Phage therapy: resurrecting a historical solution for the contemporary challenge of rising antibiotic resistance in Latin America.

INTRODUCTION: Antimicrobial resistance in Latin America is a growing concern in both human and non-human animal populations. The economic burden that is likely to be imposed through increased resistance will cause further strains on public health systems and the population at large. AREAS COVERED: We propose the rapid adoption and implementation of phage therapy as a necessary addition to the medical arsenal to help mitigate antimicrobial resistance, with an emphasis on considering the potential benefits that highly biodiverse countries such as Ecuador may have on phage discovery. However, programs may count on limited government support and/or facilitation, which could slow progress. EXPERT OPINION: We highlight the need for educational campaigns to be implemented in parallel with the development of phage therapy programs, particularly to implement these novel treatments in rural and indigenous communities.

Geographic Distribution of Human Infections with Zoonotic Ancylostoma ceylanicum and Anthropophilic Hookworms in Ecuador: A Retrospective Analysis of Archived Stool Samples.

Zoonotic human infections with Ancylostoma ceylanicum have recently been reported in the Americas. We used archived human stool samples to study the geographic distribution of human infections with A. ceylanicum and anthropophilic hookworms in different geoclimatic regions (coastal, Andean, and Amazon) of Ecuador. We analyzed retrospectively archived human stool samples from five studies previously screened for hookworm infection by microscopy, of which four included hookworm-positive samples only and one involved hookworm-negative samples to increase geographic distribution of sampling. Stools were analyzed using multi-parallel quantitative polymerase chain reaction (qPCR) assays to detect Necator americanus, Ancylostoma duodenale, A. ceylanicum, Ascaris lumbricoides, Trichuris trichiura, and Strongyloides stercoralis. Sequencing was done for the A. ceylanicum cox1 gene. A total of 132 samples were analyzed, of which 69 (52.3%) were from hookworm-positive and 63 (47.7%) from hookworm-negative individuals by microscopy. Overall, 82.6% of microscopy-positive samples and 33.3% of microscopy-negative samples were positive for hookworm by qPCR. Of microscopy-positive samples, 36.2% were A. ceylanicum, 37.7% A. duodenale, and 33.3% N. americanus, whereas equivalent proportions for microscopy-negative samples were 1.6%, 31.7%, and 1.6%, respectively. Ancylostoma duodenale was the most widely dispersed geographically, followed by N. americanus. Ancylostoma ceylanicum was least dispersed but was detected in coastal and Amazon regions. In conclusion, human infections with A. ceylanicum, A. duodenale, and N. americanus were detected in different geoclimatic regions of Ecuador. Additional studies are required to further define the epidemiology of human A. ceylanicum infections, but the potentially widespread presence of this helminth in human populations in Ecuador has implications for hookworm control strategies.

Epidemiology of giardiasis and assemblages A and B and effects on diarrhea and growth trajectories during the first 8 years of life: Analysis of a birth cohort in a rural district in tropical Ecuador.

BACKGROUND: There are limited longitudinal data on the acquisition of Giardia lamblia infections in childhood using molecular assays to detect and type assemblages, and measure effects of infections on diarrhea risk and childhood growth. METHODS: We analysed stool samples from a surveillance sample within a birth cohort in a rural district in tropical Ecuador. The cohort was followed to 8 years of age for the presence of G. lamblia in stools by quantitative PCR and A and B assemblages by Taqman assay or Sanger sequencing. We explored risk factors associated with infection using generalized estimating equations applied to longitudinal binary outcomes, and longitudinal panel data analysis to estimate effects of infection on diarrhea and growth trajectories. RESULTS: 2,812 stool samples collected between 1 month and 8 years of age from 498 children were analyzed and showed high rates of infection: 79.7% were infected at least once with peak prevalence (53.9%) at 5 years. Assemblage B was accounted for 56.8% of genotyped infections. Risk factors for infection included male sex (P = 0.001), daycare attendance (P<0.001), having a household latrine (P = 0.04), childhood (P<0.001) and maternal soil-transmitted helminth (P = 0.029) infections, and exposures to donkeys (age interaction P = 0.034). G. lamblia was associated with increased risk of diarrhea (per episode, RR 1.03, 95% CI 1.01-1.06, P = 0.011) during the first 3 years of life and a transient impairment of weight (age interaction P = 0.017) and height-for-age (age interaction P = 0.025) trajectories between 1 and 4 years of age. There was no increased risk of either assemblage being associated with outcomes. CONCLUSION: Our data show a relatively high edemicity of G. lamblia transmission during childhood in coastal Ecuador, and evidence that infection is associated with a transiently increased risk of diarrhea during the first 3 years of life and impairment of weight and height between 1 and 4 years.

Diagnostic accuracy of qPCR and microscopy for cutaneous leishmaniasis in rural Ecuador: A Bayesian latent class analysis.

BACKGROUND: Clinical and laboratory diagnosis of cutaneous leishmaniasis (CL) is hampered by under-ascertainment of direct microscopy. METHODS: This study compared the diagnostic accuracy of qPCR on DNA extracted from filter paper to the accuracy of direct smear slide microscopy in participants presenting with a cutaneous lesion suspected of leishmaniasis to 16 rural healthcare centers in the Ecuadorian Amazon and Pacific regions, from January 2019 to June 2021. We used Bayesian latent class analysis to estimate test sensitivity, specificity, likelihood ratios (LR), and predictive values (PV) with their 95% credible intervals (95%CrI). The impact of sociodemographic and clinical characteristics on predictive values was assessed as a secondary objective. RESULTS: Of 320 initially included participants, paired valid test results were available and included in the diagnostic accuracy analysis for 129 from the Amazon and 185 from the Pacific region. We estimated sensitivity of 68% (95%CrI 49% to 82%) and 73% (95%CrI 73% to 83%) for qPCR, and 51% (95%CrI 36% to 66%) and 76% (95%CrI 65% to 86%) for microscopy in the Amazon and Pacific region, respectively. In the Amazon, with an estimated disease prevalence among participants of 73%, negative PV for qPCR was 54% (95%CrI 5% to 77%) and 44% (95%CrI 4% to 65%) for microscopy. In the Pacific, (prevalence 88%) the negative PV was 34% (95%CrI 3% to 58%) and 37% (95%CrI 3% to 63%). The addition of qPCR parallel to microscopy in the Amazon increases the observed prevalence from 38% to 64% (+26 (95%CrI 19 to 34) percentage points). CONCLUSION: The accuracy of either qPCR on DNA extracted from filter paper or microscopy for CL diagnosis as a stand-alone test seems to be unsatisfactory and region-dependent. We recommend further studies to confirm the clinically relevant increment found in the diagnostic yield due to the addition of qPCR.

Comparative genome analysis of colistin-resistant Escherichia coli harboring mcr isolated from rural community residents in Ecuador and Vietnam.

The spread of colistin-resistant bacteria among rural community residents of low- and middle-income countries is a major threat to community health. Although the mechanism of the spread of colistin-resistant bacteria in communities is unknown, geographic and regional characteristics may influence it. To elucidate the spread mechanism of colistin-resistant bacteria, we analyzed the genomes of colistin-resistant Escherichia coli isolated from Vietnam and Ecuador residents, which are geographically and socially different. Stool specimens of 139 and 98 healthy residents from Ecuador and Vietnam rural communities, respectively, were analyzed for colistin-resistant E. coli with mcr. Its prevalence in the residents of all the communities assessed was high and approximately equal in both countries: 71.8% in Ecuador and 69.4% in Vietnam. A phylogenetic tree analysis revealed that the sequence type of colistin-resistant E. coli was diverse and the major sequence types were different between the two countries. The location of mcr in the isolates showed that the proportion of chromosomal mcr was 35.1% and 8.5% in the Vietnam and Ecuador isolates, respectively. Most of these chromosomal mcr genes (75%-76%) had an intact mcr-transposon Tn6330. Contrastingly, the replicon types of the mcr-carrying-plasmids were diverse in both countries, but almost all belonged to IncI2 in Ecuador and IncX1/X4 in Vietnam. Approximately 26%-45% of these mcr-plasmids had other resistance genes, which also varied between countries. These results suggest that although the overall profile of the colistin-resistant E. coli isolates is diverse in these countries, the phylogenesis of the isolates and mcr-carrying plasmids has regional characteristics. Although the contributing factors are not clear, it is obvious that the overall profile of colistin-resistant bacteria dissemination varies between countries. Such different epidemic patterns are important for establishing country-specific countermeasures against colistin-resistant bacteria.

New Primers for Detection and Differentiation between Leishmania viannia and L. leishmania Subgenera by Polymerase Chain Reaction.

Background: Leishmania is the parasitic protozoan responsible for leishmaniases, a disease that can cause a range of cutaneous, mucosal, and visceral infections. Two subgenera L. Viannia and L. Leishmania are known to infect humans in the tropics and subtropics of the Americas. The aim of the present study was to develop a new pair of primers for the two subgenera and test in clinical samples. Methods: We designed two new pairs of primers for a PCR method from two conserved genes, cysteine proteinase B (cpb) and N-acetylglucosamine-6-phosfate deacetylase-like protein (nagA), as specific markers for those two respective subgenera. Primers were tested with 16 microscopical positive clinical samples from the Amazon region of Ecuador obtained in 2010-2020 period. Results: The cpb presented a band of 172 bp and the nagA a band of 300 bp, thus clearly differentiating L. viannia from L. leishmania. Additionally, primers identified and differentiated the clinical samples in the two subgenera. Conclusion: The new primers targeting different two genes and standardized in a PCR assay could identified and differentiated Leishmania parasites at subgenus level. This protocol could be used for Leishmania genus identification and diagnosis at the subgenus level and for determining the parasite's geographical distribution where different Leishmania subgenera are found in the same area.

Multi-sensorial perceptions of risk: the aesthetics behind (muco)cutaneous leishmaniasis-related stigma in Ecuador.

Previous research on the stigma associated with cutaneous leishmaniasis, a vector-transmitted parasitic disease, focuses on aesthetic appearance affectation as the leading cause of stigmatisation. However, Indigenous populations in the hinterland of Amazonian Ecuador trigger stigma expressions by recognising (muco)cutaneous leishmaniasis, primarily through atypical smell, followed by the odd voice sound, appearance and taste. This empirical way of recognising symptoms relies on embodied forms of identifying a disease, contrasting the Western supremacy of visuality and demanding to be understood via multi-sensorial anthropology. Through ethnographic research and data retrieved from eighty-three semistructured interviews and fifteen focus groups in seven Ecuadorian ethnic groups - including six Indigenous groups in the Amazon region - this paper analyses how the sensorium is a health thermometer. Findings reveal that differentiated cultural responses to a sense of peril, contagion and social (self)rejection, understood as stigma expressions, are linked to the holistic approach to health (or well-being) shared by Indigenous populations. In forest societies, well-being is explained through successful (non-)human relationships, and disease permeates through bodies that lack balanced relations.

A Fatal Case of Disseminated Histoplasmosis by Histoplasma capsulatum var. capsulatum Misdiagnosed as Visceral Leishmaniasis-Molecular Diagnosis and Identification.

Histoplasmosis is an endemic mycosis in the Americas. However, its diagnosis is challenging due to the complexity and limited availability of conventional laboratory techniques-antigen tests, culture, and staining. Microscopic preparations often confuse with other pathogens, such as Leishmania spp. The genus Histoplasma capsulatum comprises three varieties: var. capsulatum, var. duboissi, and var. farciminosum, which cannot be distinguished using conventional techniques. An infant from a tropical region of Ecuador was hospitalized for fever, bloody diarrhea, and anemia persisting for two months. Upon admission, he received antibiotics and immunosuppressants. Histopathological examination of the lymph nodes, intestines, and bone marrow aspirate reported the presence of Leishmania-like amastigotes, and treatment was initiated with meglumine antimoniate and conventional amphotericin B. However, subsequent analysis of samples using PCR and DNA sequencing identified H. capsulatum var. capsulatum but not Leishmania. Despite fluconazole and amphotericin B, the infant succumbed to the disease. The delay in clinical and laboratory diagnosis of histoplasmosis and the use of nonspecific and ineffective drugs such as fluconazole led to disease dissemination and, ultimately, death. Implementing molecular diagnosis and antigen tests in laboratories located in endemic regions and reference hospitals is crucial.

Case report: First report on human infection by tick-borne Babesia bigemina in the Amazon region of Ecuador.

Babesiosis is a protozoan disease acquired by the bite of different species of ticks. More than 100 Babesia spp. infect wild and domestic animals worldwide, but only a few have been documented to infect humans. Generally, babesiosis is asymptomatic in immunocompetent persons; however, in immunocompromised can be life-threatening. A 13-year-old boy from the Amazon region presented with a 3-month evolution of fever, chills, general malaise, and arthralgia accompanied by anemia and jaundice. In the last 4 years was diagnosed with chronic kidney failure. By nested-PCR using 18S RNA ribosomal gene as target and DNA sequencing, the phylogenetic analysis showed Babesia bigemina as the causative agent in the blood. Treatment with oral quinine plus clindamycin for six continuous weeks was effective with no relapse occurring during 12 months of follow-up. This is the second human case in Ecuador but the first caused by the zoonotic B. bigemina which confirms the existence of active transmission that should alert public health decision-making authorities on the emergence of this zoonosis and the need for research to determine strategies to reduce tick exposure.

Leishmania species and clinical characteristics of Pacific and Amazon cutaneous leishmaniasis in Ecuador and determinants of health-seeking delay: a cross-sectional study.

BACKGROUND: Cutaneous Leishmaniasis (CL) affects up to 5.000 people in Ecuador each year. L. guyanensis and L. braziliensis are the most common of the eight CL-causing Leishmania species. Earlier CL research concentrated on the easily accessible Pacific region. This study aims to describe the Leishmania species in Pacific and Amazon ecoregions, to analyze regional differences in CL patient clinical presentation, and to identify determinants of health-seeking delay. METHODS: All cases in this cross-sectional study were diagnosed using smear slide microscopy, PCR, or both. Cytochrome B gene sequencing was used to identify the causative Leishmania species in qPCR-positive samples. RESULTS: This study included 245 patients, with 154 (63%) infected in the Pacific region and 91 (37%) infected in the Amazon. Causative Leishmania species were identified in 135 patients (73% of qPCR positives). L. guyanensis was identified in 76% (102/135) of the samples and L. braziliensis in 19% (26/135). The Pacific region had a low prevalence of 6% (5/89) of L. braziliensis. For the first time, we report L. guyanensis from the central Amazon, L. braziliensis from the northern Pacific, and L. lainsoni from both the central Amazon and northern Pacific. Amazon cases had a longer median health-seeking delay in months (2.0, IQR 3.0) than Pacific cases (1.0, IQR 1.5). Prolonged health-seeking delay was associated with older age, Amerindian ethnicity, infection at lower altitudes, non-ulcerative lesions, and lesions on the lower limbs. CONCLUSIONS: In the Pacific region, health-seeking delay is relatively short and L. braziliensis prevalence remains low. Limited access to health care and stigma might explain the prolonged health-seeking delay in the Amazon. We recommend larger studies on the distribution of Leishmania species in Amazon CL cases and additional regional research into diagnostic test accuracy. Furthermore, the determinants of health-seeking delay in Ecuador should be investigated further.

Epidemiology and clinical features of venomous snake bites in the Northern Amazon of Ecuador (2017-2021) / Epidemiología y características clínicas de las mordeduras de serpientes venenosas en el norte de la Amazonía del Ecuador (2017-2021)

Introduction: In Ecuador, poisonous snakebites are a public health problem. However, there is no recent hospital information from the Amazon. Objective: To retrospectively analyse the clinical-epidemiological characteristics of snakebites in patients admitted to a hospital in the Ecuadorian Amazon. Materials and methods: This is a cross-sectional study conducted at the Nueva Loja-Sucumbíos Provincial Hospital, bordering Colombia (2017-2021). Demographic, epidemiological and clinical variables, and condition at hospital discharge, were obtained from the epidemiological file of the Ministerio de Salud Pública. Results: In 5 years, 147 patients (29.4 per year) were hospitalized with no mortality. They corresponded to 26, 34, 32, 29 and 26 cases, in 2017, 2018, 2019, 2020 and 2021, respectively. Men with 99 (67.3%), aged 21-30 years with 28, mixed race with 94 cases, students, and farmers, were the most affected. The most affected (99; 67.3%) were men, people 21 to 30 years-old (28; 19.0%), mestizos (94; 63.9%), students, and farmers. The median age was 28 (range: 4-81) years. Prevalence was higher in April, June, and September. All accidents were caused by Viperidae snakes. Twenty (13.6%) cases were mild, (61.2%) were moderate and 37 (25.2%) were severe. The feet with 45 were the most bitten. Pre-hospitalization anti-venom serum was received by 53.1% and tourniquet by 19.8% patients. Median hospital arrival time was 5 (range 1-192) hours, mostly between 2-3 hours with 41 cases. No statistically significant differences were found considering the severity. Conclusions: A high prevalence of snakebites was evidenced in the north of the Amazon in Ecuador, with a higher incidence in the rainy season and all by Viperidae species. It is important to highlight the null mortality. Information campaigns on prevention and first aid, such as discouraging the use of tourniquets, especially among vulnerable groups.

Introducción. En Ecuador, las mordeduras de serpientes venenosas son un problema de salud pública. Sin embargo, no existe información hospitalaria reciente desde la Amazonía. Objetivo. Analizar retrospectivamente las características clínico-epidemiológicas de las mordeduras de serpientes en pacientes ingresados en un hospital de la Amazonía del Ecuador. Materiales y métodos. Se llevó a cabo un estudio transversal en el hospital provincial de Nueva Loja (Sucumbíos), que colinda con Colombia, 2017-2021. La información sobre las variables demográficas, epidemiológicas y clínicas, y la condición al egreso hospitalario, se obtuvieron de la ficha epidemiológica del Ministerio de Salud Pública. Resultados. En cinco años se hospitalizaron 147 pacientes (29,4 por año), sin que se presentaran muertes. Corresponden a 26, 34, 32, 29 y 26 casos, en el 2017, 2018, 2019, 2020 y 2021, respectivamente. Según el sexo, los más afectados fueron los hombres (n=99; 67,3 %), según el grupo etario, los de 21 a 30 años (n=28; 19,0 %) y, según la raza, los de etnia mestiza (n=94; 63,9 %), estudiantes y agricultores. La mediana de edad fue de 28 años (rango: 4 a 81). Hubo mayor prevalencia en abril, junio y septiembre. Todos los accidentes fueron causados por serpientes Viperidae. Veinte (13,6 %) casos fueron leves, 90 (61,2 %), moderados, y 37 (25,2 %), graves. La mordedura fue más frecuente en los pies (45 casos). El 53,1 % recibió suero antiofídico antes de la hospitalización y en el 19,8 % de los pacientes se colocó un torniquete. La mediana de tiempo de llegada al hospital fue de 5 horas (rango: 1-192), y lo más frecuente fue entre 2 y 3 horas (41 casos). No se encontraron diferencias estadísticamente significativas según la gravedad del caso. Conclusiones. Se evidenció una gran prevalencia de mordeduras de serpientes en el norte de la región amazónica-Ecuador, con mayor incidencia en la estación lluviosa y todas causadas por Viperidae. Es importante resaltar la mortalidad nula. Las campañas informativas sobre prevención y primeros auxilios, como la desmotivación del uso de torniquetes, serían fundamentales para reducir los casos, especialmente, en los grupos vulnerables.

Co-Harboring of Beta-Lactamases and mcr-1 Genes in Escherichia coli and Klebsiella pneumoniae from Healthy Carriers and Backyard Animals in Rural Communities in Ecuador.

Few studies have addressed drug resistance of Enterobacterales in rural communities in developing countries. This study aimed to determine the coexistence of extended-spectrum ß-lactamase (ESBL) and carbapenemase genes in Escherichia coli and Klebsiella pneumoniae strains carrying the mcr-1 gene in rural communities in Ecuador from healthy humans and their backyard animals. Sixty-two strains, thirty E. coli and thirty-two K. pneumoniae strains carrying the mcr-1 gene were selected from a previous study. PCR were performed for the presence of ESBLs and carbapenemase genes. The strains were further characterized, and the genetic relationship was studied with multi-locus sequencing typing (MLST) of seven housekeeping genes. Fifty-nine of the sixty-two mcr-1 isolates (95%) harbored at least on ß-lactam resistance gene. The most prevalent ESBL genes were the blaTEM genes (present in in 80% of the E. coli strains) and the blaSHV gene (present in 84% of the K. pneumoniae strains). MSLT analysis revealed 28 different sequence types (ST); 15 for E. coli and 12 for K. pneumoniae, with most ST never described in humans and animals. The coexistence of mcr-1 and ß-lactams resistant genes in E. coli and K. pneumoniae strains is alarming and threatens the efficacy of last-resort antibiotics. Our findings highlight backyard animals as a reservoir of mcr-1/ß-lactams resistant genes.

Leptospirosis in Ecuador: Current Status and Future Prospects.

The location of Ecuador-an equatorial nation-favors the multiplication and dispersal of the Leptospira genus both on the Pacific Coast and in the Amazon tropical ecoregions. Nevertheless, leptospirosis epidemiology has not been fully addressed, even though the disease has been recognized as a significant public health problem in the country. The purpose of this literature review is to update knowledge on the epidemiology and geographical distribution of Leptospira spp. and leptospirosis in Ecuador to target future research and develop a national control strategy. A retrospective literature search using five international, regional, and national databases on Leptospira and leptospirosis including humans, animals, and environmental isolations of the bacteria and the disease incidence in Ecuador published between 1919 and 2022 (103 years) with no restriction on language or publication date was performed. We found and analyzed 47 publications including 22 of humans, 19 of animals, and two of the environments; three of these covered more than one of these topics, and one covered all three (i.e., One Health). Most (60%) of the studies were conducted in the Coastal ecoregion. Twenty-four (51%) were published in international journals, and 27 (57%) were in Spanish. A total of 7342 human and 6314 other animal cases were studied. Leptospirosis was a frequent cause of acute undifferentiated febrile illness in the Coast and Amazon and was associated with rainfall. All three major clusters of Leptospira-pathogenic, intermediate, and saprophytic-were identified from both healthy and febrile humans, the environment, and animals; moreover, nine species and 29 serovars were recorded over the three Ecuadorian ecoregions. Leptospira infections were diagnosed in livestock, companion, and wild animals from the Amazon and the Coast regions along with sea lions from the Galápagos Islands. Microscopic-agglutination test was the diagnostic tool most widely used. Three reviews covering national data on outpatients and inpatients determined the varied annual incidence and mortality rate, with males being more commonly affected. No human cases have been reported in the Galápagos Islands. Genomic sequences of three pathogenic Leptospira were reported. No studies on clinical ground, antibiotic resistance, or treatment were reported, nor were control programs or clinical-practice guidelines found. The published literature demonstrated that leptospirosis was and still is an endemic disease with active transmission in the four geoclimatic regions of Ecuador including the Galápagos Islands. Animal infections, distributed in mainland and insular Ecuador, pose a significant health risk for humans. Nationwide epidemiological surveys-encouraging more research on the fauna and environment with appropriate sampling design on risk factors for human and animal leptospirosis, Leptospira genotyping, increased laboratory capability, and readily available official data-are required to improve our understanding of transmission patterns and to develop effective national intervention strategies with the intention of applying One Health approaches.

Colistin resistance in Escherichia coli and Klebsiella pneumoniae in humans and backyard animals in Ecuador.

Objective: Colistin is an antibiotic of last resort for treating serious Gram-negative bacterial infections. However, the misuse of colistin, especially as an animal growth promoter, has contributed to increasing antimicrobial resistance, mediated mainly through plasmid transfer of the mcr-1 gene. This study assessed the prevalence of phenotypic and molecular colistin resistance in Escherichia coli and Klebsiella pneumoniae in Ecuador in healthy humans and their chickens and pigs. Methods: Fecal samples were collected from humans and their chickens and pigs in two rural coastal and Amazon regions between April and August 2020. Gram-negative bacteria were isolated and identified using conventional techniques. Phenotypic resistance was determined using the broth microdilution technique, and the mcr-1 gene was detected using conventional polymerase chain reaction. Results: A total of 438 fecal samples were obtained from 137 humans, 147 pigs and 154 chickens. The prevalence of E. coli isolates was 86.3% (378/438) and K. pneumoniae, 37.4% (164/438). Overall, the mcr-1 gene was found in 90% (340/378) of E. coli isolates, with higher prevalences found in isolates from coastal regions (96.5%, 191/198), humans (95.6%, 111/116) and chickens (91.8%, 123/134); for K. pneumoniae, the gene was found in 19.5% (32/164) of isolates, with equal distribution between regions and hosts. Only four isolates, two E. coli and two K. pneumoniae, showed phenotypic resistance: mcr-1 was present in both E. coli strains but absent in the K. pneumoniae strains. Conclusions: Despite a low prevalence of phenotypic resistance to colistin, the high prevalence of the mcr-1 gene in E. coli is of concern. Ecuador's ban on using colistin in animal husbandry must be enforced, and continual monitoring of the situation should be implemented.

Diversity and prevalence of gastrointestinal helminths of free-roaming dogs on coastal beaches in Ecuador: Potential for zoonotic transmission.

Free-roaming dogs are highly exposed to a range of zoonotic parasites, including helminths, which can be transmitted to humans, particularly in rural tropical settings of developing countries. To evidence the diversity and prevalence of gastrointestinal helminths in the stools of free-roaming dogs on the public Pacific coastal beaches of Ecuador, a cross-sectional study was conducted from August 2021 to August 2022. The sampling beaches are located along the tropical Pacific coastal region. Stools were collected from the ground in containers with 10% formalin and processed using the Ritchie method; eggs were identified under a microscope. A total of 573 stools were examined from 20 beaches; the overall prevalence was 157 (27.4%) for one or more helminths. Ten parasites were identified, nine of which are potentially zoonotic. Ancylostoma spp. was the most prevalent (19.4%), followed by Toxocara spp. (7.2%). Trichuris spp., Dipylidium caninum, Diphyllobothrium spp., Capillaria spp., Dicrocoelium spp., Heterobilharzia americana, Hymenolepis spp. and Spirocerca spp. were also observed. Five of them are reported for the first-time infecting dogs in Ecuador. Hence, we evidenced that Ecuadorian beaches are highly contaminated with the dogs' zoonotic gastrointestinal helminths, posing a great risk to public health. Differences in the presence and prevalence were found in samples of tropical humid and dry climate zones. Therefore, based on our findings, we encourage the implementation of broad antiparasitic treatment and prevention strategies to reduce the zoonotic risk.

Colistin resistance in Escherichia coli and Klebsiella pneumoniae in humans and backyard animals in Ecuador

[ABSTRACT]. Objective. Colistin is an antibiotic of last resort for treating serious Gram-negative bacterial infections. However, the misuse of colistin, especially as an animal growth promoter, has contributed to increasing antimicrobial resistance, mediated mainly through plasmid transfer of the mcr-1 gene. This study assessed the prevalence of phenotypic and molecular colistin resistance in Escherichia coli and Klebsiella pneumoniae in Ecuador in healthy humans and their chickens and pigs. Methods. Fecal samples were collected from humans and their chickens and pigs in two rural coastal and Amazon regions between April and August 2020. Gram-negative bacteria were isolated and identified using conventional techniques. Phenotypic resistance was determined using the broth microdilution technique, and the mcr-1 gene was detected using conventional polymerase chain reaction. Results. A total of 438 fecal samples were obtained from 137 humans, 147 pigs and 154 chickens. The preva- lence of E. coli isolates was 86.3% (378/438) and K. pneumoniae, 37.4% (164/438). Overall, the mcr-1 gene was found in 90% (340/378) of E. coli isolates, with higher prevalences found in isolates from coastal regions (96.5%, 191/198), humans (95.6%, 111/116) and chickens (91.8%, 123/134); for K. pneumoniae, the gene was found in 19.5% (32/164) of isolates, with equal distribution between regions and hosts. Only four isolates, two E. coli and two K. pneumoniae, showed phenotypic resistance: mcr-1 was present in both E. coli strains but absent in the K. pneumoniae strains. Conclusions. Despite a low prevalence of phenotypic resistance to colistin, the high prevalence of the mcr-1 gene in E. coli is of concern. Ecuador’s ban on using colistin in animal husbandry must be enforced, and con- tinual monitoring of the situation should be implemented.

[RESUMEN]. Objetivo. La colistina es un antibiótico de último recurso para tratar infecciones graves por bacterias gramneg- ativas. Sin embargo, su uso indebido, especialmente para estimular el crecimiento animal, ha contribuido con el aumento de la resistencia a los antimicrobianos, mediada principalmente por la transferencia de plásmidos del gen mcr-1. En este estudio se evaluó la prevalencia de la resistencia fenotípica y molecular a la colistina de las bacterias Escherichia coli y Klebsiella pneumoniae en humanos sanos, sus pollos y cerdos en Ecuador. Métodos. Se recolectaron muestras fecales de humanos, así como de sus pollos y cerdos, en dos zonas rurales de la región costera y la región amazónica entre abril y agosto del 2020. Se aislaron las bacterias gramnegativas y se identificaron empleando técnicas convencionales. Se determinó la resistencia fenotípica mediante la técnica de microdilución en caldo y se detectó el gen mcr-1 con la técnica convencional de reac- ción en cadena de la polimerasa. Resultados. Se obtuvo un total de 438 muestras fecales de 137 humanos, 147 cerdos y 154 pollos. La preva- lencia de E. coli en las cepas aisladas fue del 86,3% (378/438) y la de K. pneumoniae, del 37,4% (164/438). En general, se detectó el gen mcr-1 en el 90% (340/378) de las cepas aisladas de E. coli y la mayor prevalencia encontrada fue en cepas aisladas de la región costera (96,5%, 191/198), humanos (95,6%, 111/116) y pollos (91,8%, 123/134); en el caso de K. pneumoniae, el gen se encontró en el 19,5% (32/164) de las cepas, con una distribución equitativa entre regiones y hospedadores. Únicamente cuatro cepas aisladas, dos de E. coli y dos de K. pneumoniae, mostraron resistencia fenotípica: el gen mcr-1 estaba presente en ambas cepas de E. coli y ausente en las cepas de K. pneumoniae. Conclusiones. Si bien hubo una baja prevalencia de resistencia fenotípica a la colistina, la alta prevalencia del gen mcr-1 en E. coli es preocupante. Es necesario hacer cumplir la prohibición del uso de colistina en la cría de animales en Ecuador, así como realizar un seguimiento continuo de la situación.

[RESUMO]. Objetivo. A colistina é um antibiótico de último recurso para o tratamento de infecções graves por bac- térias Gram-negativas. Entretanto, o uso indevido da colistina, principalmente como promotor de crescimento animal, tem contribuído para o aumento da resistência a antimicrobianos, principalmente por transferência horizontal do gene mcr-1 mediada por plasmídeos. Este estudo avaliou a prevalência de resistência fenotípica e molecular à colistina em Escherichia coli e Klebsiella pneumoniae no Equador em humanos hígidos e em galinhas e porcos por eles criados. Métodos. Entre abril e agosto de 2020, foram coletadas amostras de fezes de habitantes de duas regiões litorâneas e amazônicas do Equador e de galinhas e porcos por eles criados. Bactérias Gram-negativas foram isoladas e identificadas por meio de técnicas convencionais. A resistência fenotípica foi determinada pela técnica de microdiluição em caldo, e o gene mcr-1 foi detectado por reação em cadeia da polimerase convencional. Resultados. Foram obtidas 438 amostras fecais de 137 humanos, 147 suínos e 154 galinhas. A prevalência de isolados de E. coli foi de 86,3% (378/438), e de K. pneumoniae, 37,4% (164/438). Em geral, o gene mcr-1 foi encontrado em 90% (340/378) dos isolados de E. coli, com maiores prevalências encontradas em isola- dos de regiões litorâneas (96,5%, 191/198), humanos (95,6%, 111/116) e galinhas (91,8%, 123/134); para K. pneumoniae, o gene foi encontrado em 19,5% (32/164) dos isolados, com igual distribuição entre regiões e hospedeiros. Somente quatro isolados, dois de E. coli e dois de K. pneumoniae, demonstraram resistência fenotípica: o gene mcr-1 estava presente em ambas as cepas de E. coli, mas ausente nas de K. pneumoniae. Conclusões. Apesar da baixa prevalência de resistência fenotípica à colistina, a alta prevalência do gene mcr-1 em E. coli é preocupante. É preciso fiscalizar a proibição ao uso agropecuário de colistina no Equador e implementar o monitoramento contínuo da situação.

Epidemiología y características clínicas de las mordeduras de serpientes venenosas en el norte de la Amazonía del Ecuador (2017-2021) / Epidemiology and clinical features of venomous snake bites in the Northern Amazon of Ecuador (2017-2021)

Introducción. En Ecuador, las mordeduras de serpientes venenosas son un problema de salud pública. Sin embargo, no existe información hospitalaria reciente desde la Amazonía. Objetivo. Analizar retrospectivamente las características clínico-epidemiológicas de las mordeduras de serpientes en pacientes ingresados en un hospital de la Amazonía del Ecuador. Materiales y métodos. Se llevó a cabo un estudio transversal en el hospital provincial de Nueva Loja (Sucumbíos), que colinda con Colombia, 2017-2021. La información sobre las variables demográficas, epidemiológicas y clínicas, y la condición al egreso hospitalario, se obtuvieron de la ficha epidemiológica del Ministerio de Salud Pública. Resultados. En cinco años se hospitalizaron 147 pacientes (29,4 por año), sin que se presentaran muertes. Corresponden a 26, 34, 32, 29 y 26 casos, en el 2017, 2018, 2019, 2020 y 2021, respectivamente. Según el sexo, los más afectados fueron los hombres (n=99; 67,3 %), según el grupo etario, los de 21 a 30 años (n=28; 19,0 %) y, según la raza, los de etnia mestiza (n=94; 63,9 %), estudiantes y agricultores. La mediana de edad fue de 28 años (rango: 4 a 81). Hubo mayor prevalencia en abril, junio y septiembre. Todos los accidentes fueron causados por serpientes Viperidae. Veinte (13,6 %) casos fueron leves, 90 (61,2 %), moderados, y 37 (25,2 %), graves. La mordedura fue más frecuente en los pies (45 casos). El 53,1 % recibió suero antiofídico antes de la hospitalización y en el 19,8 % de los pacientes se colocó un torniquete. La mediana de tiempo de llegada al hospital fue de 5 horas (rango: 1-192), y lo más frecuente fue entre 2 y 3 horas (41 casos). No se encontraron diferencias estadísticamente significativas según la gravedad del caso. Conclusiones. Se evidenció una gran prevalencia de mordeduras de serpientes en el norte de la región amazónica-Ecuador, con mayor incidencia en la estación lluviosa y todas causadas por Viperidae. Es importante resaltar la mortalidad nula. Las campañas informativas sobre prevención y primeros auxilios, como la desmotivación del uso de torniquetes, serían fundamentales para reducir los casos, especialmente, en los grupos vulnerables.

Introduction: In Ecuador, poisonous snakebites are a public health problem. However, there is no recent hospital information from the Amazon. Objective: To retrospectively analyse the clinical-epidemiological characteristics of snakebites in patients admitted to a hospital in the Ecuadorian Amazon. Materials and methods: This is a cross-sectional study conducted at the Nueva Loja-Sucumbíos Provincial Hospital, bordering Colombia (2017-2021). Demographic, epidemiological and clinical variables, and condition at hospital discharge, were obtained from the epidemiological file of the Ministerio de Salud Pública. Results: In 5 years, 147 patients (29.4 per year) were hospitalized with no mortality. They corresponded to 26, 34, 32, 29 and 26 cases, in 2017, 2018, 2019, 2020 and 2021, respectively. Men with 99 (67.3%), aged 21-30 years with 28, mixed race with 94 cases, students, and farmers, were the most affected. The most affected (99; 67.3%) were men, people 21 to 30 years-old (28; 19.0%), mestizos (94; 63.9%), students, and farmers. The median age was 28 (range: 4-81) years. Prevalence was higher in April, June, and September. All accidents were caused by Viperidae snakes. Twenty (13.6%) cases were mild, (61.2%) were moderate and 37 (25.2%) were severe. The feet with 45 were the most bitten. Pre-hospitalization anti-venom serum was received by 53.1% and tourniquet by 19.8% patients. Median hospital arrival time was 5 (range 1-192) hours, mostly between 2-3 hours with 41 cases. No statistically significant differences were found considering the severity. Conclusions: A high prevalence of snakebites was evidenced in the north of the Amazon in Ecuador, with a higher incidence in the rainy season and all by Viperidae species. It is important to highlight the null mortality. Information campaigns on prevention and first aid, such as discouraging the use of tourniquets, especially among vulnerable groups.

Colistin resistance in Escherichia coli and Klebsiella pneumoniae in humans and backyard animals in Ecuador / Resistencia a la colistina de las bacterias Escherichia coli y Klebsiella pneumoniae en humanos y animales de granja en Ecuador / Resistência à colistina em Escherichia coli e Klebsiella pneumoniae em humanos e animais de quintal no Equador

ABSTRACT Objective. Colistin is an antibiotic of last resort for treating serious Gram-negative bacterial infections. However, the misuse of colistin, especially as an animal growth promoter, has contributed to increasing antimicrobial resistance, mediated mainly through plasmid transfer of the mcr-1 gene. This study assessed the prevalence of phenotypic and molecular colistin resistance in Escherichia coli and Klebsiella pneumoniae in Ecuador in healthy humans and their chickens and pigs. Methods. Fecal samples were collected from humans and their chickens and pigs in two rural coastal and Amazon regions between April and August 2020. Gram-negative bacteria were isolated and identified using conventional techniques. Phenotypic resistance was determined using the broth microdilution technique, and the mcr-1 gene was detected using conventional polymerase chain reaction. Results. A total of 438 fecal samples were obtained from 137 humans, 147 pigs and 154 chickens. The prevalence of E. coli isolates was 86.3% (378/438) and K. pneumoniae, 37.4% (164/438). Overall, the mcr-1 gene was found in 90% (340/378) of E. coli isolates, with higher prevalences found in isolates from coastal regions (96.5%, 191/198), humans (95.6%, 111/116) and chickens (91.8%, 123/134); for K. pneumoniae, the gene was found in 19.5% (32/164) of isolates, with equal distribution between regions and hosts. Only four isolates, two E. coli and two K. pneumoniae, showed phenotypic resistance: mcr-1 was present in both E. coli strains but absent in the K. pneumoniae strains. Conclusions. Despite a low prevalence of phenotypic resistance to colistin, the high prevalence of the mcr-1 gene in E. coli is of concern. Ecuador's ban on using colistin in animal husbandry must be enforced, and continual monitoring of the situation should be implemented.

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RESUMO Objetivo. A colistina é um antibiótico de último recurso para o tratamento de infecções graves por bactérias Gram-negativas. Entretanto, o uso indevido da colistina, principalmente como promotor de crescimento animal, tem contribuído para o aumento da resistência a antimicrobianos, principalmente por transferência horizontal do gene mcr-1 mediada por plasmídeos. Este estudo avaliou a prevalência de resistência fenotípica e molecular à colistina em Escherichia coli e Klebsiella pneumoniae no Equador em humanos hígidos e em galinhas e porcos por eles criados. Métodos. Entre abril e agosto de 2020, foram coletadas amostras de fezes de habitantes de duas regiões litorâneas e amazônicas do Equador e de galinhas e porcos por eles criados. Bactérias Gram-negativas foram isoladas e identificadas por meio de técnicas convencionais. A resistência fenotípica foi determinada pela técnica de microdiluição em caldo, e o gene mcr-1 foi detectado por reação em cadeia da polimerase convencional. Resultados. Foram obtidas 438 amostras fecais de 137 humanos, 147 suínos e 154 galinhas. A prevalência de isolados de E. coli foi de 86,3% (378/438), e de K. pneumoniae, 37,4% (164/438). Em geral, o gene mcr-1 foi encontrado em 90% (340/378) dos isolados de E. coli, com maiores prevalências encontradas em isolados de regiões litorâneas (96,5%, 191/198), humanos (95,6%, 111/116) e galinhas (91,8%, 123/134); para K. pneumoniae, o gene foi encontrado em 19,5% (32/164) dos isolados, com igual distribuição entre regiões e hospedeiros. Somente quatro isolados, dois de E. coli e dois de K. pneumoniae, demonstraram resistência fenotípica: o gene mcr-1 estava presente em ambas as cepas de E. coli, mas ausente nas de K. pneumoniae. Conclusões. Apesar da baixa prevalência de resistência fenotípica à colistina, a alta prevalência do gene mcr-1 em E. coli é preocupante. É preciso fiscalizar a proibição ao uso agropecuário de colistina no Equador e implementar o monitoramento contínuo da situação.

Extended-Spectrum Beta-Lactamases Producing Escherichia coli in South America: A Systematic Review with a One Health Perspective.

Purpose: Extended-spectrum beta-lactamase-producing (ESBL) Enterobacteriaceae, which includes Escherichia coli, has emerged as a global health threat. ESBL enzymes including CTX-M, TEM, and SHV are the most detected. Here, a systematic review was developed to assess the status of ESBLs in E. coli considering studies performed in the human, animal, food, and environmental realms in South America. Methods: Following PRISMA guidelines, a systematic review was performed using the PubMed database as a primary source to identify studies containing data on ESBL-producing E. coli in South America. To obtain a comprehensive sample, studies in English, Spanish, and Portuguese were included from 1990 to April 2021. Inclusion such as the reporting of sample origin and diagnostic method and exclusion criteria such as review/letter articles were established to complete data extraction steps. Results: Amongst 506 articles retrieved, 130 met the inclusion criteria. Brazil reported 65 (50%) of publications, followed by Argentina, and Ecuador with 11.5% each. According to the category of studies, human studies represented the 56%, animals the 20%, environmental the 11%, and food studies the 6%. Interestingly, studies assessing more than one category (ie, interdisciplinary) represented the 7%. Prevalence of ESBL producing E. coli in animal, food, and environmental studies was widely superior compared to human sources. In clinical studies, Brazil presented the greatest diversity in terms of ESBLs, featuring CTX-M, TEM, SHV, TOHO, OXA, and AmpC. CTX-M enzymes were the most frequent variants with 89.4% detections. Conclusion: The present One Health review of 130 studies conducted over the past 21 years found ESBLs producing E. coli distributed across human, animal, food, and environmental samples across South America. There is a need to increment studies in underrepresented countries and to strengthen multi-sectoral antimicrobial resistance research and surveillance. This information can be used as basis for subsequent implementation of monitoring programs, targeting potential critical points of transmission sources.

Accessibility Analysis of Worldwide COVID-19-Related Information Portals.

Since the beginning of the COVID-19 pandemic, communication technology has demonstrated its usefulness in sharing and receiving health data and communicating with the public. This study evaluated the accessibility of 199 websites containing official COVID-19 information related to medical schools, governments, ministries, and medical associations, obtained from the Geneva Foundation for Medical Education and Research website. We used the Web Content Accessibility Guidelines 2.1 to evaluate web accessibility, using a six-phase process with an automatic review tool. The study results reveal that the highest number of barriers encountered are concentrated in the perceivable principle with 6388 errors (77.8%), followed by operability with 1457 (17.7%), then robustness with 291 (3.5%), and finally understandability with 78 errors (0.9%). This study concludes that most COVID-19-related websites that provide information on the context of the pandemic do not have an adequate level of accessibility. This study can contribute as a guide for designing inclusive websites; web accessibility should be reviewed periodically due to technological advances and the need to adapt to these changes.