Category Archives: Microbiology

Research -Antibacterial Effect of a Mixed Natural Preservative against Listeria monocytogenes on Lettuce and Raw Pork Loin

Journal of Food Protection

ABSTRACT

A mixed natural preservative, including grapefruit seed extract (GSE), cinnamaldehyde (CA), and nisin, was investigated for the reduction of Listeria monocytogenes growth on lettuce and raw pork loin. The MIC of each natural preservative was investigated for L. monocytogenes strains tested. Following central composite design, lettuce and pork loin were inoculated with a cocktail of three strains of L. monocytogenes (ATCC 15313, H7962, and NADC 2045 [Scott A]) and treated with the mixed natural preservative that included GSE (0.64 to 7.36 ppm), CA (1.6 to 18.4 ppm), and nisin (0.48 to 5.5 ppm). The MIC of GSE was 31.25 ppm in tested L. monocytogenes strains, and of CA was 500 and 1,000 ppm in L. monocytogenes ATCC 15313 and the other L. monocytogenes strains, respectively. The MIC of nisin was 250 ppm. The R2 value of this model was more than 0.9, and the lack of fit was not significant. The mixed natural preservative showed a synergistic antimicrobial effect and reduced the growth of L. monocytogenes by 4 to 5 log CFU/g on lettuce. In addition, the reduction of L. monocytogenes on pork loin was 3 log CFU/g. The mixed natural preservative, which consisted of GSE (6 to 8 ppm), CA (15 to 20 ppm), and nisin (5 to 6 ppm), increased the antibacterial effect against L. monocytogenes. These results suggest that the use of the mixed natural preservative could reduce the economic cost of food preparation, and response surface methodology is considered effective when measuring synergy among antimicrobials.

HIGHLIGHTS
  • Antibacterial effect of a mixed preservative was tested on lettuce and pork loin.

  • The mixed preservative consists of GSE, CA, and nisin.

  • The RSM via the CCD using GSE, CA, and nisin was statistically relevant.

  • The mixed preservative was effective against L. monocytogenes on food products.

 

RASFF Alert -Escherichia coli -Pseudomonas aeruginosa – Enterococcus – Still and Sparkling Bottled Water

RASFF-Logo

RASFF – Escherichia coli, Pseudomonas aeruginosa (>100 CFU/250ml) and Enterococcus in still and sparkling bottled water from Ireland in Ireland

RASFF Alerts – Animal Feed – Salmonella – Fish Meal

RASFF-Logo

RASFF – Salmonella enterica ser. anatum (in 4 out of 5 samples /25g) in fishmeal from the United States in Greece

RASFF – Salmonella enterica ser. anatum (present /25g) ,Salmonella enterica ser. Molade (present /25g) and Salmonella enterica ser. Ochiogu (present /25g) in fishmeal from the United States in Greece

RASFF – Salmonella enterica ser. Montevideo (presence /25g) in fish meal from the United States in Greece

USA – Salmonella Pig Ear Treat Outbreak over at 154

Food Poison Journal

A total of 154 people infected with the outbreak strains of Salmonella were reported from 34 states from contact with pig ear treats.

Illnesses started on dates ranging from June 10, 2015, to September 13, 2019. Ill people ranged in age from less than 1 year to 90 years, with a median age of 40 years. Seventy (45%) ill people were female. Of 133 ill people with information available, 35 (26%) were hospitalized. No deaths were reported.

Research – Antibiofilm Efficacy of Peptide 1018 against Listeria monocytogenes and Shiga Toxigenic Escherichia coli on Equipment Surfaces

Journal of Food Protection

ABSTRACT

Listeria monocytogenes and Shiga toxigenic Escherichia coli (STEC) are important foodborne bacterial pathogens that can form biofilms on equipment surfaces at food processing facilities. Pathogens in biofilms are resistant to conventional antimicrobials and require higher antimicrobial concentrations to be inactivated. In this study, the efficacy of a synthetic innate defense regulator peptide 1018 (peptide 1018) for inactivating L. monocytogenes and STEC (O26, O111, O145, O157) biofilms on stainless steel and polycarbonate surfaces was investigated. Stainless steel and polycarbonate coupons (12 mm in diameter) were used in a Centers for Disease Control and Prevention biofilm reactor containing 400 mL of 10% tryptic soy broth (TSB) that had been inoculated with an individual strain of L. monocytogenes or STEC to obtain 6 log CFU/mL populations. The reactor was set with a constant flow rate at 50 mL/h of 10% TSB for 48 h. After 48 h, coupons were treated with peptide 1018 at 0, 10, 20, or 50 μg/mL in phosphate buffer saline (PBS) for 24 h. Surviving bacterial populations were determined by scraping off the coupons and spiral plating on selective media. Significantly higher levels of pathogens in biofilms formed by certain bacterial strains, including L. monocytogenes F6854, E. coli O157:H7 RM4407 and NADC5713, and non-O157 E. coli NADC3629, were recovered on polycarbonate surfaces than on stainless steel. Antibiofilm efficacy of peptide 1018 against pathogens was concentration-dependent and varied with the type of pathogen and material surfaces. Peptide 1018 at 50 μg/mL significantly inactivated all tested bacterial biofilms on both surfaces compared with the PBS control (P < 0.05). L. monocytogenes was the bacterium most sensitive to peptide 1018; on stainless steel surfaces treated with 50 μg/mL peptide 1018, there was a 3.7- to 4.6-log CFU/cm2 reduction in Listeria populations compared with a 1.0- to 3.5-log CFU/cm2 reduction of STEC. Results suggest that peptide 1018 may be used to inactivate L. monocytogenes and STEC biofilms on equipment surfaces.

HIGHLIGHTS
  • Bacteria attach at higher levels on polycarbonate surfaces than on stainless steel.

  • L. monocytogenes is more sensitive than STEC to peptide 1018.

  • Peptide 1018 can be used to inactivate biofilms on equipment surfaces.

Research – Outbreaks of Shiga Toxin–Producing Escherichia coli Linked to Sprouted Seeds, Salad, and Leafy Greens: A Systematic Review

Journal of Food Protection

ABSTRACT

Shiga toxin–producing Escherichia coli (STEC) outbreaks involving ready-to-eat salad products have been described in the scientific literature since 1995. These products typically do not undergo a definitive control step such as cooking to eliminate pathogens. To reduce the number of STEC infections from salad products, efforts will need to focus on preventing and reducing contamination throughout the food chain. We performed a systematic review of STEC outbreaks involving sprouted seeds, salad, or leafy green products to determine whether there were recurrent features, such as availability of microbiological evidence or identification of the contamination event, which may inform future investigations and prevention and control strategies. Thirty-five STEC outbreaks linked to contaminated leafy greens were identified for inclusion. The outbreaks occurred from 1995 to 2018 and ranged from 8 to more than 8,500 cases. Detection of STEC in the food product was rare (4 of 35 outbreaks). For the remaining outbreaks, the determination of leafy greens as the source of the outbreak mainly relied on analytical epidemiology (20 of 35) or descriptive evidence (11 of 35). The traceback investigation in 21 of 32 outbreaks was not able to identify possible routes leading to where the STEC bacteria came from or how the leaves were contaminated. Investigations in eight outbreaks found poor practice during processing that may have contributed to the outbreak, such as insufficient postharvest disinfection of the product. Six outbreak investigations were able to identify the outbreak strain in animal feces near the growing fields; two of these were also able to find it in irrigation water on the farms, providing a likely route of contamination. These results highlight the limitations of relying on microbiological confirmation as a basis to initiate investigations of upstream production to understand the source of contamination. This review also demonstrates the importance of, and difficulties associated with, food-chain traceback studies to inform control measures and future prevention.

HIGHLIGHTS
  • Systematic review identified 35 STEC outbreaks linked to contaminated leafy greens.

  • Most (20 of 35) outbreaks relied on epidemiological evidence to identify leafy greens.

  • In 21 of 35 studies, no evidence was found for how original contamination occurred.

  • In 11 studies, water was identified as the probable vector in the contaminating product.

  • Only two studies were able to identify the likely source and route of contamination.

Research – Characterization of Virulence and Persistence Abilities of Listeria monocytogenes Strains Isolated from Food Processing Premises

Journal of Food Protection

ABSTRACT

We report the characterization of 15 Listeria monocytogenes strains isolated from various food processing plants by multivirulence locus sequence typing to determine virulence types (VTs) and epidemic clones. Molecular mechanisms involved in adaptation to food processing environments and related to virulence were also studied. Phenotypic behaviors associated with various antimicrobials, biofilm formations, and invasiveness were assessed. There were 11 VTs among the 15 L. monocytogenes strains. Strains belonging to six VTs were stress survival islet 1 (SSI-1) and one strain of VT94 was SSI-2. Tn6188 was found in VT6 and VT94 strains, and bcrABC cassette genes were identified in VT21, VT60, and VT63 strains. Only one strain, in VT20, showed llxS, whereas a full-size inlA was detected in strains belonging to VT8, VT20, VT21, and VT63. VT10, VT20, VT21, VT60, and VT63 strains were the most tolerant to studied disinfectants. A VT6 strain showed the strongest biofilm formation ability in polyvinyl chloride, and strains belonging to VT10, VT11, VT20, and VT94 had moderate abilities. Antimicrobial sensitivity tests showed that all the L. monocytogenes strains were multidrug resistant. F tests revealed that only strains of VT10, VT60, and VT94 were significantly noninvasive (P < 0.05) in Caco-2 cells. Our findings illustrate how L. monocytogenes isolates exploit diverse mechanisms to adapt to adverse conditions. Consequently, detailed characterization of L. monocytogenes isolates is required for comprehensive elimination of this pathogenic bacterium in food processing environments.

HIGHLIGHTS
  • Virulence types of epidemic clones are multidrug resistant and tolerant to disinfectants.

  • Presence of SSI-1, SSI-2, Tn6188, and ability to form biofilm support persistent strains.

  • Strains unable to invade Caco-2 cells have higher tolerance to disinfectants exposure.

Research – Sanitization of Chicken Frames by a Combination of Hydrogen Peroxide and UV Light To Reduce Contamination of Derived Edible Products

Journal of Food Protection

ABSTRACT

Chicken carcass frames are used to obtain mechanically separated chicken (MSC) for use in other further processed food products. Previous foodborne disease outbreaks involving Salmonella-contaminated MSC have demonstrated the potential for the human pathogen to be transmitted to consumers via MSC. The current study evaluated the efficacy of multiple treatments applied to the surfaces of chicken carcass frames to reduce microbial loads on noninoculated frames and frames inoculated with a cocktail of Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium. Inoculated or noninoculated frames were left untreated (control) or were subjected to treatment using a prototype sanitization apparatus. Treatments consisted of (i) a sterile water rinse, (ii) a water rinse followed by 5 s of UV-C light application, or (iii) an advanced oxidation process (AOP) combining 5 or 7% (v/v) hydrogen peroxide (H2O2) with UV-C light. Treatment with 7% H2O2 and UV-C light reduced numbers of aerobic bacteria by up to 1.5 log CFU per frame (P < 0.05); reductions in aerobic bacteria subjected to other treatments did not statistically differ from one another (initial mean load on nontreated frames: 3.6 ± 0.1 log CFU per frame). Salmonella numbers (mean load on inoculated, nontreated control was 5.6 ± 0.2 log CFU per frame) were maximally reduced by AOP application in comparison with other treatments. No difference in Salmonella reductions obtained by 5% H2O2 (1.1 log CFU per frame) was detected compared with that obtained following 7% H2O2 use (1.0 log CFU per frame). The AOP treatment for sanitization of chicken carcass frames reduces microbial contamination on chicken carcass frames that are subsequently used for manufacture of MSC.

HIGHLIGHTS
  • Chicken carcass frames were sanitized using an advanced oxidation process.

  • Salmonella was reduced by 1.1 log CFU per frame with H2O2 and UV-C light.

  • Aerobic bacteria were reduced by up to 1.5 log CFU per frame with 7% H2O2 plus UV-C light.

  • Advanced oxidation processing produced greater reductions than water or UV-C light alone.

Research -Characterization of bacterial pathogens associated with milk microbiota in Egypt

Academic Journals

Abstract

Milk is a substantial source of nutrients needed by all humans across lifespan development. Given its nutritional composition, milk is considered a vehicle for various microbes including beneficial and pathogenic bacteria. In this study, 270 milk samples comprising raw cow and buffalo milk and pasteurized milk with different shelf-life durations were tested along with pasteurized organic milk for the presence of Staphylococcus aureus and Escherichia coli. Collectively, 21 E. coli and 14 S. aureus isolates were cultivated and identified from total milk samples. All E. coli and S. aureus isolates exhibited resistance to erythromycin and penicillin, respectively. Serogroups O26, O128, and O111 were the most frequently identified amongst E. coli isolates, whereas staphylococcal enterotoxins (SEs) were inconsistently produced across S. aureus isolates. The molecular profile showed clustering of 6 isolates of E. coli by harboring stx1, stx2, eaeA genes, and 5 isolates of S. aureus by mecA gene. Findings revealed the bacteriological quality of popularly consumed milk in Egypt, including raw and pasteurized milk with preference to pasteurized organic milk and 7-day shelf life (7DSL) pasteurized milk. However, raw milk and 3MSL pasteurized milk were the major sources of E. coli and S. aureus, posing a serious public health issue.

Research -Occurrence, Seasonal Distribution, and Molecular Characterization of Vibrio vulnificus, Vibrio cholerae, and Vibrio parahaemolyticus in Shellfish (Mytilus galloprovincialis and Ruditapes decussatus) Collected in Sardinia (Italy)

Journal of Food Protection

ABSTRACT

In this study, we investigated the occurrence, seasonal distribution, and molecular characterization of pathogenic vibrios in Mediterranean mussels (Mytilus galloprovincialis) and grooved carpet shells (Ruditapes decussatus) from two harvesting areas of Sardinia (Italy). Samples collected before and after depuration were submitted for qualitative and quantitative determination of Vibrio spp. Vibrio spp. isolates were presumptively identified by means of biochemical methods. Identification and virulence profile of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus were performed by molecular methods. The prevalence of Vibrio spp. in M. galloprovincialis and R. decussatus was, respectively, 96 and 77%. The averaged enumeration (mean ± standard deviation) of Vibrio spp. in samples of M. galloprovincialis and R. decussatus collected at the harvesting time was 2.04 ± 0.45 and 2.51 ± 0.65 log CFU/g, respectively. The average contamination levels in samples collected after purification were 2.28 ± 0.58 log CFU/g (M. galloprovincialis) and 2.12 ± 0.67 log CFU/g (R. decussatus). Four potentially pathogenic V. parahaemolyticus isolates (tdh+ or trh+) were recovered from grooved carpet shells samples. No isolate was tdh+/trh+. The presence of potentially pathogenic vibrios in Sardinian waters strengthens the need for rational purification practices under controlled conditions to guarantee the protection of consumers.

HIGHLIGHTS
  • Occurrence and pathogenicity characteristics of Vibrio pathogens were investigated.

  • Prevalence of Vibrio spp. in M. galloprovincialis was 96% and in R. decussatus was 77%.

  • Environmental conditions influence the occurrence of Vibrio spp.

  • Four V. parahaemolyticus isolates carried tdh or trh genes.

  • Rational purification practices are needed to guarantee the protection of consumers.