Indicators sandflies and environment associated to spatial landscaping change in Chocó Biosphere Reserve UNESCO.

BACKGROUND OBJECTIVES: Sandflies are vector insects associated with terrestrial forest ecosystems; in the Ecuadorian Andes, they participate in the transmission of human cutaneous leishmaniasis. This geographical area represents an opportunity to evaluate the role of sandflies as bioindicators of the degree of intervention of tropical humid forest ecosystems (THF) associated with changes in the ecology of the local landscape. METHODS: CDC-light traps were used for collecting adult sandflies in February 2020 in a humid tropical forest within the Chocó Biosphere Reserve. All species were identified using morphological keys. Analysis data about abundance, richness, species accumulation, diversity index, species composition communities, species sex proportion, spatial sandflies environmental, Renyi's Diversity Profile were performed to compare six spatial habitats in Mashpi locality, Ecuador. RESULTS: Sandflies were collected (n-1435); the main species are represented by Trichophoromyia reburra, Nyssomyia trapidoi, Psathyromyia aclydifera, Psychodopygus panamensis and Lutzomyia hartmanni. Only Th. reburra is associated with not intervened forest, while the other three species are associated with intervened forest within Mashpi in the Choco Biosphere Reserve. The secondary forest has major sandflies' richness, while the primary forest exhibits major abundance. INTERPRETATION CONCLUSION: Th. reburra is a sandfly restricted to the Andean Forest and is a bioindicator of the high environmental health quality of the forest, while Ny. trapidoi and Pa. aclydifera are bioindicators of environmental disturbances in the forest. Additionally, Ps. panamensis, Lu. hartmanni and Ny. trapidoi are bioindicators of human impact and the risk of leishmaniasis.

Bacterial community assessment of drinking water and downstream distribution systems in highland localities of Ecuador.

Bacterial communities in drinking water provide a gauge to measure quality and confer insights into public health. In contrast to urban systems, water treatment in rural areas is not adequately monitored and could become a health risk. We performed 16S rRNA amplicon sequencing to analyze the microbiome present in the water treatment plants at two rural communities, one city, and the downstream water for human consumption in schools and reservoirs in the Andean highlands of Ecuador. We tested the effect of water treatment on the diversity and composition of bacterial communities. A set of physicochemical variables in the sampled water was evaluated and correlated with the structure of the observed bacterial communities. Predominant bacteria in the analyzed communities belonged to Proteobacteria and Actinobacteria. The Sphingobium genus, a chlorine resistance group, was particularly abundant. Of health concern in drinking water reservoirs were Fusobacteriaceae, Lachnospiraceae, and Ruminococcaceae; these families are associated with human and poultry fecal contamination. We propose the latter families as relevant biomarkers for establishing local standards for the monitoring of potable water systems in highlands of Ecuador. Our assessment of bacterial community composition in water systems in the Ecuadorian highlands provides a technical background to inform management decisions.

Indicators sand flies and environment associated to spatial landscaping change in Choco Biosphere Reserve UNESCO.

BACKGROUND OBJECTIVES: Sand flies are insects vector associated with terrestrial forest ecosystems; in the Ecuadorian Andes, they participate in the transmission of human cutaneous leishmaniasis. This geographical area is an opportunity to evaluate the role of sand flies as bioindicators od the degree of intervention of a tropical humid forest ecosystems (THF) associated with changes in the ecology of the local landscape. METHODS: CDC-light traps were used for collecting adults' sand flies in February 2020 in a humid tropical forest within Choco Biosphere Reserve. All species were identified using morphological keys. Analysis data about abundance, richness, species accumulation, diversity index, species composition communities, species sex proportion, spatial sand flies environmental, Renyi's Diversity Profile were performed to compare six spatial habitats in Mashpi locality, Ecuador. RESULTS: Sand flies were collected (n-1435); the main species are represented by Th. reburra Ny. trapidoi, Pa. aclydifera, Py. panamensis and Lu. hartmanni. Only Th. reburra is associated with not intervened forest, while the other 3 species are associated with intervened forest within Mashpi in the Choco Biosphere Reserve. The secondary forest has the major sandflies' richness, while the primary forest exhibits the major abundance. INTERPRETATION CONCLUSION: The results indicated that Th. reburra is a sandfly restricted to the Andean Forest and is a bioindicator of the high environmental health quality of the forest, while Ny. trapidoi and Pa. aclydifera are bioindicators of environmental disturbances in the forest. Additionally, Ps. panamensis, Lu. hartmanni and Ny. trapidoi are bioindicators of human impact and the risk of leishmaniasis.

Prevalence and Risk Factors of Eimeria spp. in Broiler Chickens from Pichincha and Santo Domingo de los Tsáchilas, Ecuador.

Coccidiosis in chickens is a parasitic disease of economic importance for the poultry industry. In Ecuador, there is limited information regarding the prevalence of Eimeria spp. on commercial broiler farms. Therefore, a total of 155 poultry farms in the provinces of Pichincha and Santo Domingo de los Tsáchilas were surveyed. The analysis of fresh fecal samples was conducted to determine the parasitic load of six of the seven chicken Eimeria species (excluding E. mitis) through multiplex PCR. Additionally, an epidemiological survey was performed to assess the risk factors associated with the infection using a multivariable logistic regression model. All samples tested positive for the presence of Eimeria spp., despite the farmers having implemented prophylactic measures, and no clinical coccidiosis cases were recorded. The parasitic load varied between 25 and 69,900 oocyst per gram. The species prevalence was as follows: Eimeria spp. 100%, E. maxima 80.4%, E. acervulina 70.6%, E. praecox 55.4%, E. tenella 53.6%, E. necatrix 52.2%, and E. brunetti 30.8%. The main species combination was E. cervuline, E. maxima, E. necatrix, and E. praecox (23.90%), followed by E. tenella, as a unique species (10.69%), and then E. acervulina, E. maxima, and E. praecox (8.81%). It was observed that farms operated by independent producers had a higher amount of Eimeria spp. and higher probability of the presence of E. brunetti, E. necatrix, E. praecox, and E. tenella. Poultry houses located below 1300 m above sea level were associated with a higher parasitic load and the presence of E. brunetti. Birds younger than 35 days of age and from open-sided poultry houses (with rudimentary environmental control) had a higher probability of presenting E. maxima. Drinking water from wells increased the risk of E. praecox presence. Research aimed at designing control strategies to improve health management on poultry farms in the region would help minimize the impact of coccidiosis.

The microbiome of the ice-capped Cayambe Volcanic Complex in Ecuador.

A major challenge in microbial ecology is to understand the principles and processes by which microbes associate and interact in community assemblages. Microbial communities in mountain glaciers are unique as first colonizers and nutrient enrichment drivers for downstream ecosystems. However, mountain glaciers have been distinctively sensitive to climate perturbations and have suffered a severe retreat over the past 40 years, compelling us to understand glacier ecosystems before their disappearance. This is the first study in an Andean glacier in Ecuador offering insights into the relationship of physicochemical variables and altitude on the diversity and structure of bacterial communities. Our study covered extreme Andean altitudes at the Cayambe Volcanic Complex, from 4,783 to 5,583 masl. Glacier soil and ice samples were used as the source for 16S rRNA gene amplicon libraries. We found (1) effects of altitude on diversity and community structure, (2) the presence of few significantly correlated nutrients to community structure, (3) sharp differences between glacier soil and glacier ice in diversity and community structure, where, as quantified by the Shannon γ-diversity distribution, the meta-community in glacier soil showed more diversity than in glacier ice; this pattern was related to the higher variability of the physicochemical distribution of variables in the former substrate, and (4) significantly abundant genera associated with either high or low altitudes that could serve as biomarkers for studies on climate change. Our results provide the first assessment of these unexplored communities, before their potential disappearance due to glacier retreat and climate change.

Beta-Lactam Antibiotic Resistance Genes in the Microbiome of the Public Transport System of Quito, Ecuador.

Multidrug-resistant bacteria present resistance mechanisms against ß-lactam antibiotics, such as Extended-Spectrum Beta-lactamases (ESBL) and Metallo-ß-lactamases enzymes (MBLs) which are operon encoded in Gram-negative species. Likewise, Gram-positive bacteria have evolved other mechanisms through mec genes, which encode modified penicillin-binding proteins (PBP2). This study aimed to determine the presence and spread of ß-lactam antibiotic resistance genes and the microbiome circulating in Quito's Public Transport (QTP). A total of 29 station turnstiles were swabbed to extract the surface environmental DNA. PCRs were performed to detect the presence of 13 antibiotic resistance genes and to identify and to amplify 16S rDNA for barcoding, followed by clone analysis, Sanger sequencing, and BLAST search. ESBL genes blaTEM-1 and blaCTX-M-1 and MBL genes blaOXA-181 and mecA were detected along QPT stations, blaTEM being the most widely spread. Two subvariants were found for blaTEM-1, blaCTX-M-1, and blaOXA-181. Almost half of the circulating bacteria found at QPT stations were common human microbiota species, including those classified by the WHO as pathogens of critical and high-priority surveillance. ß-lactam antibiotic resistance genes are prevalent throughout QPT. This is the first report of blaOXA-181 in environmental samples in Ecuador. Moreover, we detected a new putative variant of this gene. Some commensal coagulase-negative bacteria may have a role as mecA resistance reservoirs.

An in-depth characterization of the entomopathogenic strain Bacillus pumilus 15.1 reveals that it produces inclusion bodies similar to the parasporal crystals of Bacillus thuringiensis.

In the present work, the local isolate Bacillus pumilus 15.1 has been morphologically and biochemically characterized in order to gain a better understanding of this novel entomopathogenic strain active against Ceratitis capitata. This strain could represent an interesting biothechnological tool for the control of this pest. Here, we report on its nutrient preferences, extracellular enzyme production, motility mechanism, biofilm production, antibiotic suceptibility, natural resistance to chemical and physical insults, and morphology of the vegetative cells and spores. The pathogen was found to be ß-hemolytic and susceptible to penicillin, ampicillin, chloramphenicol, gentamicin, kanamycin, rifampicin, tetracycline, and streptomycin. We also report a series of biocide, thermal, and UV treatments that reduce the viability of B. pumilus 15.1 by several orders of magnitude. Heat and chemical treatments kill at least 99.9 % of vegetative cells, but spores were much more resistant. Bleach was the only chemical that was able to completely eliminate B. pumilus 15.1 spores. Compared to the B. subtilis 168 spores, B. pumilus 15.1 spores were between 2.67 and 350 times more resistant to UV radiation while the vegetative cells of B. pumilus 15.1 were almost up to 3 orders of magnitude more resistant than the model strain. We performed electron microscopy for morphological characterization, and we observed geometric structures resembling the parasporal crystal inclusions synthesized by Bacillus thuringiensis. Some of the results obtained here such as the parasporal inclusion bodies produced by B. pumilus 15.1 could potentially represent virulence factors of this novel and potentially interesting strain.

Identification, sequencing and comparative analysis of pBp15.S plasmid from the newly described entomopathogen Bacillus pumilus 15.1.

The Bacillus pumilus 15.1 strain, a recently described entomopathogenic strain active against Ceratitis capitata, contains at least two extrachromosomal elements, pBp15.1S and pBp15.1B. Given that B. pumilus is not a typical entomopathogenic bacterium, the acquisition of this extrachromosomal DNA may explain why B. pumilus 15.1 is toxic to an insect. One of the plasmids present in the strain, the pBp15.1S plasmid, was sub-cloned, sequenced and analyzed using bioinformatics to identify any potential virulence factor. The pBp15.1S plasmid was found to be 7785 bp in size with a GC content of 35.7% and 11 putative ORFs. A replication module typical of a small rolling circle plasmid and a sensing and regulatory system specific for plasmids was found in pBp15.1S. Additionally, we demonstrated the existence of ssDNA in plasmid preparations suggesting that pBp15.1S replicates by the small rolling circle mechanism. A gene cluster present in plasmid pPZZ84 from a distantly isolated B. pumilus strain was also present in pBp15.1S. The plasmid copy number of pBp15.1S in exponentially growing B. pumilus cells was determined to be 33 copies per chromosome. After an extensive plasmid characterization, no known virulence factor was found so a search in the other extrachromosomal elements of the bacteria is needed.

Selection of a Bacillus pumilus strain highly active against Ceratitis capitata (Wiedemann) larvae.

Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), the Mediterranean fruit fly (medfly), is one of the most important fruit pests worldwide. The medfly is a polyphagous species that causes losses in many crops, which leads to huge economic losses. Entomopathogenic bacteria belonging to the genus Bacillus have been proven to be safe, environmentally friendly, and cost-effective tools to control pest populations. As no control method for C. capitata based on these bacteria has been developed, isolation of novel strains is needed. Here, we report the isolation of 115 bacterial strains and the results of toxicity screening with adults and larvae of C. capitata. As a result of this analysis, we obtained a novel Bacillus pumilus strain, strain 15.1, that is highly toxic to C. capitata larvae. The toxicity of this strain for C. capitata was related to the sporulation process and was observed only when cultures were incubated at low temperatures before they were used in a bioassay. The mortality rate for C. capitata larvae ranged from 68 to 94% depending on the conditions under which the culture was kept before the bioassay. Toxicity was proven to be a special characteristic of the newly isolated strain, since other B. pumilus strains did not have a toxic effect on C. capitata larvae. The results of the present study suggest that B. pumilus 15.1 could be considered a strong candidate for developing strategies for biological control of C. capitata.