Implementation of a rapid diagnostic assay package for cryptococcosis, histoplasmosis and tuberculosis in people living with HIV in Paraguay.

BACKGROUND: Opportunistic infections (OIs) are common causes of mortality among people living with HIV (PLHIV). We determined prevalence and 30-day mortality due to histoplasmosis, cryptococcosis, and TB in PLHIV with advanced HIV disease (AHD). METHODS: PLHIV 18 years and older, with a CD4 + T-cell count of less than 350 cells/mm3 newly diagnosed with HIV infection or re-engaged in care after being without ART for more than 90 days (Group A). The second group included symptomatic PLHIV regardless of ART status or CD4 + T-cell count (Group B); all followed for 30 days. Detection of Histoplasma Ag (HisAg) in urine was done by enzyme immunoassay (EIA), Cryptococcus antigen (CrAg) was detected in serum and cerebrospinal fluid (CSF) specimens by lateral flow assay (LFA), and lipoarabinomannan (LAM) detection in urine was by LFA (TB LAM) and in sputum by GeneXpert for diagnosis of Mycobacterium infections. RESULTS: From August 2021 to June 2022, 491 PLHIV were enrolled; 482 (98%) had a CD4 + T-cell result, and 381 patients (79%) were classified with AHD according to CD4 + T-cell count (< 200 CD4/mm3). Frequency of an OI was 38% (n = 145/381). Antigen test positivity rate was 16% (72/467) for TB-LAM, 9% (43/464) for HisAg, and 11% (51/484) for CrAg. Twenty-one of 34 (62%) patients receiving CSF CrAg tests were positive, confirming meningitis. Significant differences in 30-day mortality were observed in patients with an OI (16%) vs. no OI (7%) (p = 0.002). Mortality was highest in patients with histoplasmosis (25%), co-infection (22%), cryptococcosis (18% overall; 19% for cryptococcal meningitis), and TB (10%). CONCLUSIONS: TB and fungal OIs, including co-infection, were common in PLHIV in Paraguay and had high associated mortality. Laboratories and health facilities need access to CD4 + T-cell testing and rapid diagnostic assays.

Mortalidad intrahospitalaria de pacientes con COVID-19 complicados con neumonías bacterianas en asistencia respiratoria mecánica en Cuidados Intensivos de Adultos en un Hospital del Paraguay / In-hospital mortality of patients with COVID-19 complicated by bacterial pneumonia on mechanical ventilation in Adult Intensive Care in a Hospital in Paraguay

Introducción: los pacientes con COVID-19 ingresan en mayor proporción a asistencia respiratoria mecánica, aumentando: el riesgo de neumonía asociada a ventilador (NAV) las tasas de mortalidad, los días de permanencia en las unidades de terapia intensiva (UCI) y los costos sanitarios. Objetivo: determinar la Mortalidad intrahospitalaria de pacientes con COVID-19 complicados con neumonías bacterianas en asistencia respiratoria mecánica en Cuidados Intensivos de Adultos en un Hospital del Paraguay durante los años 2020 a 2021. Metodología: estudio analítico de tipo cohorte retrospectiva. Se registraron variables demográficas, comorbilidades, puntajes en scores de gravedad como el APACHE II al ingreso, la cifra más baja de oxigenación durante la internación expresado por la PaO2 / FIO2, días de ventilación, colocación en decúbito prono, traqueotomía, medidas terapéuticas farmacológicas y no farmacológicas, días de internación, así como las complicaciones y la mortalidad. Resultados: fueron incluidos 214 pacientes, 135 ingresaron a asistencia respiratoria mecánica (ARM) de los cuales 58 (42,9 %) desarrollaron NAV, con edad mediana de 52 años (40-60). Los microorganismos de NAV fueron cocos Gram negativos en 98,3 %, incluyendo Acinetobacter baumanii en 46,5 %, Klebsiella pneumoniae en 22,8 %, Pseudomona aeruginosa en 15,5 % y 5,2 % Stenotrophomona maltofilia. La mortalidad intrahospitalaria fue del 44,8 %. Los menores de 50 años tienen una sobrevida mayor que los mayores (34 días vs 22 días, con p de 0,026). Conclusión: la mortalidad intrahospitalaria fue del 44,8 %. La edad fue un factor de riesgo independiente para la mortalidad en pacientes con NAV, por lo que los profesionales de la salud deben estar atentos a la posibilidad de NAV en pacientes que requieren asistencia respiratoria mecánica, especialmente en pacientes mayores de 50 años.

Introduction: patients with COVID-19 are more likely to require mechanical ventilation, which increases the risk of ventilator-associated pneumonia (VAP), mortality rates, length of stay in intensive care units (ICUs), and healthcare costs. Objective: to determine the in-hospital mortality of patients with COVID-19 complicated by bacterial pneumonia on mechanical ventilation in Adult Intensive Care in a Hospital in Paraguay during the years 2020 to 2021. Methodology: this is a retrospective cohort analytical study. Demographic variables, comorbidities, severity scores such as APACHE II on admission, the worst oxygenation during hospitalization expressed by PaO2/FiO2, days of ventilation, prone position, tracheostomy, pharmacological and non-pharmacological therapeutic measures, days of hospitalization, as well as complications and mortality were recorded. Results: a total of 214 patients were included, 135 were admitted to mechanical ventilation (MRA), of which 58 (42.9%) developed VAP, with a median age of 52 years (40-60). VAP microorganisms were Gram-negative cocci in 98.3%, including Acinetobacter baumanii in 46.5%, Klebsiella pneumoniae in 22.8%, Pseudomona aeruginosa in 15.5%, and Stenotrophomona maltophilia in 5.2%. In-hospital mortality was 44.8%. Those under 50 years of age have a longer survival than those older (34 days vs. 22 days, with p of 0.026). Conclusion: the overall mortality rate was 44.8%. Age was an independent risk factor for mortality in patients with VAP, so healthcare professionals should be aware of the possibility of VAP in patients who require mechanical ventilation, especially in patients over 50 years of age.

Perfil de transmisión del SARS-COV-2 en una región sanitaria del Paraguay

Introducción: La transmisión viral a menudo se propaga en grupos, donde las infecciones se pueden rastrear hasta un caso índice o una ubicación geográfica, para así poder tomar medidas de prevención al respecto. Entender el perfil de transmisión del SARS-CoV-2 es esencial para desarrollar estrategias efectivas de prevención y control de la enfermedad. Objetivo: Determinar el perfil de transmisión del Sars-Cov-2, a partir de clúster con casos índices identificados, en la V región sanitaria (Caaguazu) del Paraguay, entre julio y octubre del 2020. Metodología: Se realizó un estudio observacional, descriptivo, de corte transversal, con fichas de notificación de casos de COVID-19 y resultados laboratoriales de la RT_PCR en pacientes diagnosticados con COVID-19 en la V Región Sanitaria entrejulio y octubre del 2020. Resultados: Fueron identificadas 703 personas con Sars-Cov-2 positivo. El 55,49 % de las personas no reconoció algún nexo de contagio. Con respecto al tipo de evento donde ocurrían con mayor frecuencia los contagios, ocurrieron en Eventos Sociales en 58,14 % de los casos y en el Ambiente Familiar en el 33,89 %. La transmisión secundaria se observó en el 17,40 % de los casos. Fueron identificados 58 Clústers, con una mediana de tamaño de los mismos de 3 (RIQ 2-4), y 267 personas (37,98 %) asociadas a los mismos. Discusión: La mayoría de las personas no reconocieron su nexo de contagio, sin embargo, en los que, si conocieron, se puede observar que la participación en eventos sociales fue el principal nexo, por lo que es fundamental realizar este tipo de ejercicios para hacer el seguimiento oportuno de los casos.

Introduction: Viral transmission often spreads in clusters, where infections can be traced to an index case or a geographic location, in order to take preventive measures in this regard, understanding the transmission profile of SARS-CoV-2 is essential. to develop effective disease prevention and control strategies. Objective: to determine the transmission profile of Sars-Cov-2, from clusters with identified index cases, in the V health region (Caaguazú) of Paraguay, between July and October 2020. Methodology: We carried out an observational, descriptive, cross-sectional study, with notification sheets of COVID-19 cases and laboratory results of the RT_PCR in patients diagnosed with COVID-19 in the V Sanitary Region between July and October 2020. Results: We identified 703 people with positive Sars-Cov-2. 55.49% of people did not recognize any contagion link. Regarding the type of event where infections occurred most frequently, was in Social Events in 58.14% of the cases and the Family Environment in 33.89%, secondary transmission was observed in 17.40% of the cases. 58 Clusters were identified, with a median size of 3 (IQR 2-4), and 267 people (37.98%) associated with them. Discussion: Most of the people did not recognize their contagion link, however, in those who did know, the main link was because they participated in social events, so it is essential to carry out this type of exercise to do timely follow-up. of the cases.

Intergenerational sex and early sexual debut are associated with HIV infection among transgender women in Paraguay.

INTRODUCTION: HIV prevalence among transgender women is high worldwide. The objectives of the present study were to estimate the current prevalence of HIV and identify factors associated with high HIV burden among transgender women in Paraguay. METHODS: Transgender women aged ≥15 years in four regions of Paraguay were recruited by Starfish sampling between February and March 2021. RESULTS: In total, 322 transgender women were included. Mean age was 31 years (range 15-67), and 102 had positive HIV test results (31.7%, 95% confidence interval [CI] 26.6-37.1). In multivariable analysis, factors associated with HIV infection were age at first intercourse ≤17 years (adjusted odds ratio [aOR] 5.47; 95% CI 1.05-28.42), >10 years difference in age with the last sexual partner (aOR 1.60; 95% CI 1.04-2.46), substance use (mostly cocaine) (aOR 3.00; 95% CI 1.47-6.12), higher risk perception (aOR 3.08; 95% CI 1.53-6.17), not testing for HIV (aOR 1.23; 95% CI 1.09-1.39), and accessed by a peer educator (aOR 3.86; 95% CI 1.77-8.38). CONCLUSIONS: Sexual debut as a minor and a large age difference with sexual partners are associated with high burden of HIV among transgender women in Paraguay. Our study corroborates the finding of cocaine use during sex as a risk factor for HIV. Prevention programmes must address structural and social vulnerabilities to stem the tragically high burden of HIV among transgender women.

LC-QToF-Based Metabolomics Identifies Aberrant Tissue Metabolites Associated with a Higher-Fat Diet and Their 'Reversion to Healthy' with Dietary Probiotic Supplementation.

Over 33% of Americans are labeled as obese, leading the World Health Organization to designate obesity as a major public health problem. One consequence of obesity is the development of metabolic syndrome, a condition which has been correlated to an increased risk for developing cardiovascular disease and Type 2 diabetes. Prolonged ingestion of a higher-fat diet, one cause of obesity, results in alterations to the gut microbiome. These alterations are implicated to have a profound role in the evolution and progression of obesity-linked diseases. Probiotics are associated with positive health effects such as limiting pathogen colonization, aiding in digestion, and vitamin synthesis. Using Ossabaw pigs as a model for obesity, and in conjunction with our previous research, we performed an in-depth, nontargeted, metabolomic analysis on select organs to elucidate the effects of dietary supplementation with the probiotic Lacticaseibacillus paracasei. We focused our analysis on the effects of probiotic supplementation on a higher-fat (obesogenic) diet and a nutritionally balanced diet. Notably, our findings reveal that the brain cortex is highly sensitive to dietary influencers, and with probiotic supplementation, several aberrant metabolites associated with a higher-fat diet revert to healthy levels, thus demonstrating the potential for a probiotic intervention for obesity-linked disease.

Vigilancia de la resistencia del Mycobacterium tuberculosis a los fármacos en Paraguay, 2014 a 2017 / Surveillance of drug resistance of Mycobacterium tuberculosis in Paraguay, 2014 to 2017

El objetivo del estudio fue identificar la resistencia del Mycobacterium tuberculosis a los fármacos en Paraguay, 2014 a 2017. Se realizó un estudio observacional retrospectivo. Se utilizaron los datos del Programa Nacional de Tuberculosis del Paraguay comprendidos entre los años 2014 a 2017. Se incluyeron todos los pacientes con diagnóstico de Tuberculosis que se realizaron un test de resistencia. Se extrajeron los datos en Excel y fueron analizados con Stata 17.0. Se incluyeron 3429 pacientes con tuberculosis que contaban con resultado de al menos una prueba de sensibilidad. La resistencia se encontró en 2.1% de los pacientes. La resistencia a la Rifampicina estuvo presente en el 0.3% de los casos mientras que a la Izionazida en el 0.6% de los casos. La prevalencia de resistencia fue más alta en hombres 3.4 (IC 95% 2.2 - 4.8) p=0.003, que residían en el chaco 6.0 (IC 95% 3.4 - 9.7) p=0.000, previamente tratados 2.7 (IC 95% 1.1 - 5.1) p=0.010. En el modelo se pudo observar que un paciente previamente tratado tiene mayores posibilidades de tener resistencia OR 2.62 (IC 95% 1.1 - 6.24). La prevalencia de resistencia del Mycobacterium tuberculosis a fármacos estuvo relacionada con haber sido previamente tratado

The objective of the study was to identify the resistance of Mycobacterium tuberculosis to drugs in Paraguay, 2014 to 2017. A retrospective observational study was carried out. The data from the National Tuberculosis Program of Paraguay between the years 2014 to 2017 were used. All patients with a diagnosis of Tuberculosis who underwent a resistance test were included. Data were extracted in Excel and analyzed with Stata 17.0. 3429 tuberculosis patients who had a result of at least one sensitivity test were included. Resistance was found in 2.1% of patients. Resistance to Rifampicin was present in 0.3% of cases while to Izionazide in 0.6% of cases. The prevalence of resistance was higher in men 3.4 (95% CI 2.2 - 4.8) p = 0.003, who resided in the Chaco 6.0 (95% CI 3.4 - 9.7) p = 0.000, previously treated 2.7 (95% CI 1.1 - 5.1) p = 0.010. In the model, it was observed that a previously treated patient has a greater chance of having resistance OR 2.62 (95% CI 1.1 - 6.24). The prevalence of resistance of Mycobacterium tuberculosis to drugs was related to having been previously treated

Deficiencias de cobre, hierro y zinc en niños menores de 5 años

Introducción: La determinación de niveles séricos de micronutrientes de los niños menores de 5 años, es un requerimiento básico para establecer la prevalencia de deficiencias nutricionales de la población infantil y es una estrategia útil para incentivar a Salud Pública para que tome medidas en cuanto a aplicación y/o modificación de las Políticas Alimentarias Nacionales. Objetivo: Determinar los niveles séricos de cobre, hierro y zinc en niños paraguayos menores de 5 años. Materiales y Métodos: Estudio observacional, descriptivo y de corte transversal. Se procesaron las muestras de sangre de 1.441 niños menores de 5 años de edad que provenían de los Departamentos de Alto Paraná, Caaguazú, Central y de la capital de Paraguay. Se realizaron hemogramas y se determinaron las concentraciones séricas de albúmina, cobre, ferritina y zinc, de todas las muestras. Resultados: Los niños tuvieron una edad promedio de 3 años y 3 meses, fueron 52,74% de sexo masculino y 47,26% de sexo femenino. La prevalencia de anemia fue de 45,25% y la prevalencia de hipoalbuminemia fue de 20,65%. Se detectaron deficiencias de cobre en 14,03%, de ferritina en 6,98% y de zinc en 43,62%, de los niños. Conclusión: Los resultados de este estudio avalan la necesidad de ampliar la cobertura del Programa Alimentario Nutricional Integral (PANI), de plantearse si se deben ajustar las cantidades de minerales y vitaminas de la fórmula nutricional del Programa y de mejorar el seguimiento de los controles prenatales de las madres.

Introduction: The determination of serum levels of micronutrients in children under 5 years of age is a basic requirement to establish the prevalence of nutritional deficiencies in the child population and is a useful strategy to encourage Public Health agencies to take steps regarding application. and/or modification of National Food Policies. Objective: To determine the serum levels of copper, iron and zinc in Paraguayan children under 5 years of age. Materials and Methods: This was an observational, descriptive and cross-sectional study. Blood samples from 1,441 children under 5 years of age who came from the Departments of Alto Paraná, Caaguazú, Central and the capital of Paraguay were processed. Complete blood counts were performed and the serum concentrations of albumin, copper, ferritin and zinc were determined in all samples. Results: The children in this study had an average age of 3 years and 3 months, they were 52.74% male and 47.26% female. The prevalence of anemia was 45.25% and the prevalence of hypoalbuminemia was 20.65%. Copper deficiencies were detected in 14.03%, ferritin in 6.98% and zinc in 43.62% of the children. Conclusion: The results of this study support the need to expand the coverage of the Comprehensive Nutritional Food Program (PANI program), to consider whether the amounts of minerals and vitamins in the Program's nutritional formula should be adjusted, and to improve prenatal care.

Correction: Solano-Aguilar et al. Fruit and Vegetable Supplemented Diet Modulates the Pig Transcriptome and Microbiome after a Two-Week Feeding Intervention. Nutrients 2021, 13, 4350.

There was an error in the original publication [...].

HIV prevalence and associated risk factors among men who have sex with men in three regions of Paraguay, 2020.

BACKGROUND: Our study aimed to measure HIV prevalence and associated risk factors among men who have sex with men (MSM) in three regions of Paraguay in 2020. METHODS: MSM were recruited for cross-sectional surveys in three regions of Paraguay using respondent-driven sampling. Interview were conducted face-to-face to collect demographic characteristics and risk and preventive behaviors. The analysis assessed HIV prevalence and associated risk factors in the three samples of MSM within each region. RESULTS: A total of 1,207 MSM were recruited, including 559 in Asunción-Central, 245 in Alto Paraná, and 403 in Caaguazú. HIV prevalence was 24.2% (95% CI 20.6-27.9) in Asunción-Central, 10.2% (95% CI 6.7-14.6) in Alto Paraná, and 3.2% (95% CI 1.7-5.4) in Caaguazú. In Asunción-Central, associations with HIV were age ≥25 years (1.86, 95% CI 1.15-3.00), being employed (1.82, 95% CI 1.07-3.11), self-reporting as homosexual (1.90, 95% CI 1.06-3.43), having sex with a known HIV-positive partner acquisition (4.19, 95% CI 2.37-7.43), self-perceived as being at higher risk for HIV acquisition (4.15, 95% CI 2.54-6.77), and able to access condoms and lubricants (1.82, 95% CI 1.08-3.05). In Alto Paraná, associations with HIV were self-reporting as homosexual (4.33, 95% CI 1.19-15.65) and having higher HIV knowledge (2.53, 95% CI 0.97-6.61). In Caaguazú, associations with HIV were self-reporting as homosexual (7.06, 95% CI 1.53-32.46) and being diagnosed with depression (4.68, 95% CI 0.89-24.43). CONCLUSIONS: HIV prevalence among MSM in Paraguay varied by region, being highest in the capital and major metropolitan area of Asunción-Central, followed by the border area of Alto Paraná. While being self-identified as homosexual was associated with HIV in all three regions, other associations differed, indicating prevention programs need to be tailored to the locale.

Factores asociados a la mortalidad por tuberculosis en indígenas en Paraguay, 2014 a 2019

Introducción: La tasa de mortalidad estimada de tuberculosis (TBC) en Paraguay en 2019 fue de 3,9 por 100.000 habs. Las comunidades indígenas presentan un elevado riesgo de padecer TBC. Objetivo: Determinar los factores asociados a la mortalidad en personas de origen indígena con diagnóstico de TBC en Paraguay, 2014-2019. Métodos : Se realizó un estudio observacional retrospectivo. Se utilizaron los datos del Programa Nacional de Control de la TBC del Paraguay comprendidos entre los años 2014 y 2019. Se registraron los datos sociodemográficos y factores de riesgo. Resultados: Se incluyeron 2.210 personas de origen indígena con diagnóstico de TBC, el 53,8% fue de sexo masculino, entre 20 y 39 años (32,3%) y 0 a 19 años de edad (30,2%), la localización de la TBC fue mayoritariamente pulmonar (92,3%). Durante el 2014 a 2019 se observaron 217 muertes (9,8%). Los factores asociados a mortalidad en pacientes con TBC fueron la edad (adOR = 13,95; CI: 7,07-27,55 mayor a 80 años), (adOR = 4,20; CI: 2,59-6,82 mayor a 60 años) y (adOR = 3,30; CI: 2,06-5,28 para 40 a 59 años), la co-infección VIH (adOR =), y la localización de la TBC (adOR = 3,60; CI: 1,88-6,90 para TBC diseminada). Conclusión: La mayor edad, el diagnóstico de co-infección VIH y localización de la TBC diseminada, están asociados a un mayor riesgo de muerte en población indígena con TBC.

Background: The estimated tuberculosis (TB) mortality rate in Paraguay in 2019 was 3.9 per 100,000 people. Indigenous communities are at high risk for TB. Aim: To determine the factors associated with mortality in indigenous people with a diagnosis of TB in Paraguay, 2014-2019. Methods: A retrospective observational study was done. sociodemographic data and risk factors data from the National TB Program of Paraguay between the years 2014 to 2019 were used. Results: A total of 2,210 indigenous people with a diagnosis of TB were included, 53,8% were male, between 20 to 39 (32.3%) and 0 to 19 years old (30.2%), the localization of TBC was majority pulmonary (92.3%). During 2014 to 2019 the were 217 deaths (9.8%). The factors associated to mortality in TB patients were the age (adOR = 13.95; CI: 7.07-27.55 older than 80 years), (adOR = 4.20; CI: 2.59-6.82 older than 60 years), and (adOR = 3.30; CI: 2.06 - 5.2840 to 59 years), HIV co-infection (adOR = 7.07; CI 3.74-13.87), and localization of TB (adOR = 3.60; CI: 1.88-6.90 for disseminated TB). Conclusion: Older age, HIV co-infection and disseminated localization of TBC are associated with a higher risk of death in indigenous people with TB.

A hybrid sequencing and assembly strategy for generating culture free Giardia genomes.

Giardia duodenalis is a pathogenic intestinal protozoan parasite of humans and many other animals. Giardia duodenalis is found throughout the world, and infection is known to have adverse health consequences for human and other mammalian hosts. Yet, many aspects of the biology of this ubiquitous parasite remain unresolved. Whole genome sequencing and comparative genomics can provide insight into the biology of G. duodenalis by helping to reveal traits that are shared by all G. duodenalis assemblages or unique to an individual assemblage or strain. However, these types of analyses are currently hindered by the lack of available G. duodenalis genomes, due, in part, to the difficulty in obtaining the genetic material needed to perform whole genome sequencing. In this study, a novel approach using a multistep cleaning procedure coupled with a hybrid sequencing and assembly strategy was assessed for use in producing high quality G. duodenalis genomes directly from cysts obtained from feces of two naturally infected hosts, a cat and dog infected with assemblage A and D, respectively. Cysts were cleaned and concentrated using cesium chloride gradient centrifugation followed by immunomagnetic separation. Whole genome sequencing was performed using both Illumina MiSeq and Oxford Nanopore MinION platforms. A hybrid assembly strategy was found to produce higher quality genomes than assemblies from either platform alone. The hybrid G. duodenalis genomes obtained from fecal isolates (cysts) in this study compare favorably for quality and completeness against reference genomes of G. duodenalis from cultured isolates. The whole genome assembly for assemblage D is the most contiguous genome available for this assemblage and is an important reference genome for future comparative studies. The data presented here support a hybrid sequencing and assembly strategy as a suitable method to produce whole genome sequences from DNA obtained from G. duodenalis cysts which can be used to produce novel reference genomes necessary to perform comparative genomics studies of this parasite.

HIV drug resistance in persons initiating or reinitiating first-line antiretroviral therapy in Paraguay: Results of a National Patient Survey.

Human immunodeficiency virus (HIV) drug resistance increases mortality and morbidity and antiretroviral therapy (ART) costs. We describe Paraguay's first nationally representative survey on pretreatment drug resistance (PDR) conducted among persons who initiated or reinitiated ART in 2019. ​​​​We conducted a cross-sectional survey of antiretroviral (ARV) drug resistance in Paraguay in 2019. Participants were sampled at four comprehensive care clinics where 90% of patients with HIV in Paraguay initiate ART. Patients included were adults ≥18 years old who initiated first-line ART or reinitiated the same first-line ART regimen after ≥3 months of discontinuation. Of 208 patients, 93.8% had no prior ART exposure, 3.8% reinitiated the same regimen, 2.4% had unknown prior ART exposure; and 31.3% had a CD4 count <200 cells/µl. Mutations associated with resistance were present in 15.4% of patients. Mutations associated with resistance to nonnucleoside reverse transcriptase inhibitors (NNRTI) were present in 13.0% of patients, nucleoside reverse transcriptase inhibitors in 4.3%, and integrase inhibitors in 3.4%. Mutations associated with resistance to tenofovir were present in 1.0% of patients and emtricitabine/lamivudine in 1.4%. ​​Nearly one in six patients had PDR in Paraguay's first nationally representative sample. High NNRTI PDR prevalence underscores the need to accelerate the transition to dolutegravir-based first-line ART. The low PDR prevalence of tenofovir and emtricitabine is reassuring as these ARVs are part of the World Health Organization (WHO)-recommended oral pre-exposure prophylaxis regimen. The high proportion of individuals initiating ART at a late disease stage highlights the need to improve treatment linkage strategies and implement WHO rapid ART initiation recommendations.

WHO method for estimating congenital syphilis to inform surveillance and service provision, Paraguay.

Problem: In Paraguay, incomplete surveillance data resulted in the burden of congenital syphilis being underestimated, which, in turn, led to missed opportunities for infant diagnosis and treatment. Approach: The prevalence of congenital syphilis, as defined by the World Health Organization (WHO), was estimated for Paraguay using the WHO congenital syphilis estimation tool. This tool was also used to monitor progress towards the elimination of mother-to-child transmission of syphilis. Local setting: The burden of syphilis in Paraguay has historically been high: its prevalence in pregnant women was estimated to be 3% in 2018. Relevant changes: The incidence rate of congenital syphilis estimated using the WHO tool was around nine times the reported prevalence. Subsequently, Paraguay: (i) provided training to improve diagnosis and case reporting; (ii) strengthened information systems for case monitoring and reporting; and (iii) procured additional rapid dual HIV-syphilis and rapid plasma reagin tests to increase syphilis testing capacity. In addition, the Ministry of Health prepared a new national plan for eliminating mother-to-child transmission of syphilis, with clear monitoring milestones. Lessons learnt: Health-care providers' reporting and surveillance procedures for congenital syphilis may not adequately reflect national and international case definitions. Use of the WHO congenital syphilis estimation tool in Paraguay drew attention to congenital syphilis as a national public health problem and highlighted the importance of comprehensive national surveillance systems and accurate data. Ongoing use of the WHO tool can track progress towards the elimination of mother-to-child transmission of syphilis by helping improve syphilis service coverage and national surveillance.

Identification and Distribution of Sterols, Bile Acids, and Acylcarnitines by LC-MS/MS in Humans, Mice, and Pigs-A Qualitative Analysis.

Sterols, bile acids, and acylcarnitines are key players in human metabolism. Precise annotations of these metabolites with mass spectrometry analytics are challenging because of the presence of several isomers and stereoisomers, variability in ionization, and their relatively low concentrations in biological samples. Herein, we present a sensitive and simple qualitative LC-MS/MS (liquid chromatography with tandem mass spectrometry) method by utilizing a set of pure chemical standards to facilitate the identification and distribution of sterols, bile acids, and acylcarnitines in biological samples including human stool and plasma; mouse ileum, cecum, jejunum content, duodenum content, and liver; and pig bile, proximal colon, cecum, heart, stool, and liver. With this method, we detected 24 sterol, 32 bile acid, and 27 acylcarnitine standards in one analysis that were separated within 13 min by reversed-phase chromatography. Further, we observed different sterol, bile acid, and acylcarnitine profiles for the different biological samples across the different species. The simultaneous detection and annotation of sterols, bile acids, and acylcarnitines from reference standards and biological samples with high precision represents a valuable tool for screening these metabolites in routine scientific research.

Point prevalence survey of antibiotic use in hospitals in Latin American countries.

BACKGROUND: Point prevalence surveys (PPSs) on antibiotic use are useful for understanding different aspects related to prescription patterns in hospitals. METHODS: An adaptation of the WHO methodology for a PPS on antibiotic use was applied. Hospital wards were divided into medical (MED), surgical (SUR), ICUs, gynaecology and obstetrics (GO), high-risk (HR) and mixed wards (MIX). A web application (RedCap©) through a mobile device was used for data collection. RESULTS: Between December 2018 and August 2019, 5444 patients in 33 hospitals in five countries were included (10 hospitals in Cuba, 7 in Paraguay, 6 in El Salvador, 5 in Mexico and 5 in Peru). Of these patients, 54.6% received at least one antibiotic, with variations between and within hospitals and countries. Antibiotics were more frequently used in ICUs (67.2%), SUR (64.5%) and MED wards (54.2%), with 51.2% of antibiotics prescribed for community-acquired infections (CAIs), 22.9% for healthcare-associated infections (HAIs), 11.1% for surgical prophylaxis and 6.1% for unknown reasons. Adherence to guidelines was observed in 68.6% of cases (72.8% for CAIs, 72.4% for HAIs and 44.3% for prophylaxis). Third-generation cephalosporins were the class of antibiotics most frequently used (26.8%), followed by carbapenems (10.3%) and fluoroquinolones (8%). Targeted treatments were achieved in 17.3% of cases. CONCLUSIONS: Antibiotic use was generally higher than that published in other studies. There is an urgent need to promote and strengthen the antimicrobial stewardship programmes in Latin America.

Fruit and Vegetable Supplemented Diet Modulates the Pig Transcriptome and Microbiome after a Two-Week Feeding Intervention.

A study was conducted to determine the effects of a diet supplemented with fruits and vegetables (FV) on the host whole blood cell (WBC) transcriptome and the composition and function of the intestinal microbiome. Nine six-week-old pigs were fed a pig grower diet alone or supplemented with lyophilized FV equivalent to half the daily recommended amount prescribed for humans by the Dietary Guideline for Americans (DGA) for two weeks. Host transcriptome changes in the WBC were evaluated by RNA sequencing. Isolated DNA from the fecal microbiome was used for 16S rDNA taxonomic analysis and prediction of metabolomic function. Feeding an FV-supplemented diet to pigs induced differential expression of several genes associated with an increase in B-cell development and differentiation and the regulation of cellular movement, inflammatory response, and cell-to-cell signaling. Linear discriminant analysis effect size (LEfSe) in fecal microbiome samples showed differential increases in genera from Lachnospiraceae and Ruminococcaceae families within the order Clostridiales and Erysipelotrichaceae family with a predicted reduction in rgpE-glucosyltransferase protein associated with lipopolysaccharide biosynthesis in pigs fed the FV-supplemented diet. These results suggest that feeding an FV-supplemented diet for two weeks modulated markers of cellular inflammatory and immune function in the WBC transcriptome and the composition of the intestinal microbiome by increasing the abundance of bacterial taxa that have been associated with improved intestinal health.

The Effects of Consuming White Button Mushroom Agaricus bisporus on the Brain and Liver Metabolome Using a Targeted Metabolomic Analysis.

A targeted metabolomic analysis was performed on tissues derived from pigs fed diets supplemented with white button mushrooms (WBM) to determine the effect on the liver and brain metabolome. Thirty-one pigs were fed a grower diet alone or supplemented with either three or six servings of freeze-dried WBM for six weeks. Tissue metabolomes were analyzed using targeted liquid chromatography-mass spectrometry (LC-MS) combined with chemical similarity enrichment analysis (ChemRICH) and correlated to WBM-induced changes in fecal microbiome composition. Results indicated that WBM can differentially modulate metabolites in liver, brain cortex and hippocampus of healthy pigs. Within the glycero-phospholipids, there was an increase in alkyl-acyl-phosphatidyl-cholines (PC-O 40:3) in the hippocampus of pigs fed six servings of WBM. A broader change in glycerophospholipids and sphingolipids was detected in the liver with a reduction in several lipid species in pigs fed both WBM diets but with an increase in amino acids known as precursors of neurotransmitters in the cortex of pigs fed six servings of WBM. Metabolomic changes were positively correlated with increased abundance of Cryomorphaceae, Lachnospiraceae, Flammeovirgaceae and Ruminococcaceae in the microbiome suggesting that WBM can also positively impact tissue metabolite composition.