Prevalence of Antibiotic-Resistant E. coli Strains in a Local Farm and Packing Facilities of Honeydew Melon in Hermosillo, Sonora, Mexico.

Pathogenic strains of Escherichia coli threaten public health due to their virulence factors and antibiotic resistance. Additionally, the virulence of this bacterium varies by region depending on environmental conditions, agricultural practices, and the use of antibiotics and disinfectants. However, there is limited research on the prevalence of antibiotic-resistant E. coli in agriculture. Therefore, this research aimed to determine the antibiotic resistance of E. coli isolated from the Honeydew melon production system in Hermosillo, Sonora, Mexico. Thirty-two E. coli strains were isolated from 445 samples obtained from irrigation water, harvested melons, the hands of packaging workers, boxes, and discarded melons. The resistance profile of the E. coli strains was carried out to 12 antibiotics used in antimicrobial therapeutics against this bacterium; a high level of resistance to ertapenem (100%) was detected, followed by meropenem (97%), and ampicillin (94%); 47% of the strains were classified as multidrug-resistant. It was possible to identify the prevalence of the extended-spectrum ß-lactamase (ESBLs) gene blaTEM (15.6%), as well as the non-ESBL genes qepA (3.1%) and aac(6')lb-cr (3.1%). The E. coli strains isolated from irrigation water were significantly associated with resistance to aztreonam, cefuroxime, amikacin, and sulfamethoxazole/trimethoprim. Irrigation water, packing workers' hands, and discarded melons showed a higher prevalence of antibiotic-resistant, ESBL, and non-ESBL genes of E. coli strains in a farm and packing facility of Honeydew melon in Hermosillo, Sonora.

Cactaceae plants as sources of active bioavailable phytochemicals.

Arid-land plants from the Cactaceae family are endemic to the Americas and cultivated worldwide. Cactaceous plants and their fruits contain phenolic compounds, betalains, vitamins, carotenoids, minerals, and soluble fiber. Edible cactaceous matrices can be considered functional foods since their consumption may confer health benefits. These plants could be a source of novel bioactive compounds relevant to the area of phytomedicine. However, consumption of high concentrations of active molecules is not necessarily correlated to beneficial physiological effects because phytochemicals must be released from the food matrices under physiological conditions, resist digestion-associated chemical transformations, and remain in their active state in systemic circulation until the target tissues are reached. Notably, although digestion may either increase or decrease the bioactive phytochemicals' activity and stability, non-absorbed compounds may also be relevant for human health. Additionally, food matrices' type and composition and their technological processing operations may influence the compounds' release, stability, and accessibility. Thus, this review provides insights on the feasibility of using Cactaceae plants as sources of functional compounds. It is focused on compounds' bioactivity, bioaccessibility, and overall bioavailability after their metabolic transformation. Also, it addresses the influence of food processing on bioactive compounds. Many Cactaceae species are unexplored, and our understanding of how they confer health benefits is limited. To better understand the physiological relevance, nutraceutical potential, and therapeutic feasibility of cactaceous bioactive phytochemicals, future research should focus on the metabolic stability and safety of these compounds, as well as their assimilation mechanisms (absorption, distribution, and metabolic fate).

Relevance of tracking the diversity of Escherichia coli pathotypes to reinforce food safety.

Escherichia coli is among the most prevalent food contaminant microorganisms that have evolved, generating variants based on their effects on the host; these include commensals or pathobiont strains. The last classifications of E. coli intestinal pathobionts found in this review are enteroinvasive, enterohemorrhagic, enteropathogenic, enterotoxigenic, diffusely adherent, and enteroaggregative strains. Meanwhile, the most ancestral are enteropathogenic and enteroaggregative, and the most contemporaries are the enterotoxigenic and enteroinvasive strains. These pathobionts have been proposed based on their infective mechanisms, including toxin production, adherence effects, and tissue damage. It is also evidenced that environmental stresses, including bacterial exposition to antibiotics and disinfectants, contribute to this evolution. Therefore, new antibacterial and antivirulence agents are being explored, mainly from natural sources. In this context, this review discusses the diversity of E. coli pathobionts, their participation in foodborne outbreaks, and strategies to survey and control their spread and virulence.

Antibiofilm Action of Plant Terpenes in Salmonella Strains: Potential Inhibitors of the Synthesis of Extracellular Polymeric Substances.

Salmonella can form biofilms that contribute to its resistance in food processing environments. Biofilms are a dense population of cells that adhere to the surface, creating a matrix composed of extracellular polymeric substances (EPS) consisting mainly of polysaccharides, proteins, and eDNA. Remarkably, the secreted substances, including cellulose, curli, and colanic acid, act as protective barriers for Salmonella and contribute to its resistance and persistence when exposed to disinfectants. Conventional treatments are mostly ineffective in controlling this problem; therefore, exploring anti-biofilm molecules that minimize and eradicate Salmonella biofilms is required. The evidence indicated that terpenes effectively reduce biofilms and affect their three-dimensional structure due to the decrease in the content of EPS. Specifically, in the case of Salmonella, cellulose is an essential component in their biofilms, and its control could be through the inhibition of glycosyltransferase, the enzyme that synthesizes this polymer. The inhibition of polymeric substances secreted by Salmonella during biofilm development could be considered a target to reduce its resistance to disinfectants, and terpenes can be regarded as inhibitors of this process. However, more studies are needed to evaluate the effectiveness of these compounds against Salmonella enzymes that produce extracellular polymeric substances.

Mitochondrial manganese superoxide dismutase knock-down increases oxidative stress and caspase-3 activity in the white shrimp Litopenaeus vannamei exposed to high temperature, hypoxia, and reoxygenation.

Shrimp are increasingly exposed to warmer temperatures and lower oxygen concentrations in their habitat due to climate change. These conditions may lead to oxidative stress and apoptosis. We studied the effects of high temperature, hypoxia, reoxygenation, and the combination of these factors on lipid peroxidation, protein carbonylation, and caspase-3 activity in gills of white shrimp Litopenaeus vannamei. Silencing of mitochondrial manganese superoxide dismutase (mMnSOD) was used to determine the role of this enzyme in response to the abiotic stressors described above, to avoid oxidative damage and apoptosis. In addition, mMnSOD gene expression and mitochondrial SOD activity were evaluated to determine the efficiency of silencing this enzyme. The results showed that there was no effect of the abiotic stress conditions on the thiobarbituric acid reactive substances (TBARS), but protein carbonylation increased in all the oxidative stress treatments and caspase-3 activity decreased in hypoxia at 28 °C. On the other hand, mMnSOD-silenced shrimp experienced higher oxidative stress, since TBARS, carbonylated proteins and caspase-3 activity increased in some silenced treatments. Unexpectedly, mitochondrial SOD activity increased in some of the silenced treatments as well. Altogether, these results suggest that mMnSOD has a key role in shrimp for the prevention of oxidative damage development and induction of apoptosis in response to hypoxia, reoxygenation, high temperature, and their interactions, as conditions derived from climate change.

Comparison of nutritional properties and bioactive compounds between industrial and artisan fresh tortillas from maize landraces.

Consumers are seeking for native-traditional foods to improve their intake of both nutrients and health-promoting phytochemicals. This study was designed to evaluate the difference in content of nutrients and bioactive compounds from handmade tortillas elaborated by a small-scale artisan producer and tortillas sold by a large food retailer available to consumers. All tortillas were analyzed for chemical composition, dietary fiber, calcium and phytochemical content, antioxidant capacity, and phenolic acids profile. Chemical and nutritional variation in the tortillas was estimated using principal component analysis. Data showed that artisan tortillas made from blue and white maize landraces had significantly (p < 0.05) higher content of nutritional and bioactive compounds compared to those of the supermarket. Handmade blue maize tortillas (HBMT) had a high content of free phenolics content and the highest antioxidant capacity (DPPH and ABTS methods), which was around 1.7-2.1 fold higher than that of commercially produced white maize tortillas (CWMT). Total dietary fiber was higher in HBMT (15.7 ± 1.06 g/100 g) than in CWMT (11.6 ± 0.96 g/100 g). CWMT had the lowest calcium content (42.1 ± 0.9 mg/100 g) compared to handmade tortillas (155.5 ± 4.5 mg/100 g). HPLC results indicated the presence of ferulic, p-coumaric, caffeic, syringic and 4-hydroxybenzoic acids. Interestingly, handmade tortillas from blue maize had 4.5-fold ferulic acid content compared with commercially produced white maize tortillas, consequently it can be a good source of phenolic antioxidants, particularly ferulic acid. This study showed that artisan fresh tortillas had superior nutritional-nutraceutical properties compared to CWMT.

Inhibition of Glucosyltransferase Activity and Glucan Production as an Antibiofilm Mechanism of Lemongrass Essential Oil against Escherichia coli O157:H7.

The resistance of Escherichia coli O157:H7 to disinfection is associated with its ability to form biofilms, mainly constituted by glucans produced by glucosyltransferases. Citral and geraniol, terpenes found in the essential oil of Cymbopogon citratus (EO), have proven antibacterial activity against planktonic E. coli; however, no information was found about their efficacy and mode of action against E. coli biofilms. Therefore, the inhibitory effect of C. citratus EO, citral, and geraniol on glucans production and glucosyltransferase activity as anti-biofilm mechanism against E. coli was evaluated. EO, citral, and geraniol inhibited the planktonic growth of E. coli (minimal inhibitory concentration or MIC= 2.2, 1.0, and 3.0 mg/mL, respectively) and the bacterial adhesion (2.0, 2.0, and 4.0 mg/mL, respectively) on stainless steel. All compounds decreased the glucans production; citral and geraniol acted as uncompetitive inhibitors of glucosyltransferase activity (The half maximal inhibitory concentrations or IC50 were 8.5 and 6.5 µM, respectively). The evidence collected by docking analysis indicated that both terpenes could interact with the helix finger of the glucosyltransferase responsible for the polymer production. In conclusion, C. citratus EO, citral, and geraniol inhibited glucosyltransferase activity, glucans production, and the consequent biofilm formation of E. coli O157:H7.

Postharvest application of pectic-oligosaccharides on quality attributes, activities of defense-related enzymes, and anthocyanin accumulation in strawberry.

BACKGROUND: The postharvest application of pectic-oligosaccharides (POS) as an elicitor to improve the postharvest shelf-life and nutritional quality by stimulating natural defense mechanisms in strawberries was studied. Strawberries (cv. Festival) were treated with POS (at 0, 2, 5, and 9 g L-1 ) and evaluated for firmness, weight loss, color, soluble solids, titratable acidity (TA), total phenolic and anthocyanin content, antioxidant capacity, decay, and some defense-related enzyme activity during storage at 2 ± 0.5 °C for 14 days. RESULTS: Treatment with POS significantly delayed (P < 0.05) strawberry decay, and reduced the water loss and softening of fruit during storage. Strawberries treated with POS showed a significant increase in total phenolic and anthocyanin content, and antioxidant capacity when compared with controls. Interestingly, POS induced higher activity of phenylalanine ammonia-lyase (PAL), chitinase, and ß-1,3-glucanase in strawberries. Compared to the control, the activity of enzymes was markedly increased in fruit treated with all tested POS concentrations, particularly chitinase, and ß-1,3-glucanase activities, but 5 and 9 g L-1 POS were the most effective treatments for maintaining the quality attributes and improving anthocyanin accumulation and antioxidant capacity of strawberries. CONCLUSION: These findings suggest that POS treatment could potentially be applied to maintain quality attributes, reduce decay, and further enrich the anthocyanin content and antioxidant capacity of strawberries during postharvest storage. The results also suggest that the positive effects of POS on strawberries could be associated with the rapid accumulation of chitinase and ß-1,3-glucanase activities, and the increase of PAL enzyme activity leading to the synthesis and accumulation of anthocyanins. © 2019 Society of Chemical Industry.

Cloning, expression, purification and biochemical characterization of recombinant metallothionein from the white shrimp Litopenaeus vannamei.

Metallothioneins (MTs) are cysteine rich proteins with antioxidant capacity that participate in the homeostasis and detoxification of metals and other cellular processes, and help to counteract the oxidative stress produced by Reactive Oxygen Species (ROS). The production of ROS increases during several stress conditions, including metal intoxication and hypoxia (oxygen deficiency). During hypoxia the expression of the MT gene is induced in the shrimp Litopenaeus vannamei; however, the MT protein coded by this gene has not been purified nor characterized. In this work, the coding sequence of L. vannamei MT was cloned and overexpressed in Escherichia coli as a fusion protein, containing an intein and a chitin binding domain (CBD). The MT was purified by chitin affinity chromatography and its antioxidant capacity and ability to bind cadmium (Cd) and copper (Cu) were evaluated. This MT has an antioxidant capacity of 27.23 µM equivalent to Trolox in a 100 µg/mL solution. Addition of CdCl2 to the culture media augments 273-fold the Cd content, while addition of CuCl2 increases Cu content 569-fold in the purified MT. Thus, the shrimp MT gene codes for a functional protein that has antioxidant capacity and binds Cu and Cd.

Genome-Wide Identification of Mango (Mangifera indica L.) Polygalacturonases: Expression Analysis of Family Members and Total Enzyme Activity During Fruit Ripening.

Mango (Mangifera indica L.) is an important commercial fruit that shows a noticeable loss of firmness during ripening. Polygalacturonase (PG, E.C. 3.2.1.15) is a crucial enzyme for cell wall loosening during fruit ripening since it solubilizes pectin and its activity correlates with fruit softening. Mango PGs were mapped to a genome draft using seventeen PGs found in mango transcriptomes and 48 bonafide PGs were identified. The phylogenetic analysis suggests that they are related to Citrus sinensis, which may indicate a recent evolutive divergence and related functions with orthologs in the tree. Gene expression analysis for nine PGs showed differential expression for them during post-harvest fruit ripening, MiPG21-1, MiPG14, MiPG69-1, MiPG17, MiPG49, MiPG23-3, MiPG22-7, and MiPG16 were highly up-regulated. PG enzymatic activity also increased during maturation and these results correlate with the loss of firmness observed in mango during post-harvest ripening, between the ethylene production burst and the climacteric peak. The analysis of PGs promoter regions identified regulatory sequences associated to ripening such as MADS-box, ethylene regulation like ethylene insensitive 3 (EIN3) factors, APETALA2-like and ethylene response element factors. During mango fruit ripening the action of at least these nine PGs contribute to softening, and their expression is regulated at the transcriptional level. The prediction of the tridimensional structure of some PGs showed a conserved parallel beta-helical fold related to polysaccharide hydrolysis and a modular architecture, where exons correspond to structural elements. Further biotechnological approaches could target specific softening-related PGs to extend mango post-harvest shelf life.

Effect of UV-C irradiation and low temperature storage on bioactive compounds, antioxidant enzymes and radical scavenging activity of papaya fruit.

Mature green 'Maradol' papaya fruits were exposed to ultraviolet (UV)-C irradiation (1.48 kJ·m(-2)) and stored at 5 or 14 °C. Changes in total phenols, total flavonoids, enzymatic activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), as well as the scavenging activity against 2,2-diphenyl-1picrylhydrazyl (DPPH) and 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals were investigated in peel and flesh tissues at 0, 5, 10 and 15 days of storage. UV-C irradiation increased significantly (P < 0.05) the flavonoid content (2.5 and 26 %) and ABTS radical scavenging activity (5.7 and 6 %) in flesh and peel at 14 °C respectively; and CAT activity (16.7 %) in flesh at 5 °C. Flavonoid contents, CAT and SOD activities were positively affected under low storage temperature (5 °C). DPPH and ABTS radical scavenging activities increased in both control and UV-C treated papaya peel during storage at 5 °C. UV-C irradiation effect on radical scavenging of papaya peel could be attributed to increased flavonoid content. Papaya antioxidant system was activated by UV-C and cold storage by increasing phenolic content and antioxidant enzymatic activities as a defense response against oxidative-stress.

Molecular modeling and expression analysis of a MADS-box cDNA from mango (Mangifera indica L.).

MADS-box genes are a large family of transcription factors initially discovered for their role during development of flowers and fruits. The MADS-box transcription factors from animals have been studied by X-ray protein crystallography but those from plants remain to be studied. In this work, a MADS-box cDNA from mango encoding a protein of 254 residues was obtained and compared. Based on phylogenetic analysis, it is proposed that the MADS-box transcription factor expressed in mango fruit (MiMADS1) belongs to the SEP clade of MADS-box proteins. MiMADS1 mRNA steady-state levels did not changed during mango fruit development and were up-regulated, when mango fruits reached physiological maturity as assessed by qRT-PCR. Thus, MiMADS1 could have a role during development and ripening of this fruit. The theoretical structural model of MiMADS1 showed the DNA-binding domain folding bound to a double-stranded DNA. Therefore, MiMADS1 is an interesting model for understanding DNA-binding for transcriptional regulation.

Volatiles emitted by Carya illinoinensis (Wang.) K. Koch as a prelude for semiochemical investigations to focus on Acrobasis nuxvorella Nuenzig (Lepidoptera: Pyralidae).

BACKGROUND: Plant volatiles have complex intra- and interspecific effects in the environment that include plant/herbivore interactions. Identifying the quantity and quality of volatiles produced by a plant is needed to aid the process of determining which chemicals are exerting what effects and then examining whether these effects can be manipulated to benefit society. The qualitative characterization of volatile compounds emitted by pecan, Carya illinoinensis (Wang.) K. Koch, was begun in order to establish a database for investigating how these volatiles affect Acrobasis nuxvorella Nuenzig, a monophagous pest of pecan. Headspace solid-phase microextraction combined with gas chromatography-mass spectrometry was used for the analysis of the volatile constituents of pecan during three phenological stages (dormant buds, intact new shoot growth and intact nutlets) of the Western Schley and Wichita cultivars. RESULTS: About 111 distinct compounds were identified from the two cultivars, accounting for ∼99% of the headspace volatiles. The chromatographic profiles of both varieties revealed variations in the volatile composition and proportion between cultivars, with a predominance of terpene hydrocarbons, of the sesquiterpenes class, as well as monoterpenes. CONCLUSION: The significantly higher responsiveness recorded for the larvae of A. nuxvorella to C. illinoinensis shoots indicates that the larvae may be activated by terpenes emanating from the new shoot growth. This is the first study that has examined volatiles of pecan in Mexico.

Development and characterization of edible films based on mucilage of Opuntia ficus-indica (L.).

Mucilage of Opuntia ficus-indica (OFI) was extracted and characterized by its composition and molecular weight distribution. Mucilage film-forming dispersions were prepared under different pHs (3, 4, 5.6, 7, and 8) and calcium concentration (0% and 30% of CaCl(2), with respect to mucilage's weight), and their particle size determined. Mucilage films with and without calcium (MFCa and MF, respectively) were prepared. The effect of calcium and pH on mucilage films was evaluated determining thickness, color, water vapor permeability (WVP), tensile strength (TS), and percentage of elongation (%E). The average molecular weight of the different fractions of mucilage was: 3.4 x 10(6) (0.73%), 1 x 10(5) (1.46%), 1.1 x 10(3) (45.79%), and 2.4 x 10(2) Da (52.03%). Aqueous mucilage dispersions with no calcium presented particles with an average size d(0.5) of 15.4 microm, greater than the dispersions with calcium, 13.2 microm. MFCa films showed more thickness (0.13 mm) than the MF films (0.10 mm). The addition of calcium increased the WVP of the films from 109.94 to 130.45 gmm/m(2)dkPa. Calcium and pH affected the mechanical properties of the films; the largest TS was observed on MF films, whereas the highest %E was observed on MFCa films. The highest differences among MF and MFCa films were observed at pHs 5.6 and 7 for TS and at pHs 4 and 8 for %E. No effect of pH and calcium was observed on luminosity and hue angle. Chroma values were higher for MF when compared with MFCa, and increased as pH of the films increased. Practical Application: In this study mucilage from nopal was extracted and characterized by its ability to form edible films under different pHs, and with or without the addition of calcium. Opuntia ficus-indica mucilage had the ability to form edible films. In general, it can be considered that mucilage films without modification of pH and without the addition of calcium have the best water vapor barrier properties and tensile strength. Mucilage from nopal could represent a good option for the development of edible films in countries where nopal is highly produced at low cost, constituting a processing alternative for nopal.