Determination of the effect of the bacteriocin enterocin AS-48 on the microbial loads and bacterial diversity of blueberries.

Fresh fruits have been involved in transmission of foodborne pathogens. In the present work, five different batches of blueberries were used. One aliquot from each batch was washed with sterile saline solution (SSS) and the other one with a solution of the circular bacteriocin enterocin AS-48 in SSS. Then, the surface microbiota of controls and bacteriocin-treated samples was recovered and used for microbiota analyses, both using viable counts and high-throughput amplicon sequencing. Total aerobic mesophilic loads ranged from 2.70 to 4.09 log CFU/g in most of the samples. Only two samples yielded detectable viable counts on selective media (Enterobacteriaceae, presumptive Salmonella and coliforms), with values ranging from 2.84 to 3.81 log CFU/g. The bacteriocin treatment reduced viable cell counts of total aerobic mesophiles to a range of 1.40-1.88 log CFU/g. No viable cells were detected on selective media. Amplicon sequencing indicated large batch-to-batch variations in the surface microbiota of blueberries and also an effect of the bacteriocin treatment on microbiota composition.

Effect of High Hydrostatic Pressure Processing on the Microbiological Quality and Bacterial Diversity of Sous-Vide-Cooked Cod.

High hydrostatic pressure (HP) is a promising method to improve the microbiological quality of sous-vide foods. Monitoring the composition and behavior of the microbial communities in foods is of most importance for the production of high-quality and safe products. High-throughput sequencing (HTS) provides advanced approaches to determine food's microbial community composition and structure. The aim of the present study was to determine the impact of different HP treatments on the microbial load and bacterial diversity of sous-vide Atlantic cod. Sous-vide cooking at 57.1 °C for 30 min followed by HP treatment at 500 MPa for 8 min reduced viable cell counts (total aerobic mesophiles) in the cod samples below detectable levels for 45 days of storage under refrigeration. In a second trial with cod cooked sous-vide at 52 °C for 20 min followed by HP treatments at 300 or 600 MPa (with HP treatment temperatures of 22 °C or 50 °C for 4 or 8 min, depending on treatment), only the treatments at 600 MPa delayed bacterial growth for at least 30 days under refrigeration. The optimal HP conditions to improve the microbiological quality of sous-vide cod cooked at low temperatures were obtained at 600 MPa for 4 min at a pressurization temperature of 50 °C. Bacterial diversity was studied in cod cooked sous-vide at 52 °C for 20 min by HTS. In the absence of HP treatment, Proteobacteria was the main bacterial group. A succession of Pseudomonadaceae (Pseudomonas) and Enterobacteriaceae was observed during storage. Firmicutes had low relative abundances and were represented mainly by Anoxybacillus (early storage) and Carnobacterium (late storage). The HP-treated sous-vide cod showed the greatest differences from controls during late storage, with Aerococcus and Enterococcus as predominant groups (depending on the HP conditions). The application of HTS provided new insights on the diversity and dynamics of the bacterial communities of sous-vide cod, revealing the presence of bacterial genera not previously described in this food, such as Anoxybacillus. The significance of Anoxybacillus as a contaminant of seafoods should be further investigated.

Draft Genome Sequence of a Putatively Antibiotic-Resistant Escherichia coli Strain Isolated from Calf Intestine in Boyacá, Colombia.

A draft genome of the putatively antibiotic-resistant Escherichia coli strain ANGUJ1, which was isolated from calf intestine from Boyacá, Colombia, is reported. The genome possessed genetic determinants for antibiotic resistance and multicompound resistance efflux pumps. In silico prediction analysis suggests phenotypic resistance to six classes of antibiotics plus aldehyde and peroxide.

Antimicrobial Resistance, Biocide Tolerance, and Bacterial Diversity of a Dressing Made from Coriander and Parsley after Application of Treatments Using High Hydrostatic Pressure Alone or in Combination with Moderate Heat.

The effects of high-hydrostatic pressure (HP) treatments (450 and 600 megapascals, MPa, for 5 min at temperatures of 22 °C and 50 °C) on the microbiota of a coriander and parsley dressing was studied via culture-dependent and culture-independent approaches. Samples were refrigerated for 20 days, with periodic counts of the culture media supplemented with, or without, antimicrobials. HP-treated samples showed significantly lower viable cell counts compared to untreated controls. Only the control samples yielded bacterial growth on media with antimicrobials (imipenem, cefotaxime, benzalkonium chloride), including mostly Pseudomonas and Lactobacillus. Bacillus and Paenibacillus were identified from pressurized samples. Few isolates showed higher tolerance to some of the biocides tested. Pseudomonads showed outstanding resistance to meropenem and ceftazidime. According to high-throughput sequencing analysis, the microbiota of the dressing control samples changes during storage, with a reduction in the relative abundance of Proteobacteria and an increase in Firmicutes. The composition of the residual microbiota detected during storage was highly dependent on the pressure applied, and not on the treatment temperature.

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.

Changes in the Bacterial Diversity of Human Milk during Late Lactation Period (Weeks 21 to 48).

Breast milk from a single mother was collected during a 28-week lactation period. Bacterial diversity was studied by amplicon sequencing analysis of the V3-V4 variable region of the 16S rRNA gene. Firmicutes and Proteobacteria were the main phyla detected in the milk samples, followed by Actinobacteria and Bacteroidetes. The proportion of Firmicutes to Proteobacteria changed considerably depending on the sampling week. A total of 411 genera or higher taxons were detected in the set of samples. Genus Streptococcus was detected during the 28-week sampling period, at relative abundances between 2.0% and 68.8%, and it was the most abundant group in 14 of the samples. Carnobacterium and Lactobacillus had low relative abundances. At the genus level, bacterial diversity changed considerably at certain weeks within the studied period. The weeks or periods with lowest relative abundance of Streptococcus had more diverse bacterial compositions including genera belonging to Proteobacteria that were poorly represented in the rest of the samples.

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.

Biocide tolerance and antibiotic resistance of Enterobacter spp. isolated from an Algerian hospital environment.

OBJECTIVES: In this study, 77 Enterobacter spp. isolates from a collection of 175 Gram-negative bacilli isolated from Tlemcen University Hospital Center (North-West of Algeria) were tested for antibiotic resistance, biocide tolerance and genetic determinants of antimicrobial resistance. METHODS: The isolates were identified by 16S rDNA gene sequencing. Biocide tolerance was determined by broth microdilution, and antibiotic resistance was determined by disk diffusion. Genetic determinants of resistance were studied by PCR amplification using suitable primers. RESULTS: The most common Enterobacter species was Enterobacter cloacae (58.4%), followed by Enterobacter hormaechei (24.7%). The most common antibiotic resistance was to ticarcillin either alone or in combination with clavulanic acid (70.1%), followed by cefepime (68.8%), cefotaxime (63.6%), ceftazidime (54.5%) and gentamicin (54.5%). Tobramycin was active against 87.0% of the isolates. Levels of biocide tolerance were high for hexachlorophene and to a lesser extent for benzalkonium chloride. The extended-spectrum ß-lactamase genes blaTEM and blaCTX-M were detected in 44.2% and 36.4% of isolates, respectively. Other antimicrobial resistance genes (ARGs) frequently detected were aac(6')-Ib (57.1%) and sul2 (50.6%). Multidrug-resistant isolates carrying several ARGs were common. Significant positive correlations were detected for efflux pump genes with ARGs and also between ARGs. CONCLUSION: The results of this study reveal thatEnterobacter spp. isolates from hospital settings are both resistant to clinically-used antibiotics and tolerant to biocides. Biocide tolerance could be an advantage for antibiotic-resistant strains in hospitals.

Treatment With High-Hydrostatic Pressure, Activated Film Packaging With Thymol Plus Enterocin AS-48, and Its Combination Modify the Bacterial Communities of Refrigerated Sea Bream (Sparus aurata) Fillets.

The aim of this study was to determine the impact of activated plastic films with thymol and enterocin AS-48 and high-hydrostatic pressure (HP) treatment on the bacterial load and bacterial diversity of vacuum-packaged sea bream fillets under refrigerated storage for 10 days. The activated film and the HP treatment reduced aerobic mesophiles viable counts by 1.46 and 2.36 log cycles, respectively, while the combined treatment achieved a reduction of 4.13 log cycles. HP and combined treatments resulted in longer delays in bacterial growth. Proteobacteria were the dominant phyla in sea bream fillets. The relative abundance of Firmicutes increased by the end of storage both in controls and in samples treated by HP singly or in combination with the activated films. The predominant operational taxonomic units (OTUs) found at time 0 in control samples (Listeria, Acinetobacter, Pseudomonas, Enterobacteriaceae, Chryseobacterium) rapidly changed during storage (with an increase of Vibrio, Photobacterium, and Shewanella together with Cloacibacterium and Lactobacillales by the end of storage). The activated film and the HP treatment induced drastic changes in bacterial diversity right after treatments (with Comamonadaceae, Methylobacterium, Acidovorax, and Sphingomonas as main OTUs) and also induced further modifications during storage. Bacterial diversity in activated film samples was quite homogeneous during storage (with Vibrio, Photobacterium, and Shewanella as main OTUs) and approached control samples. HP treatments (singly or in combination with activated films) determined a high relative abundance of Acinetobacter (followed by Pseudomonas and Shewanella) during early storage as well as a higher relative abundance of lactic acid bacteria by the end of storage. The results indicate that the complex dynamics of bacterial populations in the refrigerated sea bream fillets are markedly influenced by treatment and antimicrobials applied.

Analysis of potential risks from the bacterial communities associated with air-contact surfaces from tilapia (Oreochromis niloticus) fish farming.

Tilapia farming is a promising growing sector in aquaculture. Yet, there are limited studies on microbiological risks associated to tilapia farms. The aim of the present study was to analyse the bacterial communities from solid surfaces in contact with air in a tilapia farm in order to evaluate the presence of bacteria potentially toxinogenic or pathogenic to humans or animals. Samples from a local tilapia farm (tank wall, aerator, water outlets, sink and floor) were analyzed by high throughput sequencing technology. Sequences were assigned to operational taxonomic units (OTUs). Proteobacteria was the main phylum represented in most samples (except for one). Cyanobacteria were a relevant phylum in the inner wall from the fattening tank and the wet floor by the pre-fattening tank. Bacteroidetes were the second phylum in relative abundance for samples from the larval rearing tank and the pre-fattening tank and one sample from the fattening tank. Fusobacteria showed highest relative abundances in samples from the larval rearing tank and pre-fattening tank. Other phyla (Verrucomicrobia, Actinobacteria, Firmicutes, Planktomycetes, Acidobacteria, Chloroflexi, Chlorobi, Gemmatiomonadetes or Fibrobacters) had lower relative abundances. A large fraction of the reads (ranging from 43.67% to 72.25%) were assigned to uncultured bacteria. Genus Acinetobacter (mainly A. calcoaceticus/baumanni) was the predominant OTU in the aerator of the fattening tank and also in the nearby sink on the floor. The genera Cetobacterium and Bacteroides showed highest relative abundances in the samples from the larval rearing tank and the pre-fattening tank. Genera including fish pathogens (Fusobacterium, Aeromonas) were only detected at low relative abundances. Potential human pathogens other than Acinetobacter were either not detected or had very low relative abundances (< 0.01%). The results of the study suggest that the main risk factors to be monitored in tilapia farm are putative human pathogenic Acinetobacter and potential cyanotoxin-producing cyanobacteria.

Resistance to Antibiotics, Biocides, Preservatives and Metals in Bacteria Isolated from Seafoods: Co-Selection of Strains Resistant or Tolerant to Different Classes of Compounds.

Multi-drug resistant bacteria (particularly those producing extended-spectrum ß-lactamases) have become a major health concern. The continued exposure to antibiotics, biocides, chemical preservatives, and metals in different settings such as the food chain or in the environment may result in development of multiple resistance or co-resistance. The aim of the present study was to determine multiple resistances (biocides, antibiotics, chemical preservatives, phenolic compounds, and metals) in bacterial isolates from seafoods. A 75.86% of the 87 isolates studied were resistant to at least one antibiotic or one biocide, and 6.90% were multiply resistant to at least three biocides and at least three antibiotics. Significant (P < 0.05) moderate or strong positive correlations were detected between tolerances to biocides, between antibiotics, and between antibiotics with biocides and other antimicrobials. A sub-set of 30 isolates selected according to antimicrobial resistance profile and food type were identified by 16S rDNA sequencing and tested for copper and zinc tolerance. Then, the genetic determinants for biocide and metal tolerance and antibiotic resistance were investigated. The selected isolates were identified as Pseudomonas (63.33%), Acinetobacter (13.33%), Aeromonas (13.33%), Shewanella, Proteus and Listeria (one isolate each). Antibiotic resistance determinants detected included sul1 (43.33% of tested isolates), sul2 (6.66%), blaTEM (16.66%), blaCTX-M (16.66%), blaPSE (10.00%), blaIMP (3.33%), blaNDM-1 (3.33%), floR (16.66%), aadA1 (20.0%), and aac(6')-Ib (16.66%). The only biocide resistance determinant detected among the selected isolates was qacEΔ1 (10.00%). A 23.30 of the selected isolates were able to grow on media containing 32 mM copper sulfate, and 46.60% on 8 mM zinc chloride. The metal resistance genes pcoA/copA, pcoR, and chrB were detected in 36.66, 6.66, and 13.33% of selected isolates, respectively. Twelve isolates tested positive for both metal and antibiotic resistance genes, including one isolate positive for the carbapenemase gene blaNDM-1 and for pcoA/copA. These results suggest that exposure to metals could co-select for antibiotic resistance and also highlight the potential of bacteria on seafoods to be involved in the transmission of antimicrobial resistance genes.

Analysis of the microbiota of refrigerated chopped parsley after treatments with a coating containing enterocin AS-48 or by high-hydrostatic pressure.

Parsley can be implicated in foodborne illness, yet chopped parsley is used as an ingredient or garnish for multiple dishes. The aim of the present study was to determine the effect of two different treatments on the bacterial diversity of parsley: (i) coating with a pectin-EDTA solution containing the circular bacteriocin enterocin AS-48, and (ii) treatment by high hydrostatic pressure (HHP) at 600MPa for 8min. Control and treated parsley were stored in trays at 5°C for 10days. Both treatments reduced viable counts by 3.7 log cycles and retarded growth of survivors during storage. The bacterial diversity of the chopped parsley was studied by high throughput sequencing (Illumina Miseq). Bacterial diversity of control samples mainly consists of Proteobacteria (96.87%) belonging to genera Pseudomonas (69.12%), Rheinheimera (8.56%) and Pantoea (6.91%) among others. During storage, the relative abundance of Bacteroidetes (mainly Flavobacterium and Sphingobacterium) increased to 26.66%. Application of the pectin-bacteriocin-EDTA coating reduced the relative abundance of Proteobacteria (63.75%) and increased that of Firmicutes (34.70%). However, the relative abundances of certain groups such as Salmonella, Shigella and Acinetobacter increased at early storage times. Late storage was characterized by an increase in the relative abundance of Proteobacteria, mainly Pseudomonas. Upon application of HHP treatment, the relative abundance of Proteobacteria was reduced (85.88%) while Actinobacteria increased (8.01%). During early storage of HHP-treated samples, the relative abundance of Firmicutes increased. Potentially-pathogenic bacteria (Shigella) only increased in relative abundance by the end of storage. Results of the present study indicate that the two treatments had different effects on the bacterial diversity of parsley. The HHP treatment provided a safer product, since no potentially-pathogenic bacteria were detected until the end of the storage period.

Adaptation to Biocides Cetrimide and Chlorhexidine in Bacteria from Organic Foods: Association with Tolerance to Other Antimicrobials and Physical Stresses.

Chlorhexidine (CH) and quaternary ammonium compounds (QAC), such as cetrimide (CE), are widely used as disinfectants because of their broad antimicrobial spectrum. However, their frequent use for disinfection in different settings may promote bacterial drug resistance against both biocides and clinically relevant antibiotics. This study analyzes the effects of stepwise exposure to cetrimide (CE) and chlorhexidine (CH) of bacteria from organic foods and previously classified as biocide-sensitive. Gradual exposure of these strains to biocides resulted in mainly transient decreased antimicrobial susceptibility to other antibiotics and to biocides. Biocide-adapted bacteria also exhibit alterations in physiological characteristics, mainly decreased heat tolerance, or gastric acid tolerance in CE-adapted strains, while bile resistance does not seem to be influenced by biocide adaptation. Results from this study suggest that changes in membrane fluidity may be the main mechanism responsible for the acquisition of stable tolerance to biocides.

Effects of exposure to quaternary-ammonium-based biocides on antimicrobial susceptibility and tolerance to physical stresses in bacteria from organic foods.

In the present study, a collection of 76 biocide-sensitive bacterial strains isolated from organically produced food were adapted by repeated exposure to increasing concentrations of the quaternary ammonium compounds (QACs) benzalkonium chloride (BC) and hexadecylpyridinium chloride (HDP). The sensitivity of both wildtype strains and their corresponding QAC-adapted strains to other biocides and to antibiotics was studied. QAC tolerance increased in 88.2% of strains for BC and in 30.3% of strains for HDP, with increases in minimum inhibitory concentrations between 2 and over 100 fold. Adaptive resistance was stable after 20 subcultures in biocide-free medium for 7 and 5 of the BC- and HDP-adapted strains, respectively. Adaptation to BC and HDP also reduced the susceptibility to other biocides, mainly hexachlorophene (CF), didecyldimethylammonium bromide (AB), triclosan (TC) and chlorhexidine (CH). BC-adapted strains showed increased antibiotic resistance to ampicillin (AM) followed by sulfamethoxazol (SXT) and cefotaxime (CTX), and some showed increased sensitivity to ceftazidime (CAZ), CTX, AM and STX. Changes in antibiotic resistance in HDP-adapted strains were more heterogeneous and strain-dependent. Main efflux pump genes detected in QAC-adapted strains were acrB, sugE, norC, qacE and qacH, as well as antibiotic resistance genes aac(6_)-Ie-aph(2_)-Ia, aph(2_)-Ic, ant(4_)-Ia, lsa, mrsA/B, ereA, ermB and cat. Membrane anisotropy experiments revealed that QAC adaptation induced an increase in membrane rigidity in the case of BC, while response to HDP was more heterogeneous and strain-dependent. Growth capacity was significantly higher in some QAC-adapted strains and strain-dependent changes in heat tolerance were also detected in QAC-adapted strains. Gastric acid or bile resistances do not seem to be influenced by QAC adaptation.

Characterization of biocide-tolerant bacteria isolated from cheese and dairy small-medium enterprises.

A collection of 120 bacterial isolates from small medium enterprises involved in the production of cow milk and the manufacture of goat cheese were screened for sensitivity to biocides benzalkonium chloride (BC), cetrimide (CT), hexadecylpyridinium chloride (HDP), triclosan (TC), hexachlorophene (CF) and poly-(hexamethylen guanidinium) hydrochloride (PHMG). Nineteen isolates were selected according to biocide tolerance and identified by 16S rDNA sequencing as Lactococcus sp. (6) Enterococcus sp. (1), Lactobacillus sp. (4), Bacillus sp. (1) Escherichia sp. (5), Enterobacter sp. (1) and Helicobacter sp. (1). These were further characterised regarding antimicrobial resistance phenotype and genotype. Several isolates were multiply (3 or more) tolerant to biocides or resistant to antibiotics, but only two Escherichia sp. isolates and Enterobacter sp. were multiply resistant to biocides and antibiotics. Statistical analysis of biocide tolerance and antibiotic resistance revealed significant positive correlations between different biocides and between biocides and antibiotics. The biocide tolerance genes most frequently found were qacEΔ1 and qacA/B. The sulfonamide resistance gene sul1 was found in two Escherichia sp. isolates and in Enterobacter sp., all of which also carried qacEΔ1. Beta-lactam (blaCTX-M, blaPSE) and tetracycline resistance genes [tet(A), tet(C) and tet(D)] were detected. Efflux pump genes acrB and mdfA were found in most Gram-negative isolates. Results from the study suggest that exposure to biocides can indirectly select for antibiotic resistance.

The human gastrointestinal tract and oral microbiota in inflammatory bowel disease: a state of the science review.

Inflammatory bowel disease (IBD) includes a spectrum of diseases from ulcerative colitis (UC) to Crohn's disease (CD). Many studies have addressed the changes in the microbiota of individuals affected by UC and CD. A decrease in biodiversity and depletion of the phyla Bacteroidetes and Firmicutes has been reported, among others. Changes in microbial composition also result in changes in the metabolites generated in the gut from microbial activity that may involve the amount of butyrate and other metabolites such as H2 S being produced. Other factors such as diet, age, or medication need to be taken into consideration when studying dysbiosis associated with IBD. Diverse bacterial species have been associated specifically or non-specifically to IBD, but none of them have been demonstrated to be its ethiological agent. Recent studies also suggest that micro-eukaryotic populations may also be altered in IBD patients. Last, but not least, viruses, and specially bacteriophages, can play a role in controlling microbial populations in the gastrointestinal tract. This may affect both bacterial diversity and metabolism, but possible implications for IBD still remain to be solved. Dysbiosis in the oral microbiome associated with IBD remains an emerging field for future research.

Virulence factors and antimicrobial resistance in Escherichia coli strains isolated from hen egg shells.

Eggs may contain extraintestinal pathogenic (ExPEC) and diarrheogenic (DEC) Escherichia coli which in addition may carry antibiotic resistance. The wide use of biocides and disinfectants in the food industry may induce biocide tolerance in bacteria. The aim of the present study was to evaluate biocide tolerance and antibiotic resistance in E. coli from hen egg shells. A total of 27 isolates obtained from a screening of 180 eggs were studied. Seven isolates carried both eae and bfpA genes of typical enteropathogenic E. coli (EPEC) strains, while 14 isolates only carried eae associated with atypical EPEC strains. Shiga toxin genes stx and stx2 were detected in four isolates. Heat-stable and heat-labile enterotoxin genes as well as aggR were also detected. Several isolates had minimum inhibitory concentrations (MICs) that were higher than the wild-type for the biocide hexadecylpyridinium chloride (HDP, 18.52%) or the commercial disinfectant P3 oxonia (OX, 14.81%). Antibiotic resistance was detected for ampicillin (37.03%), streptomycin (37.03%), tetracycline (37.03%), chloramphenicol (11.11%), nalidixic acid (18.51%) and trimethoprim-sulfamethoxazole (14.81%). Eight isolates (29.63%) were biocide tolerant and antibiotic resistant. Efflux pump genes detected included acrB (96.29%), mdfA (85.18%) and oxqA (37.03%), in addition to quaternary ammonium compound (QAC) resistance genes qacA/B (11.11%) and qacE (7.40%). Antibiotic resistance genes detected included blaCTX-M-2 (22.22%), blaTEM (3.70%), blaPSE (3.70%), tet(A) (29.63%), tet(B) (29.63%), tet(C) (7.40%), tet(E) (11.11%), aac(6')-Ib (3.70%), sul1 (14.81%), dfrA12 (3.70%) and dfrA15 (3.70%). Most isolates (96.30%) carried more than one genetic determinant of resistance. The most frequent combinations were efflux pump components acrB and mdfA with tetracycline resistance genes (33.33% of isolates). Isolates carrying QAC resistance genes also carried between 4 and 8 of the additional antimicrobial resistance genes investigated. Regardless of biocide tolerance and antibiotic resistance, all isolates were sensitive to carvacrol (0.25%), thymol (0.125%) and trisodium phosphate (1 to 1.5%), but they exhibited a heterogeneous response to sodium lactate and lysozyme-EDTA combinations that apparently were not related with antibiotic resistance. Results from the study reveal not only a low incidence of biocide tolerance but also the presence of multiple resistance strains carrying multiple genetic determinants of resistance.

Changes in microbial diversity of brined green asparagus upon treatment with high hydrostatic pressure.

The application of high hydrostatic pressure (HHP, 600MPa, 8 min) on brined green asparagus and the changes in bacterial diversity after treatments and during storage at 4 °C (30 days) or 22 °C (10 days) were studied. HHP treatments reduced viable cell counts by 3.6 log cycles. The residual surviving population did not increase during storage at 4 °C. However, bacterial counts significantly increased at 22 °C by day 3, leading to rapid spoilage. The microbiota of green asparagus was composed mainly by Proteobacteria (mainly Pantoea and Pseudomonas), followed by Firmicutes (mainly Lactococcus and Enterococcus) and to a less extent Bacteroidetes and Actinobacteria. During chill storage of untreated asparagus, the relative abundance of Proteobacteria as well as Enterococcus and Lactococcus decreased while Lactobacillus increased. During storage of untreated asparagus at 22 °C, the abundance of Bacteroidetes decreased while Proteobacteria increased during late storage. The HHP treatment determined a reduction of the Proteobacteria both early after treatment and during chill storage. In the HHP treated samples stored at 22 °C, the relative abundance of Pseudomonas rapidly decreased at day 1, with an increase of Bacteroidetes. This was followed by a marked increase in Enterobacteriaceae (Escherichia) simultaneously with increase in viable counts and spoilage. Results from the study indicate that the effect of HHP treatments on the viability ofmicrobial populations in foods also has an impact on the dynamics of microbial populations during the storage of the treated foods.