Laboratory activity to teach about the proliferation of salmonella in vegetables

We designed a three-week laboratory experience that can complement any Microbiology teaching laboratory to expand students’ knowledge of the ecology of human enteric pathogens outside of their animal hosts. Through their participation in this laboratory activity, students learned that vegetative and reproductive plant parts could be a natural habitat for enteric bacteria such as non-typhoidal strains of Salmonella enterica. This field was recently brought to the forefront of the scientific and public interest by outbreaks of human illness linked to the consumption of fresh fruits and vegetables. Students were encouraged to develop their own testable hypotheses to compare proliferation of Salmonella enterica sv Typhimurium LT2 in different vegetables: cherry and regular size tomatoes, onions, lettuce, yellow and red bell peppers (Escherichia coli can be substituted for BSL1 laboratories). Upon completion of the laboratory experience, students were able to: 1) Develop testable hypotheses addressing the ability of a human pathogen Salmonella enterica to colonize and proliferate in vegetables; 2) Determine that different vegetables support the growth of Salmonella to a different extent; 3) Conduct statistical analysis and identify any significant differences. The teaching-learning process was assessed with a pre/post test, with an average increase in content understanding from ~15% to 85%. We also measured students’ proficiency while conducting specific technical tasks, revealing no major difficulties while conducting the experiments. Students indicated satisfaction with organization and content of the practices. 100% of the students agreed that the exercises improved their knowledge of this subject

Frontiers in Plant Breeding: Perspectives for the Selection of Vegetables Less Susceptible to Enteric Pathogens

Fresh vegetables including baby greens, microgreens, and sprouts can host human pathogens without exhibiting any visible signs of spoilage. It is clear that the vast majority of foodborne disease outbreaks associated with vegetable produce are not simply a result of an oversight by a producer, as it was shown that zoonotic pathogens fromEnterobacteriaceaecan contaminate produce through various routes throughout the entire production cycle. In this context, phenotypic and genotypic signatures have been used since early ages in agriculture to obtain better produce, and can be used today as a strategy to reduce the risk of outbreaks through plant breeding. In this mini-review, we provide an updated view and perspectives on to what extent the selection of biological markers can be used to select safer cultivars of vegetable crops such as tomato (the most studied), leafy greens and cabbage. Once this knowledge will be better consolidated, these approaches should be integrated into the development of comprehensive farm-to-fork produce safety programs

Fitness of antibiotic-resistant bacteria in the environment: a laboratory activity

In this laboratory experiment, we propose an opportunity for students to broaden their understanding of the ecology of antibiotic-resistant and sensitive waterborne bacteria. Antibiotics can be found in rivers or soil as a consequence of agricultural practices or as a result of human use. Concentrations of antibiotics in the environment may range from a few ng to μg L-1. Such concentrations can affect the selection and fitness of resistant bacteria. In this laboratory activity, students learn how to set up a fitness experiment by using an isogenic pair of antibiotic-resistant and sensitive bacteria in the presence or absence of selective pressure. Microcosms were generated by using filtered river water containing populations of resistant and sensitive bacteria. Competition of both populations was measured in the presence or absence of antibiotics. Students appreciated the use of microcosms for in vitro experiments and the extent to which the fitness of resistant and sensitive bacteria changed in the presence and/or absence of a selective pressure in river water. Student learning was measured by using different types of assessments: multiple-choice, true/false, fill in the blanks, laboratory skills observations, and laboratory reports. After the laboratory activity, the percentage of correct answers significantly rose from ~20% to ~85%. Laboratory skills were also evaluated during the exercises, showing no major issues during the experiment. Students showed proficiency in analyzing the complexity of fitness data by reaching a mean of 5.57 (standard error 0.57) over a maximum score of 7 points