Potentially pathogenic bacteria isolated from Paipa cheese and its susceptibility profiles to antibiotics and biocides.

The objective of this work was to evaluate the microbiological quality of cheese produced by formal and informal micro-enterprises in Paipa, Colombia, to isolate potentially pathogenic bacteria and to determine their prevalence and resistance to antimicrobials such as antibiotics and biocides. Sixteen micro-enterprises of the seventy existing in the region were sampled during 3 years. Viable concentrations of aerobic mesophiles, total and fecal coliforms, Salmonella sp., Listeria monocytogenes, Staphylococcus sp., yeasts, and molds were determined. Seventy-three bacterial isolates were identified by 16S rRNA gene sequencing. The susceptibility of the isolates to antibiotics and biocides was determined. The results indicated that between 98 and 100% of the cheese samples (n = 48 samples) of formal and informal micro-enterprises presented populations of total and fecal coliforms and Staphylococcus sp. above the limits established by Colombian regulations and varied according to the micro-enterprise. The results also indicated that 56% of Staphylococcus isolates were S. aureus. L. monocytogenes was positive in 38% of the samples. Salmonella sp. was not detected. The coliforms that prevailed were Escherichia coli (25%), Citrobacter freundii (14%), and Proteus mirabilis (8%). All L. monocytogenes were sensitive to ampicillin but resistant to erythromycin and trimethoprim-sulfamethoxazole. S. aureus isolates were susceptible to most antibiotics, except tetracycline and erythromycin (7% resistance). Likewise, 30% of coliforms (n = 36) were multidrug-resistant to antibiotics but susceptible to biocides.

Genetic Determinants for Metal Tolerance and Antimicrobial Resistance Detected in Bacteria Isolated from Soils of Olive Tree Farms.

Copper-derived compounds are often used in olive tree farms. In a previous study, a collection of bacterial strains isolated from olive tree farms were identified and tested for phenotypic antimicrobial resistance and heavy metal tolerance. The aim of this work was to study the genetic determinants of resistance and to evaluate the co-occurrence of metal tolerance and antibiotic resistance genes. Both metal tolerance and antibiotic resistance genes (including beta-lactamase genes) were detected in the bacterial strains from Cu-treated soils. A high percentage of the strains positive for metal tolerance genes also carried antibiotic resistance genes, especially for genes involved in resistances to beta-lactams and tetracycline. Significant associations were detected between genes involved in copper tolerance and genes coding for beta-lactamases or tetracycline resistance mechanisms. A significant association was also detected between zntA (coding for a Zn(II)-translocating P-type ATPase) and tetC genes. In conclusion, bacteria from soils of Cu-treated olive farms may carry both metal tolerance and antibiotic resistance genes. The positive associations detected between metal tolerance genes and antibiotic resistance genes suggests co-selection of such genetic traits by exposure to metals.

Impact of High-Hydrostatic Pressure Treatments Applied Singly or in Combination with Moderate Heat on the Microbial Load, Antimicrobial Resistance, and Bacterial Diversity of Guacamole.

Guacamole is an avocado sauce highly appreciated for its pleasant taste and nutritional value. The present study addressed the impact of high-hydrostatic pressure (HP) treatments on the product safety and bacterial diversity. Four HP treatments, 5 min each, were applied: (A) 450 megapascals (MPa) at 22 °C; (B) 450 MPa at 50 °C; (C) 600 MPa at 22 °C; (D) 600 MPa at 50 °C. Controls and treated samples were refrigerated stored for 50 days. The residual surviving fraction was lowest for the 600 MPa treatment at 50 °C. Bacterial growth on media supplemented with antibiotics (cefotaxime and imipenem) or the biocide benzalkonium chloride was detected only from control samples but not from HP-treated samples. High throughput sequencing analysis indicated that the bacterial diversity of control samples was dominated by members of Fam. Enterobacteriaceae, but it changed to a lactic acid microbiota during storage. HP-treated samples showed reduced relative abundances of Enterobacteriaceae and lactic acid bacteria and higher abundances of Pantoea, Ralstonia and Methylobacterium. Results from the study indicate that HP treatments of guacamole at 50 °C show higher microbial inactivation compared to 22 °C. However, all treatments reduced the levels of Enterobacteriaceae and penem-tolerant bacteria and provided product stability against acidification by lactic acid bacteria.

Analysis of the Bacterial Diversity of Paipa Cheese (a Traditional Raw Cow's Milk Cheese from Colombia) by High-Throughput Sequencing.

BACKGROUND: Paipa cheese is a traditional, semi-ripened cheese made from raw cow's milk in Colombia. The aim of this work was to gain insights on the microbiota of Paipa cheese by using a culture-independent approach. METHOD: two batches of Paipa cheese from three formal producers were sampled during ripening for 28 days. Total DNA from the cheese samples was used to obtain 16S rRNA gene sequences by using Illumina technology. RESULTS: Firmicutes was the main phylum found in the cheeses (relative abundances: 59.2-82.0%), followed by Proteobacteria, Actinobacteria and Bacteroidetes. Lactococcus was the main genus, but other lactic acid bacteria (Enterococcus, Leuconostoc and Streptococcus) were also detected. Stapylococcus was also relevant in some cheese samples. The most important Proteobacteria were Enterobacteriaceae, Aeromonadaceae and Moraxellaceae. Enterobacter and Enterobacteriaceae (others) were detected in all cheese samples. Serratia and Citrobacter were detected in some samples. Aeromonas and Acinetobacter were also relevant. Other minor genera detected were Marinomonas, Corynebacterium 1 and Chryseobacterium. The principal coordinates analysis suggested that there were producer-dependent differences in the microbiota of Paipa cheeses. CONCLUSIONS: lactic acid bacteria are the main bacterial group in Paipa cheeses. However, other bacterial groups, including spoilage bacteria, potentially toxin producers, and bacteria potentially pathogenic to humans and/or prone to carry antimicrobial resistance genes are also relevant in the cheeses.