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Journal of Dairy Science Aug 2020Due to the lack of electricity and thermostatic instruments in certain settings for convenient detection of Cronobacter species in powdered infant formula (PIF), a novel...
Due to the lack of electricity and thermostatic instruments in certain settings for convenient detection of Cronobacter species in powdered infant formula (PIF), a novel investigation was conducted to establish an electricity-free visual detection system for rapid detection of Cronobacter species in PIF. This system included a portable electricity-free heater that could use the exothermic reaction of calcium oxide and water and 3 kinds of phase change materials to supply 3 constant temperatures for immunomagnetic separation, DNA extraction, and loop-mediated isothermal amplification assay. Meanwhile, the amplified reaction combined with hydroxynaphthol blue could achieve rapid visual detection. Primers designed based on the 16S-23S ribosomal RNA internal transcribed spacer were used in loop-mediated isothermal amplification to specifically monitor Cronobacter species, and the detection limit can reach 4.2 × 10 cfu/g in PIF by an electricity-free heater in 2 h 30 min. Moreover, 2 h of pre-enrichment was necessary when the level of the PIF samples with Cronobacter spp. was 10 cfu/g. The stability of the system was evaluated in ambient temperature at 4°C, 25°C, and 37°C. The results suggested that the electricity-free heater can maintain 3 constant temperatures to support different processes. Therefore, this amplification and visual system is applicable for use in many fields for rapid and specific detection of Cronobacter species in PIF.
Topics: Cronobacter; DNA Primers; Food Microbiology; Humans; Immunomagnetic Separation; Infant; Infant Formula; Nucleic Acid Amplification Techniques; Powders
PubMed: 32505404
DOI: 10.3168/jds.2019-17661 -
Microbiology Spectrum Apr 2016The Cronobacter group of pathogens, associated with severe and potentially life-threatening diseases, until recently were classified as a single species, Enterobacter... (Review)
Review
The Cronobacter group of pathogens, associated with severe and potentially life-threatening diseases, until recently were classified as a single species, Enterobacter sakazakii. The group was reclassified in 2007 into the genus Cronobacter as a member of the Enterobacteriaceae. This chapter outlines the history behind the epidemiology, analyzes how our understanding of these bacteria has evolved, and highlights the clinical significance the Cronobacter spp. have for neonatal and elderly patient populations and treatment of the associated infections.
Topics: Animals; Anti-Bacterial Agents; Cronobacter; Enterobacteriaceae Infections; Humans
PubMed: 27227295
DOI: 10.1128/microbiolspec.EI10-0002-2015 -
Open Veterinary Journal Jan 2023sspecies are the most significant foodborne pathogen in infant milk formula (IMF). These pathogens have been incriminated in severe forms of neonatal meningitis,...
BACKGROUND
sspecies are the most significant foodborne pathogen in infant milk formula (IMF). These pathogens have been incriminated in severe forms of neonatal meningitis, sepsis, and necrotizing enterocolitis with a high mortality rate.
AIM
This study was performed to elucidate the effect of heat stress on spp. ( and ) in reconstituted IMF (RIMF).
METHODS
The reconstituted formula was inoculated with five isolates and four isolates separately. The nine isolates of spp. were heated in RIMF at 48°C, 52°C, 56°C, 60°C, 64°C, and 66°C. The - and -values were determined by using linear regression analysis.
RESULTS
The -values of all isolates of (CS1, CS3, CS4, CS5, and CS6) at 48°C, 52°C, 56°C, 60°C, 64°C, and 66°C were in the ranges 7.29-23.47, 2.77-15.50, 0.62-1.04, 0.62-1.02, 0.62-1.00, 0.62-1.00 minutes, respectively; while, the -values extended from 2.50°C to 4.28°C. The - values of isolates (CP1, CP2, CP3, CP4) were in the ranges 7.60-22.32, 1.42-8.45, 0.62-1.08, 0.62-0.78, 0.62-0.78, 0.62-0.79 minutes at 48°C, 52°C, 56°C, 60°C, 64°C, 66°C, respectively and the calculated -values ranged from 3.33°C to 4.89°C.
CONCLUSION
This study may contribute to improving the understanding of the behavior of and isolates in RIMF at various heat stress temperatures and may participate in the effective control of these pathogens in infant food production.
Topics: Animals; Cronobacter sakazakii; Milk; Food Microbiology; Infant Formula
PubMed: 36777432
DOI: 10.5455/OVJ.2023.v13.i1.11 -
Frontiers in Microbiology 2019strains harboring CRISPR-Cas systems are important foodborne pathogens that cause serious neonatal infections. CRISPR typing is a new molecular subtyping method to...
strains harboring CRISPR-Cas systems are important foodborne pathogens that cause serious neonatal infections. CRISPR typing is a new molecular subtyping method to track the sources of pathogenic bacterial outbreaks and shows a promise in typing , however, this molecular typing procedure using routine PCR method has not been established. Therefore, the purpose of this study was to establish such methodology, 257 isolates of , , and were used to verify the feasibility of the method. Results showed that 161 strains could be divided into 129 CRISPR types (CTs), among which CT15 ( = 7) was the most prevalent CT followed by CT6 ( = 4). Further, 65 strains were divided into 42 CTs and CT23 ( = 8) was the most prevalent followed by CT2, CT3, and CT13 ( = 4). Finally, 31 strains belonged to 31 CTs. There was also a relationship among CT, sequence type (ST), food types, and serotype. Compared to multi-locus sequence typing (MLST), this new molecular method has greater power to distinguish similar strains and had better accordance with whole genome sequence typing (WGST). More importantly, some lineages were found to harbor conserved ancestral spacers ahead of their divergent specific spacer sequences; this can be exploited to infer the divergent evolution of and provide phylogenetic information reflecting common origins. Compared to WGST, CRISPR typing method is simpler and more affordable, it could be used to identify sources of food-borne outbreaks, from clinical cases to food sources and the production sites.
PubMed: 31555228
DOI: 10.3389/fmicb.2019.01989 -
SpringerPlus 2015Cronobacter species are motile, non-spore forming, Gram negative emerging opportunistic pathogens mostly associated with bacteremia, meningitis, septicemia, brain... (Review)
Review
Cronobacter species are motile, non-spore forming, Gram negative emerging opportunistic pathogens mostly associated with bacteremia, meningitis, septicemia, brain abscesses and necrotizing enterocolitis in infected neonates, infants and immunocompromised adults. Members of the genus Cronobacter are previously associated with powdered infant formula although the main reservoir and routes of contamination are yet to be ascertained. This study therefore aim to summarize occurrence and prevalence of Cronobacter spp. from different food related sources. A retrospective systematic review and meta-analysis of peer reviewed primary studies reported between 2008 and 2014 for the occurrence and prevalence of Cronobacter spp. in animal and plant related sources was conducted using "Cronobacter isolation", "Cronobacter detection" and "Cronobacter enumeration" as search terms in the following databases: Web of Science (Science Direct) and ProQuest. Data extracted from the primary studies were then analyzed with meta-analysis techniques for effect rate and fixed effects was used to explore heterogeneity between the sources. Publication bias was evaluated using funnel plot. A total of 916 articles were retrieved from the data bases of which 28 articles met inclusion criteria. Cronobacter spp. could only be isolated from 103 (5.7 %) samples of animal related food while 123 (19 %) samples of plant related food samples harbors the bacteria. The result of this study shows that occurrence of Cronobacter was more prevalent in plant related sources with overall prevalence rate of 20.1 % (95 % CI 0.168-0.238) than animal originated sources with overall prevalence rate of 8 % (95 % CI 0.066-0.096). High heterogeneity (I (2) = 84) was observed mostly in plant related sources such as herbs, spices and vegetables compared to animal related sources (I (2) = 82). It could be observed from this study that plant related sources serve as reservoir and contamination routes of Cronobacter spp.
PubMed: 26435891
DOI: 10.1186/s40064-015-1324-9 -
Frontiers in Microbiology 2022This study characterized five spp. and six spp. strains that had been isolated from 155 samples of powdered infant formula (PIF) sold in Chile and manufactured in...
This study characterized five spp. and six spp. strains that had been isolated from 155 samples of powdered infant formula (PIF) sold in Chile and manufactured in Chile and Mexico in 2018-2020. Two strains of sequence type (ST) ST1 and ST31 (serotypes O:1 and O:2) and one strain of ST60 (O:1) were identified. All strains were identified as Typhimurium ST19 (serotype O:4) by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST (cgMLST). The and isolates were resistant to cephalothin, whereas the isolates were resistant to oxacillin and ampicillin. Nineteen antibiotic resistance genes were detected in the and isolates; the most prevalent were mcr-9.1, , and . In , 30 genes encoding for aminoglycoside and cephalosporin resistance were identified, including , β-, , and . In the isolates, 32 virulence-associated genes were detected by WGS and clustered as flagellar proteins, outer membrane proteins, chemotaxis, hemolysins, invasion, plasminogen activator, colonization, transcriptional regulator, survival in macrophages, use of sialic acid, and toxin-antitoxin genes. In the strains, 120 virulence associated genes were detected, adherence, magnesium uptake, resistance to antimicrobial peptides, secretion system, stress protein, toxin, resistance to complement killing, and eight pathogenicity islands. The and strains harbored I-E and I-F CRISPR-Cas systems and carried Col(pHHAD28) and IncFIB(pCTU1) plasmids, respectively. The strains harbored type I-E CRISPR-Cas systems and carried IncFII(S) plasmids. The presence of and in PIF is a health risk for infants aged less than 6 months. For this reason, sanitary practices should be reinforced for its production and retail surveillance.
PubMed: 35722296
DOI: 10.3389/fmicb.2022.884721 -
Journal of Food Protection Mar 2012Cronobacter species are opportunistic pathogens, and a mortality rate of 40 to 80% is associated with infections. This pathogen can cause a range of serious diseases... (Review)
Review
Cronobacter species are opportunistic pathogens, and a mortality rate of 40 to 80% is associated with infections. This pathogen can cause a range of serious diseases such as meningitis, septicemia, necrotizing enterocolitis, and brain abscesses and has been responsible for a variety of sequelae such as quadriplegia. Although Cronobacter can cause disease in both adults and infants, infant infections associated with powdered formula are the focus of this review. Since the first reported Cronobacter infection outbreak in 1958, powdered infant formula has been identified as a major source of these outbreaks, resulting in many recalls of powdered infant formula worldwide. This contamination has created an immense problem for the powdered infant formula industry. In this review, we discuss the taxonomy of Cronobacter species, the natural habitat of Cronobacter and its presence in foods, the physiology, pathogenicity, and virulence of Cronobacter species, and available detection methods. We also discuss reported cases of Cronobacter infection linked to powdered infant formula consumption and then focus specifically on the official World Health Organization guidelines for preparation of powdered infant formula.
Topics: Cronobacter; Cronobacter sakazakii; Food Contamination; Food Microbiology; Humans; Infant; Infant Food; Infant Formula
PubMed: 22410240
DOI: 10.4315/0362-028X.JFP-11-285 -
Foods (Basel, Switzerland) Mar 2023spp. is a food-borne pathogenic microorganism that can cause serious diseases such as meningitis, sepsis, and necrotizing colitis in infants and young children....
spp. is a food-borne pathogenic microorganism that can cause serious diseases such as meningitis, sepsis, and necrotizing colitis in infants and young children. Powdered infant formula (PIF) is one of the main contamination routes, in which the processing environment is an important source of pollution. In this investigation, 35 strains isolated from PIF and its processing environment were identified and typed by 16S rRNA sequencing and multilocus sequence typing (MLST) technology. A total of 35 sequence types were obtained, and three new sequence types were isolated for the first time. The antibiotic resistance was analyzed, showing that all isolates were resistant to erythromycin but sensitive to ciprofloxacin. Multi-drug resistant strains accounted for 68.57% of the total, among which strains with the strongest drug resistance reached 13 multiple drug resistance. Combined with transcriptomics, 77 differentially expressed genes related to drug resistance were identified. The metabolic pathways were deeply excavated, and under the stimulation of antibiotic conditions, strains can activate the multidrug efflux system by regulating the expression of chemotaxis-related genes, thus, secreting more drug efflux proteins to enhance drug resistance. The study of drug resistance of and its mechanism has important public health significance for the rational selection of existing antibacterial drugs, the development of new antibacterial drugs to reduce the occurrence of bacterial resistance, and the control and treatment of infections caused by .
PubMed: 36900599
DOI: 10.3390/foods12051084 -
Frontiers in Pediatrics 2015Cronobacter species are emerging opportunistic food-borne pathogens, which consists of seven species, including C. sakazakii, C. malonaticus, C. muytjensii, C.... (Review)
Review
Cronobacter species are emerging opportunistic food-borne pathogens, which consists of seven species, including C. sakazakii, C. malonaticus, C. muytjensii, C. turicensis, C. dublinensis, C. universalis, and C. condimenti. The organism can cause severe clinical infections, including necrotizing enterocolitis, septicemia, and meningitis, predominately among neonates <4 weeks of age. Cronobacter species can be isolated from various foods and their surrounding environments; however, powdered infant formula (PIF) is the most frequently implicated food source linked with Cronobacter infection. This review aims to provide a summary of laboratory-based strategies that can be used to identify and trace Cronobacter species. The identification of Cronobacter species using conventional culture method and immuno-based detection protocols were first presented. The molecular detection and identification at genus-, and species-level along with molecular-based serogroup approaches are also described, followed by the molecular sub-typing methods, in particular pulsed-field gel electrophoresis and multi-locus sequence typing. Next generation sequence approaches, including whole genome sequencing, DNA microarray, and high-throughput whole-transcriptome sequencing, are also highlighted. Appropriate application of these strategies would contribute to reduce the risk of Cronobacter contamination in PIF and production environments, thereby improving food safety and protecting public health.
PubMed: 26000266
DOI: 10.3389/fped.2015.00038 -
Gut Pathogens Jun 2022Cronobacter sakazakii is a foodborne pathogen that causes septicemia, meningitis, and necrotizing enterocolitis in neonates and infants. The current research details the...
Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities.
BACKGROUND
Cronobacter sakazakii is a foodborne pathogen that causes septicemia, meningitis, and necrotizing enterocolitis in neonates and infants. The current research details the full genome sequences of two extremely persistent C. sakazakii strains (H322 and GK1025B) isolated from powdered infant formula (PIF) manufacturing settings. In addition, the genetic attributes associated with five plasmids, pH322_1, pH322_2, pGK1025B_1, pGK1025B_2, and pGK1025B_3 are described.
MATERIALS AND METHODS
Using PacBio single-molecule real-time (SMRT) sequencing technology, whole genome sequence (WGS) assemblies of C. sakazakii H322 [Sequence type (ST)83, clonal complex [CC] 83) and GK1025B (ST64, CC64) were generated. Plasmids, also sequenced, were aligned with phylogenetically related episomes to determine, and identify conserved and missing genomic regions.
RESULTS
A truncated ~ 13 Kbp type 6 secretion system (T6SS) gene cluster harbored on virulence plasmids pH322_2 and pGK1025B_2, and a second large deletion (~ 6 Kbp) on pH322_2, which included genes for a tyrosine-type recombinase/integrase, a hypothetical protein, and a phospholipase D was identified. Within the T6SS of pH322_2 and pGK1025B_2, an arsenic resistance operon was identified which is in common with that of plasmids pSP291_1 and pESA3. In addition, PHASTER analysis identified an intact 96.9 Kbp Salmonella SSU5 prophage gene cluster in pH322_1 and pGK1025B_1 and showed that these two plasmids were phylogenetically related to C. sakazakii plasmids: pCS1, pCsa767a, pCsaC757b, pCsaC105731a. Plasmid pGK1025B_3 was identified as a novel conjugative Cronobacter plasmid. Furthermore, WGS analysis identified a ~ 16.4 Kbp type 4 secretion system gene cluster harbored on pGK1025B_3, which contained a phospholipase D gene, a key virulence factor in several host-pathogen diseases.
CONCLUSION
These data provide high resolution information on C. sakazakii genomes and emphasizes the need for furthering surveillance studies to link genotype to phenotype of strains from previous investigations. These results provide baseline data necessary for future in-depth investigations of C. sakazakii that colonize PIF manufacturing facility settings and genomic analyses of these two C. sakazakii strains and five associated plasmids will contribute to a better understanding of this pathogen's survival and persistence within various "built environments" like PIF manufacturing facilities.
PubMed: 35668537
DOI: 10.1186/s13099-022-00500-5