Gram-negative Foodborne Pathogens Research Articles

P.2.025 - Pharmacological stimulation of 5-HT7r rescues sensorymotor gating and memory deficits in a mouse model of Rett syndrome

Cronobacter species are Gram-negative opportunistic foodborne pathogens that may cause enterocolitis, bacteremia and meningitis in neonates and premature neonates. Lipopolysaccharide (LPS) serves as the major component of the outer membrane of cell, is a potential virulence factor for Cronobacter.Given the potential importance of this molecule in infection and virulence, SDS-PAGE of LPS, MS and TLC characterization of phospholipids and phenotypic characterization of Cronobacter spp. strains were carried out.The phospholipids from Cronobacter yielded four major peaks at m/z 719.9, 733.9, 747.9 and 773.9 in the spectrum. All Cronobacter showed O-antigen bands except C. muytjensii ATCC 51329. When Cronobacter defect O-antigen, the outer membrane permeability and cell surface hydrophobicities are increased. All Cronobacter are able to grow under pH 5.0 condition and able to grow under 6% NaCl concentration. C. dublinensis DSM 18705 has a higher infection rate to Caco-2 cells than other Cronobacter.Invasion of pathogens into a host cell is critical component to an infectious case. And C. dublinensis DSM 18705 has a higher infection rate to Caco-2 cells than other Cronobacter.

Novel bacteriocin produced by Lactobacillus alimentarius FM-MM4 from a traditional Chinese fermented meat Nanx Wudl: Purification, identification and antimicrobial characteristics

The present study aims to purify and characterize a novel bacteriocin (designated as lactocin MM4) produced by Lactobacillus alimentarius FM-MM4, which was isolated from Nanx Wudl, a traditional Chinese fermented meat. Lactocin MM4 was extracted by ethyl acetate and purified through cation exchange chromatography and semi-preparative high-performance liquid chromatography (RP-HPLC). As revealed by LC-MS/MS, lactocin MM4 has a molecular mass of 1104.58 Da with the N-terminal sequence QGVGPLGQGHR, demonstrating low homology with previously reported class II bacteriocins. Lactocin MM4 exhibited broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative food-borne pathogens, as well as several yeasts. It was considerably thermo-stable with 84.7% residual antimicrobial activity after exposure to121 °C for 15 min, and it was highly stable at acidic pH (2–5). Proteases, such as trypsin, protease K, papain and pepsin, could completely inactivate the bacteriocin, while other enzymes, such as lipase and amylase, had no effect on its antimicrobial activity. These results indicate the potential of lactocin MM4 for application in food preservation.

Mungo bean sprout microbiome and changes associated with culture based enrichment protocols used in detection of Gram-negative foodborne pathogens.

BackgroundFresh sprouted seeds have been associated with a number of large outbreaks caused by Salmonella and Shiga toxin-producing E. coli. However, the high number of commensal bacteria found on sprouted seeds hampers the detection of these pathogens. Knowledge about the composition of the sprout microbiome is limited. In this study, the microbiome of mungo bean sprouts and the impact of buffered peptone water (BPW) and Enterobacteriaceae enrichment broth (EE-broth)-based enrichment protocols on this microbiome were investigated.ResultsAssessments based on aerobic mesophilic colony counts showed similar increases in mungo bean sprout background flora levels independent of the enrichment protocol used. 16S rRNA sequencing revealed a mungo bean sprout microbiome dominated by Proteobacteria and Bacteroidetes. EE-broth enrichment of such samples preserved and increased Proteobacteria dominance while reducing Bacteroidetes and Firmicutes relative abundances. BPW enrichment, however, increased Firmicutes relative abundance while decreasing Proteobacteria and Bacteroidetes levels. Both enrichments also lead to various genus level changes within the Protobacteria and Firmicutes phyla.ConclusionsNew insights into the microbiome associated with mungo bean sprout and how it is influenced through BPW and EE-broth-based enrichment strategies used for detecting Gram-negative pathogens were generated. BPW enrichment leads to Firmicutes and Proteobacteria dominance, whereas EE-broth enrichment preserves Proteobacteria dominance in the mungo bean sprout samples. By increasing the relative abundance of Firmicutes, BPW also increases the abundance of Gram-positive organisms including some that might inhibit recovery of Gram-negative pathogens. The use of EE-broth, although preserving and increasing the dominance of Proteobacteria, can also hamper the detection of lowly abundant Gram-negative target pathogens due to outgrowth of such organisms by the highly abundant non-target Proteobacteria genera comprising the mungo bean sprout associated background flora.

전해수 생성온도에 따른 그람양성균과 그람음성균의 살균 효과

본 연구에서는 다양한 온도의 물로 제조한 전해수를 이용하여 그람양성균과 그람음성균에 대한 살균력을 확인하였다. 전해수의 물성은 물의 온도가 높아질수록 pH와 유효 잔류염소 농도 값이 높아졌으나, 산화환원전위 값은 감소하였다. 4, 22, 40°C의 물을 이용하여 생성한 전해수에 Escherichia coli O157:H7을 15분 동안 처리하였을 때 각각 1.18, 4.47, 5.46 log CFU/mL 감소하였으며, Staphylococcus aureus는 각각 0.72, 4.90, 5.54 log CFU/mL 감소하였다. 이를 통해 전해수를 생성하는 물의 온도가 증가할수록 살균 효과가 높아지는 것을 알 수 있다. 4, 40°C의 물을 이용하여 생성한 전해수에 균주를 처리한 경우, Listeria monocytogenes의 D 값(일정한 온도에서 90 %의 미생물이 사멸하는 데 걸리는 시간)이 각각 6.60, 1.57분으로 나타나 가장 낮았으며, 22°C 전해수에서는 Salmonella Enteritidis의 D 값이 2.92분으로 가장 낮은 값을 나타내었다. 그람양성균과 그람음성균을 비교하였을 때 모든 온도에서 D 값에 대한 유의적인 차이는 나타나지 않았다(P0.05). 본 연구 결과는 전해수를 제조할 경우 높은 온도의 물을 이용하여 제조하는 것이 미생물 살균 효과가 높다는 것을 보여주고 있다. 따라서 전해수를 생성할 때 물의 온도를 고려하여 높은 살균력을 나타내는 전해수를 생성할 수 있으며, 이는 물의 온도 외에도 물의 경도나 물의 종류 등을 고려하여 최적의 살균 전해수를 제조함으로써 식품산업에 적용하기 위한 자료로 활용될 것으로 기대된다.

Genomic characterization of Salmonella bacteriophages isolated from India.

Salmonella are a medically important Gram-negative foodborne pathogen. Genomic diversity of Salmonella is increasingly studied but at the same time, we have limited knowledge of Salmonella phage diversity. In this study, we have isolated Salmonella phages from sewage and river water. Genomic characterization of 12 Salmonella phages was carried out using next-generation sequencing platform. Newly sequenced phages were classified based on amino acid sequence phylogenetic analysis. In newly sequenced phages, several virulence genes, DNA metabolism genes, tRNA genes, antibiotic resistance genes and genes not having known role in the life cycle of phages were identified. Annotations of newly sequenced phage genome showed the presence of polymyxin-b resistance gene and penicillin binding protein. Annotation identified number of genes which are involved in DNA metabolism. Results suggest that most of the phages having G + C content different than their host possess DNA metabolism genes. The presence of tRNAs in the genome of Salmonella_phage38-India was identified; however, we did not observe any correlation between tRNA genes and overall codon usage in the phage genome. It is suggested that the phage-encoded tRNAs may increase fitness of phages. In summary, we isolated novel Salmonella phages, determined full genome sequences and provided phylogenetic analysis-based classification.

Lactobacillus plantarum LB95 impairs the virulence potential of Gram-positive and Gram-negative food-borne pathogens in HT-29 and Vero cell cultures.

Listeria monocytogenes, Salmonella enterica and verocytotoxigenic Escherichia coli (VTEC) are amongst the most important agents responsible for food outbreaks occurring worldwide. In this work, two Lactobacillus spp. strains (LABs), Lactobacillus plantarum (LB95) and Lactobacillus paraplantarum (LB13), previously isolated from spontaneously fermenting olive brines, and two reference probiotic strains, Lactobacillus casei Shirota and Lactobacillus rhamnosus GG, were investigated for their ability to attenuate the virulence of the aforementioned pathogens using animal cell culture assays. In competitive exclusion assays, the relative percentages of adhesion and invasion of S. enterica subsp. enterica serovar Enteritidis were significantly reduced when the human HT-29 cell line was previously exposed to LB95. The relative percentage of invasion by Listeria monocytogenes was significantly reduced when HT-29 cells were previously exposed to LB95. In the cytotoxicity assays, the cell-free supernatant of the co-culture (CFSC)of VTEC with LB95 accounted for the lowest value obtained amongst the co-cultures of VTEC with LABs, and was significantly lower than the value obtained with the co-culture of VTEC with the two probiotic reference strains. The cytotoxicity of CFSC of VTEC with both LB95 and LB13 exhibited values not significantly different from the cell-free supernatant of the nonpathogenic E. coli B strain. Our results suggested that LB95 may be able to attenuate the virulence of Gram-positive and Gram-negative food-borne pathogens; together with other reported features of these strains, our data reveal their possible use in probiotic foods due to their interesting potential in preventing enteric infections in humans.

Synergistic Role of Reuterin in Combination with Nisin against Food Borne Pathogens in Milk at Refrigerated Condition

The effect of reuterin alone and in combination with nisin on different Gram-positive and Gram-negative foodbornepathogens in milk as a model system was investigated. Reuterin (16 AU/mL) exhibited bactericidal activity against Listeria monocytogenes, whereas its bactericidal activity against Staphylococcus aureus at refrigeration temperature was more pronounced. Higher bactericidal activity was also detected against Escherichia coli O157: H7, Pseudomanasaeruginosa, Salmonella typhi. The synergism of reuterin and nisin (100 IU/mL) against L. monocytogenes and S. aureus was also found at refrigeration temperature, where the pathogens were completely inactivated. The combination of reuterin with nisin, however, did not enhance the antimicrobial effect of reuterin against Gram-negative pathogens. Refrigerated milk and milk products treated with both natural antimicrobials would mean a feasible system to control pathogenic contaminants.

Complete Genome Sequence of Campylobacter jejuni RM1285, a Rod-Shaped Morphological Variant

Campylobacter jejuni is a spiral shaped Gram-negative food-borne bacterial pathogen of humans found on poultry products. Strain RM1285 is a rod-shaped variant of this species. The genome of RM1285 was determined to be 1,635,803 bp, with a G+C content of 30.5%.

Characterisation of the antibacterial properties of a bacterial derived peptidoglycan hydrolase (LysCs4), active against C. sakazakii and other Gram-negative food-related pathogens

Illness caused by the consumption of contaminated food products continues to represent one of the main challenges facing food manufacturers worldwide. Even with current intervention technologies and increased hygiene measures, foodborne illness remains a significant threat to public health. This coupled with the increasing emergence of multidrug resistant pathogens has increased the need for the development of novel technologies for pathogen control. Bacterial derived peptidoglycan hydrolases represent a vast and highly diverse group of enzymes with potential for biocontrol of a range of Gram-positive and Gram-negative foodborne pathogens. In this study, we describe the identification, cloning, expression and purification of a peptidoglycan hydrolase (LysCs4) derived from Cronobacter sakazakii for biocontrol of the aforementioned infant formula pathogen itself. In silico analysis of LysCs4 revealed the gene to display greatest sequence similarity to a putative lysozyme encoded by the lytic Cronobacter phage ES2. Conserved domain analysis of LysCs4 revealed the presence of a single catalytic domain predicted to display O-Glycosyl hydrolase activity and to be a member of the GH24 family. The ability of this enzyme to hydrolyse the peptidoglycan of 25 Gram-negative strains, across 4 different genera, highlights its potential as a novel candidate for biocontrol of C. sakazakii and other Gram-negative food related pathogens.

Electrochemical immunosensor for Enterobacter sakazakii detection based on electrochemically reduced graphene oxide–gold nanoparticle/ionic liquid modified electrode

Abstract Enterobacter sakazakii (E. sakazakii) is a gram-negative food-borne pathogen, which can cause serious disease. Rapid screening plays an important role in the prevention and identification of E. sakazakii to cause food poisoning. In this study, a facile and sensitive approach for E. sakazakii detection has been presented. This novel electrochemical immunosensor based on electrochemically reduced graphene oxide–gold nanoparticles (ERGO–AuNP)/1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM]PF6) was developed for the detection of E. sakazakii. Firstly, a screen-printed carbon electrode (SPCE) was modified by ERGO–AuNP. Then, the mixture of [BMIM]PF6 and HRP-anti-E. sakazakii was assembled onto the modified electrode to improve the sensitivity and selectivity of the immunosensor. The surface morphologies of modified electrodes were characterized by means of AFM and SEM. The electrochemical performance of the immunosensor was evaluated by EIS and CV. The electrochemical immunosensor for E. sakazakii exhibited a linear range from 103 to 109 CFU/mL with a detection limit of 1.19 × 102 CFU/mL (S/N = 3). The good performance of the immunosensor is attributed to the good biocompatibility of ERGO–AuNP films. The proposed immunosensor also exhibits excellent long-term storage stability. In addition, this electrochemical immunosensor based on ERGO–AuNP modified SPCE could also provide many potential applications for the rapid detection of different bacteria.

Evaluation of different buffered peptone water (BPW) based enrichment broths for detection of Gram-negative foodborne pathogens from various food matrices

This study evaluated the effects of changing the composition of the pre-enrichment medium buffered peptone water (BPW) on the growth of stressed and unstressed Gram-negative foodborne pathogens in a one-broth enrichment strategy. BPW supplemented with an available iron source and sodium pyruvate, along with low levels of 8-hydroxyquinoline and sodium deoxycholate (BPW-S) improved the recovery of desiccated Cronobacter spp. from powdered infant formula. Growth of Salmonella and STEC was comparable in all BPW variants tested for different food matrices. In products with high levels of Gram-negative background flora (e.g. sprouts), the target organisms could not be reliably detected by PCR in any of the BPW variants tested unless the initial level exceeded 103cfu/10g of sprouts.Based on these results we suggest BPW-S for a one-broth enrichment strategy of stressed Gram-negative foodborne pathogens from dry products. However, a one-broth enrichment strategy based on BPW variants tested in this evaluation is not recommended for produce with a high level of Gram-negative background flora due to very high detection limits.

Regulation of oxidative stress resistance in Campylobacter jejuni, a microaerophilic foodborne pathogen.

Campylobacter jejuni is one of the leading bacterial causes of human gastroenteritis. Due to the increasing rates of human campylobacteriosis, C. jejuni is considered as a serious public health concern worldwide. C. jejuni is a microaerophilic, fastidious bacterium. C. jejuni must overcome a wide range of stress conditions during foodborne transmission to humans, such as food preservation and processing conditions, and even in infection of the gastrointestinal tracts of humans. Particularly, this microaerophilic foodborne pathogen must survive in the atmospheric conditions prior to the initiation of infection. C. jejuni possesses unique regulatory mechanisms for oxidative stress resistance. Lacking OxyR and SoxRS that are highly conserved in other Gram-negative foodborne pathogens, C. jejuni modulates the expression of genes involved in oxidative stress resistance mainly via the peroxide resistance regulator and Campylobacter oxidative stress regulator. Based on recent findings of ours and others, in this review, we described how C. jejuni regulates the expression of oxidative stress defense.

Ferrocenyl analogues of bisacodyl: Synthesis and antimicrobial activity

The antibacterial activity of bisacodyl, a drug used in therapeutic as laxative is described herein for the first time. Ten original ferrocenyl analogues have been synthesized via an efficient synthetic procedure using the McMurry coupling reaction. The antibacterial activity was investigated against Gram-positive and Gram-negative foodborne pathogens including Listeria monocytogenes, Escherichia coli, Enterococcus faecalis, Salmonella enterica, Micrococcus luteus and Staphylococcus aureus. The results showed that most of these compounds exhibit an excellent antimicrobial activity, and the bisacodyl analogues seemed to be more bactericides than bacteriostatic.

Antimicrobial activity of amurca (olive oil lees) extract against selected foodborne pathogens

The antimicrobial activity of a methanolic extract of amurca (olive oil lees) was determined against both Gram-positive (L. monocytogenes and S. aureus) and Gram-negative (E. coli O157:H7 and S. enteritidis) foodborne pathogens at 10 °C or 37 °C using microdilution and disk diffusion methods, and its relative activity was compared to selected antibiotics. Minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of amurca extract ranged from 60 to 80 µl/ml at 37 °C after 24 h against all tested strains. At 10 °C, amurca was more inhibitory with MIC and MBC values of 40 and 60 µl/ml, respectively, after 7 d against tested strains. Amurca at 40 µl/ml reduced numbers of tested pathogens by 2.5 to 3.2 log10 CFU/ml at 10 °C after 7 d, but was not inhibitory at 37 °C after 24 h. Protein prepared from amurca was not antimicrobial. The relative antimicrobial activity (inhibition zone ratio) of 80 µl/ml amurca methanolic extract compared to chloramphenicol, erythromycin, gentamycin and tetracycline ranged from 0.36 to 1.0 against Gram-negative and from 0.45 to 2.0 against Gram-positive bacteria. In addition, amurca extract inhibited E. coli O157:H7 02-0628 and S. aureus 26127 which were resistant to tetracycline and chloramphenicol, respectively.

Two putatively novel bacteriocins active against Gram-negative food borne pathogens produced by Weissella hellenica BCC 7293

Weissella hellenica BCC 7239, isolated from Thai fermented pork sausage called Nham, produced two putatively novel bacteriocins, 7293A and 7293B. Both bacteriocins had broad antimicrobial spectra and exceptionally inhibited several important Gram-negative food-borne pathogens (Pseudomonas aeruginosa, Aeromonas hydrophila, Salmonella Typhimurium and Escherichia coli). The highest amount of bacteriocin was produced in MRS and APT media at 30 °C without agitation. Bacteriocin 7293A showed relatively higher antimicrobial activity than bacteriocin 7293B. However, pH and thermal stability of bacteriocin 7293A was lower. These bacteriocins were of proteinaceous nature, in which the complete inactivation of their antimicrobial activity after treatment by proteolytic enzymes, including trypsin, α-chymotrypsin, pepsin and protease K was observed, whilst lipase and α-amylase exhibited no effect. Antimicrobial activity of both peptides was also not inactivated by organic solvents (ethanol, isopropanol, acetone, acetonitrile) and surfactants (Tween 20, Tween 80 and Triton X-100). Bacteriocin 7293A and B exhibited bactericidal effect against both Gram-positive and Gram-negative indicators without cell-lysis. According to ESI/MS analysis, the molecular masses of bacteriocin 7293A and B were determined to be 6249.302 and 6489.716 Da, respectively. Because their molecular masses were not similar to those of other known bacteriocins, both bacteriocin 7293A and B could be novel bacteriocins. Thus, both novel bacteriocins hold promise for applications in the prevention or treatment of pathogenic infections as food and feed additives to replace antibiotics for enhancing the productivity and sustainability of food animals.

Antibiotic resistance versus antimicrobial substances production by gram-negative foodborne pathogens isolated from minas frescal cheese: heads or tails?

In this study, 15 Gram-negative isolates from Minas Frescal cheese sold in commercial establishments in Rio de Janeiro, Brazil, were able to produce antimicrobial substances (AMSs). Seven, four, two, one, and one isolates identified as Yersinia, Acinetobacter, Enterobacter, Escherichia, and Hafnia genera, respectively, were considered potentially pathogenic. All 15 AMS(+) isolates were resistant to at least 1 antibiotic; however, 7 strains presented resistance to at least 3 antibiotics from different classes, exhibiting multiresistance profiles. The strains were also subjected to plasmid profile analysis. All isolates presented different plasmid forms with most ranging in size from 1 to 10 kb. Activity against various pathogens associated with food was tested and all 15 AMS(+) showed the same activity spectrum, inhibiting all Escherichia coli and Salmonella strains that were tested. Although restricted, the action spectrum of AMS-producing strains is extremely relevant to the food industry because Gram-negative bacteria such as E. coli and Salmonella spp. are most often associated with foodborne illnesses. The findings of this study reveal that even AMS produced by pathogens can have potential applications against other foodborne pathogens.

Coinfection with an intestinal helminth impairs host innate immunity against Salmonella enterica serovar Typhimurium and exacerbates intestinal inflammation in mice.

Salmonella enterica serovar Typhimurium is a Gram-negative food-borne pathogen that is a major cause of acute gastroenteritis in humans. The ability of the host to control such bacterial pathogens may be influenced by host immune status and by concurrent infections. Helminth parasites are of particular interest in this context because of their ability to modulate host immune responses and because their geographic distribution coincides with those parts of the world where infectious gastroenteritis is most problematic. To test the hypothesis that helminth infection may negatively regulate host mucosal innate immunity against bacterial enteropathogens, a murine coinfection model was established by using the intestinal nematode Heligmosomoides polygyrus and S. Typhimurium. We found that mice coinfected with S. Typhimurium and H. polygyrus developed more severe intestinal inflammation than animals infected with S. Typhimurium alone. The enhanced susceptibility to Salmonella-induced intestinal injury in coinfected mice was found to be associated with diminished neutrophil recruitment to the site of bacterial infection that correlated with decreased expression of the chemoattractants CXCL2/macrophage inflammatory protein 2 (MIP-2) and CXCL1/keratinocyte-derived chemokine (KC), poor control of bacterial replication, and exacerbated intestinal inflammation. The mechanism of helminth-induced inhibition of MIP-2 and KC expression involved interleukin-10 (IL-10) and, to a lesser extent, IL-4 and IL-13. Ly6G antibody-mediated depletion of neutrophils reproduced the adverse effects of H. polygyrus on Salmonella infection. Our results suggest that impaired neutrophil recruitment is an important contributor to the enhanced severity of Salmonella enterocolitis associated with helminth coinfection.

Effect of copper salts and reducing agents on characteristics and antimicrobial activity of copper nanoparticles

Copper nanoparticles (CuNPs) were synthesized using different types of copper salts (copper acetate, copper chloride, and copper sulfate) and reducing agents (NaOH and ascorbic acid). The resulting solutions exhibited the maximum absorption peaks between 250–350nm, which was caused by the change in the surface Plasmon resonance of CuNPs and clearly indicated the formation of CuNPs. The results of FE-SEM exhibited that the CuNPs of various shapes and size, depended upon the type of copper salts and reducing agents used. The FTIR results demonstrated the involvement of bioactive functional groups as reducing and capping agents. XRD spectra confirmed the crystalline nature of CuNPs. Furthermore, all the CuNPs exhibited strong antimicrobial activity against both, Gram-positive (Listeria monocytogenes) and Gram-negative (Escherichia coli) food-borne pathogens.

Purification of novel bacteriocin produced by Lactobacillus coryniformis MXJ 32 for inhibiting bacterial foodborne pathogens including antibiotic-resistant microorganisms

Bacteriocins produced by LAB comprise a variety of antimicrobial proteins or peptides which can be used as food biopreservatives and/or possible antibiotic alternatives. Bacteriocin-producing Lactobacillus coryniformis MXJ 32, which was identified based on its 16S rRNA gene sequence, was isolated from a traditional fermented vegetable (Jiangshui Cai) of Xixiang county, Shaanxi province, China. Thereafter bacteriocin (designated as lactocin MXJ 32A) produced by L. coryniformis MXJ 32 was purified by ammonium sulfate precipitation, dialysis and ion exchange chromatography. The molecular mass of lactocin MXJ 32A was 3520 Da determined by LC-ESI/MS and the N-terminal sequence of lactocin MXJ 32A was NH2-GEPGPMGPAGAD. Lactocin MXJ 32A had a broad antimicrobial spectrum including many Gram-positive and Gram-negative foodborne pathogens, in which even some antibiotic-resistant foodborne pathogenic strains could also be inhibited. The MIC of lactocin MXJ 32A for Staphylococcus aureus and Escherichia coli was 10 mg/mL. Lactocin MXJ 32A showed good pH-stability and heat stability and was sensitive to proteolytic enzymes. It was shown by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) that the possible mode of action of lactocin MXJ 32A was pore formation of the cytoplasmic membrane of target cells.

Loop-Mediated Isothermal Amplification Assays for DetectingYersinia pseudotuberculosisin Milk Powders

Yersinia pseudotuberculosis is a Gram-negative foodborne pathogen that causes several diseases, such as enteritis, septicemia, and reactive arthritis. Loop-mediated isothermal amplification (LAMP) assay targeting the 16S-23S rDNA internal transcribed spacer (ITS) region was developed to detect Y. pseudotuberculosis in milk powder. The DNA amplification could be completed in 1 h, and detected by produced white precipitate visible to naked eyes. The detection limit of LAMP assay was 10(0) fg/reaction for genomic DNA, and 10(0) CFU/100 g milk powder coupled with 12 h enrichment. LAMP assay is 100 times more sensitive than conventional polymerase chain reaction method for detecting Y. pseudotuberculosis, and correctly identified 18 cases of Y. pseudotuberculosis contaminations from 236 commercial milk powder products. In conclusion, the developed LAMP assay may facilitate rapid detection of Y. pseudotuberculosis contaminations in agricultural and food products. Rapid and accurate detection of Yersinia pseudotuberculosis in milk products.