Presence of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Food-Producing and Companion Animals and Wildlife on Small-Holder Farms of Floreana Island, Galápagos Islands.

Background: Antimicrobial resistance (AR) has led to increasing human and animal morbidity and mortality and negative consequences for the environment. AR among Escherichia coli (EC) is on the rise, with serious concerns about extended-spectrum ß-lactamase-producing E. coli (ESBL-EC). In the Galápagos Islands, where antimicrobials are available without a prescription, growing demands for food production can drive antimicrobial use. Food producing animals are at the interface of wildlife and environmental health on the smallest human-inhabited Galápagos Island, Floreana. We sought to determine if ESBL-EC were present in Floreana Island farm animal species and nearby wildlife and the relatedness of ESBL-EC isolates identified. Materials and Methods: During July 4-5, 2022, we visited 8 multispecies farms, representing 75% of food-producing animal production on Floreana, and collected 227 fecal samples from farm animals and wildlife. Each sample was plated on MacConkey agar supplemented with cefotaxime (4 µg/mL). Results: ESBL-EC was isolated from 20 (9%) fecal samples collected from pigs (N = 10), chickens (N = 6), wildlife (N = 3), and dog (N = 1). All ESBL-EC isolates were from samples taken at three (38%) of the eight farms. Fifteen (75%) of the ESBL-EC isolates were from a single farm. All ESBL-EC isolates were multidrug resistant. The most prevalent ESBL genes belonged to the blaCTX-M group. Among the typeable isolates from the farm with the largest proportion of ESBL-EC isolates (N = 14), we observed nine unique pulsed-field gel electrophoresis (PFGE) patterns, with identical patterns present across pig and chicken isolates. PFGE patterns in the three farms with ESBL-EC isolates were different. Conclusions: These results lend support for future routine AR monitoring activities at the livestock-wildlife interface in Galápagos to characterize potential interspecies transmission of AR bacteria and AR genes in this unique protected ecosystem, and the related human, animal, and environmental health impacts, and to formulate interventions to reduce AR spread in this setting.

Massive testing in the Galapagos Islands and low positivity rate to control SARS-CoV-2 spread during the first semester of the COVID-19 pandemic: a story of success for Ecuador and South America.

INTRODUCTION: During the first months of the COVID-19 pandemic in Latin America, countries like Ecuador, Peru and Colombia experienced chaotic scenarios with public health systems collapsing and lack of testing capacity to control the spread of the virus. In main cities like Guayaquil in Ecuador, dramatic situations such as corpses in the streets were internationally broadcasted. METHODS: While the COVID-19 pandemic was devastating South America, SARS-CoV-2 transmission was successfully managed in the Galapagos Islands due to the implementation of a massive screening strategy including hospitalized and community-dwelling populations, and travel restrictions facilitated by its geographical location (972 km from the Ecuadorian continental territory). Floreana Island was one of the few locations in the world that remained COVID-19 free during 2020. RESULTS: In this study, we retrospectively analyzed the data related to SARS-CoV-2 massive testing campaigns from April to September 2020 in the Galapagos Islands, and found this territory to have the lowest positivity rate in South America (4.8-6.7%) and the highest testing ratio among Ecuadorian provinces (9.87% of the population, which is 2480 out of 25 124 inhabitants) during the first wave of the COVID-19 pandemic. CONCLUSION: This story of success was possible because of the interinstitutional collaboration between the regional government of Galapagos Islands (Consejo de Gobierno), the local authorities (Gobiernos Autonomos Descentralizados de Santa Cruz, San Cristobal and Isabela), the regional authorities from Ecuadorian Ministry of Health, the Agencia de Regulación y Control de la Bioseguridad y Cuarentena para Galápagos and Universidad de Las Américas.

Assessing Livestock Production Practices on Small-Scale Multi-Species Farms Located on Floreana Island, Galápagos Islands.

Globally to date, established international standards for animal welfare, a priority of sustainable agriculture, have primarily focused on large-scale producers. However, across Latin America, including in Ecuador's Galápagos Islands, smallholder farms play a critical role in food safety and security. We assessed five basic animal welfare measures (feed and water access, shelter availability and housing systems, animal health management, animal behavior, and timely euthanasia) for poultry, pigs, and cattle on Floreana Island, Galápagos. Utilizing assessment standards from multiple US sources and international standards, we developed a questionnaire and used it to conduct in-depth interviews during 4-5 July 2022 with eight participating producers, representing 75% of animal agriculture on Floreana. While we identified opportunities to enhance competencies in animal health management and timely euthanasia, farms performed well in the other assessed measures. Future work should promote knowledge transfer and in-country capacity building in farm biosecurity, access to veterinary care, antimicrobial resistance surveillance, and euthanasia methods. Efforts to positively impact smallholder farm livelihoods in Galápagos-one of the most biodiverse and protected ecosystems on the planet-will sustainably support human health through the interconnected realms of animal health and welfare, wildlife and environmental health, and food safety and security.

Differential aggregation of polystyrene and titanium dioxide nanoparticles under various salinity conditions and against multiple proteins types.

The interaction of nanoplastics (NPls) and engineered nanoparticles (ENPs) with organic matter and environmental pollutants is particularly important. Therefore, their behavior should be investigated under the different salinity conditions, mimicking rivers and coastal environments, to understand this phenomenon in those areas. In this work, we analyzed the elementary characteristics of polystyrene-PS (unmodified surface and modified with amino or carboxyl groups) and titanium dioxide-TiO2 nanoparticles. The effect of salinity on their colloidal properties was studied too. Also, the interaction with different types of proteins (bovine serum albumin-BSA and tilapia proteins), as well as the formation of the BSA corona and its effect on the colloidal stability of nanoparticles, were evaluated. The morphology and dispersion of sizes were more uniform in unmodified-surface PS-NPs (70.5 ± 13.7 nm) than in TiO2-NPs (131.2 ± 125.6 nm). Likewise, Rama spectroscopy allowed recognizing peaks associated with the PS phenyl group aromatic ring in unmodified-surface PS-NPs (621, 1002, 1582, and 1602 cm-1). For TiO2-NPs, the data suggest belonging to the tetragonal form, also known as rutile (445, 610 cm-1). The elevation of salinity dose-dependently decreased NP colloid stability, with more significant variation in the PS-NPs compared to TiO2-NPs. The organic matter is also involved in this phenomenon, differentially as a function of time compared to its absence (unmodified-surface PS-NPs 30 psu/TOC 5 mgL-1/24 h: 2876.6 ± 378.03 nm; unmodified-surface PS-NPs 30 psu/24 h: 2133 ± 49.57 nm). In general, the TiO2-NPs demonstrated greater affinity with all proteins tested (0.066 g/L). It was observed that morphology, size, and surface chemical modification intervene in a relevant way in the interaction of the nanoparticles with bovine serum albumin (unmodified-surface PS-NPs 298 K: 6.08E+02; 310 K: 6.63E+02; TiO2-NPs 298 K: 8.76E+02; 310 K: 1.05E+03 L mol-1) and tilapia tissues proteins (from blood, gills, liver, and brain). Their morphology and size also determined the protein corona formation and the NPs' agglomeration. These findings can provide references during knowledge transfer between NPls and ENPs.

High sensitivity-low cost detection of SARS-CoV-2 by two steps end point RT-PCR with agarose gel electrophoresis visualization.

More than one year since Coronavirus disease 2019 (COVID-19) pandemic outbreak, the gold standard technique for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection is still the RT-qPCR. This is a limitation to increase testing capacities, particularly at developing countries, as expensive reagents and equipment are required. We developed a two steps end point RT-PCR reaction with SARS-CoV-2 Nucleocapsid (N) gene and Ribonuclease P (RNase P) specific primers where viral amplicons were verified by agarose gel electrophoresis. We carried out a clinical performance and analytical sensitivity evaluation for this two-steps end point RT-PCR method with 242 nasopharyngeal samples using the CDC RT-qPCR protocol as a gold standard technique. With a specificity of 95.8%, a sensitivity of 95.1%, and a limit of detection of 20 viral RNA copies/uL, this two steps end point RT-PCR assay is an affordable and reliable method for SARS-CoV-2 detection. This protocol would allow to extend COVID-19 diagnosis to basic molecular biology laboratories with a potential positive impact in surveillance programs at developing countries.

Corrigendum to: "One health" inspired SARS-CoV-2 surveillance: The Galapagos Islands experience [One Health 11 (2020) 100185].

[This corrects the article DOI: 10.1016/j.onehlt.2020.100185.].

Analytical and clinical comparison of Viasure (CerTest Biotec) and 2019-nCoV CDC (IDT) RT-qPCR kits for SARS-CoV2 diagnosis.

BACKGROUND: Several RT-qPCR kits are available for SARS-CoV-2 diagnosis, some of them with Emergency Use Authorization (EUA) by FDA, but most of them lacking of proper evaluation studies due to covid19 emergency. OBJECTIVE: We evaluated Viasure RT-qPCR kit (CerTest Biotec, Spain) for SARS-CoV-2 diagnosis using FDA EUA 2019-nCoV CDC kit (IDT, USA) as a gold standard. RESULTS: Although we found the lack of RNA quality control probe as the main limitation for the Viasure kit, the sensitivity was 91.9% and the specificity was 100%. The limit of detection (LOD) was 2000 copies/mL and 1000 copies/mL for Viasure and IDT kits, respectively. CONCLUSIONS: Viasure RT-qPCR kit is a reliable tool for SARS-CoV-2 diagnosis but improvement of an alternative RT-qPCR reaction for RNA extraction quality control as RNaseP is recommended.

"One health" inspired SARS-CoV-2 surveillance: The Galapagos Islands experience.

The COVID-19 pandemic has spread worldwide since the outbreak in Wuhan (China) in December 2019, currently infecting over 25 million people and causing more than 800.000 deaths. In Ecuador, up to the 30th of August 2020, overall 113.648 confirmed cases and 6.555 deaths have been declared. Besides overloading of hospital, capacity for molecular diagnosis of SARS-CoV-2 by the Ministry of Public Health was quickly overwhelmed. In this context, emergency authorization for SARS-CoV-2 RT-qPCR diagnosis was granted to other institutions, like the "Agencia de Regulación y Control para la Bioseguridad y Cuarentena de Galápagos" (ABG), that implemented "LabGal" with support of One Health Research Group from "Universidad de Las Américas" (UDLA). The previous experience of ABG and the One Health Research Group in conducting massive surveillance of zoonotic diseases on livestock was crucial to the success on the control of COVID-19 outbreak at Galapagos Islands by the end of May 2020, when Latin American countries were leading the spread of the pandemic.

"Sample pooling of RNA extracts to speed up SARS-CoV-2 diagnosis using CDC FDA EUA RT-qPCR kit".

BACKGROUND: The CDC protocol for SARS-CoV2 RT-PCR diagnosis (2019-nCoV CDC kit) is considered a gold standard worldwide; based on three different FAM probes (N1 and N2 for viral detection; RP for RNA extraction quality control), three reactions per sample are needed for SARS-CoV-2 diagnosis. RESULTS: We herein describe a sample pooling protocol: pooling 3 RNA extractions into a single PCR reaction; we tested this protocol with 114 specimens grouped in 38 pools and found no significant differences for N1 and N2 Ct values between pool and single sample PCR reaction. CONCLUSION: This pool of three protocol has a sensitivity of 100 % compared to the standard single sample protocol. For a typical 96-well plate, this pool assay allows 96 samples processing, speeding up diagnosis and reducing cost while maintaining clinical performance, particularly useful for SARS-CoV-2 diagnosis at developing countries.

Cotton-Tipped Plastic Swabs for SARS-CoV-2 RT-qPCR Diagnosis to Prevent Supply Shortages.

CDC and WHO guidelines for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis only recommend synthetic fiber swabs for nasopharyngeal (NP) sampling. We show that cotton-tipped plastic swabs do not inhibit PCR and have equivalent performance to rayon swabs. Cotton-tipped plastic swabs are massively produced worldwide and would prevent swab supply shortages under the current high SARS-CoV-2 testing demands, particularly in developing countries.

Evaluation of nCoV-QS (MiCo BioMed) for RT-qPCR detection of SARS-CoV-2 from nasopharyngeal samples using CDC FDA EUA qPCR kit as a gold standard: An example of the need of validation studies.

BACKGROUND: Several qPCR kits are available for SARS-CoV-2 diagnosis, mostly lacking of evaluation due to covid19 emergency. OBJECTIVE: We evaluated nCoV-QS (MiCo BioMed) kit using CDC kit as gold standard. RESULTS: We found limitations for nCoV-QS: 1) lower sensitivity 2) lack of RNA quality control probe. CONCLUSIONS: Validation studies should be implemented for any SARS-CoV-2 RT-qPCR commercial kit to prevent unreliable diagnosis.

Fisioterapia en la parálisis facial / Physiotherapy in the facial paralysis

Introducción: La parálisis facial involucra el VII para craneal o facial y puede ser completa o parcial presentándose en forma unilateral. Signos y síntomas: los signos y síntomas son variados y a menudo se relaciona con la exposición a una infección viral. Diagnóstico: se basa en la existencia de hallazgos en tres aspectos: clínico, topográfico y eléctrico. Tratamiento: se basa en: a) tipo de la parálisis; b) etiología y naturaleza de la lesión causal; c) lugar donde se ha producido la lesión; d) intensidad o magnitud de la lesión y de la parálisis; e) tiempo transcurrido entre la iniciación de la parálisis y el examen del enfermo. Conclusiones: El tratamiento fisioterapéutico tendrá en cuenta sensibilidad, fuerza muscular y sincinesias. Dentro de las técnicas de tratamiento usadas se encuentran: tratamiento térmico, drenaje linfático manual, acupuntura, electroterapia, estiramiento, estimulación multisensorial, vendaje neuromuscular y reeducación muscular.

Biogeography of Parasitic Nematode Communities in the Galápagos Giant Tortoise: Implications for Conservation Management.

The Galápagos giant tortoise is an icon of the unique, endemic biodiversity of Galápagos, but little is known of its parasitic fauna. We assessed the diversity of parasitic nematode communities and their spatial distributions within four wild tortoise populations comprising three species across three Galápagos islands, and consider their implication for Galápagos tortoise conservation programmes. Coprological examinations revealed nematode eggs to be common, with more than 80% of tortoises infected within each wild population. Faecal samples from tortoises within captive breeding centres on Santa Cruz, Isabela and San Cristobal islands also were examined. Five different nematode egg types were identified: oxyuroid, ascarid, trichurid and two types of strongyle. Sequencing of the 18S small-subunit ribosomal RNA gene from adult nematodes passed with faeces identified novel sequences indicative of rhabditid and ascaridid species. In the wild, the composition of nematode communities varied according to tortoise species, which co-varied with island, but nematode diversity and abundance were reduced or altered in captive-reared animals. Evolutionary and ecological factors are likely responsible for the variation in nematode distributions in the wild. This possible species/island-parasite co-evolution has not been considered previously for Galápagos tortoises. We recommend that conservation efforts, such as the current Galápagos tortoise captive breeding/rearing and release programme, be managed with respect to parasite biogeography and host-parasite co-evolutionary processes in addition to the biogeography of the host.

Sequence analysis of the ADRA2A coding region in children affected by attention deficit hyperactivity disorder.

Attention deficit hyperactivity disorder (ADHD) is a common neurobehavioral pathology characterized by distinct degrees of inattention, hyperactivity and impulsivity. Although ADHD etiology remains elusive, the ADRA2A candidate gene underlies a particular interest, since it participates in the prefrontal cortex regulation of executive function. Three SNPs located on 5' and 3'UTR regions of the gene have been extensively explored but none of them have been definitely validated as a predisposition or a causative sequence variation. In this study, in order to determine whether ADRA2A non-synonymous sequence variants, resulting in biochemical modifications of the protein, are a common cause of the disease we sequenced the complete ADRA2A coding region in a panel of ADHD children of Colombian origin. We identified the c.1138 C>A (p.Arg380Arg) silent substitution. We conclude that ADRA2A non-synonymous sequence variants do not cause ADHD in our sample population. We cannot formerly discard a potential role of this gene during ADHD pathogenesis since only the coding region was analysed. We hope that these results will encourage further researchers to sequence the promoter and coding regions of ADRA2A in large panels of ADHD patients from distinct ethnical origins.

Complejo esclerosis tuberosa / Tuberous sclerosis complex

INTRODUCCIÓN: El complejo esclerosis tuberosa (TSC) es el prototipo de malformación relacionada con alteraciones en la diferenciación y el crecimiento celular de fenotipos variables con implicaciones sistémicas. OBJETIVO: caracterizar y describir el TSC para facilitar el entendimiento de las neurodermatosis en la población Colombiana. MATERIALES Y MÉTODOS:la información fue extraída a partir de búsquedas practicadas en las bases de datos MEDLINE, en BIOSIS y en EMBASE desde 1966, 1994 y 1974, respectivamente, hasta el 1 de Marzo de 2011, usando la plataforma OVID y múltiples términos clave. Se diseñaron estrategias adicionales para las bases de datos LILACS, Best Evidence y CINHAL con criterios similares. También se recopilaron datos obtenidos de sociedades científicas relacionadas con el tema en estudio. RESULTADOS:los criterios diagnósticos han permanecido inmodificados a pesar de que el estudio molecular permite caracterizar la enfermedad. El entendimiento de la patogénesis del TSC ha favorecido el desarrollo de opciones terapéuticas contra blancos moleculares que se encuentran alterados. La vía mTOR explica la mayor parte de las alteraciones fenotípicas en pacientes con TSC. CONCLUSIÓN:El documento del Consorcio Colombiano para el Estudio de las Neurodermatosis muestra una revisión integral del conocimiento respecto del diagnóstico y tratamiento del TSC.

INTRODUCTION: The Tuberous Sclerosis Complex (TSC) is the mainstay of malformation related to alterations in cellular growth and differentiation, with highly variable phenotypes, and systemic impact. OBJETIVES: To characterize and describe TSC to facilitate understanding of the neurodermatosis in the Colombian population. MATERIALS AND METHODS: Information was obtained from searches performed in MEDLINE, BIOSIS, and EMBASE from 1966, 1994 and 1974, respectively, until March 1, 2011, using the OVID platform and multiple key terms. Additional strategies were designed for other databases like LILACS, CINAHL and Best Evidence using similar criteria. We also collected data from scientific societies related to the topic under study. RESULTS: the diagnostic criteria have not been modified instead molecular characterizations leads to a better understanding of the disease. The characterization of pathogenesis leads to the development of target therapies against those molecular alterations. mTOR pathway explains phenotypic alterations in patients suffering of TSC. CONCLUSION: the document of the Colombian Consensus for the Neurodermathosis, wants to show an integral review of the knowledge about the development, diagnosis and treatment of TSC.

Epilepsy in Colombia: epidemiologic profile and classification of epileptic seizures and syndromes.

PURPOSE: A national study was performed in Colombia to determine the general and regional prevalence of epilepsy, clinical profiles, seizure types, and clinical syndromes. METHODS: Based on the National Epidemiological Study of Neurological Diseases (EPINEURO), we evaluated and followed up for 1 year all the subjects with epilepsy from the National Sample. Clinical profiles were further assessed. Seizure types and epilepsy syndromes were established according to the international classifications. RESULTS: General prevalence was found to be 11.3 per 1,000, with little variation among regions, except the eastern region, where prevalence was 23 per 1,000; prevalence for active epilepsy was 10.1 per 1,000. Women have a slightly greater (not statistically significant) risk. Most seizures are focal (partial), frequently with secondary generalization. The most frequent epilepsy syndrome encountered was partial symptomatic/cryptogenic (80%). Epilepsy onset in Colombia occurs most frequently in childhood. CONCLUSIONS: Prevalence rates of epilepsy in Colombia are similar to those reported in nations with comparable developmental status and have diminished over time. The study presents the distribution of seizures and syndromes. The most frequent types are focal syndromes.

Síndrome de ovario poliquístico y ácido valproico / Polycystic ovary syndrome and valproic acid

Objetivo: revisar y presentar resultados actualizados sobre la relación entre epilepsia tratada con ácido valproico y el síndrome de ovario poliquístico. Desarrollo: se hace una revisión de la literatura publicada para tener un mejor entendimiento de la relación entre los trastornos endocrinos, la epilepsia y los anticonvulsivantes. Se debe atender de manera especial a aquellas pacientes obesas que reciben valproato o aquellas que experimentan un aumento desmesurado de peso. Las alteraciones aisladas de laboratorios o las imágenes diagnósticas sin síntomas, no han de conformar un trastorno endocrino de importancia clínica. Conclusiones: si se encuentra una paciente con síndrome de ovario poliquístico, debe evaluarse el tratamiento anticonvulsivante para definir el más indicado, según el tipo de crisis, y asegurarse de que el tratamiento no contribuya al trastorno endocrino. El beneficio de cambiar el tratamiento debe ser analizado frente a la efectividad en términos del control de crisis y efectos secundarios de las otras alternativas terapéuticas