Tomato Plant Microbiota under Conventional and Organic Fertilization Regimes in a Soilless Culture System.

Tomato is the main vegetable cultivated under soilless culture systems (SCSs); production of organic tomato under SCSs has increased due to consumer demands for healthier and environmentally friendly vegetables. However, organic tomato production under SCSs has been associated with low crop performance and fruit quality defects. These agricultural deficiencies could be linked to alterations in tomato plant microbiota; nonetheless, this issue has not been sufficiently addressed. Thus, the main goal of the present study was to characterize the rhizosphere and phyllosphere of tomato plants cultivated under conventional and organic SCSs. To accomplish this goal, tomato plants grown in commercial greenhouses under conventional or organic SCSs were tested at 8, 26, and 44 weeks after seedling transplantation. Substrate (n = 24), root (n = 24), and fruit (n = 24) composite samples were subjected to DNA extraction and high-throughput 16S rRNA gene sequencing. The present study revealed that the tomato core microbiota was predominantly constituted by Proteobacteria, Actinobacteria, and Firmicutes. Remarkably, six bacterial families, Bacillaceae, Microbacteriaceae, Nocardioidaceae, Pseudomonadaceae, Rhodobacteraceae, and Sphingomonadaceae, were shared among all substrate, rhizosphere, and fruit samples. Importantly, it was shown that plants under organic SCSs undergo a dysbiosis characterized by significant changes in the relative abundance of Bradyrhizobiaceae, Caulobacteraceae, Chitinophagaceae, Enterobacteriaceae, Erythrobacteraceae, Flavobacteriaceae, Nocardioidaceae, Rhodobacteraceae, and Streptomycetaceae. These results suggest that microbial alterations in substrates, roots, and fruits could be potential factors in contributing to the crop performance and fruit quality deficiencies observed in organic SCSs.

A Comprehensive Evaluation of Enterobacteriaceae Primer Sets for Analysis of Host-Associated Microbiota.

Enterobacteriaceae is one of the most important bacterial groups within the Proteobacteria phylum. This bacterial group includes pathogens, commensal and beneficial populations. Numerous 16S rRNA gene PCR-based assays have been designed to analyze Enterobacteriaceae diversity and relative abundance, and, to the best of our knowledge, 16 primer pairs have been validated, published and used since 2003. Nonetheless, a comprehensive performance analysis of these primer sets has not yet been carried out. This information is of particular importance due to the recent taxonomic restructuration of Enterobacteriaceae into seven bacterial families. To overcome this lack of information, the identified collection of primer pairs (n = 16) was subjected to primer performance analysis using multiple bioinformatics tools. Herein it was revealed that, based on specificity and coverage of the 16S rRNA gene, these 16 primer sets could be divided into different categories: Enterobacterales-, multi-family-, multi-genus- and Enterobacteriaceae-specific primers. These results highlight the impact of taxonomy changes on performance of molecular assays and data interpretation. Moreover, they underline the urgent need to revise and update the molecular tools used for molecular microbial analyses.

A Molecular Tool for Rapid Detection and Traceability of Cyclospora cayetanensis in Fresh Berries and Berry Farm Soils.

Due to recent outbreaks of cyclosporiasis associated with consumption of fresh berries, producers are demanding modern microbiological tools for the rapid and accurate identification of the human pathogen Cyclospora cayetanensis in berries and environmental samples. The aim of the present work was to develop a molecular tool based on a PCR approach for the rapid and accurate detection of C. cayetanensis. A nested PCR assay was validated for the amplification of a 294 bp size region of the 18S rRNA gene from C. cayetanensis. The limit of detection for the nested PCR assay was validated using 48 berry samples spiked with ~0, 10, 100, and 1000 oocyst per gram of sample. With this assay, it was possible to detect as few as 1 oocyst per gram of berry, in a 50 g sample. Sanger DNA sequencing and phylogenetic analysis were carried out to confirm the presence of C. cayetanensis in berry (n = 17) and soil (n = 5) samples. The phylogenetic analysis revealed that the C. cayetanensis sequences obtained from Mexico clustered within a group recovered from China, Peru, Guatemala-Haiti, and Japan. The PCR protocol designed in the present study could be an important tool for the rapid and accurate detection of this human pathogen in environmental and food samples.

Three-Year Longitudinal Study: Prevalence of Salmonella Enterica in Chicken Meat is Higher in Supermarkets Than Wet Markets from Mexico.

Worldwide, chicken meat is considered one of the main sources of Salmonella enterica in humans. To protect consumers from this foodborne pathogen, international health authorities recommend the establishment of continuous Salmonella surveillance programs in meat. However, these programs are scarce in many world regions; thus, the goal of the present study was to perform a longitudinal surveillance of S. enterica in chicken meat in Mexico. A total of 1160 samples were collected and analyzed monthly from 2016 to 2018 in ten chicken meat retailers (supermarkets and wet markets) located in central Mexico. The isolation and identification of S. enterica was carried out using conventional and molecular methods. Overall, S. enterica was recovered from 18.1% (210/1160) of the chicken meat samples. Remarkably, during the three years of evaluation, S. enterica was more prevalent (P < 0.0001) in supermarkets (27.2%, 158/580) than in wet markets (9.0%, 52/580). The study was 3.8 times more likely (odds ratio = 3.8, P < 0.0001) to recover S. enterica from supermarkets than wet markets. Additionally, a higher prevalence (P < 0.05) of this pathogen was observed during the spring, summer, autumn, and winter in supermarkets compared with wet markets. Moreover, the recovery rate of S. enterica from supermarkets showed a gradual increase from 20.78% to 42% (P < 0.0001) from 2016 to 2018. Interestingly, no correlation (P > 0.05) was observed between the S. enterica recovery rate in chicken meat and reported cases of Salmonella infections in humans. Higher levels of S. enterica in chicken meat retailed in supermarkets are not unusual; this phenomenon has also been reported in some European and Asian countries. Together, these results uncover an important health threat that needs to be urgently addressed by poultry meat producers and retailers.