Natural Bacterial Populations Research Articles

Juvenile Hormone as a contributing factor in establishing midgut microbiota for fecundity and fitness enhancement in adult female Aedes aegypti

Understanding the factors influencing mosquitoes’ fecundity and longevity is important for designing better and more sustainable vector control strategies, as these parameters can impact their vectorial capacity. Here, we address how mating affects midgut growth in Aedes aegypti, what role Juvenile Hormone (JH) plays in this process, and how it impacts the mosquito’s immune response and microbiota. Our findings reveal that mating and JH induce midgut growth. Additionally, the establishment of a native bacterial population in the midgut due to JH-dependent suppression of the immune response has important reproductive outcomes. Specific downregulation of AMPs with an increase in bacteria abundance in the gut results in increased egg counts and longer lifespans. Overall, these findings provide evidence of a cross-talk between JH response, gut epithelial tissue, cell cycle regulation, and the mechanisms governing the trade-offs between nutrition, immunity, and reproduction at the cellular level in the mosquito gut.

Occurrence of Antimicrobial-Resistant Bacteria in Intestinal Contents of Wild Marine Fish in Chile.

Antimicrobial-resistant bacteria (ARB) from the intestinal contents of wild fish may have a relevant ecological significance and could be used as indicators of antimicrobial-resistance dissemination in natural bacterial populations in water bodies impacted by urban contamination. Thus, the occurrence of ARB in the intestinal contents of pelagic and demersal wild fishes captured in anthropogenic-impacted Coquimbo Bay in Chile was studied. Culturable counts of total and antimicrobial-resistant bacteria were determined by a spread plate method using Trypticase soy agar and R2A media, both alone and supplemented with the antimicrobials amoxicillin, streptomycin, florfenicol, oxytetracycline and ciprofloxacin, respectively. Heterotrophic plate counts of pelagic and demersal fishes ranged from 1.72 × 106 CFU g-1 to 3.62 × 109 CFU g-1, showing variable proportions of antimicrobial resistance. Representative antimicrobial-resistant isolates were identified by 16S rRNA gene sequencing, and isolates (74) from pelagic fishes mainly belonged to Pseudomonas (50.0%) and Shewanella (17.6%) genera, whereas isolates (68) from demersal fishes mainly belonged to Vibrio (33.8%) and Pseudomonas (26.5%) genera. Antimicrobial-resistant isolates were tested for susceptibility to 12 antimicrobials by an agar disk diffusion method, showing highest resistance to streptomycin (85.2%) and amoxicillin (64.8%), and lowest resistance to oxytetracycline (23.2%) and ciprofloxacin (0.7%). Only furazolidone and trimethoprim/sulfamethoxazole were statistically different (p < 0.05) in comparisons between isolates from pelagic and demersal wild fishes. Furthermore, an important number of these isolates carried plasmids (53.5%) and produced Extended-Spectrum-β-lactamases (ESBL) (16.9%), whereas the detection of Metallo-β-Lactamases and class 1-integron was rare. This study provides evidence that wild fish are important reservoirs and spreading-vehicles of ARB, carrying plasmids and producing ESBLs in Chilean marine environments.

Towards estimating the number of strains that make up a natural bacterial population

What a strain is and how many strains make up a natural bacterial population remain elusive concepts despite their apparent importance for assessing the role of intra-population diversity in disease emergence or response to environmental perturbations. To advance these concepts, we sequenced 138 randomly selected Salinibacter ruber isolates from two solar salterns and assessed these genomes against companion short-read metagenomes from the same samples. The distribution of genome-aggregate average nucleotide identity (ANI) values among these isolates revealed a bimodal distribution, with four-fold lower occurrence of values between 99.2% and 99.8% relative to ANI >99.8% or <99.2%, revealing a natural “gap” in the sequence space within species. Accordingly, we used this ANI gap to define genomovars and a higher ANI value of >99.99% and shared gene-content >99.0% to define strains. Using these thresholds and extrapolating from how many metagenomic reads each genomovar uniquely recruited, we estimated that –although our 138 isolates represented about 80% of the Sal. ruber population– the total population in one saltern pond is composed of 5,500 to 11,000 genomovars, the great majority of which appear to be rare in-situ. These data also revealed that the most frequently recovered isolate in lab media was often not the most abundant genomovar in-situ, suggesting that cultivation biases are significant, even in cases that cultivation procedures are thought to be robust. The methodology and ANI thresholds outlined here should represent a useful guide for future microdiversity surveys of additional microbial species.

Signatures of kin selection in a natural population of the bacteria Bacillus subtilis.

Laboratory experiments have suggested that bacteria perform a range of cooperative behaviors, which are favored because they are directed toward relatives (kin selection). However, there is a lack of evidence for cooperation and kin selection in natural bacterial populations. Molecular population genetics offers a promising method to study natural populations because the theory predicts that kin selection will lead to relaxed selection, which will result in increased polymorphism and divergence at cooperative genes. Examining a natural population of Bacillus subtilis, we found consistent evidence that putatively cooperative traits have higher polymorphism and greater divergence than putatively private traits expressed at the same rate. In addition, we were able to eliminate alternative explanations for these patterns and found more deleterious mutations in genes controlling putatively cooperative traits. Overall, our results suggest that cooperation is favored by kin selection, with an average relatedness of r = .79 between interacting individuals.

Antagonistic bacteria against Fusarium spp. isolated from sclerotia of Claviceps gigantea in maize (Zea mays)

Fungal sclerotia house native bacterial populations of interest for biological control of plant pathogenic fungi. The objectives of this research were: i) to explore the bacterial populations associated with Claviceps gigantea sclerotia in maize from six locations in the State of Mexico, ii) to evaluate the in vitro antagonism of these bacterial populations against three species of Fusarium phytopathogens of maize, iii) to molecularly identify the more efficient antagonists and characterize the in vitro production of metabolites. Bacterial populations in the sclerotia were calculated by direct plate count; in vitro antagonism against Fusarium graminearum, F. sublgutinans, and F. verticillioides was evaluated by confrontation in Waksman agar medium. Antagonistic bacteria were identified by partial sequencing of 16S rRNA gene and evaluated in vitro for the production of indole-3 acetic acid, siderophores, lipolytic and proteolytic activity, and mineral phosphate solubilization. The bacterial density ranged from 2.023 to 2.397 Log10UFC g-1 of sclerotia. Twenty-two strains showed in vitro antagonism against at least one Fusarium species and were identified as members of the genera Bacillus, Delftia, Micromonospora, Pseudomonas, Sphingobacterium, Staphylococcus, and Stenotrophomonas. The 22 antagonists showed in vitro lipolytic, proteolytic, siderophore-producing and phosphate-solubilizing activity; only 12 (55%) produced indole-3-acetic acid. Bacillus subtilis (BA1), Pseudomonas syringae (BA2), and Bacillus amyloliquefaciens (BA18) strains were antagonists against the three Fusarium species and produced all the metabolites evaluated.

Tailoring Heat Transfer and Bactericidal Response in Multifunctional Cotton Composites.

Through the execution of scientific innovations, "smart materials" are shaping the future of technology by interacting and responding to changes in our environment. To make this a successful reality, proper component selection, synthesis procedures, and functional active agents must converge in practical and resource-efficient procedures to lay the foundations for a profitable and sustainable industry. Here we show how the reaction time, temperature, and surface stabilizer concentration impact the most promising functional properties in a cotton-based fabric coated with silver nanoparticles (AgNPs@cotton), i.e., the thermal and bactericidal response. The coating quality was characterized and linked to the selected synthesis parameters and correlated by a parallel description of "proof of concept" experiments for the differential heat transfer (conversion and dissipation properties) and the bactericidal response tested against reference bacteria and natural bacterial populations (from a beach, cenote, and swamp of the Yucatan Peninsula). The quantification of functional responses allowed us to establish the relationship between (i) the size and shape of the AgNPs, (ii) the collective response of their agglomerates, and (iii) the thermal barrier role of a surface modifier as PVP. The procedures and evaluations in this work enable a spectrum of synthesis coordinates that facilitate the formulation of application-modulated fabrics, with grounded examples reflected in "smart packaging", "smart clothing", and "smart dressing".

Community interactions drive the evolution of antibiotic tolerance in bacteria.

The emergence of antibiotic tolerance (prolonged survival against exposure) in natural bacterial populations is a major concern. Since it has been studied primarily in isogenic populations, we do not yet understand how ecological interactions in a diverse community impact the evolution of tolerance. To address this, we studied the evolutionary dynamics of a synthetic bacterial community composed of two interacting strains. In this community, an antibiotic-resistant strain protected the other, susceptible strain by degrading the antibiotic ampicillin in the medium. Surprisingly, we found that in the presence of antibiotics, the susceptible strain evolved tolerance. Tolerance was typified by an increase in survival as well as an accompanying decrease in the growth rate, highlighting a trade-off between the two. A simple mathematical model explained that the observed decrease in the death rate, even when coupled with a decreased growth rate, is beneficial in a community with weak protective interactions. In the presence of strong interactions, the model predicted that the trade-off would instead be detrimental, and tolerance would not emerge, which we experimentally verified. By whole genome sequencing the evolved tolerant isolates, we identified two genetic hot spots which accumulated mutations in parallel lines, suggesting their association with tolerance. Our work highlights that ecological interactions can promote antibiotic tolerance in bacterial communities, which has remained understudied.

A Dietary Source of High Level of Fluoroquinolone Tolerance in mcr-Carrying Gram-Negative Bacteria.

The emergence of antibiotic tolerance, characterized by the prolonged survival of bacteria following antibiotic exposure, in natural bacterial populations, especially in pathogens carrying antibiotic resistance genes, has been an increasing threat to public health. However, the major causes contributing to the formation of antibiotic tolerance and underlying molecular mechanisms are yet poorly understood. Herein, we show that potassium sorbate (PS), a widely used food additive, triggers a high level of fluoroquinolone tolerance in bacteria carrying mobile colistin resistance gene mcr. Mechanistic studies demonstrate that PS treatment results in the accumulation of intracellular fumarate, which activates bacterial two-component system and decreases the expression level of outer membrane protein OmpF, thereby reducing the uptake of ciprofloxacin. In addition, the supplementation of PS inhibits aerobic respiration, reduces reactive oxygen species production and alleviates DNA damage caused by bactericidal antibiotics. Furthermore, we demonstrate that succinate, an intermediate product of the tricarboxylic acid cycle, overcomes PS-mediated ciprofloxacin tolerance. In multiple animal models, ciprofloxacin treatment displays failure outcomes in PS preadministrated animals, including comparable survival and bacterial loads with the vehicle group. Taken together, our works offer novel mechanistic insights into the development of antibiotic tolerance and uncover potential risks associated with PS use.

NATIVE BACTERIA IN RASPBERRY CROWN GALL REDUCE THE SEVERITY OF Agrobacterium tumefaciens

Native bacterial populations in crown galls caused by Agrobacterium tumefaciens may harbour bacteria of interest for biocontrol of this pathogen. In this study, we explored the density of native bacterial populations in crown galls of raspberry (Rubus ideaus) and evaluated their in vitro and in vivo antagonism against A. tumefaciens. Bacteria morphologically similar to A. tumefaciens were isolated from six gall samples and identified by virD2 gene sequencing. Bacterial population density was calculated by direct plate count on nutrient agar and R2A media. The in vitro antagonism efficiency index against A. tumefaciens of the most frequent bacteria was evaluated by dual confrontation on nutrient agar medium, and in vivo by inoculation of 1.5 x 108 UFC mL-1 in the root of tomato (Solanum lycopersiucm) plants under greenhouse conditions. By direct sequencing and biovar characterization, it was identified as A. tumefaciens biovar 1 in raspberry galls. Native bacterial populations in galls have variable density and their diversity is limited. By partial amplification of the 16S rRNA gene, 13 strains were identified with the highest frequency in the genera Pseudomonas (61.5 %), Bacillus (15.3 %), Alcaligenes (15.3 %) and Delftia (7.6 %). Among these, Alcaligenes faecalis showed the highest in vitro antagonism index (p ≤ 0.05) against A. tumefaciens, followed by Delftia sp. and Pseudomonas citronellolis. In vivo inoculation of tomato plants with these antagonists against Agrobacterium tumefaciens did not prevent infection; however, Alcaligenes faecalis significantly reduced (p ≤ 0.05) the severity of plant stem tumours. A. faecalis is the most efficient antagonist in vitro and in vivo against A. tumefaciens.

Plasmid carriage and the natural complexity of bacterial populations contributes to plasmid persistence

Introduction: Plasmids carry and transport genes that assist their hosts to survive in many environments. Many studies have examined the conditions for plasmid persistence in bacterial populations. A limitation includes that a majority of the mathematical models for examining plasmid persistence only included bacteria from similar colonies. However, most bacterial cells inhabit complex communities where plasmids disseminate between varied bacterial host cells. Thus, there is a gap in knowledge concerning the persistence of plasmids in natural bacterial populations. To address a few of these gaps in knowledge, the present study attempted to examine the effects of plasmid carriage on intrinsic stages of bacterial populations in Bacillus subtilis co-cultures. Material and methods: B. subtilis cells were transformed with CRISPR-hCas-9 plasmid vectors where the natural phases of bacterial growth, biofilm production, and antibiotic resistance were examined in relation to plasmid carriage. These three natural phases were measured in relation to plasmid carriage through in vitro co-culture assays. Results: After calculating the CFU/mL, bacterial growth in the B. subtilis-Carrier with Escherichia coli (B. sub-C-E. coli) and Vibrio harveyi (B. sub-C-VH) co-cultures significantly decreased with a paired-t-test two-tailed P=0. The WT B. subtilis-V.H samples, the B. subtilis Carrier-V.H co-cultures, and the controls each scored a total of 40, 47, and 46 of crystal violet (CV) intensity of biofilm, respectively. Biofilm formation decreased after co-culturing E. coli with the B. subtilis-Carrier, yielding a P&lt;0.001. The antibiotic resistance levels of the co-cultures increased by 3% for the B. sub-C-V.H samples while the B. sub-C-E. coli co-cultures decreased in antibiotic sensitivity by approximately 1.5%. Conclusions: Plasmid carriage contributes to plasmid persistence via altering the natural phases of bacterial populations.

Kin selection for cooperation in natural bacterial populations

Bacteria produce a range of molecules that are secreted from the cell and can provide a benefit to the local population of cells. Laboratory experiments have suggested that these "public goods" molecules represent a form of cooperation, favored because they benefit closely related cells (kin selection). However, there is a relative lack of data demonstrating kin selection for cooperation in natural populations of bacteria. We used molecular population genetics to test for signatures of kin selection at the genomic level in natural populations of the opportunistic pathogen Pseudomonas aeruginosa We found consistent evidence from multiple traits that genes controlling putatively cooperative traits have higher polymorphism and greater divergence and are more likely to harbor deleterious mutations relative to genes controlling putatively private traits, which are expressed at similar rates. These patterns suggest that cooperative traits are controlled by kin selection, and we estimate that the relatedness for social interactions in P. aeruginosa is r = 0.84. More generally, our results demonstrate how molecular population genetics can be used to study the evolution of cooperation in natural populations.

Pan-Genome Portrait of Bacillus mycoides Provides Insights into the Species Ecology and Evolution.

ABSTRACTBacillus mycoides is poorly known despite its frequent occurrence in a wide variety of environments. To provide direct insight into its ecology and evolutionary history, a comparative investigation of the species pan-genome and the functional gene categorization of 35 isolates obtained from soil samples from northeastern Poland was performed. The pan-genome of these isolates is composed of 20,175 genes and is characterized by a strong predominance of adaptive genes (∼83%), a significant amount of plasmid genes (∼37%), and a great contribution of prophages and insertion sequences. The pan-genome structure and phylodynamic studies had suggested a wide genomic diversity among the isolates, but no correlation between lineages and the bacillus origin was found. Nevertheless, the two B. mycoides populations, one from Białowieża National Park, the last European natural primeval forest with soil classified as organic, and the second from mineral soil samples taken in a farm in Jasienówka, a place with strong anthropogenic pressure, differ significantly in the frequency of genes encoding proteins enabling bacillus adaptation to specific stress conditions and production of a set of compounds, thus facilitating their colonization of various ecological niches. Furthermore, differences in the prevalence of essential stress sigma factors might be an important trail of this process. Due to these numerous adaptive genes, B. mycoides is able to quickly adapt to changing environmental conditions.IMPORTANCE This research allows deeper understanding of the genetic organization of natural bacterial populations, specifically, Bacillus mycoides, a psychrotrophic member of the Bacillus cereus group that is widely distributed worldwide, especially in areas with continental cold climates. These thorough analyses made it possible to describe, for the first time, the B. mycoides pan-genome, phylogenetic relationship within this species, and the mechanisms behind the species ecology and evolutionary history. Our study indicates a set of functional properties and adaptive genes, in particular, those encoding sigma factors, associated with B. mycoides acclimatization to specific ecological niches and changing environmental conditions.

Nationwide genomic atlas of soil-dwelling Listeria reveals effects of selection and population ecology on pangenome evolution.

Natural bacterial populations can display enormous genomic diversity, primarily in the form of gene content variation caused by the frequent exchange of DNA with the local environment. However, the ecological drivers of genomic variability and the role of selection remain controversial. Here, we address this gap by developing a nationwide atlas of 1,854 Listeria isolates, collected systematically from soils across the contiguous United States. We found that Listeria was present across a wide range of environmental parameters, being mainly controlled by soil moisture, molybdenum and salinity concentrations. Whole-genome data from 594 representative strains allowed us to decompose Listeria diversity into 12 phylogroups, each with large differences in habitat breadth and endemism. 'Cosmopolitan' phylogroups, prevalent across many different habitats, had more open pangenomes and displayed weaker linkage disequilibrium, reflecting higher rates of gene gain and loss, and allele exchange than phylogroups with narrow habitat ranges. Cosmopolitan phylogroups also had a large fraction of genes affected by positive selection. The effect of positive selection was more pronounced in the phylogroup-specific core genome, suggesting that lineage-specific core genes are important drivers of adaptation. These results indicate that genome flexibility and recombination are the consequence of selection to survive in variable environments.

Biosensor Chip for Monitoring of Water Pollution as Novel Test Tool in Public Health: Proof of Principle

Humans are generally exposed to a variety of pollutions present in the air they breathe, the food they eat or in the water they drink. Some of the most dangerous pollutions are metals and heavy metals. These are naturally occurring substances which are harmless when present in the environment at low levels. However, due to many pollutants such as industry processes or war activities, the heavy metal concentration can exceed the limit of tolerance and become very toxic for the natural environment and living organisms in it, including humans. Unlike organic pollutants, the heavy metals (as ions and as particulate matter) once introduced into the environment cannot be biodegraded and remain there indefinitely. By rainfall these pollutants can be partially transferred from air or soil into the rivers and drinking water sources, where they accumulate in even higher toxic levels. The high concentrations of heavy metals in contaminated natural water reservoirs have an impact on the microbial community composition which resides there. This type of water pollution can cause the changes in life cycles of natural bacterial populations, influencing their metabolic processes and proliferation. The presence of pathogens in water is normally indirectly determined by the testing for “indicator organism” such as coliform bacteria. Coliforms are usually present in larger numbers in contaminated water and at the same time they are indicators of whether other pathogenic bacteria are present, too. In crisis situations, like war or some natural disasters, where trusted sources of drinking water are not available anymore, the military and residents of affected areas are forced to use some alternative water resources that cannot be tested for their microbial or metal contamination properly. Therefore, the existence of some fast test that would detect not only dangerous bacterial pathogens in water, but also the presence of metals and heavy metals as well, would be of great help and importance for the human health. Even though the number of pathogens can be drastically reduced by the boiling of water, the heavy metals are not destroyed by high temperature. Hence the main objective of our work was to optimize the biosensor chip for microbial detection in contaminated water that would serve at the same time as an indicator for the chemical composition of the water, such as presence of metals and heavy metals, with potential to be used as a novel test tool in public health.

The Effect of Inoculation of a Diazotrophic Bacterial Consortium on the Indigenous Bacterial Community Structure of Sugarcane Apoplast Fluid.

The extracellular space in plants, termed the apoplast, has a pH and sugar content that enables bacterial growth and represents a possible niche for the establishment of endophytic bacteria. Previous studies have investigated the effects of diazotrophic bacteria inoculation in sugarcane varieties, but it has not yet been analyzed how the microbial community of apoplast fluid of sugarcane is affected. High-throughput next generation sequencing of the 16S rRNA gene was used throughout this study to determine the effect of inoculation with a diazotrophic bacteria consortium, previously isolated from sugarcane, on the native bacterial population of sugarcane variety RB867515 grown in the field. The analyses were carried out 450days after inoculation. The results revealed the presence of 22 phyla, with predominance of Proteobacteria phylum. It was observed that the inoculated consortium changed the indigenous bacterial community structure of sugarcane apoplast fluid by decreasing diversity and evenness, interfering in the composition of rare species. Microbial community composition analysis revealed differences between treatments. The differential abundance test showed there were 43 amplicon sequence variants (ASVs) which were relatively more abundant in the inoculated treatment, with predominance of the Sphingomonas genus. The predicted functions of the most abundant ASVs revealed the presence of genera related to plant growth promotion and protection against phytopathogens. Analysis to evaluate the occurrence of inoculated strains in the recovered data was not conclusive since the ASVs taxonomically close to the inoculated bacteria were observed in low abundance. The present study is the first report to elucidate the bacterial community in sugarcane apoplast fluid using a culture-independent approach. It demonstrated that the diazotrophic bacterial consortium interferes in the natural bacterial community in sugarcane variety RB867515.

Effective Microorganisms in Producing Eco-Enzyme from Food Waste for Wastewater Treatment

The problem of food wastage is becoming an increasingly worrying topic as the world is heading towards sustainability to preserve the quality and the perseverance of life on Earth. Such food waste is inclusive of fresh fruit and vegetable (FFV) which make up a substantial 60% of all food wasted around the globe. Hence the purpose of this study is to valorize the FFV waste into eco-enzymes for the purpose of wastewater treatment, as having clean water sources and cleaning greywater and blackwater before discharge back into the environment are an important factors in preserving public health worldwide. Only a total of 1% of the world's freshwater is accessible for human consumption, therefore it is important to save and recycle this precious resource. The objective of this study was to fully utilize the food waste in wastewater treatment. The production of eco-enzyme is done via the process of fermentation of FFV. In this study, eco-enzymes were generated from waste derived from vegetable and fruit. The vegetable eco-enzyme was fermented via S. cerevisiae for a total of 7 days. The fruit-produced eco-enzyme was fermented with brown sugar for three months with the native bacterial population present within the fruit as the fermentation agent. The eco-enzymes produced were used to treat the water samples collected from Menjalara Lake (3°11'42.2"N 101°37'39.7"E) and Keroh River (3°12'25.0"N 101°38'27.9"E). In addition, household rice-rinsed water was obtained from Kajang (3°07'38.0"N 101°51'83.0"E). After the fermentation, the eco-enzymes solutions were acidic with a pH range of 4.1-5.24, which could be due to the metabolization of carbohydrates into volatile and organic acids. The effectiveness of the eco-enzyme treatments was assessed via measuring Ca2+, Na+, K+, NO3- and pH. It was found that the fruit-produced eco-enzyme was effective in reducing NO3-, in all three water samples. But both the fruit-produced eco-enzyme and vegetable-produced eco-enzyme were not effective in reducing the concentration of Ca2+, Na+ and K+. Further improvements that can be done are the prolonging of treatment duration from five days to 30 days. Other than that, experimentation with different concentrations to reduce the introduction of excess metal ions into the treated water samples.

Improvement of Raw Milk Cheese Hygiene through the Selection of Starter and Non-Starter Lactic Acid Bacteria: The Successful Case of PDO Pecorino Siciliano Cheese.

This review article focuses on the technological aspects and microbiological critical points of pressed-cooked cheeses processed from raw ewe’s milk without the inoculation of starter cultures, in particular “Pecorino” cheese typology produced in Italy. After showing the composition of the biofilms adhering to the surface of the traditional dairy equipment (mainly wooden vat used to collect milk) and the microbiological characteristics of PDO Pecorino Siciliano cheese manufactured throughout Sicily, this cheese is taken as a case study to develop a strategy to improve its hygienic and safety characteristics. Basically, the natural lactic acid bacterial populations of fresh and ripened cheeses were characterized to select an autochthonous starter and non-starter cultures to stabilize the microbial community of PDO Pecorino Siciliano cheese. These bacteria were applied at a small scale level to prove their in situ efficacy, and finally introduced within the consortium for protection and promotion of this cheese to disseminate their performances to all dairy factories. The innovation in PDO Pecorino Siciliano cheese production was proven to be respectful of the traditional protocol, the final cheeses preserved their typicality, and the general cheese safety was improved. An overview of the future research prospects is also reported.

Evaluation of the bacterial populations present in Spring Mix salad and their impact on the behavior of Escherichia coli O157:H7

The native bacterial populations present in three brands of Spring Mix salads (brands A, B, and C) were analyzed by plating and 16S rRNA gene sequencing during 15 days of storage at 4 °C. While the traditional plating method did not show significant differences among three brands or between before and after storage, 16S rRNA gene sequencing showed that the “brand” factor significantly impacted the diversity and abundance of native bacterial populations. In order to evaluate the interaction between native bacteria and Escherichia coli O157:H7 (ECO157), day-0 and day-15 salad homogenates were inoculated with ECO157 (a final concentration of 5.5 log CFU/ml). Inoculated samples were stored at 4 °C for additional 15 days and the behavior of inoculated ECO157 was monitored by plating samples on CHROMagar™ O157 agar on days 0, 5, 10 and 15. The top four phyla of bacteria present in all three brands of salad were Proteobacteria, Bacterioidetes, Actinobacteria, and Firmicutes. Changes in the relative abundance and diversity of bacterial populations were significantly altered by inoculation of ECO157. The inoculation remarkably decreased the beta diversity (Jaccard distance) of the bacterial populations (P = 0.005). During the additional 15-day storage, culturable ECO157 cells decreased over the entire storage time. However, fluctuations of culturable ECO157 cells were observed on days 5 and 10. In day-0 brand A and day-15 brands A and C, culturable ECO157 cells were significantly higher on day 10 than on day 5. Microbiome analysis results showed that the relative abundance of Planomicrobium, Aeromonas, Carnobacterium, and Fusobacterium were negatively correlated with the decreasing numbers of culturable ECO157 cells from day 5 to day 10 after inoculation and the relative abundance of genera Methylotenera, Yersinia, Shewanella, Dyadobacter, and Brochothrix were positively correlated with the increasing numbers of culturable ECO157 cells from day 5 to day 10. This study provides in-depth information on the microbiome changes in bagged salad and their impact on food safety, which can assist with the risk assessment of produce products of different shelf life and facilitate the development of new intervention strategies for protecting the microbial safety of fresh produce.

Bioremediation of hexavalent chromium-contaminated wastewater by Bacillus thuringiensis and Staphylococcus capitis isolated from tannery sediment

The widespread use of hexavalent chromium Cr(VI) in the leather industry causes substantial environmental problems when effluents are left untreated. Therefore, the present work attempts to assess the ability of Bacillus thuringiensis (V45) and Staphylococcus capitis (S21), isolated from tannery industry sediment, to detoxify Cr(VI) by reducing the oxidation state. Initially, the minimum tolerance of chromium by both bacteria was found up to 1000 μg/mL. V45 could tolerate Cr(VI) (520 μg/mL), and S21 could also tolerate hexavalent Cr(VI) (340 μg/mL). Similarly, both bacteria were able to tolerate other metals such as Hg2+ (40 μg/mL), Cu2+ (30 μg/mL), Ni2+ (60 μg/mL), Zn2+ (40 μg/mL), and Pb2+ (30 μg/mL). V45 and S21 could decrease Cr(VI) at a primary concentration of 50 μg/mL up to 86.42% and 97.34%, respectively. In optimization experiments, the best temperature to decrease Cr(VI) was shown to be 35 °C with pH 7 for 96 h. The occurrence of Cu2+ and Na+ slightly increased during the decrease of hexavalent Cr(VI) by V45, while the isolate S21 exhibited the same effects with Cu2+, Mn2+, and Na+. The carboxylate and amino conjugates in the biomass are intricate in the bioreduction of Cr(VI), as confirmed by FTIR spectroscopy. In addition, SEM imagery revealed the accumulation of Cr(VI) around both types of bacterial cells. The occurrence of other elements was evident from SEM-EDS spectroscopy. This study demonstrated the ability of native bacterial populations (V45 and S21) in tannery sediment to reduce Cr(VI) compounds.

Cryptic roles of tetrathionate in the sulfur cycle of marine sediments: microbial drivers and indicators

Abstract. To explore the potential role of tetrathionate in the sedimentary sulfur cycle, population ecology of microorganisms capable of metabolizing this polythionate was revealed at 15–30 cm resolution along two, ∼3 m long, cores collected from 530 and 580 m below the sea level, off India's west coast, within the oxygen minimum zone (OMZ) of the Arabian Sea. Metagenome analysis along the cores revealed widespread occurrence of genes involved in the formation, oxidation, and reduction of tetrathionate; high diversity and relative abundance were also detected for bacteria that are known to render these metabolisms in vitro. Results of slurry culture of the sediment samples in thiosulfate- or tetrathionate-containing microbial growth media, data obtained via pure-culture isolation, and finally metatranscriptome analyses corroborated the in situ functionality of the tetrathionate-forming, tetrathionate-oxidizing, and tetrathionate-reducing microorganisms. Ion chromatography of pore waters revealed the presence of up to 11.1 µM thiosulfate in the two cores, whereas tetrathionate remained undetected in spectroscopic assay based on its reaction with cyanide. While thiosulfate oxidation by chemolithotrophic bacteria prevalent in situ is the apparent source of tetrathionate in this ecosystem, high biochemical and geochemical reactivity of this polythionate could be instrumental in its cryptic status in the sulfur cycle. Potential abiotic origin of tetrathionate in the sediment horizon explored could neither be ruled out nor confirmed from the geochemical information available. On the other hand, tetrathionate potentially present in the system can be either oxidized to sulfate or reduced back to thiosulfate/sulfide via chemolithotrophic oxidation and respiration by native bacterial populations, respectively. Up to 2.01 mM sulfide present in the sediment cores may also reduce tetrathionate abiotically to thiosulfate and elemental sulfur. However, in the absence of measured data for O2 or other oxyanions having possibilities of serving as electron acceptors, the biogeochemical modalities of the oxidative half of the tetrathionate cycle remained unresolved.