Surveillance and characterisation of Cronobacter spp. in Czech retail food and environmental samples

Mozrová, Věra; Břeňová, N.; Mrázek, Jakub; Lukešová, D.; Marounek, M.

doi:10.1007/s12223-013-0266-2

Cited by 20 publications

(15 citation statements)

References 27 publications

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“…The predominant Cronobacter species cultured from the four PIF production environments studied was C. sakazakii. This finding is in agreement with others, including Müller et al (32), who reported on the microbial ecology of a Swiss PIF facility, Mozrová et al (43), who studied a dairy farm and its environment in the Czech Republic, and Pan et al (20), who reported on Cronobacter species found in commercially available PIF products.…”

Section: Discussionsupporting

confidence: 92%

Comparative Genotypic and Phenotypic Analysis of Cronobacter Species Cultured from Four Powdered Infant Formula Production Facilities: Indication of Pathoadaptation along the Food Chain

Yan

Wang

Gangiredla

et al. 2015

Appl Environ Microbiol

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cCronobacter species are opportunistic pathogens commonly found in the environment. Among the seven Cronobacter species, Cronobacter sakazakii sequence type 4 (ST-4) is predominantly associated with recorded cases of infantile meningitis. This study reports on a 26-month powdered infant formula (PIF) surveillance program in four production facilities located in distinct geographic regions. The objective was to identify the ST(s) in PIF production environments and to investigate the phenotypic features that support their survival. Of all 168 Cronobacter isolates, 133 were recovered from a PIF production environment, 31 were of clinical origin, and 4 were laboratory type strains. Sequence type 1 (n ‫؍‬ 84 isolates; 63.9%) was the dominant type in PIF production environments. The majority of these isolates clustered with an indistinguishable pulsotype and persisted for at least an 18-month period. Moreover, DNA microarray results identified two phylogenetic lineages among ST-4 strains tested. Thereafter, the ST-1 and -4 isolates were phenotypically compared. Differences were noted based on the phenotypes expressed by these isolates. The ST-1 PIF isolates produced stronger biofilms at both 28°C and 37°C, while the ST-4 clinical isolates exhibited greater swimming activity and increased binding to Congo red dye. Given the fact that PIF is a low-moisture environment and that the clinical environment provides for an interaction between the pathogen and its host, these differences may be consistent with a form of pathoadaptation. These findings help to extend our current understanding of the epidemiology and ecology of Cronobacter species in PIF production environments. The epidemiological link between Cronobacter infection in neonates and contaminated powdered infant formula (PIF) has been previously established (4, 5), with C. sakazakii sequence type 4 (ST-4) being linked to cases of meningitis (6). Outbreaks have been associated with contaminated food products and the presence of this bacterium in PIF production environments (5, 7).In order to rapidly and accurately characterize Cronobacter species in PIF and its associated environments, several molecularbased protocols have been developed, which include direct target gene detection and subtyping methods (8-20). Pulsed-field gel electrophoresis (PFGE) is an accepted method for tracking isolates across the food chain, and this approach is generally considered suitable for epidemiological studies (12,(21)(22)(23)(24)(25)(26)(27)(28). A multilocus sequence typing (MLST) scheme for Cronobacter species was developed, which focuses on single nucleotide polymorphisms associated with seven housekeeping genes (including atpD, fusA, glnS, gltB, gyrB, infB, and pps) and identifies their associated alleles (29). This protocol has been used to describe some of the diversity related to the genus (6, 29-31). Both PFGE and MLST have been widely applied to study the genomic diversity of Cronobacter isolated from manufacturing facilities, commercial PIF, and follow-up formula, as we...

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Section: Discussionsupporting

confidence: 92%

Comparative Genotypic and Phenotypic Analysis of Cronobacter Species Cultured from Four Powdered Infant Formula Production Facilities: Indication of Pathoadaptation along the Food Chain

Yan

Wang

Gangiredla

et al. 2015

Appl Environ Microbiol

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“…strains isolated from samples of retail foods and 29 strains isolated from dust samples obtained from vacuum cleaners in homes, offices, and nurseries were further characterized. The strains were isolated from October 2010 to October 2012 in accordance with the reference standard Č SN P ISO/TS 22964 (Mozrová et al, 2014). The phenotypic characteristics of the isolates were determined using an ENTEROtest 24 commercial kit and the program TNW ProAuto 7.0 (Erba Lachema Ltd., Brno, Czech Republic) according to the manufacturer's instructions.…”

Section: Bacterial Strains and Phenotypic Characteristicsmentioning

confidence: 99%

“…In our previous study, 20 Cronobacter spp. strains were isolated from 291 food samples, and 52 strains were isolated from dust samples obtained from homes, restaurants, bars, and hotels (Mozrová et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

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Multilocus Sequence Typing of Cronobacter Strains Isolated from Retail Foods and Environmental Samples

Killer

Skřivanová

Hochel

et al. 2015

Foodborne Pathogens and Disease

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Cronobacter spp. are bacterial pathogens that affect children and immunocompromised adults. In this study, we used multilocus sequence typing (MLST) to determine sequence types (STs) in 11 Cronobacter spp. strains isolated from retail foods, 29 strains from dust samples obtained from vacuum cleaners, and 4 clinical isolates. Using biochemical tests, species-specific polymerase chain reaction, and MLST analysis, 36 strains were identified as Cronobacter sakazakii, and 6 were identified as Cronobacter malonaticus. In addition, one strain that originated from retail food and one from a dust sample from a vacuum cleaner were identified on the basis of MLST analysis as Cronobacter dublinensis and Cronobacter turicensis, respectively. Cronobacter spp. strains isolated from the retail foods were assigned to eight different MLST sequence types, seven of which were newly identified. The strains isolated from the dust samples were assigned to 7 known STs and 14 unknown STs. Three clinical isolates and one household dust isolate were assigned to ST4, which is the predominant ST associated with neonatal meningitis. One clinical isolate was classified based on MLST analysis as Cronobacter malonaticus and belonged to an as-yet-unknown ST. Three strains isolated from the household dust samples were assigned to ST1, which is another clinically significant ST. It can be concluded that Cronobacter spp. strains of different origin are genetically quite variable. The recovery of C. sakazakii strains belonging to ST1 and ST4 from the dust samples suggests the possibility that contamination could occur during food preparation. All of the novel STs and alleles for C. sakazakii, C. malonaticus, C. dublinensis, and C. turicensis determined in this study were deposited in the Cronobacter MLST database available online ( http://pubmlst.org/cronobacter/).

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“…sakazakii is an opportunistic pathogen causing life-threatening infections in neonates. The pathogen can be isolated from clinical, environmental, and a wide range of food sources, predominantly from PIF and its associated production environments (21,29,45,46).…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies

Scharinger

Dietrich

Kleinsteuber

et al. 2016

Appl Environ Microbiol

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Cronobacter sakazakii is a foodborne pathogen associated with rare but often lethal infections in neonates. Powdered infant formula (PIF) represents the most frequent source of infection. Out of the identified serotypes (O1 to O7), O1, O2, and O3 are often isolated from clinical and PIF samples. Serotype-specific monoclonal antibodies (MAbs) suitable for application in enzyme immunoassays (EIAs) for the rapid detection of C. sakazakii have not yet been developed. In this study, we created specific MAbs with the ability to bind to C. sakazakii of serotypes O1, O2, and O3. Characterization by indirect EIAs, immunofluorescence, motility assays, and immunoblotting identified lipopolysaccharide (LPS) and exopolysaccharide (EPS) as the antigenic determinants of the MAbs. The established sandwich EIAs were highly sensitive and were able to detect between 2 ؋ 10 3 and 9 ؋ 10 6 CFU/ml. Inclusivity tests confirmed that 93% of serotype O1 strains, 100% of O2 strains, and 87% of O3 strains were detected at low cell counts. No cross-reactivity with >100 strains of Cronobacter spp. and other Enterobacteriaceae was observed, except for that with C. sakazakii serotype O3 and Cronobacter muytjensii serotype O1. Moreover, the sandwich EIAs detected C. sakazakii in PIF samples artificially contaminated with 1 to 10 bacterial cells per 10 g of sample after 15 h of preenrichment. The use of these serotype-specific MAbs not only allows the reliable detection of C. sakazakii strains but also enables simultaneous serotyping in a simple sandwich EIA method. Cronobacter spp. are Gram-negative opportunistic foodborne pathogens of the family Enterobacteriaceae that cause rare but severe infections in patients of all age groups. In adults, Cronobacter spp. are often associated with nosocomial infections, including pneumonia, septicemia, wound infections, and osteomyelitis, while causing invasive disease in young infants and neonates (1-4). Among the seven identified Cronobacter species, C. sakazakii, C. malonaticus, C. muytjensii, C. turicensis, C. dublinensis, C. condimenti, and C. universalis (5-8), C. sakazakii plays a prominent role due to it causing life-threatening infections in neonates (9-11). Clinically manifested infections present as necrotizing enterocolitis, sepsis, and meningitis, with a mortality rate as high as 80% (1, 12, 13). Although C. sakazakii has been isolated from a variety of different plant-and animal-based food products (14, 15), the presence in powdered infant formula (PIF) seems crucial in the infection of neonates (9, 12, 16). According to an established O-antigen serotyping scheme based on rabbit antisera and a PCRbased serotyping method (17-21), seven serotypes (O1 to O7) have been identified for C. sakazakii. Serotypes O1 and O2 seem to be most prevalent in PIF samples and in clinical cases, whereas serotype O3 has been isolated quite frequently from PIF but not as often from clinical cases (19,(22)(23)(24).Today, the contamination of PIF by C. sakazakii is being detected using conventional microbiological m...

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Surveillance and characterisation of Cronobacter spp. in Czech retail food and environmental samples

Cited by 20 publications

References 27 publications

Comparative Genotypic and Phenotypic Analysis of Cronobacter Species Cultured from Four Powdered Infant Formula Production Facilities: Indication of Pathoadaptation along the Food Chain

Comparative Genotypic and Phenotypic Analysis of Cronobacter Species Cultured from Four Powdered Infant Formula Production Facilities: Indication of Pathoadaptation along the Food Chain

Multilocus Sequence Typing of Cronobacter Strains Isolated from Retail Foods and Environmental Samples

Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies

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