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Abstract Background In Ecuador, cervical cancer is the third most common cancer among women and the second most common cause of cancer-related death in women. Although HPV represents a serious public health problem worldwide, the information about its prevalence and genotypes in remote communities of Ecuador is limited. The aim of this study was to determine the distribution of HPV genotypes among ethnic minority women from two remote communities of the northwestern region of Ecuador: Afro-Ecuadorians and Chachis (Amerindian group). Methods We included 291 women who responded to a short survey and were screened for HPV by the amplification of the L1 gene and a nested multiplex PCR to detect 14 high risk (HR) genotypes. A survey collected information on ethnicity, age, community access, and sexual and gynecological history. We identified risk factors associated with HPV infection and co-infection using multivariate logistic regression to calculate odds ratio (OR) and Wald 95% confidence intervals (CI). Results Overall, HPV prevalence in the study cohort was 56%, and the five most common HR-HPV genotypes were HPV-58, -16, -68, -39, and -43; however, the distribution of HPV genotypes varied according to ethnicity. We identified lineage A (European variant) for HPV-16 and sublineage A2 for HPV-58 in both ethnic groups. Adjusting for ethnicity, age, community access, and number of sexual partners, we found that Afro-Ecuadorian women were less likely to have an HPV infection than Chachi women (OR: 0.49, 95% CI: 0.25, 0.96), and that participants in communities only accessible by river had 64% less chances of an HPV infection when compared to women in communities accessible by road (OR: 0.36, 95% CI: 0.19, 0.71), and women with 2 to 3 sexual partners had 2.47 times the odds of HPV infection than participants with 0–1 partners (OR: 2.47, 95% CI: 1.32, 4.6). Similar associations were observed with prevalence of co-infection. Conclusions This study provides baseline knowledge regarding the prevalence and distribution of HPV genotypes in ethnic groups of the northwestern coastal Ecuador and essential information for the implementation of appropriate HPV testing and vaccination program to prevent cervical cancer.

ABSTRACT Chagas disease, leishmaniasis, and malaria are major parasitic diseases disproportionately affecting the underprivileged population in developing nations. Finding new, alternative anti-parasitic compounds to treat these diseases is crucial because of the limited number of options currently available, the side effects they cause, the need for long treatment courses, and the emergence of drug-resistant parasites. Anti-microbial peptides (AMPs) derived from amphibian skin secretions are small bioactive molecules capable of lysing the cell membrane of pathogens while having low toxicity against human cells. Here, we report the anti-parasitic activity of five AMPs derived from skin secretions of three Ecuadorian frogs: cruzioseptin-1, cruzioseptin-4 (CZS-4), and cruzioseptin-16 from Cruziohyla calcarifer; dermaseptin-SP2 from Agalychnis spurrelli; and pictuseptin-1 from Boana picturata. These five AMPs were chemically synthesized. Initially, the hemolytic activity of CZS-4 and its minimal inhibitory concentration against Escherichia coli, Staphylococcus aureus, and Candida albicans were determined. Subsequently, the cytotoxicity of the synthetic AMPs against mammalian cells and their anti-parasitic activity against Leishmania mexicana promastigotes, erythrocytic stages of Plasmodium falciparum and mammalian stages of Trypanosoma cruzi were evaluated in vitro. The five AMPs displayed activity against the pathogens studied, with different levels of cytotoxicity against mammalian cells. In silico molecular docking analysis suggests this bioactivity may occur via pore formation in the plasma membrane, resulting in microbial lysis. CZS-4 displayed anti-bacterial, anti-fungal, and anti-parasitic activities with low cytotoxicity against mammalian cells. Further studies about this promising AMP are required to gain a better understanding of its activity.IMPORTANCEChagas disease, malaria, and leishmaniasis are major tropical diseases that cause extensive morbidity and mortality, for which available treatment options are unsatisfactory because of limited efficacy and side effects. Frog skin secretions contain molecules with anti-microbial properties known as anti-microbial peptides. We synthesized five peptides derived from the skin secretions of different species of tropical frogs and tested them against cultures of the causative agents of these three diseases, parasites known as Trypanosoma cruzi, Plasmodium falciparum, and Leishmania mexicana. All the different synthetic peptides studied showed activity against one of more of the parasites. Peptide cruzioseptin-4 is of special interest since it displayed intense activity against parasites while being innocuous against cultured mammalian cells, which indicates it does not simply hold general toxic properties; rather, its activity is specific against the parasites.

Carbapenem-resistant Acinetobacter spp. is associated with nosocomial infections in intensive care unit patients, resulting in high mortality. Although Acinetobacter spp. represent a serious public health problem worldwide, there are a few studies related to the presence of carbapenemases in health care facilities and other environmental settings in Ecuador. The main aim of this study was to characterize the carbapenem-resistant Acinetobacter spp. isolates obtained from four hospitals (52) and from five rivers (27) close to Quito. We used the disc diffusion and EDTA sinergy tests to determine the antimicrobial susceptibility and the production of metallo β-lactamases, respectively. We carried out a multiplex PCR of gyrB gene and the sequencing of partial rpoB gene to bacterial species identification. We performed molecular screening of nine carbapenem-resistant genes (blaSPM, blaSIM, blaGIM, blaGES, blaOXA-23, blaOXA-24, blaOXA-51, blaOXA-58, and blaOXA-143) by multiplex PCR, followed by identification using sequencing of blaOXA genes. Our findings showed that carbapenem-resistant A. baumannii were the main species found in health care facilities and rivers. Most of the clinical isolates came from respiratory tract samples and harbored blaOXA-23, blaOXA-366, blaOXA-72, blaOXA-65, blaOXA-70, and blaOXA-143-like genes. The river isolates harbored only the blaOXA-51 and probably blaOXA-259 genes. We concluded that the most predominant type of carbapenem genes among isolates were both blaOXA-23 and blaOXA-65 among A. baumannii clinical isolates.

Listeria monocytogenes is a foodborne pathogen that can produce serious, even fatal, infections. Among other foods, it can be found in unpasteurized dairy and ready-to-eat products. Surveillance of L. monocytogenes is of great interest since sources of infection are difficult to determine due to the long incubation period, and because the symptoms of listeriosis are similar to other diseases. We performed a genomic study of L. monocytogenes isolated from fresh cheeses and clinical samples from Ecuador. Sixty-five isolates were evaluated and sequenced, 14 isolates from cheese samples and 20 from clinical listeriosis cases from the National Institute of National Institute of Public Health Research, and 31 isolates from artisanal cheese samples from 8 provinces. All isolates exhibited heterogeneous patterns of the presence of pathogenicity islands. All isolates exhibited at least 4 genes from LIPI-1, but all references (26 L. monocytogenes closed genomes available in the NCBI database) showed the complete island, which encompasses 5 genes but is present in only two Ecuadorian isolates. Most isolates lacked gene actA. Genes from LIPI-2 were absent in all isolates. LIPI-3 and LIPI-4 were present in only a few references and isolates. With respect to the stress survival islets, our samples either presented SSI-1 or SSI-F2365, except for one isolate that presented SSI-F2365 and also one gene from SSI-1. None of the samples presented SSI-2. The predominant ST (sequence type) was ST2 (84.62% 55/65), and the only ST found in food (93.33% 42/45) and clinical samples (65% 13/20). Isolates were not grouped according to their sampling origin, date, or place in a phylogenetic tree obtained from the core alignment. The presence of ST2 in food and clinical samples, with high genomic similarity, suggests a foodborne infection risk linked to the consumption of fresh cheeses in Ecuador.

Abstract Background Understanding the natural microbiome and resistome of wildlife from remote places is necessary to monitor the human footprint on the environment including antimicrobial use (AU). Marine iguanas are endemic species from the Galapagos Islands where they are highly affected by anthropogenic factors that can alter their microbiota as well as their abundance and diversity of antimicrobial-resistant genes (ARGs). Thus, this study aims to apply culture-independent approaches to characterize the marine iguana’s gut metagenomic composition of samples collected from the uninhabited islands Rabida (n = 8) and Fernandina (Cabo Douglas, n = 30; Punta Espinoza, n = 30). Fresh feces from marine iguanas were analyzed through SmartChip RT-PCR, 16S rRNA, and metagenomic next-generation sequencing (mNGS) to identify their microbiome, microbial-metabolic pathways, resistome, mobilome, and virulome. Results The marine iguana’s gut microbiome composition was highly conserved despite differences in ecological niches, where 86% of taxa were shared in the three locations. However, site-specific differences were mainly identified in resistome, mobilome, virulorome, and metabolic pathway composition, highlighting the existence of factors that induce microbial adaptations in each location. Functional gut microbiome analyses revealed its role in the biosynthesis and degradation of vitamins, cofactors, proteinogenic amino acids, carbohydrates, nucleosides and nucleotides, fatty acids, lipids, and other compounds necessary for the marine iguanas. The overall bacterial ARG abundance was relatively low (0.006%); nevertheless, the presence of genes encoding resistance to 22 drug classes was identified in the iguana’s gut metagenome. ARG-carrying contig and co-occurrence network analyses revealed that commensal bacteria are the main hosts of ARGs. Taxa of public health interest such as Salmonella, Vibrio, and Klebsiella also carried multidrug-resistance genes associated with MGEs which can influence the dissemination of ARGs through horizontal gene transfer. Conclusion Marine iguanas depend on the gut microbiome for the biosynthesis and degradation of several compounds through a symbiotic relationship. Niche-specific adaptations were evidenced in the pool of microbial accessory genes (i.e., ARGs, MGEs, and virulence) and metabolic pathways, but not in the microbiome composition. Culture-independent approaches outlined the presence of a diverse resistome composition in the Galapagos marine iguanas from remote islands. The presence of AR pathogens in marine iguanas raises concerns about the dispersion of microbial-resistant threats in pristine areas, highlighting wildlife as sentinel species to identify the impact of AU.

Helminths are not usually considered important pathogens for birds of prey. There is a single published report of mortality in raptors due to an air sac trematode infection. We report a well-documented death case from massive infection by an air sac trematode of the family Cyclocoelidae in a wild-caught, juvenile male Snail Kite (Rostrhamus sociabilis) in Ecuador. The necropsy of a Snail Kite revealed more than 200 trematodes among air sacs, lungs, heart, gizzard, proventriculus, and liver. Within air sacs and lungs, mature flukes were associated with sacculitis, bronchitis, pneumonia, and atelectasis. Using an integrative taxonomic approach with morphological and molecular data, we identified the parasites as Bothrigaster variolaris (Trematoda: Cyclocoelidae: Ophthalmophaginae). This case provides the first evidence for the pathologic presence of air sac trematodes associated with morbidity in birds of prey in South America. Our results suggest that cyclocoelids may cause debilitation and significant clinical lesions in birds of prey, with potentially fatal consequences.

This study explored the composition of the mycobiome in the rhizosphere of Inga seedlings in two different but neighboring forest ecosystems in the undisturbed tropical Amazon rainforest at the Tiputini Biodiversity Station in Ecuador. In terra firme plots, which were situated higher up and therefore typically outside of the influence of river floods, and in várzea plots, the lower part of the forest located near the riverbanks and therefore seasonally flooded, tree seedlings of the genus Inga were randomly collected and measured, and the rhizosphere soils surrounding the root systems was collected. Members of the Fabaceae family and the genus Inga were highly abundant in both forest ecosystems. Inga sp. seedlings collected in terra firme showed a lower shoot to root ratio compared to seedlings that were collected in várzea, suggesting that Inga seedlings which germinated in várzea soils could invest more resources in vegetative growth with shorter roots. Results of the physical-chemical properties of soil samples indicated higher proportions of N, Mo, and V in terra firme soils, whereas várzea soils present higher concentrations of all other macro- and micronutrients, which confirmed the nutrient deposition effect of seasonal flooding by the nearby river. ITS metabarcoding was used to explore the mycobiome associated with roots of the genus Inga. Bioinformatic analysis was performed using Qiime 2 to calculate the alpha and beta diversity, species taxonomy and the differential abundance of fungi and arbuscular mycorrhizal fungi. The fungal community represented 75% of the total ITS ASVs, and although present in all samples, the subphylum Glomeromycotina represented 1.42% of all ITS ASVs with annotations to 13 distinct families, including Glomeraceae (72,23%), Gigasporaceae (0,57%), Acaulosporaceae (0,49%). AMF spores of these three AMF families were morphologically identified by microscopy. Results of this study indicate that AMF surround the rhizosphere of Inga seedlings in relatively low proportions compared to other fungal groups but present in both terra firme and várzea Neotropical ecosystems.

Introducción: el presente artículo tuvo como objetivo analizar el estatus laboral de los adultos mayores en América Latina con énfasis en el impacto producido por la COVID-19 en 2020. Método: se caracterizó la dinámica laboral de las personas mayores de 60 años con encuestas de hogares de siete países de Latinoamérica, las cuales se resumieron con un pseudopanel. Se realizó un análisis de varianza (ANOVA) para identificar las diferencias entre países, sexo y regiones urbanas y rurales. Se estimaron regresiones por MCO para cuantificar el efecto diferencial existente y, particularmente, el cambio producido en 2020. Resultados: se identificó una correlación negativa (β = -0.75, p = 0.00) entre la cobertura del ingreso pensional y la participación de las personas mayores en el mercado laboral. Por lo cual, las deficiencias en la seguridad social estarían motivando la permanencia en el trabajo de las personas mayores. Dicha participación se redujo 11 puntos porcentuales (p.p.) entre 2016 y 2020, principalmente Costa Rica, Colombia y Chile, así como en las zonas urbanas y entre los hombres. El desempleo aumentó 1 p.p. de manera especial en las zonas urbanas, y el subempleo se mantuvo invariante. Conclusiones: las diferencias en la participación laboral están mediadas por roles de género y la estructura económica de los países y de las zonas urbanas y rurales. Se evidencia un deterioro en el estatus socioeconómico que no se compensa con el leve incremento en la cobertura del ingreso pensional. Las mujeres parecen enfrentarse a una mayor exclusión y autoexclusión del mercado laboral.

Toxoplasma gondii is a zoonotic intracellular parasite of particular concern in the conservation of wildlife due to its ability to infect all homeotherms and potentially cause acute fatal disease in naive species. In the Galapagos (Ecuador), an archipelago composed of more than a hundred islets and islands, the presence of T. gondii can be attributed to human-introduced domestic cats, but little is known about its transmission in wildlife populations. We compared the prevalence of antibodies against T. gondii in sympatric Galapagos wild bird species that differ in diet and contact with oocyst-contaminated soil to determine the relative importance of trophic habits as an exposure factor. Plasma samples were obtained from 163 land birds inhabiting Santa Cruz, one of the cat-inhabited islands, and from 187 seabirds breeding in cat-free surrounding islands (Daphne Major, North Seymour, and South Plaza). These samples were tested for the presence of T. gondii antibodies using the modified agglutination test (MAT ≥ 1:10). All seven species of land birds and 4/6 species of seabirds presented seropositive results. All great frigatebirds (Fregata minor) (N = 25) and swallow-tailed gulls (Creagrus furcatus) (N = 23) were seronegative. Prevalence ranged from 13% in Nazca boobies (Sula granti) to 100% in Galapagos mockingbirds (Mimus parvulus). It decreased from occasional carnivores (63.43%) to granivores-insectivores (26.22%), and strict piscivores (14.62%). These results indicate that the consumption of tissue cysts poses the highest risk of exposure to T. gondii for Galapagos birds, followed by the ingestion of plants and insects contaminated by oocysts as important transmission pathways.

Summary The use of wastewater for irrigation and animal manure as fertilizer can cause transmission of intestinal pathogens, conditions frequently observed in low‐ and middle‐income countries (LMICs). Here, we tested the ability of Salmonella to grow in the faecal matter. We inoculated freshly isolated Salmonella strains (from chickens) in chicken faecal matter and incubated for 1 to 12 days, under aerobic and anaerobic conditions. We found that both Salmonella and Escherichia coli multiplied massively in faecal matter outside a host and significantly higher in aerobic conditions. Our results have critical implications in waste management, as we demonstrate that aerobic treatments may not be the best to reduce the number of Salmonella in the environment.

Salmonella enterica is one of the most important foodborne pathogens around the world. In the last years, S. enterica serovar Infantis has become an important emerging pathogen in many countries, often as multidrug resistant clones. To understand the importance of S. enterica in the broiler industry in Ecuador, we performed a study based on phenotypic and WGS data of isolates from poultry farms, chicken carcasses and humans. We showed a high prevalence of S. enterica in poultry farms (41.4%) and chicken carcasses (55.5%), but a low prevalence (1.98%) in human samples. S. Infantis was shown to be the most prevalent serovar with a 98.2, 97.8, and 50% in farms, foods, and humans, respectively, presenting multidrug resistant patterns. All sequenced S. Infantis isolates belonged to ST32. For the first time, a pESI-related megaplasmid was identified in Ecuadorian samples. This plasmid contains genes of antimicrobial resistance, virulence factors, and environmental stress tolerance. Genomic analysis showed a low divergence of S. Infantis strains in the three analyzed components. The results from this study provide important information about genetic elements that may help understand the molecular epidemiology of S. Infantis in Ecuador.

Oropouche virus (OROV) is responsible for outbreaks of Oropouche fever in parts of South America. We recently identified and isolated OROV from a febrile Ecuadorian patient, however, a previously published qRT-PCR assay did not detect OROV in the patient sample. A primer mismatch to the Ecuadorian OROV lineage was identified from metagenomic sequencing data. We report the optimisation of an qRT-PCR assay for the Ecuadorian OROV lineage, which subsequently identified a further five cases in a cohort of 196 febrile patients. We isolated OROV via cell culture and developed an algorithmically-designed primer set for whole-genome amplification of the virus. Metagenomic sequencing of the patient samples provided OROV genome coverage ranging from 68-99%. The additional cases formed a single phylogenetic cluster together with the initial case. OROV should be considered as a differential diagnosis for Ecuadorian patients with febrile illness to avoid mis-diagnosis with other circulating pathogens.

Fast and accurate identification of biting midges is crucial in the study of Culicoides-borne diseases. In this work, we propose a two-stage method for automatically analyzing Culicoides (Diptera: Ceratopogonidae) species. First, an image preprocessing task composed of median and Wiener filters followed by equalization and morphological operations is used to improve the quality of the wing image in order to allow an adequate segmentation of particles of interest. Then, the segmentation of the zones of interest inside the biting midge wing is made using the watershed transform. The proposed method is able to produce optimal feature vectors that help to identify Culicoides species. A database containing wing images of C. obsoletus, C. pusillus, C. foxi, and C. insignis species was used to test its performance. Feature relevance analysis indicated that the mean of hydraulic radius and eccentricity were relevant for the decision boundary between C. obsoletus and C. pusillus species. In contrast, the number of particles and the mean of the hydraulic radius was relevant for deciding between C. foxi and C. insignis species. Meanwhile, for distinguishing among the four species, the number of particles and zones, and the mean of circularity were the most relevant features. The linear discriminant analysis classifier was the best model for the three experimental classification scenarios previously described, achieving averaged areas under the receiver operating characteristic curve of 0.98, 0.90, and 0.96, respectively.

We report identification of an Oropouche virus strain in a febrile patient from Ecuador by using metagenomic sequencing and real-time reverse transcription PCR. Virus was isolated from patient serum by using Vero cells. Phylogenetic analysis of the whole-genome sequence showed the virus to be similar to a strain from Peru.

Protected areas recently created in Argentina often include previously degraded lands, such as sheep ranches in the Patagonian deserts. We show the results of a 14-year monitoring program of three formerly persecuted carnivores, the culpeo fox (Lycalopex culpaeus), the South American grey fox (Lycalopex griseus) and the puma (Puma concolor), in two abandoned sheep ranches that were incorporated into a Patagonian national park approximately 25 years ago. The culpeo fox population underwent an average annual decline of 10–23%, whereas the grey fox and puma populations increased at an average annual rate of 7% and 19%, respectively. The grey fox’s increasing trends were strongly correlated with the decline of the culpeo fox, whereas the correlations between the fox and puma trends were weaker. Culpeo fox decline was stronger in the ranch where sheep and predator controls had been removed earlier. These relationships between species trends support the competitive release hypothesis, assuming that puma competition with the culpeo fox for trophic resources is stronger than competition with the grey fox, and that the puma can exclude culpeo foxes through interference. Species trends suggest a competitive hierarchy between fox species, with grey fox being the inferior competitor. However, mechanisms other than competition could not be discounted. Our study illustrates how long-term monitoring of interacting species allows a better understanding of ecological processes and wildlife ecology.

Abstract Background Rapid, accurate and high-throughput identification of vector arthropods is of paramount importance in surveillance programmes that are becoming more common due to the changing geographic occurrence and extent of many arthropod-borne diseases. Protein profiling by MALDI-TOF mass spectrometry fulfils these requirements for identification, and reference databases have recently been established for several vector taxa, mostly with specimens from laboratory colonies. Methods We established and validated a reference database containing 20 phlebotomine sand fly (Diptera: Psychodidae, Phlebotominae) species by using specimens from colonies or field-collections that had been stored for various periods of time. Results Identical biomarker mass patterns (‘superspectra’) were obtained with colony- or field-derived specimens of the same species. In the validation study, high quality spectra (i.e. more than 30 evaluable masses) were obtained with all fresh insects from colonies, and with 55/59 insects deep-frozen (liquid nitrogen/-80 °C) for up to 25 years. In contrast, only 36/52 specimens stored in ethanol could be identified. This resulted in an overall sensitivity of 87 % (140/161); specificity was 100 %. Duration of storage impaired data counts in the high mass range, and thus cluster analyses of closely related specimens might reflect their storage conditions rather than phenotypic distinctness. A major drawback of MALDI-TOF MS is the restricted availability of in-house databases and the fact that mass spectrometers from 2 companies (Bruker, Shimadzu) are widely being used. We have analysed fingerprints of phlebotomine sand flies obtained by automatic routine procedure on a Bruker instrument by using our database and the software established on a Shimadzu system. The sensitivity with 312 specimens from 8 sand fly species from laboratory colonies when evaluating only high quality spectra was 98.3 %; the specificity was 100 %. The corresponding diagnostic values with 55 field-collected specimens from 4 species were 94.7 % and 97.4 %, respectively. Conclusions A centralized high-quality database (created by expert taxonomists and experienced users of mass spectrometers) that is easily amenable to customer-oriented identification services is a highly desirable resource. As shown in the present work, spectra obtained from different specimens with different instruments can be analysed using a centralized database, which should be available in the near future via an online platform in a cost-efficient manner.

Objectives: Burkholderia cepacia has been linked to healthcare-associated infections and colonization caused by contamination of alcohol-free mouthwash used in patients undergoing mechanical ventilation. The purpose of our study was to establish the source of a clustering of healthcare-associated B. cepacia isolates in patients on mechanical ventilation in the intensive care unit (ICU). Methods: During April 2012 the Infection Control Committee became concerned when B. cepacia was isolated from tracheal aspirate cultures of three ICU patients. The medical records for the years 2011 and 2012 were reviewed to identify further cases. Cultures of potential reservoirs were done. Isolates from patients and an alcohol-free mouthwash were submitted to multilocus sequence typing (MLST) analysis and antimicrobial resistance testing. Results: Four patients with positive cultures for B. cepacia were identified before the review of the medical records for the years 2011 and 2012. Nine further cases were identified in the review, defined as a patient with pneumonia who had a culture of respiratory secretions that was positive for B. cepacia. Three were cases of infection and 10 were colonizations. All of the isolates from patients (J, K, L, and M) and mouthwash samples (B19, B20, and B21) were genetically identical by MLST analysis. Conclusions: Our findings strongly suggest that alcohol-free mouthwash solution intrinsically contaminated with B. cepacia was the source of these colonizations and infections involving adults in the ICU.

Although Tadarida brasiliensis is one of the most widely distributed species of bats in Argentina, it is nevertheless extremely scarce in Patagonia. We report a second record of T. brasiliensis for Santa Cruz province, which fills a gap in the geographical distribution of this species in that province and is also the third southernmost record for the species and for any molossid bat in the world.

Indigenous beers (chicha) are part of the indigenous culture in Ecuador. The fermentation process of these beers probably relies on microorganisms from fermented substrates, environment and human microbiota. We analyzed the microbiota of artisanal beers (including a type of beer produced after chewing boiled cassava) using bacterial culture and 16S ribosomal RNA (rRNA) gene-based tag-encoded FLX amplicon pyrosequencing (bTEFAP). Surprisingly, we found that Streptococcus salivarius and Streptococcus mutans (part of the human oral microbiota) were among the most abundant bacteria in chewed cassava and in non-chewed cassava beers. We also demonstrated that S. salivarius and S. mutans (isolated from these beers) could proliferate in cassava mush. Lactobacillus sp. was predominantly present in most types of Ecuadorian chicha.

Water kefir is a homemade fermented drink, to which certain probiotic properties are attributed. In this study, changes in acidity and oBrix of Ecuadorian homemade water kefir, elaborated with three different types of sweeteners (brown sugar, honey and granulated sugar) and at different fermentation times (24, 48 and 72 hours), were determined. The treatments arranged in a completely randomized block design (factorial 32) was applied. A focus group evaluated and selected three prototypes due to their sensory characteristics, also acidity and oBrix were measured for chemical specifications. According to the weighting of the variable of response for microbiological count, two treatments were chosen. The treatment with the greatest acceptance due to their chemical, microbiological and sensory characteristics was the sample fermented with honey at 48 h. A phenotypic and genotypic characterization (16S DNAr) of microbiota showed the presence of Leuconostoc holzapfelii, Leuconostoc pseudomesenteroides and Saccharomyces cerevisiae, which could be attributed to water kefir probiotic properties.

Ninety one samples of traditional fermented foods were collected from 20 Ecuadorian provinces. In order to identify strains with potential use as starter cultures in fermented foods, 119 lactic acid bacteria (LAB) strains were isolated and characterized by phenotypic tests and 16S ribosomal DNA (rDNA) sequencing. Forty seven percent of isolates were identified as facultative heterofermentative Lactobacillus sp. the next predominant groups (31%) were homofermentative cocci belonging to the genera Pediococcus, Lactococcus and Enterococcus; Finally, 11% were obligate heterofermentative Lactobacillus sp. and 11% heterofermentative cocci (Weissella and Leuconostoc). Several isolates showed Exopolysaccharide and bacteriocin production as well as caseinolytic activity. These properties could have interesting applications in food industry.