Salmonella spp. is one of the most important causal agents of food-borne illness in developed countries and its presence in irrigation water poses a risk to public health. Its detection in environmental samples is not easy when culture methods are used, and molecular techniques such as PCR or ribosomal rRNA probe hybridization (Fluorescent in situ Hybridization, FISH) are outstanding alternatives. The aim of this work was to determine the environmental risk due to the presence of Salmonella spp. in wastewater by culture, PCR and FISH. A new specific rDNA probe for Salmonella was designed and its efficiency was compared with the rest of methods Serotype and antibiotic resistance of isolated strains were determined. Forty-five wastewater samples (collected from two secondary wastewater treatment plants) were analysed. Salmonella strains were isolated in 24 wastewater samples (53%), two of them after disinfection treatment. Twenty-three Salmonella strains exhibited resistance to one or more antimicrobial agent. Analysis of wastewater samples yielded PCR positive results for Salmonella in 28 out of the 45 wastewater samples (62%). FISH analysis allowed for the detection of Salmonella in 27 (60%) samples. By using molecular methods, Salmonella was detected in four samples after disinfection treatment. These results show the prevalence of Salmonella in reclaimed wastewater even after U.V. disinfection, what is a matter of public health concern, the high rates of resistance to antibiotics and the adequacy of molecular methods for its rapid detection. FISH method, with SA23 probe developed and assayed in this work provides a tool for detecting Salmonella in water within few hours, with a high rate of effectiveness.

Fresh fruits and vegetables are potential reservoirs for antimicrobial resistance determinants, but few studies have focused specifically on organic vegetables. The present study aimed to determine the presence of third-generation cephalosporin (3GC)- and carbapenem-resistant Gram-negative bacteria on fresh organic vegetables produced in the city of Valencia (Spain). Main expanded spectrum beta-lactamase (ESBL)- and carbapenemase-encoding genes were also detected in the isolates. One hundred and fifteen samples were analyzed using selective media supplemented with cefotaxime and meropenem. Resistance assays for twelve relevant antibiotics in medical use were performed using a disc diffusion test. A total of 161 isolates were tested. Overall, 33.5% presented multidrug resistance and 16.8% were resistant to all β-lactam antibiotics tested. Imipenem resistance was observed in 18% of isolates, and low resistance levels were found to ceftazidime and meropenem. Opportunistic pathogens such as Acinetobacter baumannii, Enterobacter spp., Raoultella sp., and Stenotrophomonas maltophilia were detected, all presenting high rates of resistance. PCR assays revealed bla_VIM to be the most frequently isolated ESBL-encoding gene, followed by bla_TEM and bla_OXA-48. These results confirm the potential of fresh vegetables to act as reservoirs for 3GC- and carbapenem-producing ARB. Further studies must be carried out to determine the impact of raw organic food on the spread of AMRs into the community.

The present work describes an alternative method for detecting and identifying Listeria monocytogenes in food samples by developing a nanophotonic biosensor containing bioreceptors and optical transducers. The development of photonic sensors for the detection of pathogens in the food industry involves the implementation of procedures for selecting probes against the antigens of interest and the functionalization of the sensor surfaces on which the said bioreceptors are located. As a previous step to functionalizing the biosensor, an immobilization control of these antibodies on silicon nitride surfaces was carried out to check the effectiveness of in plane immobilization. On the one hand, it was observed that a Listeria monocytogenes-specific polyclonal antibody has a greater binding capacity to the antigen at a wide range of concentrations. A Listeria monocytogenes monoclonal antibody is more specific and has a greater binding capacity only at low concentrations. An assay for evaluating selected antibodies against particular antigens of Listeria monocytogenes bacteria was designed to determine the binding specificity of each probe using the indirect ELISA detection technique. In addition, a validation method was established against the reference method for many replicates belonging to different batches of meat-detectable samples, with a medium and pre-enrichment time that allowed optimal recovery of the target microorganism. Moreover, no cross-reactivity with other nontarget bacteria was observed. Thus, this system is a simple, highly sensitive, and accurate platform for L. monocytogenes detection.

Changes undergone by tomato-antioxidants during gastrointestinal digestion of raw and fried tomato, with or without presence of the probiotic Lactobacillus reuteri ATCC 55730, were studied. Frying process enhanced the extractability of antioxidant compounds, being their content higher in fried than in raw tomato. In vitro digestion led to a significant loss of antioxidant activity (65 and 75% losses for raw and fried tomato, respectively), and total lycopene (60 and 50% losses for raw and fried tomato, respectively); and promoted trans-cis lycopene isomerization initiated during frying. Bioaccessibility of the antioxidant compounds was within 10% and 30%, being higher for phenolic compounds in raw tomato but lower for total lycopene. Finally, although the presence of Lactobacillus reuteri ATCC 55730 reduced the bioaccessibility of antioxidant compounds, the results suggests that the tomato’s antioxidant compounds could have a protective effect against the loss of viability of the probiotic.

To benefit the health of consumers, bioactive compounds must reach an adequate concentration at the end of the digestive process. This involves both an effective release from the food matrix where they are contained and a high resistance to exposure to gastrointestinal conditions. Accordingly, this study evaluates the impact of trehalose addition (10% w/w) and homogenization (100 MPa), together with the structural changes induced in vacuum impregnated apple slices (VI) by air-drying (AD) and freeze-drying (FD), on Lactobacillus salivarius spp. salivarius (CECT 4063) survival and the bioaccessibility of antioxidants during in vitro digestion. Vacuum impregnated apple slices conferred maximum protection to the lactobacillus strain during its passage through the gastrointestinal tract, whereas drying with air reduced the final content of the living cells to values below 10 cfu/g. The bioaccessibility of antioxidants also reached the highest values in the VI samples, in which the release of both the total phenols and total flavonoids to the liquid phase increased with in vitro digestion. The addition of trehalose and homogenization at 100 MPa increased the total bioaccessibility of antioxidants in FD and AD apples and the total bioaccessibility of flavonoids in the VI samples. Homogenizing at 100 MPa also increased the survival of L. salivarius during in vitro digestion in FD samples.