Uropathogenic E. coli and Hybrid Pathotypes in Mexican Women with Urinary Tract Infections: A Comprehensive Molecular and Phenotypic Overview.

Uropathogenic Escherichia coli (UPEC) is the main cause of urinary tract infections (UTIs) and carries virulence and resistance factors often found in mobilizable genetic elements, such as plasmids or pathogenicity islands (PAIs). UPEC is part of the extraintestinal pathogenic E. coli (ExPEC), but hybrid strains possessing both diarrheagenic E. coli (DEC) and ExPEC traits, termed "hypervirulent", present a significant health threat. This study assessed the prevalence of UPEC PAIs, ExPEC sequence types (ST), DEC genes, carbapenemase and extended-spectrum ß-lactamase (ESBL) phenotypes, resistance genotypes, and plasmids in 40 clinical isolates of UPEC. Results showed that 72.5% of isolates had PAIs, mainly PAI IV536 (53%). ESBL phenotypes were found in 65% of ß-lactam-resistant isolates, with 100% of carbapenem-resistant isolates producing carbapenemase. The predominant ESBL gene was blaCTX-M-2 (60%), and the most common resistance gene in fluoroquinolone and aminoglycoside-resistant isolates was aac(6')Ib (93%). Plasmids were present in 57% of isolates, and 70% belonged to the ST131 clonal group. Molecular markers for DEC pathotypes were detected in 20 isolates, with 60% classified as hybrid pathotypes. These findings indicate significant pathogenic potential and the presence of hybrid pathotypes in E. coli UTI clinical isolates in the Mexican population.

First Report of the Emerging Pathogen Kodamaea ohmeri in Honduras.

Kodamaea ohmeri is an environmental yeast considered a rare emerging pathogen. In clinical settings, the correct identification of this yeast is relevant because some isolates are associated with resistance to antifungals. There is a lack of available data regarding the geographical distribution, virulence, and drug resistance profile of K. ohmeri. To contribute to the knowledge of this yeast, this study aimed to describe in depth three isolates of K. ohmeri associated with fungemia in Honduras. The identification of the isolates was carried out by sequencing the ribosomal ITS region. In addition, the susceptibility profile to antifungals was determined, and some properties associated with virulence were evaluated (exoenzyme production, biofilm formation, cell adhesion, and invasion). The isolates showed strong protease, phospholipase, and hemolysin activity, in addition to being biofilm producers. Adherence and invasion capacity were evident in the HeLa and Raw 264.7 cell lines, respectively. This study expands the understanding of the underlying biological traits associated with virulence in K. ohmeri, and it is the first report of the detection and identification of K. ohmeri in Honduras as a cause of human infection.

PCR-RFLP assays for the identification of Anopheles (Diptera: Culicidae) species circulating in Honduras.

BACKGROUND: Vector populations are a key target for malaria control and elimination. In Honduras, there are at least 12 reported anopheline species, however, the definitive number of species remains uncertain. Due to the inherent limitations of morphological identification of Anopheles species, molecular approaches have been developed to provide accurate identification and robust surveillance of local malaria vectors. The aim of this study was to design and assess three PCR-RFLP assays to identify anopheline species known to presently occur in Honduras. METHODS: Mosquitoes captured between 2018 and 2022 in seven malaria-endemic and non-endemic departments in Honduras were analysed. The ITS2 ribosomal region and three restriction enzyme-based assays were evaluated in silico and experimentally. RESULTS: A total of 132 sequences from 12 anopheline species were analysed. The ITS2 marker showed length polymorphisms that generated products between 388 and 592 bp and no relevant intraspecies polymorphisms were found. Furthermore, the three PCR-RFLP assays were able to differentiate 11 species with sufficient precision and resolution. CONCLUSION: The ITS2 region was shown to be a useful molecular marker for identifying local Anopheles species. In addition, the PCR-RFLP assays evaluated here proved to be capable of discriminating most of the anopheline species present in Honduras. These methods provide alternatives to improve entomological surveillance of Anopheles in Honduras and other Mesoamerican countries.

A PCR-RFLP Technique to Assess the Geographic Origin of Plasmodium falciparum Strains in Central America.

The elimination of malaria requires strengthening diagnosis and offering adequate and timely treatment. Imported cases of falciparum malaria represent a major challenge for pre-elimination areas, such as Central America, where chloroquine and primaquine continue to be used as first-line treatment. The pfs47 gene has been previously described as a precise molecular marker to track the geographic origin of the parasite. The aim of this study was to design a simple and low-cost technique using the polymorphic region of pfs47 to assess the geographic origin of P. falciparum strains. A PCR-RFLP technique was developed and evaluated using the MseI enzyme that proved capable of discriminating, with reasonable precision, the geographical origin of the parasites. This method could be used by national surveillance laboratories and malaria elimination programs in countries such as Honduras and Nicaragua in cases of malaria where an origin outside the Central American isthmus is suspected.

Genetic Diversity among Four Populations of Aedes aegypti (Diptera: Culicidae) from Honduras as Revealed by Mitochondrial DNA Cytochrome Oxidase I.

Aedes aegypti is a hematophagous and highly anthropophilic mosquito with a wide distribution, particularly in tropical and subtropical regions of the world. Ae. aegypti is the main vector of several febrile diseases called arboviruses (dengue, yellow fever, chikungunya, and zika viruses), which represent an important public health problem. Populations of this mosquito were nearly eliminated from the Americas in the mid-20th century; however, after the abandonment of control measures, mosquito populations have been recovering territory, have expanded by anthropogenic mechanisms, and have been joined by new populations reintroduced from other continents. The objective of this pilot study was to determine the genetic variability of Aedes aegypti collected in four cities located along the so-called logistics corridor of Honduras, which connects the Caribbean Sea to the Pacific Ocean. We studied the sequences of two molecular markers: the cytochrome c oxidase 1 (COI mtDNA) gene and the internal transcribed spacer 2 (ITS2 rDNA) of 40 mosquitoes. Phylogenetic analyzes show two separate clades with a low number of nucleotide differences per site, three haplotypes, and low haplotype diversity. These results suggest a low genetic diversity in the populations of Ae. aegypti in Honduras in relation to that reported in other countries of the Central American isthmus.

Field Evaluation of a Hemozoin-Based Malaria Diagnostic Device in Puerto Lempira, Honduras.

The diagnosis of malaria in Honduras is based mainly on microscopic observation of the parasite in thick smears or the detection of parasite antigens through rapid diagnostic tests when microscopy is not available. The specific treatment of the disease depends exclusively on the positive result of one of these tests. Given the low sensitivity of conventional methods, new diagnostic approaches are needed. This study evaluates the in-field performance of a device (Gazelle™) based on the detection of hemozoin. This was a double-blind study evaluating symptomatic individuals with suspected malaria in the department of Gracias a Dios, Honduras, using blood samples collected from 2021 to 2022. The diagnostic performance of Gazelle™ was compared with microscopy and nested 18ssr PCR as references. The sensitivity and specificity of Gazelle™ were 59.7% and 98.6%, respectively, while microscopy had a sensitivity of 64.9% and a specificity of 100%. The kappa index between microscopy and Gazelle™ was 0.9216 using microscopy as a reference. Both methods show similar effectiveness and predictive values. No statistical differences were observed between the results of the Gazelle™ compared to light microscopy (p = 0.6831). The turnaround time was shorter for Gazelle™ than for microscopy, but the cost per sample was slightly higher for Gazelle™. Gazelle™ showed more false-negative cases when infections were caused by Plasmodium falciparum compared to P. vivax. Conclusions: The sensitivity and specificity of Gazelle™ are comparable to microscopy. The simplicity and ease of use of the Gazelle™, the ability to run on batteries, and the immediacy of its results make it a valuable tool for malaria detection in the field. However, further development is required to differentiate Plasmodium species, especially in those regions requiring differentiated treatment.

First Molecular Identification of Three Clinical Isolates of Fungi Causing Mucormycosis in Honduras.

Mucormycoses are rare but serious opportunistic fungal infections caused by filamentous organisms of the order Mucorales. Here we report the first molecular identification of Rhizopus oryzae (heterotypic synonym Rhizopus arrhizus), R. delemar, and Apophysomyces ossiformis as the etiological agents of three cases of severe mucormycosis in Honduras. Conventional microbiological cultures were carried out, and DNA was extracted from both clinical samples and axenic cultures. The ITS ribosomal region was amplified and sequenced. Molecular tools are suitable strategies for diagnosing and identifying Mucorales in tissues and cultures, especially in middle-income countries lacking routine diagnostic strategies.

Influence of mobility restrictions on air quality in the historic center of Cuenca city and its inference on the Covid-19 rate infections.

At the end of 2019 in Wuhan China city, the outbreak of the virus called SARS-CoV 2 was originated, which later became a pandemic. In Ecuador, patient zero arrived on February 14, 2020 and the first mobility restriction imposed by the Government occurred on Tuesday, March 17 of the same year. Throughout the confinement, vehicle mobility restrictions have been modified by government entities depending on the number of infected people. This article presents an air quality study in the historic center of Cuenca city as consequence of mobility changes caused by Covid-19, where a comparison of concentration levels of polluting gases of the first semester of 2018, 2019 and 2020 is made, that allow differentiating and identifying the influence of vehicular flow on air quality. It can also be verified how the decrease in vehicle mobility restrictions influenced the increase in the rate of daily infections. For the study, air quality data published by the public mobility company of the city of Cuenca (EMOV EP) and the communications issued by the Emergency Operations Committee (COE), before and during the confinement, were collected. The acquisition, classification, analysis and interpretation of the data obtained through machine learning techniques was carried out. It can be concluded that while mobility restrictions were more severe, air quality improved and infections rate of decrease. Obtaining that polluting gases such as NO2 and CO produced by vehicular traffic show correlations of 61% and 60% respectively, which means that after 15 days of lifting the restrictive measures, the pollutants increased as well as the number of infected.

Candida species isolated from clinical samples in a tertiary hospital in Honduras: Where is Candida auris?

Background and Purpose: Infections by emerging and multiresistant Candida species are becoming more frequent throughout the world. This study aimed to describe Candida species in different wards of a tertiary hospital in Honduras. Materials and Methods: The prevalence of species within the C. albicans complex was estimated using a molecular approach, and C. auris was investigated using a yeast pool-based DNA extraction method. In total, 328 yeast isolates were identified using phenotypic approaches. For the identification of species within the C. albicans complex, a molecular approach based on the size polymorphisms of the hpw1 gene was used. In addition, a technique was optimized based on DNA extraction in pools for the rapid identification of C. auris. Results: A total of 11 species of Candida were identified in the hospital wards. C. albicans showed the highest number of isolates (52.4%). Within the C. albicans complex, C. albicans sensu stricto was the most common, followed by C. dubliniensis. However, C. auris was not found. Conclusion: Reports on the distribution of Candida species in Honduras are limited; accordingly, the data from this study are of importance for a better understanding of their epidemiology. Moreover, a simple method was offered for the detection of C. auris that could help in its detection in low-resource settings.

High-Elevation Populations of Montane Grasshoppers Exhibit Greater Developmental Plasticity in Response to Seasonal Cues.

Populations of insects can differ in how sensitive their development, growth, and performance are to environmental conditions such as temperature and daylength. The environmental sensitivity of development can alter phenology (seasonal timing) and ecology. Warming accelerates development of most populations. However, high-elevation and season-limited populations can exhibit developmental plasticity to either advance or prolong development depending on conditions. We examine how diurnal temperature variation and daylength interact to shape growth, development, and performance of several populations of the montane grasshopper, Melanoplus boulderensis, along an elevation gradient. We then compare these experimental results to observed patterns of development in the field. Although populations exhibited similar thermal sensitivities of development under long-day conditions, development of high-elevation populations was more sensitive to temperature under short-day conditions. This developmental plasticity resulted in rapid development of high elevation populations in short-day conditions with high temperature variability, consistent with their observed capacity for rapid development in the field when conditions are permissive early in the season. Notably, accelerated development generally did not decrease body size or alter body shape. Developmental conditions did not strongly influence thermal tolerance but altered the temperature dependence of performance in difficult-to-predict ways. In sum, the high-elevation and season-limited populations exhibited developmental plasticity that enables advancing or prolonging development consistent with field phenology. Our results suggest these patterns are driven by the thermal sensitivity of development increasing when days are short early in the season compared to when days are long later in the season. Developmental plasticity will shape phenological responses to climate change with potential implications for community and ecosystem structure.

Publication Trends in Neglected Tropical Diseases of Latin America and the Caribbean: A Bibliometric Analysis.

(1) Background: Neglected tropical diseases (NTDs) have been overlooked on the global health agenda and in the priorities of national systems in low- and middle-income countries (LMICs). In 2012, the Sustainable Development Goals (SDGs) were created to ensure healthy lives and promoting well-being for all. This roadmap set out to accelerate work to overcome the global impact of NTDs. Almost a decade has passed since NTDs were re-launched as a global priority. Investment in research and development, as well as the production of scientific literature on NTDs, is expected to have increased significantly. (2) Methods: A bibliometric analysis of the scientific production of Latin America and the Caribbean (LAC) was carried out in relation to 19 endemic NTDs. These data were compared with the scientific production in malaria, tuberculosis, and HIV/AIDS. The database available from Thomson Reuters Web of Science (WoS) was used. In addition, the average annual growth percentage was calculated for each disease. (3) Results: In the last decade, the NTDs with the highest number of publications in the world were dengue and leishmaniasis. The United States was the most prolific country in the world in 15 out of 19 NTDs analyzed. In the LAC region, Brazil was the largest contributor for 16 of the 19 NTDs analyzed. Arboviral diseases showed the highest average annual growth. The number of publications for malaria, tuberculosis and HIV/AIDS was considerably higher than for NTDs. The contribution of most LAC countries, especially those considered to be LMICs, is inadequate and does not reflect the relevance of NTDs for the public health of the population. (4) Conclusions: This is the first bibliometric analysis to assess the trend of scientific documents on endemic NTDs in LAC. Our results could be used by decision makers both to strengthen investment policies in research and development in NTDs.

Identification of Cryptic Species of Four Candida Complexes in a Culture Collection.

Candida spp. are one of the most common causes of fungal infections worldwide. The taxonomy of Candida is controversial and has undergone recent changes due to novel genetically related species. Therefore, some complexes of cryptic species have been proposed. In clinical settings, the correct identification of Candida species is relevant since some species are associated with high resistance to antifungal drugs and increased virulence. This study aimed to identify the species of four Candida complexes (C. albicans, C. glabrata, C. parapsilosis, and C. haemulonii) by molecular methods. This is the first report of six cryptic Candida species in Honduras: C. dubliniensis, C. africana, C. duobushaemulonii, C. orthopsilosis, and C. metapsilosis, and it is also the first report of the allele hwp1-2 of C. albicans sensu stricto. It was not possible to demonstrate the existence of C. auris among the isolates of the C. haemulonii complex. We also propose a simple method based on PCR-RFLP for the discrimination of the multi-resistant pathogen C. auris within the C. haemulonii complex.

Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital.

Candida species are one of the most important causes of human infections, especially in hospitals and among immunocompromised patients. The correct and rapid etiological identification of yeast infections is important to provide adequate therapy, reduce mortality, and control outbreaks. In this study, Candida species were identified in patients with suspected fungal infection, and phenotypic and genotypic identification methods were compared. A total of 167 axenic fungal cultures and 46 clinical samples were analyzed by HardyCHROM®, MicroScan®(Omron Microscan Systems Inc, Renton, WA, USA), and PCR-RFLP (Restriction Fragment Length Polymorphisms). The species of the C. albicans complex were the most frequent, followed by C. tropicalis and C. glabrata. Less common but clinically relevant species of Candida were also isolated. The comparison between the three methods was concordant, especially for the most common Candida species. Fungal DNA amplification was successful in all clinical samples.

A herpesvirus transactivator and cellular POU proteins extensively regulate DNA binding of the host Notch signaling protein RBP-Jκ to the virus genome.

Reactivation of Kaposi's sarcoma-associated herpesvirus (KSHV) from latency requires the viral transactivator Rta to contact the host protein Jκ recombination signal-binding protein (RBP-Jκ or CSL). RBP-Jκ normally binds DNA sequence-specifically to determine the transcriptional targets of the Notch-signaling pathway, yet Notch alone cannot reactivate KSHV. We previously showed that Rta stimulates RBP-Jκ DNA binding to the viral genome. On a model viral promoter, this function requires Rta to bind to multiple copies of an Rta DNA motif (called "CANT" or Rta-c) proximal to an RBP-Jκ motif. Here, high-resolution ChIP/deep sequencing from infected primary effusion lymphoma cells revealed that RBP-Jκ binds nearly exclusively to different sets of viral genome sites during latency and reactivation. RBP-Jκ bound DNA frequently, but not exclusively, proximal to Rta bound to single, but not multiple, Rta-c motifs. To discover additional regulators of RBP-Jκ DNA binding, we used bioinformatics to identify cellular DNA-binding protein motifs adjacent to either latent or reactivation-specific RBP-Jκ-binding sites. Many of these cellular factors, including POU class homeobox (POU) proteins, have known Notch or herpesvirus phenotypes. Among a set of Rta- and RBP-Jκ-bound promoters, Rta transactivated only those that also contained POU motifs in conserved positions. On some promoters, POU factors appeared to inhibit RBP-Jκ DNA binding unless Rta bound to a proximal Rta-c motif. Moreover, POU2F1/Oct-1 expression was induced during KSHV reactivation, and POU2F1 knockdown diminished infectious virus production. Our results suggest that Rta and POU proteins broadly regulate DNA binding of RBP-Jκ during KSHV reactivation.

Genetic variability of Plasmodium falciparum histidine-rich proteins 2 and 3 in Central America.

BACKGROUND: Malaria is an important disease in many tropical countries. Rapid diagnostic tests (RDTs) are valuable tools for diagnosing malaria in remote areas. The majority of RDTs used for the diagnosis of Plasmodium falciparum are based on the detection of the specific histidine-rich proteins (PfHRP2 and PfHRP3). During the last decade, the threat posed by the lack of expression of these antigens and the variability of the proteins on the diagnosis of malaria has been widely discussed. The aim of this study was to evaluate the genetic diversity of pfhrp2 and pfhrp3 of P. falciparum isolates collected in three Central American countries. METHODS: DNA samples were amplified and sequenced to assess the diversity of nucleotides and amino acids. A search for known epitopes within the amino acid sequence was carried out, and the sensitivity of the sequences was evaluated according to a predictive model. A phylogenetic analysis was carried out including homologous sequences from different regions of the world. Protein structures were predicted in silico. RESULTS: Five different patterns for PfHRP2 and one pattern for PfHRP3 were identified. Isolates from Central America show a high level of genetic diversity in pfhrp2; however, the amino acid sequences seem to contain enough motifs to be detected by the RDTs currently available. CONCLUSION: It is unlikely that the variability of the pfhrp2 and pfhrp3 genes has a significant impact on the ability of the RDTs to detect the PfHRP antigens in Central America.

Deletions of pfhrp2 and pfhrp3 genes of Plasmodium falciparum from Honduras, Guatemala and Nicaragua.

BACKGROUND: Malaria remains a public health problem in some countries of Central America. Rapid diagnostic tests (RDTs) are one of the most useful tools to assist in the diagnosis of malaria in remote areas. Since its introduction, a wide variety of RDTs have been developed for the detection of different parasite antigens. PfHRP2 is the most targeted antigen for the detection of Plasmodium falciparum infections. Genetic mutations and gene deletions are important factors influencing or affecting the performance of rapid diagnostic tests. METHODS: In order to demonstrate the presence or absence of the pfhrp2 and pfhrp3 genes and their flanking regions, a total of 128 blood samples from patients with P. falciparum infection from three Central American countries were analysed through nested or semi-nested PCR approaches. RESULTS: In total, 25.8 and 91.4% of the isolates lacked the region located between exon 1 and exon 2 of pfhrp2 and pfhrp3 genes, respectively. Parasites from the three countries showed deletions of one or both genes. The highest proportion of pfhrp2 deletions was found in Nicaragua while the isolates from Guatemala revealed the lowest number of pfhrp2 deletions. Parasites collected from Honduras showed the highest proportion of phfrp3 absence (96.2%). Twenty-one percent of isolates were double negative mutants for the exon 1-2 segment of both genes, and 6.3% of isolates lacked the full-length coding region of both genes. CONCLUSIONS: This study provides molecular evidence of the existence of P. falciparum isolates lacking the pfhrp2 and pfhrp3 genes, and their flanking regions, in Honduras, Guatemala and Nicaragua. This finding could hinder progress in the control and elimination of malaria in Central America. Continuous evaluation of RDTs and molecular surveillance would be recommended.

Molecular identification of Candida species from urinary infections in Honduras / Identificación molecular de especies de Candida de infecciones urinarias en Honduras

Background: Candiduria is a common infection among hospitalised patients. Although the clinical relevance of yeasts in urine is not clearly defined, fungal urinary tract infections have increased significantly in the last decades, becoming a growing public health problem. Candida albicans is the most commonly reported species in urinary infections, although other species of the genus are becoming particularly important, because some of them are linked with resistance to antifungal drugs. Aims: This study aimed to evaluate the frequency of Candida species causing candiduria in a hospital in Honduras. Methods: A simple and cost-effective PCR-RFLP approach was used, by amplifying a partial sequence of the ribosomal ITS1-ITS2 region and a subsequent digestion with the enzyme MspI. Results: During 2016, an analysis was performed on 73 urine samples from patients of different ages. Seven species were found. Candida albicans/dubliniensis was the most frequent species (30%); Candida glabrata (28.8%) was the most isolated among the rest of the species. Candida kefyr was the least frequent species found (2.5%). Conclusions: This study shows, for the first time in Honduras, the frequency of the Candida species isolated from urine using PCR-RFLP for their identification. This approach could be applied in future epidemiological studies at local and national level

Antecedentes: La candiduria es una infección común entre pacientes hospitalizados. Aunque la relevancia clínica de las levaduras en orina aun no está del todo esclarecida, el número de infecciones urinarias por hongos se ha incrementado en las últimas décadas, convirtiéndose en un creciente problema de salud pública. Candida albicans es la levadura más común en las infecciones urinarias, si bien otras especies del género están adquiriendo importancia debido a la resistencia a las drogas antifúngicas asociada a algunas especies. Objetivos: Este estudio pretendió evaluar la frecuencia de especies de Candida responsables de candiduria en un hospital nacional de Honduras. Métodos: Se utilizó un análisis in silico y PCR-RFLP de una región parcial de la secuencia ribosomal ITS1-ITS2, con posterior digestión mediante la enzima MspI. Resultados: Se analizaron 73 muestras de orina de pacientes de diferentes edades durante el año 2016. Se encontraron siete especies diferentes. Candida albicans/dubliniensis fue la especie más común (30%); Candida glabrata (28,8%) fue la más frecuente entre el resto de especies. Candida kefyr fue la especie menos encontrada (2,5%). Conclusiones: Este estudio muestra la frecuencia de especies de Candida aisladas a partir de orina, e identificadas mediante la técnica PCR-RFLP por primera vez en Honduras. Este enfoque podría ser aplicado para futuros estudios epidemiológicos a nivel local y nacional

Molecular identification of Candida species from urinary infections in Honduras.

BACKGROUND: Candiduria is a common infection among hospitalised patients. Although the clinical relevance of yeasts in urine is not clearly defined, fungal urinary tract infections have increased significantly in the last decades, becoming a growing public health problem. Candida albicans is the most commonly reported species in urinary infections, although other species of the genus are becoming particularly important, because some of them are linked with resistance to antifungal drugs. AIMS: This study aimed to evaluate the frequency of Candida species causing candiduria in a hospital in Honduras. METHODS: A simple and cost-effective PCR-RFLP approach was used, by amplifying a partial sequence of the ribosomal ITS1-ITS2 region and a subsequent digestion with the enzyme MspI. RESULTS: During 2016, an analysis was performed on 73 urine samples from patients of different ages. Seven species were found. Candida albicans/dubliniensis was the most frequent species (30%); Candida glabrata (28.8%) was the most isolated among the rest of the species. Candida kefyr was the least frequent species found (2.5%). CONCLUSIONS: This study shows, for the first time in Honduras, the frequency of the Candida species isolated from urine using PCR-RFLP for their identification. This approach could be applied in future epidemiological studies at local and national level.