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In this study, seed-surface-associated bacteria from fresh fruits of Capsicum spp. were analyzed to explore potential isolates for biocontrol of phytopathogenic fungal strains. A total of 76 bacterial isolates were obtained from three different species of chili pepper (C. annuum L., C. pubescens R. & P., and C. chinense Jacq.), and two isolates were selected via mycelial growth inhibition assays based on their production of volatile organic compounds (VOCs) against six fungal strains. Genomic analysis identified these isolates as Bacillus altitudinis CH05, with a chromosome size of 3,687,823 bp and with 41.25% G+C, and Bacillus tropicus CH13, with a chromosome size of 5,283,706 bp and with 35.24% G+C. Both bacterial strains showed high mycelial growth inhibition capacities against Sclerotium rolfsii, Sclerotinia sp., Rhizoctonia solani, and Alternaria alternata but lower inhibition capacities against Colletotrichum gloesporoides and Fusarium oxysporum. VOC identification was carried out after 24 h of fermentation with 64 VOCs for B. altitudinis CH05 and 53 VOCs for B. tropicus CH13. 2,5-Dimethyl pyrazine and acetoin had the highest relative abundance values in both bacterial strains. Our findings revealed that seed-surface-associated bacteria on Capsicum spp. have the metabolic ability to produce VOCs for biocontrol of fungal strains and have the potential to be used in sustainable agriculture.

Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), also known as the fall armyworm, is an economically important and widespread polyphagous pest. Microorganisms associated to this insect during life cycle play important ecological roles. We report 3 metagenome-assembled bacterial genomes reconstructed from a metagenome dataset obtained from S. frugiperda larvae F3 3rd-instar reared using artificial diet under laboratory conditions. Genome data for Enterococcus casseliflavus indicated a genome length of 3,659,8333 bp and GC content of 42.54%. Genome data for E. mundtii indicated a genome length of 2,921,701 bp and GC content of 38.37%. Finally, genome data for Lactiplantibacillus plantarum indicated a genome length of 3,298,601 bp, GC content of 44.31%. Genome analysis allowed us to identify genus-specific protein families (PLFams), transporters and antibiotic resistance-related genes among others. DNA sequences were deposited in National Center for Biotechnology Information (https://www.ncbi.nlm.nih.gov/) as Bioproject accession PRJNA899064.

The competitive colonization of bacteria on similar ecological niches has a significant impact during their establishment. The synthesis speeds of different chemical classes of molecules during early competitive colonization can reduce the number of competitors through metabolic effects. In this work, we demonstrate for the first time that Kosakonia cowanii Cp1 previously isolated from the seeds of Capsicum pubescens R. P. produced volatile organic compounds (VOCs) during competitive colonization against Pectobacterium aroidearum SM2, affecting soft rot symptoms in serrano chili (Capsicum annuum L.). The pathogen P. aroidearum SM2 was isolated from the fruits of C. annuum var. Serrano with soft rot symptoms. The genome of the SM2 strain carries a 5,037,920 bp chromosome with 51.46% G + C content and 4925 predicted protein-coding genes. It presents 12 genes encoding plant-cell-wall-degrading enzymes (PCDEWs), 139 genes involved in five types of secretion systems, and 16 genes related to invasion motility. Pathogenic essays showed soft rot symptoms in the fruits of C. annuum L., Solanum lycopersicum, and Physalis philadelphica and the tubers of Solanum tuberosum. During the growth phases of K. cowanii Cp1, a mix of VOCs was identified by means of HS-SPME-GC-MS. Of these compounds, 2,5-dimethyl-pyrazine showed bactericidal effects and synergy with acetoin during the competitive colonization of K. cowanii Cp1 to completely reduce soft rot symptoms. This work provides novel evidence grounding a better understanding of bacterial interactions during competitive colonization on plant tissue, where VOC synthesis is essential and has a high potential capacity to control pathogenic microorganisms in agricultural systems.

Chili powder is an important condiment around the world. However, according to various reports, the presence of pathogenic microorganisms could present a public health risk factor during its consumption. Therefore, microbiological quality assessment is required to understand key microbial functional traits, such as antibiotic resistance genes (ARGs). In this study, metagenomic next-generation sequencing (mNGS) and bioinformatics analysis were used to characterize the comprehensive profiles of the bacterial community and antibiotic resistance genes (ARGs) in 15 chili powder samples from different regions of Mexico. The initial bacterial load showed aerobic mesophilic bacteria (AMB) ranging between 6 × 103 and 7 × 108 CFU/g, sporulated mesophilic bacteria (SMB) from 4.3 × 103 to 2 × 109 CFU/g, and enterobacteria (En) from 6 CFU/g. The most representative families in the samples were Bacillaceae and Enterobacteriaceae, in which 18 potential pathogen-associated species were detected. In total, the resistome profile in the chili powder contained 68 unique genes, which conferred antibiotic resistance distributed in 13 different classes. Among the main classes of antibiotic resistance genes with a high abundance in almost all the samples were those related to multidrug, tetracycline, beta-lactam, aminoglycoside, and phenicol resistance. Our findings reveal the utility of mNGS in elucidating microbiological quality in chili powder to reduce the public health risks and the spread of potential pathogens with antibiotic resistance mechanisms.

El tratamiento biológico es el más utilizado para las aguas residuales domésticas. El reactor de lecho fijo es una biopelícula de microorganismos capaces de degradar la materia orgánica, y es una alternativa eficiente en comparación con los sistemas convencionales, ya que aumenta la superficie de contacto entre el microorganismo y la materia orgánica. El presente trabajo se centra en la adaptación, aclimatación y uso de lodo activado para la formación de una biopelícula en un biorreactor para el tratamiento de efluentes domésticos con un soporte PET; se comprobó que dicho material es factible para ser implementado como medio de soporte. El sistema de lecho fijo se mantuvo en operación durante 150 días, utilizando agua residual doméstica como sustrato, y evaluando la eficiencia del reactor por medio de parámetros de calidad de agua. La diversidad microbiana se evaluó por análisis de ARNr 16S. Las eficiencias de remoción de la Demanda Química de Oxigeno (DQO) y Demanda Bioquímica de Oxigeno (DBO) fueron mayores del 90 % y se logró la neutralización del efluente con características alcalinas. El tiempo de tratamiento fue de 4 h. Se identificaron 11 Phylum bacterianos correspondientes para biopelícula y 8 en suspensión, donde los principales grupos bacterianos fueron Proteobacterias, Bacteroidetes y Firmicutes. El tiempo de tratamiento disminuyó en un 50 % respecto a los sistemas convencionales. La identificación de los Phylum permite comprender mejor la complejidad de las estructuras bacterianas en aguas residuales y como interviene en la eficiencia de los procesos del tratamiento de aguas residuales.

The Kosakonia cowanii Cp1 strain was isolated from seeds of Capsicum pubescens R. & P. cultivated in Michoacan, Mexico. Genetic and ecological role analyses were conducted for better characterization. The results show that genome has a length of 4.7 Mbp with 56.22% G + C and an IncF plasmid of 128 Kbp with 52.51% G + C. Furthermore, pathogenicity test revealed nonpathogenic traits confirmed by the absence of specific virulence-related genes. Interestingly, when fungal inhibitory essays were carried out, the bacterial synthesis of volatile organic compounds (VOCs) with antifungal activity showed that Sclerotinia sp. and Rhizoctonia solani were inhibited by 87.45% and 77.24%, respectively. Meanwhile, Sclerotium rolfsii, Alternaria alternata, and Colletotrichum gloeosporioides demonstrated a mean radial growth inhibition of 52.79%, 40.82%, and 55.40%, respectively. The lowest inhibition was by Fusarium oxysporum, with 10.64%. The VOCs’ characterization by headspace solid–phase microextraction combined with gas chromatography–mass spectrometry (HS-SPME-GC–MS) revealed 65 potential compounds. Some of the compounds identified with high relative abundance were ketones (22.47%), represented by 2-butanone, 3-hydroxy (13.52%), and alcohols (23.5%), represented by ethanol (5.56%) and 1-butanol-3-methyl (4.83%). Our findings revealed, for the first time, that K. cowanii Cp1 associated with C. pubescens seeds possesses potential traits indicating that it could serve as an effective biocontrol.

Kosakonia cowanii strain Ch1 was isolated from Mexican chili powder, and the genome was sequenced. The genome was 4,765,544 bp in length, with an average G + C content of 56.22%, and a plasmid (pCh1) of 128,063 bp with an average G + C content of 52.50%. A phylogenetic analysis revealed a close relation with pathogenic strains; nevertheless, some virulence-related genes were absent, and this genetic characteristic may explain the fact that K. cowanii Ch1 behaved as a non-pathogenic strain when infection assays were performed on the leaves and fruits of Capsicum annuum L. Surprisingly, we observed that this bacterial strain had the ability to spread throughout serrano pepper seeds. Furthermore, K. cowanii Ch1 was evaluated for the production of volatile organic compounds (VOCs) against fungal pathogens, and the results showed that Alternaria alternata and Sclerotium rolfsii were inhibited in a radial mycelial growth assay by a mean rate of 70% and 64%, while Fusarium oxysporum was inhibited by only approximately 10%. Based on the headspace solid-phase microextraction combined with the gas chromatography mass spectrometry (HS-SPME-GC-MS), 67 potential VOCs were identified during the fermentation of K. cowanii Ch1 in TSA medium. From these VOCs, nine main compounds were identified based on relative peak area: dodecanoic acid; 3-hydroxy ethanol; 1-butanol-3-methyl; acetaldehyde; butanoic acid, butyl ester; cyclodecane; 2-butanone, 3-hydroxy; disulfide, dimethyl and pyrazine-2,5-dimethyl. Our findings show the potential of K. cowanii Ch1 for the biocontrol of fungal pathogens through VOCs production and reveal additional abilities and metabolic features as beneficial bacterial specie.

Chili powder is the most frequently consumed spice in Mexican diets. Thus, the dissemination of microorganisms associated with chili powder derived from Capsicum annuum L. is significant during microbial quality analysis, with special attention on detection of potential pathogens. The results presented here describe the initial characterization of bacterial community structure in commercial chili powder samples. Our results demonstrate that, within the domain Bacteria, the most abundant family was Bacillaceae, with a relative abundance of 99% in 71.4% of chili powder samples, while 28.6% of samples showed an average relative abundance of 60% for the Enterobacteriaceae family. Bacterial load for aerobic mesophilic bacteria (AMB) ranged from 104 to 106 cfu/g, while for sporulated mesophilic bacteria (SMB), the count ranged from 102 to 105 cfu/g. Bacillus cereus sensu lato (s.l.) was observed at ca. ˂600 cfu/g, while the count for Enterobacteriaceae ranged from 103 to 106 cfu/g, Escherichia coli and Salmonella were not detected. Fungal and yeast counts ranged from 102 to 105 cfu/g. Further analysis of the opportunistic pathogens isolated, such as B. cereus s.l. and Kosakonia cowanii, using antibiotic-resistance profiles and toxinogenic characteristics, revealed the presence of extended-spectrum β-lactamases (ESBLs) and Metallo-β-lactamases (MBLs) in these organisms. These results extend our knowledge of bacterial diversity and the presence of opportunistic pathogens associated with Mexican chili powder and highlight the potential health risks posed by its use through the spread of antibiotic-resistance and the production of various toxins. Our findings may be useful in developing procedures for microbial control during chili powder production.

Background: In spore-forming bacteria, the molecular mechanisms of accumulation of transfer RNA (tRNA) during sporulation must be a priority as tRNAs play an essential role in protein synthesis during spore germination and outgrowth. However, tRNA processing has not been extensively studied in these conditions, and knowledge of these mechanisms is important to understand long-term stress survival. Methods:To gain further insight into tRNA processing during spore germination and outgrowth, the expression of the single copy tRNACys gene was analyzed in the presence and absence of 1.2 M NaCl in Bacillus subtilis using RNA-Seq data obtained from the Gene Expression Omnibus (GEO) database. The CLC Genomics work bench 12.0.2 (CLC Bio, Aarhus, Denmark, https://www.qiagenbioinformatics.com/) was used to analyze reads from the tRNACys gene. Results:The results show that spores store different populations of tRNACys-related molecules. One such population, representing 60% of total tRNACys, was composed of tRNACys fragments. Half of these fragments (3´-tRF) possessed CC, CCA or incorrect additions at the 3´end. tRNACys with correct CCA addition at the 3´end represented 23% of total tRNACys, while with CC addition represented 9% of the total and with incorrect addition represented 7%. While an accumulation of tRNACys precursors was induced by upregulation of the rrnD operon under the control of σA -dependent promoters under both conditions investigated, salt stress produced only a modest effect on tRNACys expression and the accumulation of tRNACys related species. Conclusions:The results demonstrate that tRNACys molecules resident in spores undergo dynamic processing to produce functional molecules that may play an essential role during protein synthesis.

The potential presence of spore-forming bacteria related to the Bacillus cereus group in Mexican chili powder elaborated from Capsicum annuum L. is of commercial and clinical interest, because chili powder is an essential spice in the Mexican diet and in diets around the globe. To facilitate detection and isolation of members of this group of spore-forming bacteria from Mexican chili powder samples, we identified colonies that grew on agar medium selective for Bacillus cereus sensu lato, supplemented with polymyxin B (10 µg/mL) and ampicillin (10 to 100 µg/mL). The presumptive B. cereus (s.l.) isolates were tested using a tRNACys-PCR-based approach and the results identified species related phylogenetically to B. cereus, B. thuringiensis, and B. toyonensis. Their toxigenic potential was assessed by serological tests to detect enterotoxins (Nhe and Hbl) and by PCR targeting the hemolysin BL (hbl) component C (hblC) and non-hemolytic enterotoxin component A (nheA). The antibiotic profiles of the isolates showed a high resistance to β-lactams (100% of the isolates), trimethoprim-sulfamethoxazole (100%), tetracycline (90%), erythromycin (77%), clindamycin (74%), and chloramphenicol (42%). Our results indicate the presence of B. cereus s.l. with toxigenic characteristics in Mexican chili powder. Because of the potential for these organisms to cause disease through their production of various toxins, and resistance to antibiotics, we recommend that a microbiological risk assessment must be considered in the Mexican regulatory requirements.

Sustainable processing from native vegetation such as agave, nopal and aloe, is mainly centered on the components of lignin, cellulose, and hemicellulose, defined generally as lignocellulosic. Cytotoxicity of lignocellulosic biopolymer membranes (LBMs) of mucilages of Opuntia ficus-indica, Aloe barbadensis Miller, fructans, and residual Agave tequilana Weber fibers at different concentrations (30, 50 and 70%), was evaluated in contact with human fibroblast cells (ATCC® PCS-201-010), cultured in vitro. Long and short LMBs were formed from the fibrous material. The interaction showed mild cytotoxicity according to ISO 10993-5: 2009. Cytotoxic activity demonstrated in the presence of fibroblasts implied a maximum cytotoxicity of close to 46% and a minimum of 7% in LBMs. UV light absorption results of the large fiber LMBs showed the highest prevention of passage of light, having a protective effect mechanism of delaying the response of hypersensitivity as a result of exposure to UV radiation. The morphology and structure were characterized using SEM, FT-IR, and image texture analysis. According to the LBMs reactivity measured, the feasible uses of these membranes can be suggested as materials for biomedical applications over a short period of time, with feasibility as a support to provide some beneficial solutions.

Endospore-forming bacteria related to the Bacillus cereus group produce toxins that cause illnesses in organisms from invertebrates to mammals, including foodborne illnesses in humans. As commercial bee pollen can be contaminated with these bacteria, a comprehensive microbiological risk assessment of commercial bee pollen must be incorporated into the relevant regulatory requirements, including those that apply in Mexico. To facilitate detection of members of this group of bacteria, we have developed a PCR strategy that is based on the amplification of the single-copy tRNACys gene and specific genes associated with tRNACys to detect Bacillus cereus sensu lato (B. cereus s.l.). This tRNACys-PCR-based approach was used to examine commercial bee pollen for endospore-forming bacteria. Our analysis revealed that 3% of the endospore-forming colonies isolated from a commercial source of bee pollen were related to B. cereus s.l., and this result was corroborated by phylogenetic analysis, bacterial identification via MALDI-TOF MS, and detection of enterotoxin genes encoding the HBL and NHE complexes. The results show that the isolated colonies are closely related phylogenetically to B. cereus, B. thuringiensis, and B. bombysepticus. Our results indicate that the tRNACys-PCR, combined with other molecular tools, will be a useful approach for identifying B. cereus s.l. and will assist in controlling the spread of potential pathogens.

Background: In spore-forming bacteria, the molecular mechanisms of accumulation of transfer RNA (tRNA) during sporulation must be a priority as tRNAs play an essential role in protein synthesis during spore germination and outgrowth. However, tRNA processing has not been extensively studied in these conditions, and knowledge of these mechanisms is important to understand long-term stress survival. Methods:To gain further insight into tRNA processing during spore germination and outgrowth, the expression of the single copy tRNA Cys gene was analyzed in the presence and absence of 1.2 M NaCl in Bacillus subtilis using RNA-Seq data obtained from the Gene Expression Omnibus (GEO) database. The CLC Genomics work bench 12.0.2 (CLC Bio, Aarhus, Denmark, https://www.qiagenbioinformatics.com/) was used to analyze reads from the tRNA Cys gene. Results:The results show that spores store different populations of tRNA Cys-related molecules. One such population, representing 60% of total tRNA Cys, was composed of tRNA Cys fragments. Half of these fragments (3´-tRF) possessed CC, CCA or incorrect additions at the 3´end. tRNA Cys with correct CCA addition at the 3´end represented 23% of total tRNA Cys, while with CC addition represented 9% of the total and with incorrect addition represented 7%. While an accumulation of tRNA Cys precursors was induced by upregulation of the rrnD operon under the control of σ A -dependent promoters under both conditions investigated, salt stress produced only a modest effect on tRNA Cys expression and the accumulation of tRNA Cys related species. Conclusions:The results demonstrate that tRNA Cys molecules resident in spores undergo dynamic processing to produce functional molecules that may play an essential role during protein synthesis.

Klebsiella pneumoniae Ch1-39 was isolated from chili powder elaborated at San Luis Potosí, México. This microorganism can be found in diverse ecological niches as water, soil, air, plants and hospital setting, it is considered as a relevant opportunistic pathogen causing several diseases and showing increasingly multi-resistance to antibiotics. The genome was sequenced on the Illumina NovaSeq platform and bioinformatic analyses were made at the Bacterial and Viral Bioinformatics Resource Center (BV-BRC). The genome consisted of 72 contigs with a total size of 5,410,125 bp, 5,361 protein coding sequences (CDS), a total of 6 rRNA and 76 tRNA with an average G + C content of 57.22 %. The genome data was deposited at National Center for Biotechnology Information (NCBI) under accession number Bioproject ID PRJNA1062060, Bio Sample ID SAMN40269967. The genome accession number was JBAWUH000000000.

In this study, we tested the insectistatic and insecticidal effects of the ethanolic extract of stems and bark of Ficus petiolaris Kunth (Moraceae) in laboratory bioassays with an artificial diet against newly hatched larvae of the fall armyworm, Spodoptera frugiperda Smith (Lepidoptera: Noctuidae). The extract was evaluated at five different concentrations (500, 1000, 1500, 2000, and 2500 ppm). The 2500 ppm extract had the strongest inhibitory effects on larval (89%) and pupal (20%) weight as well as the highest mortality (80%). The positive control, methyl parathion (Methyl Parathion®) at 1%, eliminated 100% of the fall armyworm, and the negative control (artificial diet) had a mortality of only 5%. Chemical fractionation of F. petiolaris stem and bark extract produced five fractions of FpR1-5, each of which was evaluated at 250, 500, 750, 1000, and 1250 ppm. FpR4 presented the strongest inhibitory effect, reducing the weight of the larva and pupa by 35% and 18%, while FpR2 had the strongest insecticidal effect, with 90% mortality at 1250 ppm. The pure 8-Methoxypsoralen compound extracted from this fraction was even more effective, with 100% S. frugiperda mortality at 100 ppm. The 50% lethal concentration (LC50) of 8-Methoxypsoralen was 67.68 ppm. Our results indicate that the F. petiolaris extract showed toxic activity against the fall armyworm, and its compound 8-Methoxypsoralen showed strong insecticidal activity at low concentrations.