New Approaches on Quantification of Campylobacter jejuni in Poultry Samples: The Use of Digital PCR and Real-time PCR against the ISO Standard Plate Count Method.

Abstract:

:Campylobacteriosis is the most frequently reported bacterial food-borne illness in the European Union and contaminated broiler meat is considered the most important source of infection in humans. The aim of the present study was to evaluate real-time PCR (qPCR) and digital PCR (dPCR) for quantification of Campylobacter jejuni in 75 broiler neck-skin samples collected from a poultry slaughterhouse, and to compare them with the ISO 10272-2 standard plate count method. For qPCR standard curve, C. jejuni-negative neck-skin samples were spiked with C. jejuni suspension with a known number of bacterial cells. The observed CFU/g values by qPCR correlated greatly with the expected values and qPCR showed good performance with the reliable limit of detection (rLOD) and limit of quantification (LOQ) of three and 31 target copies per reaction, respectively. However, both rLOD (1219 CFU/g) and LOQ (12,523 CFU/g) were beyond the EFSA-proposed critical limit of 500-1,000 CFU/g of neck skin. Although C. jejuni cell counts were ≤1,000 CFU/g in only 7/75 samples by plate counting, they were ≤LOQ in 60/75 and ≤rLOD in 26/75 (≤1,000 CFU/g in 24/75) samples by qPCR. A strong and statistically significant correlation was observed between qPCR and dPCR. Both PCR-based methods correlated significantly with the plate count method; however, the correlation was moderate. Using the Bland-Altman analysis, an average agreement was noted between all three methods, although with a large standard deviation. A significant bias toward overestimation in dPCR was observed, probably due to the relatively high number of false positive calls. The linear dynamic range was comparable in both PCR-based methods; however, qPCR proved to be more suitable for routine use. In the future, the establishment of a reliable molecular quantification of C. jejuni in poultry samples showing a wide range of contamination levels down to the proposed critical limit is needed to enable time- and cost-effective surveillance throughout all stages in the food production chain. As both rLOD and LOQ were beyond this limit, a modification of the procedure is suggested to include less sample dilution prior to DNA extraction to enable PCR-based quantification of C. jejuni at the proposed microbiological criteria.

journal_name

Front Microbiol

authors

Papić B,Pate M,Henigman U,Zajc U,Gruntar I,Biasizzo M,Ocepek M,Kušar D

doi

10.3389/fmicb.2017.00331

subject

Has Abstract

pub_date

2017-03-02 00:00:00

pages

331

issn

1664-302X

journal_volume

8

pub_type

杂志文章
  • Bacterial Composition and Diversity in Breast Milk Samples from Mothers Living in Taiwan and Mainland China.

    abstract::Human breast milk is widely recognized as the best source of nutrients for healthy growth and development of infants; it contains a diverse microbiota. Here, we characterized the diversity of the microbiota in the breast milk of East Asian women and assessed whether delivery mode influenced the microbiota in the milk ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2017.00965

    authors: Li SW,Watanabe K,Hsu CC,Chao SH,Yang ZH,Lin YJ,Chen CC,Cao YM,Huang HC,Chang CH,Tsai YC

    更新日期:2017-05-30 00:00:00

  • Genome Sequencing Reveals the Potential of Achromobacter sp. HZ01 for Bioremediation.

    abstract::Petroleum pollution is a severe environmental issue. Comprehensively revealing the genetic backgrounds of hydrocarbon-degrading microorganisms contributes to developing effective methods for bioremediation of crude oil-polluted environments. Marine bacterium Achromobacter sp. HZ01 is capable of degrading hydrocarbons ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2017.01507

    authors: Hong YH,Ye CC,Zhou QZ,Wu XY,Yuan JP,Peng J,Deng H,Wang JH

    更新日期:2017-08-09 00:00:00

  • Analysis of genes that are differentially expressed during the Sclerotinia sclerotiorum-Phaseolus vulgaris interaction.

    abstract::The fungus Sclerotinia sclerotiorum (Lib.) de Bary, one of the most important plant pathogens, causes white mold on a wide range of crops. Crop yield can be dramatically decreased due to this disease, depending on the plant cultivar and environmental conditions. In this study, a suppression subtractive hybridization c...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2015.01162

    authors: Oliveira MB,de Andrade RV,Grossi-de-Sá MF,Petrofeza S

    更新日期:2015-10-26 00:00:00

  • Perspectives on the Impact of Sampling Design and Intensity on Soil Microbial Diversity Estimates.

    abstract::Soil bacterial communities have long been recognized as important ecosystem components, and have been the focus of many local and regional studies. However, there is a lack of data at large spatial scales, on the biodiversity of soil microorganisms; national or more extensive studies to date have typically consisted o...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2019.01820

    authors: Hermans SM,Buckley HL,Lear G

    更新日期:2019-08-07 00:00:00

  • Assessment of Ruminal Bacterial and Archaeal Community Structure in Yak (Bos grunniens).

    abstract::The aim of this study was to determine the microbial community composition in the rumen of yaks under different feeding regimes. Microbial communities were assessed by sequencing bacterial and archaeal 16S ribosomal RNA gene fragments obtained from yaks (Bos grunniens) from Qinghai-Tibetan Plateau, China. Samples were...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2017.00179

    authors: Zhou Z,Fang L,Meng Q,Li S,Chai S,Liu S,Schonewille JT

    更新日期:2017-02-07 00:00:00

  • The Addition of ViriditecTM Aqueous Ozone to Peracetic Acid as an Antimicrobial Spray Increases Air Quality While Maintaining Salmonella Typhimurium, Non-pathogenic Escherichia coli, and Campylobacter jejuni Reduction on Whole Carcasses.

    abstract::Currently, the most utilized antimicrobial in processing facilities is peracetic acid, PAA; however, this chemical is increasingly recognized as a hazard to human health. Preliminary evidence suggests that ozone, when introduced in a specific manner, can reduce the noxious nature of PAA. Therefore, the objective of th...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.03180

    authors: Dittoe DK,Feye KM,Peyton B,Worlie D,Draper MJ,Ricke SC

    更新日期:2019-01-08 00:00:00

  • Controls on soil microbial community stability under climate change.

    abstract::Soil microbial communities are intricately linked to ecosystem functioning because they play important roles in carbon and nitrogen cycling. Still, we know little about how soil microbial communities will be affected by disturbances expected with climate change. This is a significant gap in understanding, as the stabi...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2013.00265

    authors: de Vries FT,Shade A

    更新日期:2013-09-05 00:00:00

  • Few Differences in Metabolic Network Use Found Between Salmonella enterica Colonization of Plants and Typhoidal Mice.

    abstract::The human enteric pathogen Salmonella enterica leads a cross-kingdom lifestyle, actively colonizing and persisting on plants in between animal hosts. One of the questions that arises from this dual lifestyle is how S. enterica is able to adapt to such divergent hosts. Metabolic pathways required for S. enterica animal...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.00695

    authors: Kwan G,Plagenz B,Cowles K,Pisithkul T,Amador-Noguez D,Barak JD

    更新日期:2018-05-08 00:00:00

  • PVP-SVM: Sequence-Based Prediction of Phage Virion Proteins Using a Support Vector Machine.

    abstract::Accurately identifying bacteriophage virion proteins from uncharacterized sequences is important to understand interactions between the phage and its host bacteria in order to develop new antibacterial drugs. However, identification of such proteins using experimental techniques is expensive and often time consuming; ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.00476

    authors: Manavalan B,Shin TH,Lee G

    更新日期:2018-03-16 00:00:00

  • Viral Lysis of Photosynthesizing Microbes As a Mechanism for Calcium Carbonate Nucleation in Seawater.

    abstract::Removal of carbon through the precipitation and burial of calcium carbonate in marine sediments constitutes over 70% of the total carbon on Earth and is partitioned between coastal and pelagic zones. The precipitation of authigenic calcium carbonate in seawater, however, has been hotly debated because despite being in...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2016.01958

    authors: Lisle JT,Robbins LL

    更新日期:2016-12-09 00:00:00

  • 13C-DNA-SIP Distinguishes the Prokaryotic Community That Metabolizes Soybean Residues Produced Under Different CO2 Concentrations.

    abstract::The amendment of crop residues produced under elevated CO2 (eCO2) may alter soil microbial community structure and their functions on residue decomposition and carbon (C) cycling in soil. The key to understanding this process is to elucidate the structure of prokaryotic communities that metabolize crop residues derive...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2019.02184

    authors: Wang Y,Yu Z,Li Y,Wang G,Tang C,Liu X,Liu J,Xie Z,Jin J

    更新日期:2019-09-24 00:00:00

  • Novel plant inputs influencing Ralstonia solanacearum during infection.

    abstract::Ralstonia solanacearum is a soil and water-borne pathogen that can infect a wide range of plants and cause the devastating bacterial wilt disease. To successfully colonize a host, R. solanacearum requires the type III secretion system (T3SS), which delivers bacterial effector proteins inside the plant cells. HrpG is a...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2013.00349

    authors: Zuluaga AP,Puigvert M,Valls M

    更新日期:2013-11-20 00:00:00

  • The Road to Infection: Host-Microbe Interactions Defining the Pathogenicity of Streptococcus bovis/Streptococcus equinus Complex Members.

    abstract::The Streptococcus bovis/Streptococcus equinus complex (SBSEC) comprises several species inhabiting the animal and human gastrointestinal tract (GIT). They match the pathobiont description, are potential zoonotic agents and technological organisms in fermented foods. SBSEC members are associated with multiple diseases ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章,评审

    doi:10.3389/fmicb.2018.00603

    authors: Jans C,Boleij A

    更新日期:2018-04-10 00:00:00

  • The Peroxygenase Activity of the Aspergillus flavus Caleosin, AfPXG, Modulates the Biosynthesis of Aflatoxins and Their Trafficking and Extracellular Secretion via Lipid Droplets.

    abstract::Aflatoxins (AF) are highly detrimental to human and animal health. We recently demonstrated that the Aspergillus flavus caleosin, AfPXG, had peroxygenase activity and mediated fungal development and AF accumulation. We now report the characterization of an AfPXG-deficient line using reference strain NRRL3357. The resu...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.00158

    authors: Hanano A,Alkara M,Almousally I,Shaban M,Rahman F,Hassan M,Murphy DJ

    更新日期:2018-02-06 00:00:00

  • Host Matters: Medicinal Leech Digestive-Tract Symbionts and Their Pathogenic Potential.

    abstract::Digestive-tract microbiota exert tremendous influence over host health. Host-symbiont model systems are studied to investigate how symbioses are initiated and maintained, as well as to identify host processes affected by resident microbiota. The medicinal leech, Hirudo verbana, is an excellent model to address such qu...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章,评审

    doi:10.3389/fmicb.2016.01569

    authors: Marden JN,McClure EA,Beka L,Graf J

    更新日期:2016-10-13 00:00:00

  • Intestinal Microbiota as a Host Defense Mechanism to Infectious Threats.

    abstract::The intestinal microbiota is a complex microbial community, with diverse and stable populations hosted by the gastrointestinal tract since birth. This ecosystem holds multiple anti-infectious, anti-inflammatory, and immune modulating roles decisive for intestinal homeostasis. Among these, colonization resistance refer...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章,评审

    doi:10.3389/fmicb.2018.03328

    authors: Iacob S,Iacob DG,Luminos LM

    更新日期:2019-01-23 00:00:00

  • Three Capsular Polysaccharide Synthesis-Related Glucosyltransferases, GT-1, GT-2 and WcaJ, Are Associated With Virulence and Phage Sensitivity of Klebsiella pneumoniae.

    abstract::Klebsiella pneumoniae (K. pneumoniae) spp. are important nosocomial and community-acquired opportunistic pathogens, which cause various infections. We observed that K. pneumoniae strain K7 abruptly mutates to rough-type phage-resistant phenotype upon treatment with phage GH-K3. In the present study, the rough-type pha...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2019.01189

    authors: Cai R,Wang G,Le S,Wu M,Cheng M,Guo Z,Ji Y,Xi H,Zhao C,Wang X,Xue Y,Wang Z,Zhang H,Fu Y,Sun C,Feng X,Lei L,Yang Y,Ur Rahman S,Liu X,Han W,Gu J

    更新日期:2019-05-28 00:00:00

  • Virulence and Antibiotic Resistance Profiles of Cronobacter sakazakii and Enterobacter spp. Involved in the Diarrheic Hemorrhagic Outbreak in Mexico.

    abstract::Cronobacter spp. are bacterial pathogens that cause neonatal meningitis, septicemia, and necrotizing enterocolitis in infants with a lethality rate of 40-80%. Powdered infant formulas (PIF) have been implicated as the main vehicles of transmission. This pathogen can also cause infection through contaminated expressed ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.02206

    authors: Parra-Flores J,Aguirre J,Juneja V,Jackson EE,Cruz-Córdova A,Silva-Sanchez J,Forsythe S

    更新日期:2018-09-27 00:00:00

  • Origin, diversity, and maturation of human antiviral antibodies analyzed by high-throughput sequencing.

    abstract::Our understanding of how antibodies are generated and function could help develop effective vaccines and antibody-based therapeutics against viruses such as HIV-1, SARS coronavirus (SARS CoV), and Hendra and Nipah viruses (henipaviruses). Although broadly neutralizing antibodies (bnAbs) against the HIV-1 were observed...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2012.00277

    authors: Prabakaran P,Zhu Z,Chen W,Gong R,Feng Y,Streaker E,Dimitrov DS

    更新日期:2012-08-02 00:00:00

  • Citrus Leprosis Virus C Encodes Three Proteins With Gene Silencing Suppression Activity.

    abstract::Citrus leprosis virus C (CiLV-C) belongs to the genus Cilevirus, family Kitaviridae, and is considered the most devastating virus infecting citrus in Brazil, being the main viral pathogen responsible for citrus leprosis (CL), a severe disease that affects citrus orchards in Latin America. Here, proteins encoded by CiL...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2020.01231

    authors: Leastro MO,Castro DYO,Freitas-Astúa J,Kitajima EW,Pallás V,Sánchez-Navarro JÁ

    更新日期:2020-06-09 00:00:00

  • Out of the Abyss: Genome and Metagenome Mining Reveals Unexpected Environmental Distribution of Abyssomicins.

    abstract::Natural products have traditionally been discovered through the screening of culturable microbial isolates from diverse environments. The sequencing revolution allowed the identification of dozens of biosynthetic gene clusters (BGCs) within single bacterial genomes, either from cultured or uncultured strains. However,...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2020.00645

    authors: Iglesias A,Latorre-Pérez A,Stach JEM,Porcar M,Pascual J

    更新日期:2020-04-15 00:00:00

  • Insight Into the Diversity and Possible Role of Plasmids in the Adaptation of Psychrotolerant and Metalotolerant Arthrobacter spp. to Extreme Antarctic Environments.

    abstract::Arthrobacter spp. are coryneform Gram-positive aerobic bacteria, belonging to the class Actinobacteria. Representatives of this genus have mainly been isolated from soil, mud, sludge or sewage, and are usually mesophiles. In recent years, the presence of Arthrobacter spp. was also confirmed in various extreme, includi...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.03144

    authors: Romaniuk K,Golec P,Dziewit L

    更新日期:2018-12-18 00:00:00

  • Modulation of Plant Defense System in Response to Microbial Interactions.

    abstract::At different stages throughout their life cycle, plants often encounter several pathogenic microbes that challenge plant growth and development. The sophisticated innate plant immune system prevents the growth of harmful microbes via two interconnected defense strategies based on pathogen perception. These strategies ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章,评审

    doi:10.3389/fmicb.2020.01298

    authors: Nishad R,Ahmed T,Rahman VJ,Kareem A

    更新日期:2020-07-03 00:00:00

  • Experimental Evolution on a Wild Mammal Species Results in Modifications of Gut Microbial Communities.

    abstract::Comparative studies have shown that diet, life history, and phylogeny interact to determine microbial community structure across mammalian hosts. However, these studies are often confounded by numerous factors. Selection experiments offer unique opportunities to validate conclusions and test hypotheses generated by co...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2016.00634

    authors: Kohl KD,Sadowska ET,Rudolf AM,Dearing MD,Koteja P

    更新日期:2016-05-04 00:00:00

  • The Atypical Guanylate Kinase MoGuk2 Plays Important Roles in Asexual/Sexual Development, Conidial Septation, and Pathogenicity in the Rice Blast Fungus.

    abstract::Guanylate kinases (GKs), which convert guanosine monophosphate into guanosine diphosphate (GDP), are important for growth and mannose outer chain elongation of cell wall N-linked glycoproteins in yeast. Here, we identified the ortholog of Saccharomyces cerevisiae GK Guk1, named MoGuk1 and a novel family of fungal GKs ...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2017.02467

    authors: Cai X,Zhang X,Li X,Liu M,Liu X,Wang X,Zhang H,Zheng X,Zhang Z

    更新日期:2017-12-11 00:00:00

  • Catabolism of Alkylphenols in Rhodococcus via a Meta-Cleavage Pathway Associated With Genomic Islands.

    abstract::The bacterial catabolism of aromatic compounds has considerable promise to convert lignin depolymerization products to commercial chemicals. Alkylphenols are a key class of depolymerization products whose catabolism is not well-elucidated. We isolated Rhodococcus rhodochrous EP4 on 4-ethylphenol and applied genomic an...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2019.01862

    authors: Levy-Booth DJ,Fetherolf MM,Stewart GR,Liu J,Eltis LD,Mohn WW

    更新日期:2019-08-20 00:00:00

  • Variability of Gut Microbiota Across the Life Cycle of Grapholita molesta (Lepidoptera: Tortricidae).

    abstract::Grapholita molesta, the oriental fruit moth, is a serious global pest of many Rosaceae fruit trees. Gut microorganisms play important roles in host nutrition, digestion, detoxification, and resistance to pathogens. However, there are few studies on the microbiota of G. molesta, particularly during metamorphosis. Here,...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2020.01366

    authors: Wang X,Sun S,Yang X,Cheng J,Wei H,Li Z,Michaud JP,Liu X

    更新日期:2020-06-30 00:00:00

  • Transmissible ST3-IncHI2 Plasmids Are Predominant Carriers of Diverse Complex IS26-Class 1 Integron Arrangements in Multidrug-Resistant Salmonella.

    abstract::Diverse mobile genetic elements (MGEs) including plasmids, insertion sequences, and integrons play an important role in the occurrence and spread of multidrug resistance (MDR) in bacteria. It was found in previous studies that IS26 and class 1 integrons integrated on plasmids to speed the dissemination of antibiotic-r...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.02492

    authors: Zhao H,Chen W,Xu X,Zhou X,Shi C

    更新日期:2018-10-23 00:00:00

  • Transcriptional Regulation of the Type VI Secretion System 1 Genes by Quorum Sensing and ToxR in Vibrio parahaemolyticus.

    abstract::Vibrio parahaemolyticus, the leading cause of seafood-associated gastroenteritis, harbors two separate T6SSs on chromosomes 1 and 2, i.e., T6SS1 (VP1386-1420) and T6SS2 (VPA1025-1046). T6SS1 contains at least 7 putative operons: VP1386-1387, VP1388-1390, VP1392-1391, VP1393-1406, VP1400-1406, VP1409-1407, and VP1410-1...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2017.02005

    authors: Zhang Y,Gao H,Osei-Adjei G,Zhang Y,Yang W,Yang H,Yin Z,Huang X,Zhou D

    更新日期:2017-10-16 00:00:00

  • Interplay Between the Phenotype and Genotype, and Efflux Pumps in Drug-Resistant Strains of Riemerella anatipestifer.

    abstract::The number of multidrug-resistant strains of Riemerella anatipestifer continues to increase, and new strategies for the treatment of associated infections are necessary. Recently, numerous studies have shown that efflux pumps (EPs) play key roles in universal bacterial mechanisms that contribute to antibiotic resistan...

    journal_title:Frontiers in microbiology

    pub_type: 杂志文章

    doi:10.3389/fmicb.2018.02136

    authors: Chen Q,Gong X,Zheng F,Ji G,Li S,Stipkovits L,Szathmary S,Liu Y

    更新日期:2018-10-01 00:00:00