Effectiveness of Individual or Combined Sanitizer Treatments for Inactivating Salmonella spp. on Smooth Surface, Stem Scar, and Wounds of Tomatoes

ABSTRACT: Unwaxed, green tomatoes ('Florida 47' cultivar) were contaminated with Salmonella and then treated with aqueous solutions of sodium hypochlorite (HOCl; 200 ppm), acidified sodium chlorite (ASC; 1200 ppm), peroxyacetic acid (PAA; 87 ppm), or chlorine dioxide gas (ClO₂; total 100 mg). Additionally, a combined treatment of immersion in HOCl, followed by immersion in ASC and then exposure to ClO₂ gas was investigated. Tomatoes were spot inoculated with a 5-strain Salmonella cocktail on smooth surfaces, stem scar tissue, or puncture wounds. A 3 replicate set of each of the sample groups was stored at 20 °C and 95% relative humidity (RH) and retested after 5 d. Greater than 4.0-log unit reductions of Salmonella spp. inoculated on the smooth surface of the tomatoes were seen for all aqueous sanitizer treatments, with Salmonella populations below the detection limit after 5 d of storage. All aqueous treatment groups showed > 1.0-log unit reductions in Salmonella at the stem scar and >2.0-log unit reduction at puncture wounds. The ClO₂ gas treatment reduced Salmonella to undetectable levels at the stem scar, but had no apparent effect on populations inoculated in puncture wounds. The combined treatment resulted in a 3.0-log unit reduction of inoculated Salmonella at puncture wounds. In all cases except for treatment with chlorine, surviving Salmonella populations did not increase after the 5 d of storage. Results of this study suggest the combined treatment was most effective for minimizing the risk of Salmonella contaminated on tomatoes.     

Effects of Water,Sodium Hypochlorite,Peroxyacetic Acid,and Acidified Sodium Chlorite on In‐Shell Hazelnuts Inoculated with Salmonella Enterica Serovar Panama

Recent foodborne disease outbreaks involving minimally processed tree nuts have generated a need for improved sanitation procedures. Chemical sprays and dips have shown promise for reducing pathogens on fresh produce, but little research has been conducted for in‐shell hazelnuts. This study analyzed the effectiveness of 3 chemical sanitizers for reducing Salmonella on in‐shell hazelnuts. Treatments of water, sodium hypochlorite (NaOCl; 25 and 50 ppm), peroxyacetic acid (PAA; 80 and 120 ppm), and acidified sodium chlorite (ASC; 450, 830, and 1013 ppm) were sprayed onto hazelnut samples inoculated with Salmonella enterica serovar Panama. Hazelnut samples were immersed in liquid cultures of S. Panama for 24 h, air‐dried, and then sprayed with water and chemical treatments. Inoculation achieved S. Panama populations of approximately 8.04 log CFU/hazelnut. Surviving S. panama populations were evaluated using a nonselective medium (tryptic soy agar), incubated 3 h, and then overlaid with selective media (xylose lysine deoxycholate agar). All of the chemical treatments significantly reduced S. Panama populations (P ≤ 0.0001). The most effective concentrations of ASC, PAA, and NaOCl treatments reduced populations by 2.65, 1.46, and 0.66 log units, respectively. ASC showed the greatest potential for use as a postharvest sanitation treatment.     

Acidified Sodium Chlorite as an Alternative to Chlorine for Elimination of Salmonella on Alfalfa Seeds

ABSTRACT:  The health and environmental hazard associated with the use of chlorine for food processing has been documented previously. This study was conducted to determine if acidified sodium chlorite (ASC) could be used to replace calcium hypochlorite (Ca[OCl]₂) for disinfection of alfalfa seeds. Contaminated seeds containing approximately 1.5 × 10⁷ CFU/g of Salmonella were treated with ASC or Ca(OCl)₂ at different concentrations and for different periods of time. Results showed that the efficacy of ASC and Ca(OCl)₂ for elimination of Salmonella on contaminated seeds could be improved greatly by extending the treatment time from the traditional 15 to 45 min. Treatment of seeds with 800 ppm of ASC for 45 min reduced the number of Salmonella by 3.9 log units, approximately 1.2 log units higher than that treated with 20000 ppm of Ca(OCl)₂. Treatment of seeds with a lower concentration (100 to 400 ppm) of ASC for 45 min reduced the number of Salmonella by 1.3 to 2.2 log units. Soaking alfalfa seeds in 800 ppm of ASC for 45 min did not affect seed germination. However, soaking seeds in 20000 ppm of Ca(OCl)₂ for 45 min reduced seed germination by 20%. Unlike Ca(OCl)₂, antimicrobial efficiency of ASC was not affected by pre-exposure to alfalfa seeds. Data presented also showed that Salmonella on newly inoculated seeds that had been stored at 4 °C for less than 7 d were more sensitive to sanitizer treatment than those on seeds that had been stored for 4 wk or longer.     

Evaluation of overhead spray-applied sanitizers for the reduction of Salmonella on tomato surfaces

Abstract: Efficacy of sanitizers in an overhead spray and brush roller system was examined for reducing Salmonella on unwaxed, mature green tomatoes. Surface inoculated tomatoes were treated in the overhead spray system for 5, 15, 30, and 60 s. A sodium hypochlorite (NaOCl) study tested NaOCl (25, 50, and 100 mg/L) against a water control. A sanitizer study examined NaOCl (100 mg/L), chlorine dioxide (ClO₂; 5 mg/L), peroxyacetic acid (PAA; 80 mg/L), and water. The overhead spray system was also compared to a scale‐model flume. All NaOCl concentrations were significantly more effective at removing Salmonella than water and achieved at least a 3‐log₁₀ CFU/mL reduction at different treatment times (P < 0.05). NaOCl (100 mg/L) achieved a 4 ± 1.8 log₁₀ CFU/mL reduction at 15 s. In the sanitizer study, NaOCl, ClO₂, and PAA achieved at least a 3‐log₁₀ CFU/mL reduction at 15 s and between 3.9 and 5.5 log₁₀ CFU/mL reductions at 30 to 60 s. NaOCl (100 mg/L) in the overhead spray system significantly reduced more Salmonella than in the flume at 15 to 60 s. NaOCl flume treatment only reached a 1.3 ± 1.1 log₁₀ CFU/mL reduction at 15 s. Results of this study demonstrate the ability of sanitizers in the laboratory model overhead spray system to reduce Salmonella on tomato surfaces. An overhead spray system could be implemented instead of flumes to achieve higher pathogen reduction with less water and sanitizer use, thereby lowering packing costs. Practical Application: The use of a non‐recirculating, overhead spray brush roller system could offer a cost effective and efficacious way of washing tomatoes. The use large communal dump tanks in tomato processing has been suspected as a source of contamination in the tomato processing process. If effective, the brush roller system could augment or possible replace currently used dump tanks.     

Thermal Inactivation of Salmonella spp. in Chicken Broth, Beef, Pork, Turkey, and Chicken: Determination of D- and Z-values

ABSTRACT: The heat resistance of 35 Salmonella strains was determined at 55 to 65°C. No correlation between the heat resistance and the origin of the Salmonella spp. could be established. D-values in chicken broth, using a linear regression, of an 8 Salmonella serotype cocktail were 4.87, 2.72, 1.30, and 0.41 min at 55, 58, 60 and 62°C, respectively. Using a linear model, the D-values ranged from 4.86 min at 55°C to 0.38 min at 62°C. When the 8 Salmonella serotype cocktail was heated in meat, D-values at the common test temperatures of 58 and 60°C calculated by both approaches were significantly higher (p < 0.05) than those observed in chicken broth.     

Comparison of the Antimicrobial and Sanitizer Resistance of Salmonella Isolates from Chicken Slaughter Processes in Korea

Salmonella is a foodborne pathogen worldwide. Outbreaks of Salmonella are commonly associated with consumption of contaminated foods such as poultry products. Therefore, the objective of this study was to determine the occurrence, biofilm formation, antibiotic resistance, and sanitizer resistance of Salmonella enterica isolated from chicken carcasses. A total of 318 samples were collected from 15 chicken slaughterhouses in 8 provinces of Korea. They were then examined for Salmonella contamination. S. enterica isolates were tested for their susceptibilities to 15 antimicrobials by broth microdilution method. Their biofilm formation ability and resistance to sanitizers were also evaluated. Eighty‐two isolates of S. enterica were obtained from the 318 samples. There were 14 serotypes and 2 untypable isolates. Fifty‐seven (69.5%) isolates were resistant to at least one antibiotic while 30 (36.6%) isolates were resistant to 5 or more antibiotics. Two S. Senftenberg and 3 S. Montevideo isolates exhibited considerable biofilm formation ability (A₆₀₀>0.2) following incubation in Luria‐Bertani (LB) broth for 48 h. Biofilm cell survival and recovery growth assay after sanitization showed that most isolates were highly susceptible to 2.5% lactic acid and 0.1% cetylpyridinium chloride. Therefore, lactic acid and cetylpyridinium chloride might be alternatively or additionally used in addition to chlorine‐based sanitizers that are frequently used to reduce Salmonella contamination of chicken carcasses. Our results provide basic information on the distribution of Salmonella serotypes in chicken slaughterhouses. This study also highlights the necessity to improve farming practices and use antimicrobial agents cautiously. This study also suggests that sanitization during the slaughtering process might be necessary to reduce Salmonella contamination of chicken carcasses.     

肉禽及其制品中沙门氏菌失活建模的研究进展

沙门氏菌是常见的食源性致病菌,可引起人类的胃肠炎、败血症等疾病,严重者会危及生命安全。肉禽及其制品是消费者餐桌上必不可少的食材,然而在肉禽类的生长、运输、屠宰及其制品的加工、贮藏、销售过程中,极易受到沙门氏菌的污染,威胁消费者舌尖上的安全。预测微生物学中的失活模型能反映出传统的热处理、新兴的杀菌技术对肉禽及其制品中沙门氏菌的灭活效果。该研究首先对肉禽及其制品中沙门氏菌传统热失活、非热失活模型的最新研究进行论述;其次,考虑了亚致死损伤沙门氏菌的存在对失活模型建立的影响;再次,对单细胞失活的概率模型进行讨论;最后,对沙门氏菌在失活模型中的研究进行总结并提出建议,以期对未来制定食品质量和安全的管理方法提供理论参考。     

Thermal Inactivation of Salmonella spp. and Listeria innocua in the Chicken Breast Patties Processed in a Pilot-Scale Air-Convection Oven

ABSTRACT Salmonella spp. or Listeria innocua containing chicken patties were processed via a pilot-scale air-convection oven to a final temperature of 55 to 80 °C. Thermal processing was conducted at an air temperature of 149 °C, an air velocity of 7.1 to 12.7 m3/min, and a wet bulb temperature of 39 to 98 °C. The cooking time was correlated with cooking conditions using a standardized least square regression model. The models were developed to correlate cooking time and the thermal inactivation of Salmonella or Listeria with cooking conditions. The thermal lethality of Salmonella and Listeria increased with increasing product temperature and wet bulb temperature. This study bridged the gap between laboratory studies and commercial applications and will help commercial processors to evaluate and validate their thermal processes.     

Thermal Inactivation of Salmonella spp., Salmonella typhimurium DT104, and Escherichia coli 0157:H7 in Ground Beef

ABSTRACT: In 19.1% fat ground beef, Escherichia coli 0157:H7 was less heat- resistant at ≥58°C than the Salmonella typhimurium DT104 and Salmonella senftenberg , but at 55°C the D value was similar to DT104 strains and higher than an eight-strain Salmonella cocktail. Inactivation of E. coli 0157:H7 was more temperature-dependent than the cocktail and DT104 strains. E. coli and DT104 strains were more heat-resistant in beef containing 19% fat than 4.8% fat. The cocktail was more thermally stable in stationary as compared to log phase. Freezing of inoculated raw meat decreased heat resistance of the cocktail. The pathogenic strain, growth phase of the organism, state of the meat (fresh or frozen) and meat composition must be considered when designing protocols to verify thermal processes.     

Multiplex PCR for the simultaneous detection of Salmonella spp. and Salmonella Enteritidis in food

Multiplex PCR assay (mPCR) for the detection of Salmonella spp. and S. Enteritidis was developed in this study using artificially contaminated chicken carcasses. The assay showed 100% specificity to detect approximately 1 CFU of Salmonella in 10 g of chicken skin after non‐selective enrichment. The mPCR was evaluated in Minas cheese, fresh pork sausage and chicken carcasses commercially available. Salmonella spp. was detected in nine of sixty‐six chicken carcasses, five of fifty‐two cheese samples, and five of fifty‐two sausage samples. The serovar Enteritidis was detected in two samples of contaminated sausage. The mPCR results were confirmed by conventional culture and biochemical identification of the isolates. Serotyping confirmed the presence of S. Enteritidis in sausage samples and showed contamination by serovars Schwarzengrund and Montevideo in chicken carcasses.     

SD-PMA-qPCR快速检测冷冻肉制品中活性沙门氏菌的研究

为实现冷冻肉制品中活性沙门氏菌的快速检测与控制,本文利用PMA（叠氮溴化丙锭）和SD（脱氧胆酸钠）消除死菌和损伤菌的影响,建立了运用SD-PMA-qPCR法检测活性沙门氏菌的反应体系和反应条件,并进行人工染菌样品检测试验。SD的最佳浓度确定为0.1%,最佳孵育时间为20 min。对-20 ℃低温冷冻处理3~4 d的沙门氏菌处理液,使用平板计数法、qPCR、PMA-qPCR和SD-PMA-qPCR进行计数,结果发现qPCR与PMA-qPCR二者检测值接近且明显高于平板计数值,而SD-PMA-qPCR检测值与平板计数值相近,这表明SD和PMA的联合使用能有效的消除死菌和损伤菌的影响。人工染菌样品试验表明,沙门氏菌检出限在102 CFU/g,同时106 CFU/g大肠杆菌O157:H7的存在不会影响检测结果。本研究所建立的SD-PMA-qPCR检测方法特异性好、灵敏度高,有望成为快速检测冷冻肉制品中活性沙门氏菌的新方法,具有很好的研究价值和应用前景。     

PCR Detection and Microbiological Isolation of Salmonella spp. from Fresh Beef and Cantaloupes

ABSTRACT:  Species belonging to the genus Salmonella are an important cause of enteric fevers, gastroenteritis, and septicemia, and the pathogens are commonly transmitted through contaminated food. In this study, polymerase chain reaction (PCR) amplification of a 287-bp region of the invA gene was compared to a microbiological technique to determine the presence of Salmonella in retail beef and in cantaloupe rinse samples. Both methods showed the same level of sensitivity, detecting 1 CFU/25 g of meat after enrichment for 24 h at 42 °C. The presence of Salmonella was determined in 50 commercial top sirloin beef samples that were not artificially inoculated. Three samples were positive by the microbiological method, and these samples and an additional sample were positive by the PCR. Both methods were also used to test surface rinses of cantaloupes collected from 4 farms in Nayarit, Mexico. Salmonella was detected by the microbiological method in 9 of 20 samples (45%), whereas the pathogen was detected by the PCR in 11 samples (55%). This study demonstrates the utility of the PCR targeting the invA gene to determine the presence of Salmonella spp. in beef and cantaloupe samples.     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium,and Listeria monocytogenes on Stored Iceberg Lettuce by Aqueous Chlorine Dioxide Treatment

ABSTRACT: Inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes in iceberg lettuce by aqueous chlorine dioxide (ClO₂) treatment was evaluated. Iceberg lettuce samples were inoculated with approximately 7 log CFU/g of E. coli O157:H7, S. typhimurium, and L. monocytogenes. Iceberg lettuce samples were then treated with 0, 5, 10, or 50 ppm ClO₂ solution and stored at 4 °C. Aqueous ClO₂ treatment significantly decreased the populations of pathogenic bacteria on shredded lettuce (P < 0.05). In particular, 50 ppm ClO₂ treatment reduced E. coli O157:H7, S. typhimurium, and L. monocytogenes by 1.44, 1.95, and 1.20 log CFU/g, respectively. The D₁₀‐values of E. coli O157:H7, S. typhimurium, and L. monocytogenes in shredded lettuce were 11, 26, and 42 ppm, respectively. The effect of aqueous ClO₂ treatment on the growth of pathogenic bacteria during storage was evaluated, and a decrease in the population size of these pathogenic bacteria was observed. Additionally, aqueous ClO₂ treatment did not affect the color of lettuce during storage. These results suggest that aqueous ClO₂ treatment can be used to improve the microbial safety of shredded lettuce during storage.     

肉鸡屠宰加工过程中沙门氏菌的污染情况及其耐药性分析

目的：了解四川某肉鸡屠宰加工过程不同环节沙门氏菌的污染情况、耐药性和耐药谱,为食品安全和临床用药提供理论依据。方法：根据GB 4789.4-2010《食品安全国家标准 食品微生物学检验 沙门氏菌检验》、沙门氏菌科玛嘉显色培养基筛选疑似沙门氏菌,并针对沙门氏菌invA和hut基因的二重PCR方法对疑似沙门氏菌鉴定,再用纸片扩散法对其分离株进行10 种抗菌药物（组合）的药敏实验,参考临床和实验室标准化研究所（Clinical andLaboratory Standards Institute,CLSI）标准（2010）判定药敏结果。结果：从1 350 份样品中,分离鉴定出156 株沙门氏菌;肉鸡屠宰前沙门氏菌的污染率为13.53%;烫毛脱毛、开肛、净膛、冲淋4 个环节鸡胴体和分割鸡肉、冷冻鸡肉沙门氏菌的污染率分别为0、7.23%、9.80%、11.54%、14.50%、9.33%。沙门氏菌分离株对萘啶酸（100.00%）和氨苄西林（85.90%）的耐药率最高,对甲氧苄啶/磺胺甲噁唑（44.23%）、庆大霉素（39.10%）、四环素（35.26%）的耐药率较高,对头孢曲松敏感,多重耐药率为53.85%,共有39 种耐药谱,从肉鸡屠宰前到分割鸡肉沙门氏菌分离株的耐药谱型先下降再上升。结论：四川某肉鸡屠宰生产链中沙门氏菌的污染率及其耐药情况比较严重,且可能存在从上游向下游生产链传播的情况,需要加强卫生和抗菌药物使用监督。     

食品能力验证中沙门氏菌的分离和鉴定

目的为了提升实验室食品中沙门氏菌检测能力,增强实验室竞争能力,本实验室参加了中国食品药品检定研究院组织的沙门氏菌检测能力验证活动。方法依据中华人民共和国国家标准GB 4789.4-2010,采用传统分离方法和血清学鉴定,联合全自动微生物生化鉴定系统(VITEK2-compact)对分离出的疑似菌进行生化鉴定。结果编号为CODE1样品+CODE1奶粉混合样检出纽波特沙门氏菌和科林德尔沙门氏菌,CODE3样品+CODE3奶粉混合样检出维普拉沙门氏菌、鼠伤寒沙门氏菌和埃科沙门氏菌,其余8个样品未检出。结论顺利完成本次能力验证活动。     

四川省猪肉源大肠杆菌和沙门氏菌的分离与耐药性分析

采集四川省鲜猪肉126份,选择性培养基分离大肠杆菌、沙门氏菌,划线纯化后,通过形态特性及16S rDNA序列对菌株进行鉴定,利用K-B法(CLSI)药敏试验检测大肠杆菌和沙门氏菌对17种抗生素的耐药性。结果表明：共获得大肠杆菌105株,沙门氏菌18株,检出率分别为83.33%和14.28%。大肠杆菌对各种抗生素的耐药率在0%～55.3%,对四环素、氨苄西林耐药率最高,分别为55.3%、52.3%,对其他抗菌药物的耐药率依次为卡那霉素(26.7%)＞链霉素(19.1%)＞环丙沙星(17.1%)＞诺氟沙星(14.3%)＞头孢噻吩(11.5%)＞氨苄西林/舒巴坦(4.8%)＞头孢曲松、头孢噻肟、氨曲南、庆大霉素(2.9%)＞头孢他啶(1.9%)>阿莫西林/克拉维酸(0.8%)＞丁胺卡那霉素(0.1%)＞亚胺培南、哌拉西林/他唑巴坦(0%),共产生28种耐药谱,氨苄西林、四环素及氨苄西林/四环素谱型占优势。沙门氏菌对各种抗生素的耐药率在0%～55.5%,其中对四环素、氨苄西林、链霉素的耐药率最高,依次是55.5%、38.9%、27.8%,对其他抗菌药物的耐药率依次是：氨苄西林/舒巴坦、阿莫西林/克拉维酸(11.1%)＞头孢噻肟、卡那霉素、头孢噻吩、庆大霉素、环丙沙星、诺氟沙星(5.6%)＞头孢曲松、头孢他啶、亚胺培南、氨曲南、哌拉西林/他唑巴坦、丁胺卡那霉素(0%),共产生13种耐药谱。四川省肉源大肠杆菌、沙门氏菌对各类抗生素已表现出一定的耐药性,虽耐药率相对较低,但多重耐药已经成为普遍现象,应引起足够重视,并加强对其监测。     

江苏省沙门氏菌全基因组序列中外排泵耐药基因分布及其特征研究

目的 了解江苏省沙门氏菌的耐药及其耐药相关基因的分布状况。方法 根据美国实验室标准化临床协会的操作规范推荐的肉汤稀释法测定沙门氏菌的药物敏感性,应用在线数据库分析基于多个位点序列型、耐药基因的种类和数量。结果 至少一半以上的沙门氏菌对链霉素、氨苄西林、复方磺胺甲恶唑、四环素、大观霉素、利福平耐药。尤其是对大观霉素和利福平的耐药率高达90%以上。D群沙门氏菌是主要的血清群。ST11型为主要的序列型。与外排泵耐药相关的9种MFS耐药基因均可以检出,最多的沙门氏菌携带8种基因。结论 江苏沙门氏菌的耐药问题比较严重,在临床用药和畜牧业养殖中,应加强抗生素合理使用。