for detection of Helicobacter pylori from stool sample: current options and developments.

Accurate detection of Helicobacter pylori infection and determination of antibiotics have significant meaning in clinical practice. The detection methods can be categorized into two types, invasive and non-invasive, but nowadays we use the urease breath test most frequently which is non-invasive. However, many developing countries cannot meet the requirements for having specialized equipment and they lack trained personnel. Also, for the children, it is difficult to make them cooperate for the test. Methods that detect Helicobacter pylori from stool sample can be a promising alternative for detection used in children and mass screening. Stool antigen tests have several advantages such as rapidity, simplicity, and cheapness, though their results may be influenced by the heterogenicity of antigens, the nature of biochemical techniques, and the amount of antigen presented in the stool. PCR-based methods can specifically detect Helicobacter pylori infection and antibiotic resistance by targeting specific gene sequence, but they also are limited by the requirements of facilities and experts, the existence of inhibitory substance, and interference from the dead bacteria. Some novel methods also deserve our attention. Here we summarized the results of researches about methods using stool sample and we hope our work can help clinicians choose the appropriate test in clinical practice.

Geographic differences in the distribution of parasitic infections in children of Bolivia.

BACKGROUND: A high percentage of the population in Latin America lives with intestinal parasitic infections, a neglected tropical disease frequently not treated. Intestinal parasitism is associated with other disorders, but information about the epidemiological situation in countries like Bolivia is scarce. Environmental conditions play a role in the prevalence of certain parasites. The main objective was to know the current situation of parasitic infections among children under 12 years old from different geographical areas of Cochabamba - Bolivia. METHODS: We analysed the laboratory reports of four second-line hospitals in different areas and the Tertiary Care Hospital. Results of stool examinations performed between 2011 and 2015 in children under 12 years of age were collected. RESULTS: We gathered the results of 23,221 examinations. The 89% of children were less than five years old. Pathogenic parasites were found in 31%. Entamoeba histolytica and Giardia lamblia were the two most prevalent parasites in all areas. Helminths were 19% of positive samples and Ascaris lumbricoides was the most prevalent. Parasitic infections are more frequent in tropical areas where helminths are highly concentrated. Pre-school age children (OR: 5.296; 95% CI: 4.81-5.83) and semi-tropical area (OR: 3.26; 95% CI: 2.90-3.66) were strongly associated to the presence of pathogenic parasites. CONCLUSIONS: Parasitic infections in children are still very prevalent in Bolivia. Protozoan infections are a major problem, while the prevalence of helminths seems to be decreasing. The most vulnerable population is still concentrated in semi-tropical and tropical areas, where the risk of parasitic infection is probably increased due to poor environmental conditions. Our results could allow reconsideration of more effective parasitic disease control policies, taking into account regional characteristics.

Plasma and stool metabolomics to identify microbiota derived-biomarkers of metabolic dysfunction-associated fatty liver disease: effect of PNPLA3 genotype.

INTRODUCTION: Non-invasive biomarkers are needed for metabolic dysfunction-associated fatty liver disease (MAFLD), especially for patients at risk of disease progression in high-prevalence areas. The microbiota and its metabolites represent a niche for MAFLD biomarker discovery. However, studies are not reproducible as the microbiota is variable. OBJECTIVES: We aimed to identify microbiota-derived metabolomic biomarkers that may contribute to the higher MAFLD prevalence and different disease severity in Latin America, where data is scarce. METHODS: We compared the plasma and stool metabolomes, gene patatin-like phospholipase domain-containing 3 (PNPLA3) rs738409 single nucleotide polymorphism (SNP), diet, demographic and clinical data of 33 patients (12 simple steatosis and 21 steatohepatitis) and 19 healthy volunteers (HV). The potential predictive utility of the identified biomarkers for MAFLD diagnosis and progression was evaluated by logistic regression modelling and ROC curves. RESULTS: Twenty-four (22 in plasma and 2 in stool) out of 424 metabolites differed among groups. Plasma triglyceride (TG) levels were higher among MAFLD patients, whereas plasma phosphatidylcholine (PC) and lysoPC levels were lower among HV. The PNPLA3 risk genotype was related to higher plasma levels of eicosenoic acid or fatty acid 20:1 (FA(20:1)). Body mass index and plasma levels of PCaaC24:0, FA(20:1) and TG (16:1_34:1) showed the best AUROC for MAFLD diagnosis, whereas steatosis and steatohepatitis could be discriminated with plasma levels of PCaaC24:0 and PCaeC40:1. CONCLUSION: This study identified for the first time MAFLD potential non-invasive biomarkers in a Latin American population. The association of PNPLA3 genotype with FA(20:1) suggests a novel metabolic pathway influencing MAFLD pathogenesis.

Stool microbiome dataset of the critically endangered Puerto Rican parrot (Amazona vittata).

The Puerto Rican parrot (Amazona vittata), endemic to Puerto Rico, is the only native parrot in the United States and is classified as a critically endangered species. There are two captive populations of A. vittata in Puerto Rico located in the Iguaca Aviary in El Yunque National Rainforest and the José L. Vivaldi Aviary in the Río Abajo Forest. To characterize the microbial communities of A. vittata's stool, 21 stool samples from captive birds were collected, DNA extracted and sequenced using Illumina MiSeq. Sequences were processed by removing host sequences (A. vittata genome) and low-quality reads. Taxonomic and functional profiles were generated using MG-RAST. The most abundant domain was Bacteria (96%), followed by Virus (3%), and Eukaryota (0.6%). Among the functions in the microbiome, the most abundant was related to carbohydrates (14%), followed by clustering-based subsystems (12%), protein metabolism (8%), and amino acids and derivatives (7%). This dataset describes the stool microbiome of A. vittata using a metagenomics approach. Data can be used to develop holistic conservation strategies for A. vittata and other endangered birds, as well as to search for bioprospects with potential biomedical and biotechnological applications.

Critical review on microplastics in fecal matter: Research progress, analytical methods and future outlook.

There has been an ever-increasing attention over years for investigating microplastics in feces of lower to higher trophic organisms from diverse environments. Focusing on the standardization of methodologies for reliable generation and comparison of data is one of the important aspects in microplastic area. This first review, comprising 20 studies in total, critically summarizes and compares the methodological approaches for the determination of microplastics in feces as well outlines the levels and characteristics of microplastics detected in feces worldwide. Contaminations and QA/QC measures are also discussed. Despite variations among the approaches, most studies (n = 12) described herein rely on the digestion processes involving H2O2 (n = 7) and KOH (n = 6) for the separation of microplastics, whereas very few included wet sieving (n = 5), density separation using NaCl (n = 3) and NaI (n = 1) and enzymatic digestion (n = 2). Microscopical sorting and spectroscopic methods such as infrared and Raman were combined for identification and characterization of microplastics. The detected microplastics varied by size, shape, color and polymer types and the differences in reporting units of microplastic abundance make comparison across studies difficult. Taking advantage of the current knowledge, our review identified analytical challenges and suggested appropriate methods on research into microplastic contamination in feces. This work will serve as a valuable information of available analytical methods for examining microplastics in feces and will stimulate further research to advance our understanding of microplastics from feces.