多重PCR法检测产志贺毒素性大肠杆菌的 4个血清型

目的建立多重PCR法检测产志贺毒素性大肠杆菌(shiga toxin-producing Escherichia coli, STEC)主要血清型O26、O145、O45和O121的分析方法。方法根据GenBank登录序列,设计扩增O抗原翻转酶(wzx)基因的引物,建立PCR方法,并以STECO26、O145、O45和O121基因组为模板,检验多重PCR的灵敏度和特异性。使用建立的PCR,检测牛胴体表面拭子,阳性扩增条带送测序,以验证PCR扩增的可靠性。同时将阳性扩增样品,涂布显色平板,分离靶标血清型细菌。结果本研究成功建立STEC O26、O145、O45和O121的多重PCR方法, PCR循环参数中退火温度为60℃,扩增片段分别为249、353、890和587 bp。多重PCR直接检测O26、O145、O45和O121时,最低检测限介于10~3～10~4 CFU/mL,而增菌后再检测,最低检测限均为1CFU/m L。多重PCR用于其他血清型STEC,和非大肠杆菌扩增时,均未扩增出目的条带,只有O26、O145、O45和O121能够扩增出相应条带。当使用多重PCR直接检测胴体擦拭子时,阳性率为5.45%(3/55),主要为O26、O145血清型;增菌后检测阳性率为7.27%(4/55),主要为O26、O145和O121血清型。阳性PCR扩增样品,成功分离到O26两株、O145和O121各一株。分离菌株具有典型大肠杆菌的生化特性,携带STEC代表性毒力因子志贺毒素和紧密素,且具有多重耐药性。结论以STECO26、O145、O45和O121的wzx基因为检测靶标,成功建立多重PCR方法,灵敏度和特异性良好,与细菌分离联合使用,可减少工作量,精准分离目的病原菌。     

Heat Resistance of Salmonella spp., Listeria monocytogenes, Escherichia coli 0157:H7, and Listeria innocua M1, a Potential Surrogate for Listeria monocytogenes, in Meat and Poultry: A Review

ABSTRACT: The heat-resistance data in meat and poultry for various strains of Salmonella, Listeria monocytogenes , and Escherichia coli O157:H7 as well as Listeria innocua M1 are summarized. Heat resistance of these organisms is affected by many factors. Different strains of the same organism have different responses to heat. Heat resistance can also be influenced by the age of the culture, growth conditions, pH, and numerous other factors. Data from this review may prove useful to processors in validating their times and temperatures for thermal processing of meat and poultry. The obvious gaps in the data will provide researchers opportunities to fill those gaps. In addition, it will encourage the development of surrogates, whether biological or otherwise, that will be able to be used in an actual processing environment.     

餐饮食品中6病原菌叠氮丙啶-定量聚合酶链反应检测方法开发与检验数据分析

目的建立叠氮丙啶-定量聚合酶链反应法(propidiummonoazide-quantitativepolymerasechain reaction,PMA-qPCR)快速检测餐饮食品中6病原菌的方法。方法开发6种病原菌前增菌通用型培养基,采用热裂解方法提取样品DNA,采用PMA技术鉴别活菌与死菌,运用分子生物学技术检测样品中目标菌的特异性基因片段,建立病原菌的PMA-qPCR检测方法,开展餐饮食品样品病原菌筛查检测。结果本方法特异性良好,灵敏度较高, 1533份餐饮食品样品中检出病原菌71株,检出率为4.63%(71/1533)。结论本研究运用PMA-qPCR开展病原菌活菌检测技术研究,所建立研究方法可广泛应用于餐饮食品及相关食品中病原菌的快速筛查检测,具有较好的应用前景和现实意义。     

大黄抑制食源性致病菌的活性成分研究

研究了大黄对大肠杆菌、沙门氏菌和金黄色葡萄球菌的抑菌效果,初筛的结果显示大黄醇提物对这3种菌均有较强的抑菌作用,并发现其中抑菌作用最强的物质分布在大黄氯仿段萃取物中。利用UPLC-MS-MS分析大黄氯仿段萃取物,发现大黄氯仿段萃取物的主要成分分别是大黄素、芦荟大黄素、大黄酸、大黄素甲醚和大黄酚。在这五种化合物中,大黄酸对大肠杆菌和沙门氏菌具有最强的抑菌效果,对大肠杆菌和沙门氏菌的最小抑菌浓度分别为125μg/m L和250μg/m L,最小杀菌浓度分别为250μg/m L和500μg/m L,大黄素对于金黄色葡萄球菌具有较强的抑菌效果,最小抑菌浓度和最小杀菌浓度分别为62.5μg/m L和125μg/m L。      

实时荧光聚合酶链式反应技术与国标方法检测致病菌的应用与研究

目的比较实时荧光聚合酶链式反应技术(real-time polymerase chain reaction,RT-PCR)与国标方法在食品致病菌检测中的异同。方法应用RT-PCR法及国标GB 4789系列对采集的60件畜产品、禽产品、水产品和乳及乳制品中的沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特氏菌以及副溶血性弧菌同时进行检测。结果除了金黄色葡萄球菌检测结果均为阴性外,其他3种致病菌2种方法都有检出,但RT-PCR方法的阳性率均高于国标方法。对于沙门氏菌,国标方法阳性率为1.67%,RT-PCR方法阳性率3.33%;单核细胞增生李斯特氏菌国标方法阳性率为7.50%,RT-PCR方法阳性率为15.00%;副溶血性弧菌国标方法阳性率为8.33%,RT-PCR方法阳性率为11.67%。结论在本实验条件下,RT-PCR技术的阳性检测结果多于国标GB4789系列,并且结果可完全覆盖国标GB4789。各企业实验室甚至政府主导的食品风险监测项目可根据产品特点,合理应用RT-PCR技术以减少人员工作量,方便产品放行并提高工作效率。     

Biofilm Formation of O157 and Non‐O157 Shiga Toxin‐Producing Escherichia coli and Multidrug‐Resistant and Susceptible Salmonella Typhimurium and Newport and Their Inactivation by Sanitizers

This study compared biofilm formation by 7 serogroups of pathogenic Escherichia coli and 2 or 3 phenotypes of Salmonella (susceptible, multidrug‐resistant [MDR], and/or multidrug resistant with ampC gene [MDR‐AmpC]). One‐week mature biofilms were also exposed to water, quaternary ammonium compound‐based (QAC), and acid‐based (AB) sanitizers. Seven groups (strain mixture) of above‐mentioned pathogens were separately spot‐inoculated onto stainless steel coupons surfaces for target inoculation of 2 log CFU/cm², then stored statically, partially submerged in 10% nonsterilized meat homogenate at 4, 15, and 25 °C. Biofilm cells were enumerated on days 0, 1, 4, and 7 following submersion in 30 mL for 1 min in water, QAC, and AB. Counts on inoculation day ranged from 1.6 ± 0.4 to 2.4 ± 0.6 log CFU/cm² and changed to 1.2 ± 0.8 to 1.9 ± 0.8 on day 7 at 4 °C with no appreciable difference among the 7 pathogen groups. After treatment with QAC and AB on day 7, counts were reduced (P < 0.05) to less than 0.7 ± 0.6 and 1.2 ± 0.5, respectively, with similar trends among pathogens. Biofilm formation at higher temperatures was more enhanced; E. coli O157:H7, as an example, increased (P < 0.05) from 1.4 ± 0.6 and 2.0 ± 0.3 on day 0 to 4.8 ± 0.6 and 6.5 ± 0.2 on day 7 at 15 and 25 °C, respectively. As compared to 4 °C, after sanitation, more survivors were observed for 15 and 25 °C treatments with no appreciable differences among pathogens. Overall, we observed similar patterns of growth and susceptibility to QAC and AB sanitizers of the 7 tested pathogen groups with enhanced biofilm formation capability and higher numbers of treatment survivors at higher temperatures.     

Survival and Growth of Vibrio cholerae O1, Salmonella typhi, and Escherichia coli O157:H7 in Alfalfa Sprouts

The survival and growth of Vibrio cholerae O1, Salmonella typhi , and Escherichia coli O157:H7 during germination and sprouting of disinfected alfalfa seeds and alfalfa sprouts was determined. All pathogens showed ability to grow during germination and sprouting, reaching counts of approximately 6.0 log₁₀ CFU/g after 24 h. No growth was observed for any pathogen when the sprouts were inoculated after 24 h of seed germination. At this time, the background microflora was abundant. Numbers of pathogens inoculated on alfalfa sprouts decreased less than 1 log10 CFU/g over 15 d of refrigeration. Alfalfa sprouts can be an important factor contributing to the endemicity for typhoid fever and cholera in México.     

Inhibition of Three Pathogens on Bologna and Summer Sausage Using Antimicrobial Edible Films

Whey protein isolate (WPI) films (pH 5.2) containing 0.5 to 1.0% p‐aminobenzoic acid (PABA) and/or sorbic acid (SA) were assessed for antimicrobial and mechanical properties while in contact with sliced bologna and summer sausage that were inoculated with Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica subsp. enterica serovar Typhimurium DT104. WPI films containing SA or PABA decreased L. monocytogenes, E. coli, and S. Typhimurium populations by 3.4 to 4.1,3.1 to 3.6, and 3.1 to 4.1 logs, respectively, on both products after 21 d at 4 °C. Background flora was inhibited compared with controls. Film tensile strength decreased while % elongation remained unchanged following 72 h of product contact.     

Thermal inactivation of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in breaded pork patties

ABSTRACT:  Decimal reduction times ( D -values) and thermal resistance constants ( z -values) for 3 foodborne pathogenic bacteria in formulated ready-to-eat breaded pork patties were determined with thermal inactivation studies. Meat samples, inoculated with Escherichia coli O157:H7, Salmonella , and Listeria monocytogenes cultures or uninoculated controls, were packaged in sterile bags, immersed in circulated water bath, and held at 55, 57.5, 60, 62.5, 65, 67.5, and 70 °C for different durations of time. The D - and z -values were determined by using a linear regression model. Average calculated D -values for E. coli O157:H7, Salmonella , and L . monocytogenes at a temperature range of 55 to 70 °C were 32.11 to 0.08 min, 69.48 to 0.29 min, and 150.46 to 0.43 min, respectively. Calculated z -values for E. coli O157:H7, Salmonella , and L. monocytogenes were 5.4, 6.2, and 5.9 °C, respectively. The results of this study will be useful to food processors to validate thermal lethality of the studied foodborne pathogens in ready-to-eat breaded pork patties.     

Emerging chemical and physical disinfection technologies of fruits and vegetables: a comprehensive review

Abstract

With a growing demand for safe, nutritious, and fresh-like produce, a number of disinfection technologies have been developed. This review comprehensively examines the working principles and applications of several emerging disinfection technologies. The chemical treatments, including chlorine dioxide, ozone, electrolyzed water, essential oils, high-pressure carbon dioxide, and organic acids, have been improved as alternatives to traditional disinfection methods to meet current safety standards. Non-thermal physical treatments, such as UV-light, pulsed light, ionizing radiation, high hydrostatic pressure, cold plasma, and high-intensity ultrasound, have shown significant advantages in improving microbial safety and maintaining the desirable quality of produce. However, using these disinfection technologies alone may not meet the requirement of food safety and high product quality. Several hurdle technologies have been developed, which achieved synergistic effects to maximize lethality against microorganisms and minimize deterioration of produce quality. The review also identifies further research opportunities for the cost-effective commercialization of these technologies.     

基于叠氮丙啶-实时荧光聚合酶链式反应PMA-qPCR 结合与高温裂解法高通量测序技术检测鉴定检测冷链食品中5多种 病原菌并构建金黄色葡萄球菌分子进化树完成病原菌分子进化树构建与溯源分析

目的 将PMA-qPCR、高通量测序以及分子进化树构建技术相结合,应用于冷链食品中多种病原菌的快速检测、种属鉴定、亲缘关系确定与溯源分析工作。 建立冷链食品中多种病原菌的快速检测技术并完成金黄色葡萄球菌的溯源分子进化分析工作。方法 以冷链食品中的沙门氏菌、金黄色葡萄球菌、蜡样芽胞杆菌、副溶血性弧菌、单核细胞增生李斯特氏菌等等5种病原菌5种病原菌为作为研究对象, 应用PMA-qPCRPMA-qPCR技术结合高温裂解法作为冷链食品中病原菌检测初筛手段,应用微生物培养法以及生化鉴定仪器法进行方法比对验证qPCR结果,运用高通量测序以及分子进化树构建作为冷链食品中所分离病原菌的种属地位确证方法。研发病原菌活菌快速检测方法并开展冷链食品中病原菌的抽样调查。在建立稳定高效的检测方法后,为了获得冷链食品中病原菌污染水平、分布数据以及污染来源,研究开展冷链食品中病原菌的抽样调查检测工作。在检测出金黄色葡萄球菌等并分离病原菌后,进一步通过多位点采样以及16S rRNA测序构建了葡萄球菌属、弧菌属以及李斯特菌属分子生物进化树,完成菌株种属鉴定采用构建分子进化树的方法以及完成金黄色葡萄球菌等病原菌的溯源分析工作。结果 本研究应用建立的PMA-qPCR成功扩增了冷链食品中生活状态病原菌的特征性核酸片段,排除了死亡细菌以及阴性对照菌的干扰感染,病原菌最低检测浓度可达到1×103 CFU/mL,一次反应可检测42份试样,可以在16小时内完成检测工作。在冷链食品中病原菌抽样检测调查中,本研究共从随机采集的751份冷链食品中检出6162株病原菌,病原菌总体检出率为8.13% (6162/751)。通过后续的16S rRNA测序以及葡萄球菌属、弧菌属以及李斯特菌属分子进化树的构建成功溯源了金黄色葡萄球菌的污染来源并完成病原菌种属定位。通过分子进化树的构建成功溯源了金黄色葡萄菌的污染来源并完成菌种种属定位。本研究还注意到经微生物检验一株疑似单核细胞增生李斯特氏菌阳性菌株,该菌株后续通过测序、序列比对以及分子进化树构建确定为英诺克李斯特菌,这充分说明高通量测序的重要性以及方法验证的必要性。结论 研究在国内率先将PMA-qPCR、高通量测序与分子进化树构建技术相结合,应用于应用本研究方法可以快速有效检测5种冷链食品中的冷链食品中多种病原菌检测分析与种属鉴定,并完成金黄色葡萄球菌的溯源分析,方法特异性好、灵敏度高、检测通量高,研究方法与成果可以为冷链食品及相关食品中病原菌的精确检测与溯源分析提供全新的思路与方法。     

GC-MS based metabolomics for rapid simultaneous detection of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Muenchen, and Salmonella Hartford in ground beef and chicken

A metabolomic-based method for rapid detection of Escherichia coli O157:H7, Salmonella Hartford, Salmonella Typhimurium, and Salmonella Muenchen in nonselective media was developed. All pathogenic bacteria were grown in tryptic soy broth (TSB) at 37 °C followed by metabolite quantification at 2-h intervals for 24 h. Results were compared with the metabolite profiles similarly obtained with E. coli K12, Pseudomonas aeruginosa, Staphylococcus aureus, Saccharomyces cereviseae, and Aspergillus oryzae grown individually or as a cocktail under the same conditions. Principal component analysis (PCAS) discriminated pathogenic microorganisms grown in TSB. Metabolites responsible of PCAS classification were dextrose, cadaverine, the aminoacids L-histidine, glycine, and L-tyrosine, as well as the volatiles 1-octanol, 1-propanol, 1butanol, 2-ethyl-1-hexanol, and 2,5-dimethyl-pyrazine. Partial least square (PLS) models based on the overall metabolite profile of each bacteria were able to detect the presence of Escherichia coli O157:H7 and Salmonella spp. at levels of approximately 7 ± 2 CFU/25 g of ground beef and chicken within 18 h.     

Reduction of E. coli, Salmonella typhimurium, coliforms, aerobic bacteria, and improvement of ground beef color using trisodium phosphate or cetylpyridinium chloride before grinding

The impact of 10% trisodium phosphate (TSP) or 0.5% cetylpyridinium chloride (CPC) applied to beef trimmings either aerobically or under vacuum before grinding on Salmonella typhimurium (ST), Escherichia coli (EC), coliform (CO), aerobic plate count (APC), color and sensory attributes of ground beef through display was studied. For this, beef trimmings were inoculated with ST and EC then treated with either TSP or CPC in vacuum or aerobic conditions. Trimmings were ground, packaged, displayed under simulated retail conditions and sampled on days 0, 1, 2, 3 and 7 for microbial, instrumental color, and sensory color and odor characteristics. Aerobic and vacuum antimicrobial application methods were equally effective (P>0.05) for reducing microorganisms in ground beef. Trisodium phosphate and CPC reduced (P<0.05) all bacterial types monitored. In addition, TSP and CPC improved (P>0.05) ground beef redness (a*), oxymyoglobin stability (630 nm/580 nm) and sensory overall color throughout display without adversely affecting odor characteristics.     

Activity of ε–Polylysine Against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes

ABSTRACT: ε–polylysine is a homopolymer of L-lysine, an essential amino acid, with a reportedly wide antimicrobial spectrum. This study evaluated the antimicrobial activity of ε–polylysine, as compared with known preservatives and organic acids, against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes , in culture broth. The compounds tested included ε–polylysine (0.0025% to 0.05%), sodium diacetate (0.25%), sodium lactate (3.0%), lactic acid (0.1%), and acetic acid (0.1%), alone, as well as in combination with ε– polylysine (0.0025% to 0.03%); all treatments were evaluated in tryptic soy broth supplemented with 0.6% yeast extract. Treatments were inoculated (approximately 2 log colony-forming units [CFU]/mL) with 5-strain ( E. coli O157:H7, S. Typhimurium) or 10-strain ( L. monocytogenes ) mixtures of the pathogens. Survival/growth of the inoculated bacteria was periodically monitored during incubation at 4 °C (30 d) and 24 °C (48 h). Bactericidal effects of ε–polylysine were obtained against E. coli O157:H7 and S. Typhimurium at 4 °C. At the same temperature (4 °C), ε–polylysine alone or in combination with other compounds tested inhibited growth or was bactericidal against L. monocytogenes. All 3 pathogens were inhibited by ε–polylysine at 24 °C; however, L. monocytogenes was the most sensitive and S. Typhimurium the most resistant. The antimicrobial activity of ε–polylysine against E. coli O157:H7 and S. Typhimurium was enhanced ( P < 0.05) when tested in combination with sodium diacetate or acetic acid. Combination treatments with sodium lactate resulted in loss of ε–polylysine activity by the end of the incubation period. Overall, under the conditions of this study, ε–polylysine exhibited antimicrobial effects against the 3 pathogens tested.     

Impact of irradiation on the safety and quality of poultry and meat products: a review

For more than 100 years research on food irradiation has demonstrated that radiation will make food safer and improve the shelf life of irradiated foods. Using the current food safety technology, we may have reached the point of diminishing returns even though recent figures from the CDC show a significant drop in the number of foodborne illnesses. However, too many people continue to get sick and die from eating contaminated food. New and under utilized technologies such as food irradiation need to be re-examined to achieve new levels of safety for the food supply. Effects of irradiation on the safety and quality of meat and poultry are discussed. Irradiation control of the principle microbial pathogens including viruses, the differences among at-risk sub-populations, factors affecting the diminished rate of improvement in food safety and published D values for irradiating raw meat and poultry are presented. Currently permitted levels of irradiation are probably not sufficient to control pathogenic viruses. Typical gram-negative spoilage organisms are very sensitive to irradiation. Their destruction leads to a significant increase in the acceptable shelf life. In addition, the destruction of these normal spoilage organisms did not provide a competitive growth advantage for irradiation injured food pathogens. Another of the main focuses of this review is a detailed compilation of the effects of most of the food additives that have been proposed to minimize the negative quality effect of irradiation. Most of the antimicrobials and antioxidants used singly or in combination produced an increased lethality of irradiation and a decrease in oxidation by-products. Combinations of dosage, temperature, dietary and direct additives, storage temperature and packaging atmosphere can produce meats that the average consumer will find indistinguishable from non-irradiated meats. A discussion of the production of unique radiological by-products is also included.     

Synergy Between Irradiation and Chlorination in Killing of Salmonella, Escherichia Coli O157:H7, and Listeria Monocytogenes

ABSTRACT:  The objectives of the studies were to determine if radiation and chlorine acted in a simple additive process or if a nonadditive increase in inactivation occurred when chlorination followed γ-irradiation both in vitro and in situ. Separate studies evaluated the effects of γ-irradiation (0.2 kGy at 20 °C), chlorination (0.5 ppm for 10 min), or irradiation followed by chlorination of Salmonella , Escherichia coli O157:H7 Ent C9490, or Listeria monocytogenes cells suspended on membrane filters. Cocktails of Salmonella enterica serovars: Anatum, Infantis, Newport, and Stanley or of L. monocytogenes isolates: ATCC 7644, 15313, 43256, and 49594 (Scott A) were used. In each case, greater inactivation was observed from irradiation followed by chlorination than was predicted from the sum of the inactivation of the 2 treatments when applied separately. Inactivation of Salmonella cells was determined also with the pathogens adsorbed onto alfalfa seeds. Analysis of results of 3 sets of experiments with Salmonella adsorbed onto alfalfa seeds also led to the conclusion that the combination treatments were synergistic and produced a greater inactivation than were expected from the sums of the treatments although experiments with larger number of seeds gave less evidence of synergy. The effectiveness of both interventions against Salmonella was significantly reduced when the pathogen was on alfalfa seeds.     

Outbreaks and factors influencing microbiological contamination of fresh produce

Fresh fruits and vegetables are nutritionally well‐recognised as healthy components in diets. The microbiological foodborne outbreaks associated with the consumption of fresh produce have been increasing. Salmonella spp., Escherichia coli O157:H7, Staphylococcus aureus, Campylobacter spp. and Listeria monocytogenes are the most common pathogens that contaminate fresh produce. This review discusses recent foodborne outbreaks linked to fresh produce, factors that affect microbiological contamination and measures that could be adopted to reduce the foodborne illnesses. © 2016 Society of Chemical Industry     

大肠杆菌O157:H7耐酸性及酸胁迫下菌体形态变化的初步研究

目的 对大肠杆菌O157:H7 耐酸性进行初步探讨。方法 通过稀释涂布平板计数法观察大肠杆菌O157:H7 在不同的pH 生长条件下的存活能力和利用电镜扫描法观察其在酸胁迫下菌体形态的变化。结果 当pH 为5.0 到7.0 之间时, 大肠杆菌O157:H7 生长状况良好, 当pH 小于4.0 时, 其生长受到抑制, 特别是pH 降到2.0 以下时, 大肠杆菌O157:H7 完全不能生长, 由此可以说明大肠杆菌O157:H7 耐酸性能力较强, 可以抵御酸性环境的影响。扫描电镜图显示大肠杆菌O157:H7 在不同pH 的环境条件下其菌体形态会作出相应的变化,随着培养基的酸性增强, 其细胞形态从长杆菌体形态变成了短杆状或钝圆形态。结论 本研究测定不同酸性条件下大肠杆菌O157:H7 的生存和繁殖能力的研究, 有利于帮助人们进一步了解其耐酸性, 从而为制定防治方案和措施提供参考。     

Reduction of Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7 During Processing of Country-cured Hams

ABSTRACT: The country-cured ham process, including curing, equalization, cold-smoked or nonsmoked, and aging up to 6 mo, was validated and showed its effectiveness in achieving a 6-log reduction of Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7. The viable counts of L. monocytogenes populations decreased to below detection levels after 206 d, Salmonella populations required 122 d, and E. coli O157:H7 required 66 d. However, L. monocytogenes -inoculated hams were positive and Salmonella spp-inoculated and E. coli O157:H7-inoculated hams were negative following enrichment procedures at the end of the aging process. Therefore, the survival of L. monocytogenes on country-cured ham represents a risk.     

Cold plasma reduction of Salmonella and Escherichia coli O157:H7 on almonds using ambient pressure gases

Contamination of raw nuts, including almonds, is a food safety concern. Cold plasma is a novel antimicrobial intervention that can eliminate foodborne pathogens. The objective of this work was to evaluate the efficacy of rapid cold plasma treatments in eliminating Salmonella and Escherichia coli O157:H7 from dry almonds. Three isolates of Salmonella (S. Anatum F4317, S. Stanley H0558, and S. Enteritidis PT30) and 3 isolates of E. coli O157:H7 (C9490, ATCC 35150, and ATCC 43894) were separately grown and spot-inoculated (10 μL) onto whole almonds and allowed to dry for 10 min. Inoculated almonds were treated with a cold plasma jet, with treatment variables evaluated in a factorial design for each isolate: time, distance, and feed gas. Treatment time was 0 s (control), 10 s, or 20 s. Distance from the emitter was 2, 4, or 6 cm. Feed gas was dry air or nitrogen. After treatment, the almonds were sampled using swabs. Survivors were enumerated on tryptic soy agar (TSA) plates. Cold plasma significantly reduced both pathogens on almonds. The greatest reduction observed was 1.34 log cfu/mL reduction of E. coli O157:H7 C9490 after 20 s treatment at 6 cm spacing. The interaction of treatment time with distance from plasma emitter head was complex, and isolate-dependent. Longer duration of treatment did not always result in enhanced reductions. In general, nitrogen as a feed gas resulted in a reduced antimicrobial efficacy compared to dry air. These results indicate that short pulses of atmospheric pressure cold plasma can significantly reduce Salmonella and E. coli O157:H7 on almonds.