Impact of the ABO and RhD Blood Groups on the Evaluation of the Erythroprotective Potential of Fucoxanthin, ß-Carotene, Gallic Acid, Quercetin and Ascorbic Acid as Therapeutic Agents against Oxidative Stress.

Previous studies detail that different blood groups are associated with incidence of oxidative stress-related diseases such as certain carcinomas. Bioactive compounds represent an alternative for preventing this oxidative stress. The aim of this study was to elucidate the impact of blood groups on the erythroprotective potential of fucoxanthin, ß-Carotene, gallic acid, quercetin and ascorbic acid as therapeutic agents against oxidative stress. The impact of ABO blood groups on the erythroprotective potential was evaluated via the antioxidant capacity, blood biocompatibility, blood susceptibility and erythroprotective potential (membrane stabilization, in vitro photostability and antihemolytic activity). All tested antioxidants exhibited a high antioxidant capacity and presented the ability to inhibit ROO•-induced oxidative stress without compromising the cell membrane, providing erythroprotective effects dependent on the blood group, effects that increased in the presence of antigen A. These results are very important, since it has been documented that antigen A is associated with breast and skin cancer. These results revealed a probable relationship between different erythrocyte antigens with erythroprotective potential, highlighting the importance of bio-targeted drugs for groups most susceptible to certain chronic-degenerative pathologies. These compounds could be applied as additive, nutraceutical or encapsulated to improve their bioaccessibility.

Genomic Insights into Listeria monocytogenes: Organic Acid Interventions for Biofilm Prevention and Control.

Listeria monocytogenes is an important pathogen that has been implicated in foodborne illness. The aim of the present study was to investigate the diversity of virulence factors associated with the mechanisms of pathogenicity, persistence, and formation of biofilm L. monocytogenes by tandem analysis of whole-genome sequencing. The lineages that presented L. monocytogenes (LmAV-2, LmAV-3, and LmAV-6) from Hass avocados were lineages I and II. Listeria pathogenicity island 1 (LIPI-1) and LIPI-2 were found in the isolates, while LIPI-3 and Listeria genomic island (LGI-2) only was in IIb. Stress survival island (SSI-1) was identified in lineage I and II. In the in silico analysis, resistance genes belonging to several groups of antibiotics were detected, but the bcrABC and transposon Tn6188 related to resistance to quaternary ammonium salts (QACs) were not detected in L. monocytogenes. Subsequently, the anti-L. monocytogenes planktonic cell effect showed for QACs (MIC = 6.25 ppm/MBC = 100 ppm), lactic acid (MBC = 1 mg/mL), citric acid (MBC = 0.5 mg/mL) and gallic acid (MBC = 2 mg/mL). The anti-biofilm effect with organic acids (22 °C) caused a reduction of 4-5 log10 cfu/cm2 after 10 min against control biofilm L. monocytogenes formed on PP than SS. This study is an important contribution to understanding the genomic diversity and epidemiology of L. monocytogenes to establish a control measure to reduce the impact on the environment and the consumer.

Genetic and compositional analysis of biofilm formed by Staphylococcus aureus isolated from food contact surfaces.

Introduction: Staphylococcus aureus is an important pathogen that can form biofilms on food contact surfaces (FCS) in the dairy industry, posing a serious food safety, and quality concern. Biofilm is a complex system, influenced by nutritional-related factors that regulate the synthesis of the components of the biofilm matrix. This study determines the prevalence of biofilm-associated genes and evaluates the development under different growth conditions and compositions of biofilms produced by S. aureus. Methods: Biofilms were developed in TSB, TSBG, TSBNaCl, and TSBGNaCl on stainless-steel (SS), with enumeration at 24 and 192 h visualized by epifluorescence and scanning electron microscopy (SEM). The composition of biofilms was determined using enzymatic and chemical treatments and confocal laser scanning microscopy (CLSM). Results and discussion: A total of 84 S. aureus (SA1-SA84) strains were collected from 293 dairy industry FCS (FCS-stainless steel [n = 183] and FCS-polypropylene [n = 110]) for this study. The isolates harbored the genes sigB (66%), sar (53%), agrD (52%), clfB/clfA (38%), fnbA/fnbB (20%), and bap (9.5%). 99. In particular, the biofilm formed by bap-positive S. aureus onto SS showed a high cell density in all culture media at 192 h in comparison with the biofilms formed at 24 h (p < 0.05). Epifluorescence microscopy and SEM revealed the metabolically active cells and the different stages of biofilm formation. CLSM analysis detected extracellular polymeric of S. aureus biofilms on SS, such as eDNA, proteins, and polysaccharides. Finally, the level of detachment on being treated with DNase I (44.7%) and NaIO 4(42.4%) was greater in the biofilms developed in TSB compared to culture medium supplemented with NaCl at 24 h; however, there was no significant difference when the culture medium was supplemented with glucose. In addition, after treatment with proteinase K, there was a lower level of biomass detachment (17.7%) of the biofilm developed in TSBNaCl (p < 0.05 at 24 h) compared to that in TSB, TSBG, and TSBGNaCl (33.6, 36.9, and 37.8%, respectively). These results represent a deep insight into the composition of S. aureus biofilms present in the dairy industry, which promotes the development of more efficient composition-specific disinfection strategies.

Efficacy of Novel Bacteriophages against Escherichia coli Biofilms on Stainless Steel.

Biofilm formation by E. coli is a serious threat to meat processing plants. Chemical disinfectants often fail to eliminate biofilms; thus, bacteriophages are a promising alternative to solve this problem, since they are widely distributed, environmentally friendly, and nontoxic to humans. In this study, the biofilm formation of 10 E. coli strains isolated from the meat industry and E. coli ATCC BAA-1430 and ATCC 11303 were evaluated. Three strains, isolated from the meat contact surfaces, showed adhesion ability and produced extracellular polymeric substances. Biofilms of these three strains were developed onto stainless steel (SS) surfaces and enumerated at 2, 12, 24, 48, and 120 h, and were visualized by scanning electron microscopy. Subsequently, three bacteriophages showing podovirus morphology were isolated from ground beef and poultry liver samples, which showed lytic activity against the abovementioned biofilm-forming strains. SS surfaces with biofilms of 2, 14, and 48 h maturity were treated with mixed and individual bacteriophages at 8 and 9 log10 PFU/mL for 1 h. The results showed reductions greater than 6 log10 CFU/cm2 as a result of exposing SS surfaces with biofilms of 24 h maturity to 9 log10 PFU/mL of bacteriophages; however, the E. coli and bacteriophage strains, phage concentration, and biofilm development stage had significant effects on biofilm reduction (p < 0.05). In conclusion, the isolated bacteriophages showed effectiveness at reducing biofilms of isolated E. coli; however, it is necessary to increase the libraries of phages with lytic activity against the strains isolated from production environments.

Combination of Sorbitol and Glycerol, as Plasticizers, and Oxidized Starch Improves the Physicochemical Characteristics of Films for Food Preservation.

The aim of this work was to use glycerol (Gly) and sorbitol (Sor) as plasticizers with oxidized starch potato (OS) to produce biodegradable and environmentally friendly films, and to demonstrate the resulting physicochemical and functional viability without subtracting the organoleptic characteristics of the food. Analyses by water vapor permeability (WVP), attenuated total reflection Fourier transform infrared spectra (ATR-FTIR), scanning electron microscopy (SEM), tensile strength (TS), and transparency (UV) showed that the best film result was with 1.5 g of Gly and 2.0 g of Sor, conferred shine, elasticity 19.42 ± 6.20%, and mechanical support. The starch oxidized to 2.5%, contributing a greater transparency of 0.33 ± 0.12 and solubility of 78.90 ± 0.94%, as well as less permeability to water vapor 6.22 ± 0.38 gmm-2 d-1 kPa-1. The films obtained provide an alternative for use in food due to their organic compounds, excellent visual presentation, and barrier characteristics that maintain their integrity and, therefore, their functionality.

Conditions of In Vitro Biofilm Formation by Serogroups of Listeria monocytogenes Isolated from Hass Avocados Sold at Markets in Mexico.

Listeria monocytogenes is an important pathogen that has been implicated in foodborne illnesses and the recall of products such as fruit and vegetables. This study determines the prevalence of virulence-associated genes and serogroups and evaluates the effects of different growth media and environmental conditions on biofilm formation by L. monocytogenes. Eighteen L. monocytogenes isolates from Hass avocados sold at markets in Guadalajara, Mexico, were characterized by virulence-associated genes and serogroup detection with PCR. All isolates harbored 88.8% actA, 88.8% plcA, 83.3% mpl, 77.7% inlB, 77.7% hly, 66.6% prfA, 55.5% plcB, and 33.3% inlA. The results showed that 38.8% of isolates harbored virulence genes belonging to Listeria pathogenicity island 1 (LIPI-1). PCR revealed that the most prevalent serogroup was serogroup III (1/2b, 3b, and 7 (n = 18, 66.65%)), followed by serogroup IV (4b, 4d-4e (n = 5, 27.7%)) and serogroup I (1/2a-3a (n = 1, 5.5%)). The assessment of the ability to develop biofilms using a crystal violet staining method revealed that L. monocytogenes responded to supplement medium TSBA, 1/10 diluted TSBA, and TSB in comparison with 1/10 diluted TSB (p < 0.05) on polystyrene at 240 h (p < 0.05). In particular, the biofilm formation by L. monocytogenes (7.78 ± 0.03-8.82 ± 0.03 log10 CFU/cm2) was significantly different in terms of TSBA on polypropylene type B (PP) (p < 0.05). In addition, visualization by epifluorescence microscopy, scanning electron microscopy (SEM), and treatment (DNase I and proteinase K) revealed the metabolically active cells and extracellular polymeric substances of biofilms on PP. L. monocytogenes has the ability to develop biofilms that harbor virulence-associated genes, which represent a serious threat to human health and food safety.

Removal of Mixed-Species Biofilms Developed on Food Contact Surfaces with a Mixture of Enzymes and Chemical Agents.

Sanicip Bio Control (SBC) is a novel product developed in Mexico for biofilms' removal. The aims of this study were to evaluate (i) the removal of mixed-species biofilms by enzymatic (protease and α-amylase, 180 MWU/g) and chemical treatments (30 mL/L SBC, and 200 mg/L peracetic acid, PAA) and (ii) their effectiveness against planktonic cells. Mixed-species biofilms were developed on stainless steel (SS) and polypropylene B (PP) in whole milk (WM), tryptic soy broth (TSB) with meat extract (TSB+ME), and TSB with chicken egg yolk (TSB+EY) to simulate the food processing environment. On SS, all biofilms were removed after treatments, except the enzymatic treatment that only reduced 1-2 log10 CFU/cm2, whereas on PP, the reductions ranged between 0.59 and 5.21 log10 CFU/cm2, being the biofilms developed in TSB+EY being resistant to the cleaning and disinfecting process. Higher reductions in microbial load on PP were reached using enzymes, SBC, and PAA. The employed planktonic cells were markedly more sensitive to PAA and SBC than were the sessile cells. In conclusion, biofilm removal from SS can be achieved with SBC, enzymes, or PAA. It is important to note that the biofilm removal was strongly affected by the food contact surfaces (FCSs) and surrounding media.

Nosocomial pathogen biofilms on biomaterials: Different growth medium conditions and components of biofilms produced in vitro.

BACKGROUND/PURPOSE (S): Nosocomial pathogens can develop biofilms on hospital surfaces and medical devices; however, few studies have focused on the evaluation of mono-and dual-species biofilms developed by nosocomial pathogens under different growth conditions. METHODS: This study investigated biofilm development by nosocomial pathogens (Acinetobacter baumannii, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa) on biomaterials in different culture media and their components of the extracellular matrix biofilm. RESULTS: The mono-species biofilms showed cell densities from 7.50 to 9.27 Log10 CFU/cm2 on natural rubber latex type I (NLTI) and from 7.58 to 8.79 Log10 CFU/cm2 on stainless steel (SS). Dual-species biofilms consisted of S. aureus + P. aeruginosa (7.87-8.27 Log10 CFU/cm2 in TSBP and TSBME onto SS; p < 0.05), E. coli + P. aeruginosa (8.32-8.86 Log10 CFU/cm2 in TSBME onto SS and TSBP onto NLTI; p < 0.05), and S. aureus + E. coli (7.82 Log10 CFU/cm2 in TSBME onto SS; p < 0.05). Furthermore, biofilm detachment after proteinase K treatment was 5.54-32.81% compared to 7.95-24.15% after DNase I treatment in the mono-dual species biofilm matrix. Epifluorescence microscopy and scanning electron microscopy (SEM) enabled visualizing the bacteria and extracellular polymeric substances of biofilms on SS and NLTI. CONCLUSION: Nosocomial pathogens can develop biofilms on biomaterials. Mono-species biofilms of Gram-negative bacteria showed lower densities than dual-species biofilms in TSBME and TSBP. Additionally, dual-species biofilms showed a higher concentration of proteins and eDNA in the extracellular matrix.

Kinetics of biofilm formation by pathogenic and spoilage microorganisms under conditions that mimic the poultry, meat, and egg processing industries.

Pathogens and spoilage microorganisms can develop multispecies biofilms on food contact surfaces; however, few studies have been focused on evaluated mixed biofilms of these microorganisms. Therefore this study investigated the biofilm development by pathogenic (Bacillus cereus, Escherichia coli, Listeria monocytogenes, and Salmonella enterica Enteritidis and Typhimurium serotypes) and spoilage (Bacillus cereus and Pseudomonas aeruginosa) microorganisms onto stainless-steel (SS) and polypropylene B (PP) coupons; under conditions that mimic the dairy, meat, and egg processing industry. Biofilms were developed in TSB with 10% chicken egg yolk (TSB + EY), TSB with 10% meat extract (TSB + ME) and whole milk (WM) onto SS and PP. Each tube was inoculated with 25 µL of each bacteria and then incubated at 9 or 25 °C, with enumeration at 1, 48, 120, 180 and 240 h. Biofilms were visualized by epifluorescence and scanning electron microscopy (SEM). Biofilm development occurred at different phases, depending on the incubation conditions. In the reversible adhesion, the cell density of each bacteria was between 1.43 and 6.08 Log10 CFU/cm2 (p < 0.05). Moreover, significant reductions in bacteria appeared at 9 °C between 1 and 48 h of incubation. Additionally, the constant multiplication of bacteria in the biofilm occurred at 25 °C between 48 and 180 h of incubation, with increments of 2.08 Log10 CFU/cm2 to S. Typhimurium. Population establishment was observed between 48 and 180 h and 180-240 h incubation, depending on the environmental conditions (25 and 9 °C, respectively). For example, in TSB + ME at 25 °C, S. Typhimurium, P aeruginosa, and L. monocytogenes showed no statistical differences in the amounts between 48 and 180 h incubation. The dispersion phase was identified for L. monocytogenes and B. cereus at 25 °C. Epifluorescence microscopy and SEM allowed visualizing the bacteria and extracellular polymeric substances at the different biofilm stages. In conclusion, pathogens and spoilage microorganisms developed monospecies with higher cellular densities than multiespecies biofilms. In multispecies biofilms, the time to reach each biofilm phase varied is depending on environmental factors. Cell count decrements of 1.12-2.44 Log10 CFU/cm2 occurred at 48 and 240 h and were most notable in the biofilms developed at 9 °C. Additionally, cell density reached by each microorganism was different, P. aeruginosa and Salmonella were the dominant microorganisms in the biofilms while B. cereus showed the lower densities until undetectable levels.

Biofilm Formation and Detection of Fluoroquinolone- and Carbapenem-Resistant Genes in Multidrug-Resistant Acinetobacter baumannii.

Acinetobacter baumannii is an important opportunistic pathogen that shows resistance to cephalosporins, penicillins, carbapenems, fluoroquinolones, and aminoglycosides, the multiresistance being associated with its ability to form biofilms in clinical environments. The aim of this study was to determine biofilm formation and its potential association with genes involved in antibiotic resistance mechanisms of A. baumannii isolates of different clinical specimens. We demonstrated 100% of the A. baumannii isolates examined to be multidrug resistant (MDR), presenting a 73.3% susceptibility to cefepime and a 53.3% susceptibility to ciprofloxacin. All A. baumannii isolates were positive for bla OXA-51, 33.3% being positive for bla OXA-23 and ISAba1, and 73.3% being positive for gyrA. We found 86.6% of A. baumannii strains to be low-grade biofilm formers and 13.3% to be biofilm negative; culturing on Congo red agar (CRA) plates revealed that 73.3% of the A. baumannii isolates to be biofilm producers, while 26.6% were not. These properties, combined with the role of A. baumannii as a nosocomial pathogen, increase the probability of A. baumannii causing nosocomial infections and outbreaks as a complication during therapeutic treatments and emphasize the need to control A. baumannii biofilms in hospital environments.

Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite

Abstract The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37 °C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p > 0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61 ± 0.13 Log10 CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91 ± 0.44 Log10 CFU/cm2 (p < 0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500 ppm), reducing by more than 5 Log10 CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p < 0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Resistance of pathogenic and spoilage microorganisms to disinfectants in the presence of organic matter and their residual effect on stainless steel and polypropylene.

OBJECTIVES: The effectiveness of disinfectants can vary according to the microorganism, type of residues and surface. The aim of this study was to determine the effectiveness of four disinfectants in the presence of organic matter and their residual effect on stainless steel grade 304 (SS) and polypropylene B (PP-B). METHODS: The effectiveness of the disinfectants in the presence of meat extract, egg yolk and whole milk was determined according to AOAC and UNE-EN 1040:2015, and the residual effect was determined according to UNE-EN 13697:2015 using approved strains. RESULTS: The effectiveness of the disinfectants was affected to different degrees depending on the organic matter present. SANICIP Q5 [400µg/mL; fifth-generation quaternary ammonium compound (QAC)] was most effective in the presence of 10% meat extract, whilst SANICIP PAA (200µg/mL; peracetic acid) showed better activity in the presence of 10% egg yolk and whole milk. In the evaluation of residual effect on SS and PP-B, the QAC had a better effect, reducing Listeria monocytogenes ATCC 19111 by 6 Log10 CFU/mL at 24h after its application. Conversely, the disinfectants had no residual effect against Pseudomonas aeruginosa ATCC 15442. CONCLUSIONS: The antimicrobial activity of the disinfectants tested against pathogenic and spoilage microorganisms was affected according to the type of organic matter present. SANICIP Q5 had a greater residual effect than the other disinfectants evaluated. Moreover, the residual effect of a disinfectant is greater on SS than on PP-B and is dependent on the microorganism tested.

Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite.

The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37°C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p>0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61±0.13Log10CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91±0.44Log10CFU/cm2 (p<0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500ppm), reducing by more than 5Log10CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p<0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Antimicrobial activity of disinfectants commonly used in the food industry in Mexico.

OBJECTIVES: Disinfectants are widely used in food processing environments for microorganism control; their activity can vary according the microorganism and their used in the appropriated concentrations is vital. Hence, the aim of this study was determined the effectiveness and minimum inhibitory concentration (MIC) of 15 disinfectants commonly used in the food industry in Mexico. METHODS: The antimicrobial activity and the MIC were determined according to AOAC and CLSI, respectively, with approved strains. RESULTS: Most disinfectants reduced 99.999% of microorganisms in suspension after 30s of contact, so reduction rate corresponded at least 5 Log10. Only for Pseudomonas aeruginosa ATCC 15442, did all disinfectants have 99.999% effectiveness. For the MIC, only the third generation quaternary ammonium compounds (QACs) in acid medium did not have values within the range in which is used in the food industry for Staphylococcus aureus ATCC 25923. In addition, for all disinfectants the MIC at 5min was two to four times greater than the concentration with the same effect at 10min; moreover, in most cases there was no difference in the MIC at 10 and 15min (p>0.05). CONCLUSIONS: At recommended concentrations, disinfectants had bactericidal activity for at least three of the six microorganisms evaluated. However, the MIC was affected by the exposure time: it was bigger at 5min than at 10min; moreover, in the majority of the cases, it was equal at 10 and 15min; with the results we could have a better understanding of disinfectants use in food processing environments.