Ecotoxicological effects of ketoprofen and fluoxetine and their mixture in an aquatic microcosm.

The effects of fluoxetine (antidepressant) and ketoprofen (analgesic) on aquatic ecosystems are largely unknown, particularly as a mixture. This work aimed at determining the effect of sublethal concentrations of both compounds individually (0.050 mg/L) and their mixture (0.025 mg/L each) on aquatic communities at a microcosm scale for a period of 14 d. Several physicochemical parameters were monitored to estimate functional alterations in the ecosystem, while model organisms (Daphnia magna, Lemna sp., Raphidocelis subcapitata) and the sequencing of 16S/18S rRNA genes permitted to determine effects on specific populations and changes in community composition, respectively. Disturbances were more clearly observed after 14 d, and overall, the microcosms containing fluoxetine (alone or in combination with ketoprofen) produced larger alterations on most physicochemical and biological variables, compared to the microcosm containing only ketoprofen, which suffered less severe changes. Differences in nitrogen species suggest alterations in the N-cycle due to the presence of fluoxetine; similarly, all pharmaceutical-containing systems decreased the brood rate of D. magna, while individual compounds inhibited the growth of Lemna sp. No clear trends were observed regarding R. subcapitata, as indirectly determined by chlorophyll quantification. The structure of micro-eukaryotic communities was altered in the fluoxetine-containing systems, whereas the structure of bacterial communities was affected to a greater extent by the mixture. The disruptions to the equilibrium of the microcosm demonstrate the ecological risk these compounds pose to aquatic ecosystems.

Highly abundant bacteria in the gut of Triatoma dimidiata (Hemiptera: Reduviidae) can inhibit the growth of Trypanosoma cruzi (Kinetoplastea: Trypanosomatidae).

Chagas disease, caused by the protozoan Trypanosoma cruzi, is a zoonosis primarily found in rural areas of Latin America. It is considered a neglected tropical disease, and Triatoma dimidiata is the main vector of the parasite in Central America. Despite efforts, Chagas disease continues to be a public health concern, and vector control remains a primary tool to reduce transmission. In this study, we tested the hypothesis that highly abundant bacteria in the gut of T. dimidiata inhibit the growth of T. cruzi. To achieve this, bacterial diversity in the gut of T. dimidiata specimens from Costa Rica was characterized by metabarcoding of the 16S rRNA, microbial isolation was performed, and the effect of freeze-dried supernatants of the isolates on T. cruzi was investigated. Metabarcoding showed that the most abundant genera in the gut were Corynebacterium, Tsukamurella, Brevibacterium, and Staphylococcus. Barcoding and sequences comparison confirmed that 8 of the 30 most abundant amplicon sequence variants (ASVs) were isolated, and 2 of them showed an inhibitory effect on the growth of T. cruzi epimastigotes. These bacteria correspond to isolates of Tsukamurella and Brevibacterium, which were respectively the second and sixth most abundant ASVs in the gut of T. dimidiata. Notably, only the isolate of Brevibacterium showed a significant difference in growth inhibition against epimastigotes of both T. cruzi strains tested. These findings suggest that the gut microbiota of T. dimidiata may play an active role in modulating parasite development.

Microplastics in feed cause sublethal changes in the intestinal microbiota and a non-specific immune response indicator of the freshwater crayfish Procambarus clarkii (Decapoda: Cambaridae).

Microplastics (MP) are a hazardous pollutant of global concern that threatens aquatic ecosystems and public health. We used the invasive, cosmopolitan, and environmentally versatile red swamp crayfish Procambarus clarkii as a model to study the effects of MP on the intestinal microbiome. Crayfish collected from the environment were compared with specimens exposed to recycled Polyethylene terephthalate (rPET) MP in feed (30%) for 96 h in the laboratory and a control group. We analyzed the 16S rRNA of the intestinal bacteria by PCR-DGGE and high-throughput sequencing. MP exposure caused dysbiosis of the intestinal microbiota, with an increase in Alphaproteobacteria and Actinobacteria. We detected higher abundance of opportunistic genera such as Klebsiella, Acinetobacter, Hydromonas, Pseudomonas, Gemmobacter, and Enterobacter on MP fed organisms. Moreover, MP exposure reduced the abundance of Clostridia and Bateroidetes, which are important for immune system development and pathogen prevention. Furthermore, MP exposure decreased the phenoloxidase (PO) immune response in crayfish. There was a significant difference in the richness of intestinal bacterial communities after consumption of food contaminated with MP, likely increasing the abundance of opportunistic bacteria in the intestinal microbiota. Our results suggest that MP alter the gut microbial composition and impair the health of P. clarkii.

Differences in fungal communities in the fur of two- and three-toed sloths revealed by ITS metabarcoding.

Sloths have dense fur on which insects, algae, bacteria and fungi coexist. Previous studies using cultivation-dependent methods and 18S rRNA sequencing revealed that the fungal communities in their furs comprise members of the phyla Ascomycota and Basidiomycota. In this note, we increase the resolution and knowledge of the mycobiome inhabiting the fur of the two- (Choloepus hoffmanni) and three-toed (Bradypus variegatus) sloths. Targeted amplicon metagenomic analysis of ITS2 nrDNA sequences obtained from 10 individuals of each species inhabiting the same site revealed significant differences in the structure of their fungal communities and also in the alpha-diversity estimators. The results suggest a specialization by host species and that the host effect is stronger than that of sex, age and animal weight. Capnodiales were the dominant order in sloths' fur and Cladosporium and Neodevriesia were the most abundant genera in Bradypus and Choloepus, respectively. The fungal communities suggest that the green algae that inhabit the fur of sloths possibly live lichenized with Ascomycota fungal species. The data shown in this note offer a more detailed view of the fungal content in the fur of these extraordinary animals and could help explain other mutualistic relationships in this complex ecosystem.

Importancia del microbioma en la salud humana y aplicaciones médicas / Importance of the microbiome in human health and medical implications

Recientes investigaciones han relacionado la microbiota intestinal con la salud humana en múltiples aspectos. La evolución de los estilos de vida ha determinado un cambio en la composición de las bacterias intestinales, así como la implicación que la comunidad de estas ejerce sobre la salud. Actualmente, se conoce que la mayoría de las bacterias presentes en el sistema gastrointestinal pertenecen principalmente a los fila Firmicutes y Bacterioidetes, aunque también se encuentran otros grupos tales como proteobacterias y actinobacterias. A medida que se avanza en el tracto gastrointestinal predominan algunos géneros de bacterias. Los efectos de la microbiota pueden ser directos e indirectos, además, dependen de muchos factores tales como la edad de la persona, el grupo etario, la genética del individuo, la dieta y el estilo de vida. Durante los últimos años, la accesibilidad a tecnologías de secuenciación ha permitido tener un acercamiento más estrecho a la microbiota intestinal. Esto, sumado a herramientas bioinformáticas, ha permitido establecer relaciones microbiales entre la cantidad y estructura poblacional y las manifestaciones clínicas en el ser humano. Algunas de las afecciones estudiadas y que tienen relación con la microbiota intestinal son: la obesidad, la diabetes, el cáncer, las enfermedades relacionadas con el cerebro, las enfermedades cardiovasculares y las enfermedades gastrointestinales. De acuerdo con lo mencionado, se hizo una recopilación de información de carácter científico en cuanto a estudios relevantes que describen la relación microbiota-salud humana y casos donde se observa compromiso del organismo, al mismo tiempo que se describen opciones terapéuticas propuestas y un abordaje de perspectivas futuras.

Recent research has linked gut microbiota to human health in multiple ways. The evolution of lifestyles has determined a change in the composition of intestinal bacteria, as well as the implications that they exert on health. Currently, it is known that most of the bacteria present in the gastrointestinal sector belong mainly to the phylum Firmicutes and Bacterioidetes, although there are also other groups such as proteobacteria and actinobacteria. As it progresses through the gastrointestinal tract, some genera of bacteria and species predominate. The effects of the microbiota can be direct and indirect, and also depend on many factors such as the age of the person, the age group, the individual's genetics, diet, and lifestyle. In recent years, accessibility to sequencing technologies has allowed for a closer approach to the intestinal microbiota. This, added to bioinformatic tools has allowed establishing microbial relationships in terms of quantity and population structure with clinical manifestations in humans. Some of the pathologies studied that are related to intestinal microbiota are obesity, diabetes, cancer, brain-related diseases, cardiovascular diseases, and gastrointestinal diseases. A compilation of scientific information is made regarding relevant studies that describe the microbiota-human health relationship, cases where the organism is affected, as well as proposed therapeutic options and an approach to future perspectives

Presence of microplastics in six bivalve species (Mollusca, Bivalvia) commercially exploited at the Pacific coast of Costa Rica, Central America.

Microplastic pollution represents a serious threat to coastal marine systems. We evaluated the presence of microplastics (MPs) in 168 specimens of six commercially exploited bivalve species in the Golfo de Nicoya, Costa Rica. MPs were present in 100 % of the specimens of Leukoma asperrima and L. ecuadoriana, 74 % of Mytella strigata, 82 % of M. guyanensis, 84 % of Asthenometis asthenodon, and 82 % of Tagelus affinis. When considering all specimens, we found on average 4.8 ±â€¯8.0 pieces per individual and 2.1 ±â€¯3.3 items g-1 of wet tissue. There were significant differences between species with L. asperrima and L. ecuadoriana containing the highest number of pieces. Fibers and particles comprised 92 % and 8 %, respectively. This is the first study from the Central America's Pacific region showing the presence of MPs in bivalve species.

Antibiotic-producing Micrococcales govern the microbiome that inhabits the fur of two- and three-toed sloths.

Sloths have a dense coat on which insects, algae and fungi coexist in a symbiotic relationship. This complex ecosystem requires different levels of controls; however, most of these mechanisms remain unknown. We investigated the bacterial communities inhabiting the hair of two- (Choloepus Hoffmanni) and three-toed (Bradypus variegatus) sloths and evaluated their potential for producing antibiotic molecules capable of exerting control over the hair microbiota. The analysis of 16S rRNA amplicon sequence variants revealed that the communities in both host species are dominated by Actinobacteriota and Firmicutes. The most abundant genera were Brevibacterium, Kocuria/Rothia, Staphylococcus, Rubrobacter, Nesterenkonia and Janibacter. Furthermore, we isolated nine strains of Brevibacterium and Rothia capable of producing substances that inhibited the growth of common mammalian pathogens. The analysis of the biosynthetic gene clusters of these nine isolates suggests that the pathogen-inhibitory activity could be mediated by the presence of siderophores, terpenes, beta-lactones, Type III polyketide synthases, ribosomally synthesized and post-translationally modified peptides, non-alpha poly-amino acids like e-Polylysine, ectoine or non-ribosomal peptides. Our data suggest that Micrococcales that inhabit sloth hair could have a role in controlling microbial populations in that habitat, improving our understanding of this highly complex ecosystem.

Methylotrophs and Hydrocarbon-Degrading Bacteria Are Key Players in the Microbial Community of an Abandoned Century-Old Oil Exploration Well.

In this work, we studied the microbial community and the physicochemical conditions prevailing in an exploratory oil well, abandoned a century ago, located in the Cahuita National Park (Costa Rica). According to our analysis, Cahuita well is characterized by a continuous efflux of methane and the presence of a mixture of hydrocarbons including phenanthrene/anthracene, fluoranthene, pyrene, dibenzothiophene, tricyclic terpanes, pyrene, sesquiterpenes, sterane, and n-alkanes. Based on the analysis of 16S rRNA gene amplicons, we detected a significant abundance of methylotrophic bacteria such as Methylobacillus (6.3-26.0% of total reads) and Methylococcus (4.1-30.6%) and the presence of common genera associated with hydrocarbon degradation, such as Comamonas (0.8-4.6%), Hydrogenophaga (1.5-3.3%) Rhodobacter (1.0-4.9%), and Flavobacterium (1.1-6.5%). The importance of C1 metabolism in this niche was confirmed by amplifying the methane monooxygenase (MMO)-encoding gene (pmo) from environmental DNA and the isolation of two strains closely related to Methylorubrum rhodesianum and Paracoccus communis with the ability to growth using methanol and formate as sole carbon source respectively. In addition, we were able to isolated 20 bacterial strains from the genera Pseudomonas, Acinetobacter, and Microbacterium which showed the capability to grow using the hydrocarbons detected in the oil well as sole carbon source. This work describes the physicochemical properties and microbiota of an environment exposed to hydrocarbons for 100 years, and it not only represents a contribution to the understanding of microbial communities in environments with permanently high concentrations of these compounds but also has biotechnological implications for bioremediation of petroleum-polluted sites.

Variation of bacterial communities along the vertical gradient in Lake Issyk Kul, Kyrgyzstan.

We explored the diversity and community composition of bacteria along a vertical gradient in Lake Issyk Kul, Kyrgyzstan, one of the world's largest and deepest brackish lakes. We identified 4904 bacterial amplicon sequence variants based on the 16S rRNA gene analysis and determined significant changes in the composition, responding mainly to depth and salinity. A higher abundance of Proteobacteria and Bacteroidetes was observed in the surface waters and lake tributaries. Cyanobacteria were more abundant in the deep chlorophyll maximum from 28.5 to 128 m, while Planctomycetes and Chloroflexi were dominant in the deepest layers, from 128 to 600 m. According to our machine learning analyses, depth and temperature were the most critical environmental factors, with strong effects on Proteobacteria, Planctomycetes and Chloroflexi, while oxygen was associated with the variations in Cyanobacteria. We also observed that with increasing depth, the alpha diversity values increased. The dominance of Planctomycetes and Chloroflexi in the deepest layers can only be seen in a few lakes of the world. However, the lake is facing increasing anthropogenic and climatic pressure. There is an urgent need to understand better the ecological role and function of these unique deep-water microbial communities.

Temperature and elemental sulfur shape microbial communities in two extremely acidic aquatic volcanic environments.

Aquatic environments of volcanic origin provide an exceptional opportunity to study the adaptations of microorganisms to early planet life conditions. Here, we characterized the prokaryotic communities and physicochemical properties of seepage sites at the bottom of the Poas Volcano crater and the Agrio River, two geologically related extremely acidic environments located in Costa Rica. Both locations hold a low pH (1.79-2.20) and have high sulfate and iron concentrations (Fe = 47-206 mg/L, SO42- = 1170-2460 mg/L), but significant differences in their temperature (90.0-95.0 ºC in the seepages at Poas Volcano, 19.1-26.6 ºC in Agrio River) and in the elemental sulfur content. Based on the analysis of 16S rRNA gene sequences, we determined that Sulfobacillus spp. represented more than half of the sequences in Poas Volcano seepage sites, while Agrio River was dominated by Leptospirillum and members of the archaeal order Thermoplasmatales. Both environments share some chemical characteristics and part of their microbiota, however, the temperature and the reduced sulfur are likely the main distinguishing features, ultimately shaping their microbial communities. Our data suggest that in the Poas Volcano-Agrio River system there is a common metabolism but with specialization of species that adapt to the physicochemical conditions of each environment.

Fungal Communities in Sediments Along a Depth Gradient in the Eastern Tropical Pacific.

Deep waters represent the largest biome on Earth and the largest ecosystem of Costa Rica. Fungi play a fundamental role in global biogeochemical cycling in marine sediments, yet, they remain little explored. We studied fungal diversity and community composition in several marine sediments from 16 locations sampled along a bathymetric gradient (from a depth of 380 to 3,474 m) in two transects of about 1,500 km length in the Eastern Tropical Pacific (ETP) of Costa Rica. Sequence analysis of the V7-V8 region of the 18S rRNA gene obtained from sediment cores revealed the presence of 787 fungal amplicon sequence variants (ASVs). On average, we detected a richness of 75 fungal ASVs per sample. Ascomycota represented the most abundant phylum with Saccharomycetes constituting the dominant class. Three ASVs accounted for ca. 63% of all fungal sequences: the yeast Metschnikowia (49.4%), Rhizophydium (6.9%), and Cladosporium (6.7%). We distinguished a cluster composed mainly by yeasts, and a second cluster by filamentous fungi, but we were unable to detect a strong effect of depth and the overlying water temperature, salinity, dissolved oxygen (DO), and pH on the composition of fungal communities. We highlight the need to understand further the ecological role of fungi in deep-sea ecosystems.

Microbial Community Structure Along a Horizontal Oxygen Gradient in a Costa Rican Volcanic Influenced Acid Rock Drainage System.

We describe the geochemistry and microbial diversity of a pristine environment that resembles an acid rock drainage (ARD) but it is actually the result of hydrothermal and volcanic influences. We designate this environment, and other comparable sites, as volcanic influenced acid rock drainage (VARD) systems. The metal content and sulfuric acid in this ecosystem stem from the volcanic milieu and not from the product of pyrite oxidation. Based on the analysis of 16S rRNA gene amplicons, we report the microbial community structure in the pristine San Cayetano Costa Rican VARD environment (pH = 2.94-3.06, sulfate ~ 0.87-1.19 g L-1, iron ~ 35-61 mg L-1 (waters), and ~ 8-293 g kg-1 (sediments)). San Cayetano was found to be dominated by microorganisms involved in the geochemical cycling of iron, sulfur, and nitrogen; however, the identity and abundance of the species changed with the oxygen content (0.40-6.06 mg L-1) along the river course. The hypoxic source of San Cayetano is dominated by a putative anaerobic sulfate-reducing Deltaproteobacterium. Sulfur-oxidizing bacteria such as Acidithiobacillus or Sulfobacillus are found in smaller proportions with respect to typical ARD. In the oxic downstream, we identified aerobic iron-oxidizers (Leptospirillum, Acidithrix, Ferrovum) and heterotrophic bacteria (Burkholderiaceae bacterium, Trichococcus, Acidocella). Thermoplasmatales archaea closely related to environmental phylotypes found in other ARD niches were also observed throughout the entire ecosystem. Overall, our study shows the differences and similarities in the diversity and distribution of the microbial communities between an ARD and a VARD system at the source and along the oxygen gradient that establishes on the course of the river.

Resistencia a desinfectantes y su relación con la resistencia a los antibióticos / Resistance to disinfectants and their relationship with antibiotic resistance

Resumen Objetivo: Sistematizar los principales mecanismos de acción de los desinfectantes y describir mecanismos de resistencia comunes entre biocidas y antibióticos. Métodos: Se realizó una revisión bibliográfica de artículos científicos entre 2000 y julio de 2019, sobre la relación entre la resistencia a biocidas y a los antibióticos, utilizando como palabras clave "antibiotic", "biocide", "resistance" y "bacteria", en las bases de datos PubMed y Google Scholar. Conclusiones: Existe numerosa evidencia científica que indica la relación entre bacterias resistentes a los desinfectantes y la adquisición de resistencia a los antibióticos, lo que implica un replanteamiento del uso de esas sustancias en diferentes contextos clínicos, con el fin de minimizar el impacto que puedan tener en la selección de microorganismos multirresistentes a los antibióticos.

Abstract Aim: To summarize the main mechanisms of action of disinfectants and to describe some common resistance mechanisms shared between biocides and antibiotics. Methods: A bibliographic review of scientific articles was carried out between 2000 and July 2019 on the relationship between resistance to biocides and antibiotics, using as keywords "antibiotic", "biocide", "resistance" and "bacteria" in the data bases of PubMed and Google Scholar. Conclusions: Numerous scientific studies indicate a relationship between disinfectant-resistant bacteria and the acquisition of antibiotic resistance. It is necessary to analyze the use of these substances in the clinical context in order to minimize the impact that they may have on the selection of antibiotic-resistant microorganisms.

Isolation of Fungi and Bacteria Associated with the Guts of Tropical Wood-Feeding Coleoptera and Determination of Their Lignocellulolytic Activities.

The guts of beetle larvae constitute a complex system where relationships among fungi, bacteria, and the insect host occur. In this study, we collected larvae of five families of wood-feeding Coleoptera in tropical forests of Costa Rica, isolated fungi and bacteria from their intestinal tracts, and determined the presence of five different pathways for lignocellulolytic activity. The fungal isolates were assigned to three phyla, 16 orders, 24 families, and 40 genera; Trichoderma was the most abundant genus, detected in all insect families and at all sites. The bacterial isolates were assigned to five phyla, 13 orders, 22 families, and 35 genera; Bacillus, Serratia, and Pseudomonas were the dominant genera, present in all the Coleopteran families. Positive results for activities related to degradation of wood components were determined in 65% and 48% of the fungal and bacterial genera, respectively. Our results showed that both the fungal and bacterial populations were highly diverse in terms of number of species and their phylogenetic composition, although the structure of the microbial communities varied with insect host family and the surrounding environment. The recurrent identification of some lignocellulolytic-positive inhabitants suggests that particular microbial groups play important roles in providing nutritional needs for the Coleopteran host.

Study of the diversity of culturable actinomycetes in the North Pacific and Caribbean coasts of Costa Rica.

In this study, 137 actinomycetes were isolated from subtidal marine sediments in the North Pacific and Caribbean coasts of Costa Rica. Bioinformatics analysis of the 16S rRNA gene sequences assigned the isolates to 15 families and 21 genera. Streptomyces was the dominant genus while the remaining 20 genera were poorly represented. Nearly 70% of the phylotypes presented a coastal-restricted distribution whereas the other 30% were common inhabitants of both shores. The coastal tropical waters of Costa Rica showed a high diversity of actinomycetes, both in terms of the number of species and phylogenetic composition, although significant differences were observed between and within shores. The observed pattern of species distribution might be the result of several factors including the characteristics of the ecosystems, presence of endemic species and the influence of terrestrial runoff.

Dry-season leaf flushing of Enterolobium cyclocarpum (ear-pod tree): above- and below-ground phenology and water relations.

Above- and belowground phenology and water relations of Enterolobium cyclocarpum Jacq. trees in the dry forest of Santa Rosa National Park, Costa Rica were studied during two consecutive phenological cycles, from November 1998 to June 2000. Aboveground phenological activity, including leaf shedding, growth and maturation of dormant fruits, new leaf flushing and flowering, occurred during the dry season. Measurements of leaf water potential, stomatal conductance and sap flow indicated that stomata of newly flushed leaves remained essentially closed until the onset of the first rains, suggesting that the main factor accounting for the favorable water balance of dry-season flushed leaves was their capacity to restrict water loss. Evidence of a contribution from stem and root water stores to shoot expansion was mixed because only the first dry-season flushing episode monitored was accompanied by a marked decrease in stem and root water potentials. Fine root production did not precede leaf flushing, occurred only after the onset of the rainy season and stopped under drought conditions, suggesting that soil water content was the most important variable controlling fine root dynamics in this species.