Chander Shekhar Sharma

Synergistic activity between lauric arginate and carvacrol in reducing Salmonella in ground turkey

In the present study, low concentrations of carvacrol (0.025 to 0.2%) and lauric arginate (LAE; 25 to 200 ppm) were tested at 4, 22, and 45°C in a broth model, and higher concentrations of carvacrol (0.1 to 5%) and LAE (200 to 5,000 ppm) were tested individually and in combination at 4°C in 3 different ground turkey samples (with 15, 7, and 1% fat content) for their effectiveness against a 3-strain mixture of Salmonella. A low concentration of 25 ppm of LAE or 0.025% carvacrol had no effect on Salmonella in a broth model, but their mixture showed a synergistic action by reducing 6 log cfu/mL Salmonella counts to a nondetectable level within 30 min of exposure. The US Food and Drug Administration-recommended 200 ppm of LAE was not sufficient for Salmonella reductions in ground turkey when applied internally. High concentrations of 2,000 to 5,000 ppm of LAE or 1 to 2% carvacrol were needed to reduce Salmonella counts by 2 to 5 log cfu/g in ground turkey by internal application. No specific relationship existed between fat content and LAE or carvacrol concentrations for Salmonella reductions. For example, 2,000 ppm of LAE could reduce Salmonella counts by 4 log cfu/g in 1% fat-containing turkey samples but very similar ~1.5 log cfu/g reductions in both 7 and 15% fat-containing ground turkey samples. For the total microbial load, about 2,000 ppm of LAE or 2% of carvacrol treatments were needed to achieve 2 to 3 log (P ≤ 0.05) cfu/g reductions in different turkey samples. A mixture of 1% carvacrol and 2,000 ppm of LAE exhibited a synergistic action in ground turkey containing 7% fat by reducing the Salmonella counts by 4 log cfu/g, which was not found with individual antimicrobial treatments.

The combined efficacy of carvacrol and modified atmosphere packaging on the survival of Salmonella, Campylobacter jejuni and lactic acid bacteria on Turkey breast cutlets

The primary objective of this study was to determine the efficacy of carvacrol in combination with modified atmosphere packaging (MAP) in reducing Salmonella on turkey breast cutlets stored at 4 °C. In experiment I, carvacrol (0.5, 1, and 2% v/v) was applied as surface treatment and samples were stored under aerobic condition or as surface and dip treatments followed by storage in an environment of 100% carbon dioxide. The findings of the experiment I revealed the synergistic activity of carvacrol with carbon dioxide in reducing Salmonella when used as dip treatment compared to the surface treatment. In experiment II, turkey breast cutlets were dip treated with carvacrol (0.25, 0.5, and 1% v/v) for 30 s and stored under MAP (95% carbon dioxide and 5% oxygen) to evaluate the efficacy against Salmonella, Campylobacter jejuni and lactic acid bacteria on turkey breast cutlets. In experiment II, the combined application of carvacrol and MAP resulted in 1.0-2.0 log CFU/g reduction (P ≤ 0.05) of both Salmonella and Campylobacter on turkey breast cutlets for 7 d storage at 4 °C. MAP alone and in combination with carvacrol reduced lactic acid bacteria (P ≤ 0.05) on cutlets stored at 4 °C for 21 d period. There was no difference (P ≤ 0.05) in meat color among treatments and controls except for an increased paleness of meat (P ≤ 0.05) observed for the 1% carvacrol treated cutlets stored under MAP after 21 d of storage. The high concentration of carbon dioxide and carvacrol treatments did not cause any alteration in meat pH (P ≤ 0.05). In conclusion, carvacrol was effective at a low concentration of 0.25% (v/v) in reducing Salmonella and C. jejuni by ∼1.0 log CFU/g when stored under MAP.

Reduction of Salmonella on chicken meat and chicken skin by combined or sequential application of lytic bacteriophage with chemical antimicrobials.

The effectiveness of recently approved Salmonella lytic bacteriophage preparation (SalmoFresh™) in reducing Salmonella in vitro and on chicken breast fillets was examined in combination with lauric arginate (LAE) or cetylpyridinium chloride (CPC). In another experiment, a sequential spray application of this bacteriophage (phage) solution on Salmonella inoculated chicken skin after a 20s dip in chemical antimicrobials (LAE, CPC, peracetic acid, or chlorine) was also examined in reducing Salmonella counts on chicken skin. The application of phage in combination with CPC or LAE reduced S. Typhimurium, S. Heidelberg, and S. Enteritidis up to 5 log units in vitro at 4 °C. On chicken breast fillets, phage in combination with CPC or LAE resulted in significant (p<0.05) reductions of Salmonella ranging from 0.5 to 1.3 log CFU/g as compared to control up to 7 days of refrigerated storage. When phage was applied sequentially with chemical antimicrobials, all the treatments resulted in significant reductions of Salmonella. The application of chlorine (30 ppm) and PAA (400 ppm) followed by phage spray (10(9)PFU/ml) resulted in highest Salmonella reductions of 1.6-1.7 and 2.2-2.5l og CFU/cm(2), respectively. In conclusion, the surface applications of phage in combination with LAE or CPC significantly reduced Salmonella counts on chicken breast fillets. However, higher reductions in Salmonella counts were achieved on chicken skin by the sequential application of chemical antimicrobials followed by phage spray. The sequential application of chlorine, PAA, and phage can provide additional hurdles to reduce Salmonella on fresh poultry carcasses or cut up parts.

Reduction of Salmonella in skinless chicken breast fillets by lauric arginate surface application

Lauric arginate (LAE) has been found to be effective against various foodborne pathogens. In this study, the antimicrobial efficacy of LAE against Salmonella and mesophilic organisms was evaluated in fresh, skinless, boneless, uncooked chicken breast fillets. The effect of LAE treatments on pH and color of breast fillets was also assessed. Chicken breast fillets were inoculated with a 4-strain Salmonella cocktail (Salmonella Enteritidis ATCC 4931, Salmonella Heidelberg ATCC 8326, Salmonella Kentucky ATCC 9263, and Salmonella Typhimurium ATCC 14028) and then treated with sterile dionized water (positive control) and 200 ppm and 400 ppm of LAE. The chicken breast fillets were stored at 4 ± 1°C and analyzed on d 0, 1, 3, 5, and 7 for Salmonella, total aerobes, color, and pH. The fillets destined for color analysis were not inoculated with Salmonella cocktail and stored under conditions simulating the retail display. The fillets treated with 400 ppm LAE had lower (P < 0.05) Salmonella counts compared with the positive control from d 0 through d 7 of storage except on d 3, when no effect of LAE was observed. Treating fillets with 200 ppm of LAE caused a significant reduction in Salmonella counts (P < 0.1) on d 0, 1, and 7. Reductions in Salmonella spp. were 0.7 log cfu/g and 0.7 to 1.0 log cfu/g for 200 and 400 ppm treatments, respectively. Lauric arginate did not exhibit any treatment effect on the growth of mesophilic microorganisms, pH, and color of chicken breast fillets (P > 0.05) when applied at 200 and 400 ppm concentrations. These results indicate that surface application of LAE in chicken breast fillets significantly reduces Salmonella during refrigerated aerobic storage without negatively affecting the color of chicken breast fillets.

Sodium metasilicate affects growth of Salmonella Typhimurium in fresh, boneless, uncooked chicken breast fillets stored at 4°C for 7 days

The objectives of this study were to determine the antimicrobial effects of sodium metasilicate (SMS) against Salmonella and psychrotrophic organisms in fresh, boneless, uncooked chicken breast fillets and to ascertain the effects of SMS treatments on pH. Chicken breast fillets were inoculated with Salmonella enterica serovar Typhimurium treated with 0% SMS and no inoculum (negative control), 0% SMS and inoculum (positive control), 1% SMS, or 2% SMS solutions and stored at 4 ± 1°C. All samples were analyzed after 0, 1, 3, 5, and 7 d for Salmonella, psychrotrophic organisms, and pH. The fillets that were treated with 1 or 2% SMS had lower (P < 0.05) Salmonella counts as compared with those of the positive control at 3 through 7 d. Reductions in Salmonella Typhimurium were 0.83 to 0.91 log cfu/g and 1.04 to 1.16 log cfu/g for 1 and 2% SMS treatments, respectively. The psychrotrophic counts were similar (P > 0.05) for all treatments. The pH values for samples treated with 1 or 2% SMS were higher (P < 0.05) when compared with those of the negative and positive controls. This study revealed that SMS could restrict the growth of pathogenic Salmonella Typhimurium in fresh poultry.

Reduction of Salmonella on chicken breast fillets stored under aerobic or modified atmosphere packaging by the application of lytic bacteriophage preparation SalmoFreshTM

The present study evaluated the efficacy of recently approved Salmonella lytic bacteriophage preparation (SalmoFresh™) in reducing Salmonella on chicken breast fillets, as a surface and dip application. The effectiveness of phage in combination with modified atmosphere packaging (MAP) and the ability of phage preparation in reducing Salmonella on chicken breast fillets at room temperature was also evaluated. Chicken breast fillets inoculated with a cocktail of Salmonella Typhimurium, S. Heidelberg, and S. Enteritidis were treated with bacteriophage (10(9) PFU/mL) as either a dip or surface treatment. The dip-treated samples were stored at 4°C aerobically and the surface-treated samples were stored under aerobic and MAP conditions (95% CO2/5% O2) at 4°C for 7 d. Immersion of Salmonella-inoculated chicken breast fillets in bacteriophage solution reduced Salmonella (P < 0.05) by 0.7 and 0.9 log CFU/g on d 0 and d 1 of storage, respectively. Surface treatment with phage significantly (P < 0.05) reduced Salmonella by 0.8, 0.8, and 1 log CFU/g on d 0, 1, and 7 of storage, respectively, under aerobic conditions. Higher reductions in Salmonella counts were achieved on chicken breast fillets when the samples were surface treated with phage and stored under MAP conditions. The Salmonella counts were reduced by 1.2, 1.1, and 1.2 log CFU/g on d 0, 1, and 7 of storage, respectively. Bacteriophage surface application on chicken breast fillets stored at room temperature reduced the Salmonella counts by 0.8, 0.9, and 0.4 log CFU/g after 0, 4, and 8 h, respectively, compared to the untreated positive control. These findings indicate that lytic phage preparation was effective in reducing Salmonella on chicken breast fillets stored under aerobic and modified atmosphere conditions.

Evaluation of USDA approved antimicrobials on the reduction of Salmonella and Campylobacter in ground chicken frames and their effect on meat quality

&NA; The main objective of this study was to examine the efficacy of USDA approved antimicrobials in reducing Salmonella Heidelberg (S. H.) and Campylobacter jejuni (C. j.) in ground chicken frames and to determine the treatment effects on total aerobic counts and meat color. Six antimicrobials (0.1% peracetic acid [PAA], 0.6% cetylpyridinium chloride [CPC], 0.005% sodium hypochlorite, 1.5% acidified lactic acid [ALA], 0.3% propionic acid, and 0.1% lauric arginate [LAE]) applied as dip treatments were evaluated in their efficacy in reducing S. Heidelberg and C. jejuni. Fresh chicken frames were spot inoculated with nalidixic acid resistant S. H. and C. j. (ATCC 33291) to achieve a recovery level of ca. 3 log CFU/g in the ground product. Frames were dipped for 10 s in each antimicrobial solution and each treatment was replicated on 3 frames. Three separate replications were conducted for this experiment. Frames were blended, and ground samples similar to mechanically separated chicken (MSC) were obtained and stored at 4°C for 24 hours. Samples were analyzed after grinding on d zero (2 h) and d one (24 h) to determine reduction in S. H. and C. j. counts in MSC. PAA and LAE treatments had the highest reductions on Salmonella counts (P ≤ 0.05), both treatments resulting in 0.9 log CFU/g reduction as compared to control on d zero. PAA and CPC reduced Salmonella counts by 1.4 and 0.9 log CFU/g, respectively, on d one; PAA, propionic acid, ALA, and LAE resulted in one log CFU/g reductions of C. j. as compared to control on d one. There was no significant difference among the treatments in their ability to reduce C. j. on d one. The treatments had no effect on total aerobic counts. The findings from the study indicate that PAA, CPC, and LAE can reduce S. H. in ground chicken frames, whereas all the antimicrobials tested in the study, except chlorine, have the ability to reduce C. j. in ground chicken frames, a product similar to commercial MSC.

Induction and stability of oxidative stress adaptation in Listeria monocytogenes EGD (Bug600) and F1057 in sublethal concentrations of H2O2 and NaOH.

Food processing and food handling environments may contain residual levels of sanitizers or cleaners which may trigger oxidative stress adaptation in Listeria monocytogenes. The aim of this study was to determine the induction and stability of oxidative stress adaptation in L. monocytogenes EGD (Bug600) (serotype 1/2a) and F1057 (serotype 4b) at different concentrations and times of sublethal oxidative stress induced by H2O2 or sublethal alkali stress induced by NaOH at 37°C. Both L. monocytogenes Bug600 and F1057 strains showed significantly higher survival in lethal oxidative stress (1000ppm H2O2) after pre-exposure to 50ppm H2O2 for 30min compared to control cells (no pre-exposure to H2O2). When the cells were pre-exposed to sublethal alkali stress by NaOH, the oxidative stress adaptation was induced within 5min in L. monocytogenes. The survival of both L. monocytogenes strains was increased by 2 to 4.5 logs in lethal oxidative stress when the cells were pre-exposed to sublethal alkali stress at pH9 from 5 to 120min by NaOH compared to control cells (no pre-exposure to sublethal alkali pH). Two other alkali reagents tested (KOH and NH4OH) also induced oxidative stress adaptation in L. monocytogenes. For both L. monocytogenes strains, the oxidative stress adaptation induced by sublethal H2O2 was reversible in 30min and that induced by sublethal alkali stress was reversible within 60min at 37°C in the absence of such sublethal stress. These findings show that sublethal oxidative or alkali stress conditions can induce oxidative stress adaptation that may increase the risk of survival of L. monocytogenes cells in lethal oxidative stress.

Sodium metasilicate affects growth of Campylobacter jejuni in fresh, boneless, uncooked chicken breast fillets stored at 4 degrees Celsius for 7 days

The objectives of this study were to determine the antimicrobial effects of sodium metasilicate (SMS) treatments against Campylobacter jejuni in fresh, boneless, uncooked chicken breast fillets and to ascertain the effects of SMS treatments on pH. The fillets were inoculated with C. jejuni, treated with 0% SMS and no inoculum (negative control), 0% SMS and inoculum (positive control), 1 and 2% SMS solutions, and stored at 4 ± 1°C. All samples were analyzed after 0, 1, 3, 5, and 7 d storage for C. jejuni, psychrotrophic organisms, and pH. Campylobacter jejuni and psychrotrophic counts for samples treated with 1 and 2% SMS solutions were similar (P > 0.05) to the positive control on all storage days. The pH values for 2% SMS marinade treatments were higher (P < 0.05) when compared with the negative and positive controls through 7 d of storage. Based on the findings in this study, a second study was conducted to determine the level of SMS necessary to reduce C. jejuni by at least 1 log cfu/g. The treatments were the same as previously discussed, except SMS was used at levels of 1 and 2% of the weight of the meat instead of percentage of the solution. Chicken fillets treated with 1 and 2% SMS (by weight of meat) resulted in 1.12 to 1.26 and 3.27 to 3.79 log cfu/g reductions in C. jejuni, respectively, when compared with the positive control. Except for d 0, psychrotrophic counts for samples treated with 2% SMS were lower (P < 0.05) than negative and positive controls on all storage days. The pH values were higher (P < 0.05) for all SMS treatments when compared with the negative and positive controls. This study revealed that SMS, when used at elevated levels in excess of the USDA Food Safety and Inspection Service 2% approved level, could function to control Campylobacter jejuni and extend the shelf life of raw poultry by retarding the growth of psychrotrophic bacteria.

Homologous stress adaptation, antibiotic resistance, and biofilm forming ability of Salmonella enterica serovar Heidelberg ATCC8326 on different food-contact surfaces following exposure to sublethal chlorine concentrations1

Abstract Salmonella enterica serovar Heidelberg (American Type Culture Collection; ATCC 8326) was examined for the ability to adapt to the homologous stress of chlorine through exposure to increasing chlorine concentrations (25 ppm daily increments) in tryptic soy broth (TSB). The tested strain exhibited an acquired tolerance to chlorine in TSB with the tolerant cells growing in concentrations up to 400 ppm. In addition, the chlorine stressed cells displayed rugose morphology on tryptic soy agar (TSA) plates at 37°C. The minimum inhibitory concentration (MIC) of chlorine for adapted (rugose and smooth) cells was determined to be 550 ppm and 500 ppm, respectively whereas the MIC for the control was 450 ppm. The biofilm forming ability of the adapted and control cells were examined on both plastic and stainless steel surface at room temperature and 37°C. The rugose variant, in contrast to the smooth (adapted and control) showed the ability to form strong biofilms (P ≤ 0.05) on a plastic surface at room temperature and 37°C. Rugose cells compared to smooth and control attached more (P ≤ 0.05) to steel surfaces as well. The possibility of cross‐adaptation was examined by exposing the adapted and control cells to different antibiotics according to the Clinical & Laboratory Standards Institute guidelines. Adapted cells exhibited reduced susceptibility to some of the antibiotics tested as compared to control. The findings of this study suggest that exposure to sublethal chlorine concentration during the sanitization procedure can result in tolerant Salmonella cells. Chlorine may confer cross‐protection that aids in the survival of the tolerant population to other environmental stresses.