Treatment of winery effluent with upflow anaerobic sludge blanket (UASB) – granular sludges enriched with<i>Enterobacter sakazakii</i>

Keyser, M.; Witthuhn, R. Corli; Ronquest, L.-C.; Britz, T. J.

doi:10.1023/b:bile.0000003978.72266.96

Cited by 70 publications

(49 citation statements)

References 11 publications

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“…PCR was performed on 16S rRNA gene specific for Cronobacter sakazakii (20). Genomic DNA was extracted from overnight cultures.…”

Section: Pcr Assaymentioning

confidence: 99%

“…PCR reactions were set in 25µL final volumes consisting of 5 µL of the bacterial genome DNA, 1 × buffer, 1.5 U Taq polymerase (Fermentas, Lithuania), 1.5 mM MgCl 2 , 0.8 mM dNTPs and 20 pmol primers each. Primers, Esak2-5 -CCC GCA TCT CTG CAG GAT TCT C-3 and Esak3 -5 -CTA ATA CCG CAT AAC GTC TAC G-3 were used (59°C annealing temperature) in PCR to amplify an 854 bp fragment to 16S rRNA gene (20). The PCR was carried out with initiation at 94°C for 5 minutes, then 30 cycles of denaturation at 94°C for 30 seconds, annealing at 59°C for 30 seconds and extension at 72°C for 2 minutes, followed by a terminal extension step of 72°C for 5 minutes.…”

Section: Pcr Assaymentioning

confidence: 99%

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Study of Cronobacter sakazakii Strains Isolated from Powdered Milk Infant Formula by Phenotypic and Molecular Methods in Iran

Mardaneh

Dallal

2016

Arch Pediatr Infect Dis

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Background: Cronobacter is a genus within the family Enterobacteriaceae and was previously known as Enterobacter sakazakii. Cronobacter sakazakii is an opportunistic ubiquitous bacterium and is identified throughout the world as an emerging food-borne pathogen. This pathogen causes meningitis, necrotizing enterocolitis (NEC), and sepsis (septicemia) in patients hospitalized in neonatal intensive care units and has a high mortality and morbidity rate. Contamination of powdered infant milk formula occurs more easily because it is a nonsterilized product. Objectives: The aims of the present study were as follows: Isolation of Cronobacter sakazakii from powdered infant formula milk (PIF), confirmation of isolates by biochemical tests and polymerase chain reaction (PCR) molecular method, and characterization of an antibiotic susceptibility profile. Methods: A cross-sectional study was performed on 125 powdered infant formula milk (PIF) samples purchased from hospital drug stores between June 2014 and March 2015. Cronobacter sakazakii was recovered according to FDA protocol. For the final confirmation of isolates, different biochemical tests embedded in the API-20E kit system and manual biochemical tests were used according to the directions of the manufacturer. Antibiotic susceptibility testing was performed on Mueller-Hinton agar by disk diffusion method according to CLSI recommendations. For molecular confirmation of isolates confirmed by biochemical tests, PCR was performed on 16S rRNA gene specific for Cronobacter sakazakii. Results: Out of the 125 PIF samples investigated, nine (7.2%) of the samples were positive for Cronobacter sakazakii. All C. sakazakii strains isolated from PIF samples were uniformly susceptible to ticarcillin, chloramphenicol, levofloxacin, ciprofloxacin, moxifloxacin, cefepime, piperacillin-tazobactam, and colistin. All isolates were confirmed by PCR. Conclusions: Results showed that PIF that is being consumed in Iran is contaminated with this pathogen and can cause disease in infants; especially those hospitalized in neonatal intensive care units (NICUs) and fed PIF. The number of reported cases of Cronobacter infections is very low, but nevertheless there has been a slight increase recently. While the reported cases worldwide are few, it needs to be noted that the number of infections may be underestimated since not all clinical analysis laboratories carry out research on other bacterial pathogens, and not all countries have a system for reporting diseases.

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“…PCR was performed on 16S rRNA gene specific for Cronobacter sakazakii (20). Genomic DNA was extracted from overnight cultures.…”

Section: Pcr Assaymentioning

confidence: 99%

Section: Pcr Assaymentioning

confidence: 99%

Study of Cronobacter sakazakii Strains Isolated from Powdered Milk Infant Formula by Phenotypic and Molecular Methods in Iran

Mardaneh

Dallal

2016

Arch Pediatr Infect Dis

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“…Four PCR primer pairs were used as given below, using 2.5 U GoTaq Flexi DNA polymerase enzyme (Promega Corporation, WI), 5x Green GoTaq Flexi buffer (Promega Corporation, WI), and a Genius thermocycler (Techne Ltd., UK). Keyser et al [19] primer pairs Esak2 and Esak3 were used to amplify an 850-bp PCR product the 16S rDNA gene. Lehner et al [20] primer pairs Esakf and Esakr were used to amplify 929-bp PCR product from the 16S rDNA gene.…”

Section: Polymerase Chain Reaction (Pcr) Probes For Cronobacter Spp mentioning

confidence: 99%

Comparison of methods for the microbiological identification and profiling of Cronobacter species from ingredients used in the preparation of infant formula

Cetinkaya

Joseph

Ayhan

et al. 2013

Molecular and Cellular Probes

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Cronobacter spp. (formerly Enterobacter sakazakii) can be isolated from a wide range of foods and environments, and its association with neonatal infections has drawn considerable attention from regulatory authorities. The principle route of neonatal infection has been identified as the ingestion of contaminated infant formula. A number of methods have been developed to identify Cronobacter spp., however these were before the most recent (2012) taxonomic revision of the genus into seven species. In this study, phenotyping, protein profiling and molecular methods were used to identify Cronobacter strains which had been recently isolated from ingredients used in the preparation of infant formula.Pulsed field gel electrophoresis revealed that different Cronobacter strains had been recovered from the same food products. All isolates were identified as C sakazakii according to four genus specific PCR probes and protein profiling using MALDI-TOF analysis.However, 16S rDNA sequence analyses and fusA allele sequencing gave more accurate identification: four strains were C sakazakii, one strain was C malonaticus and the remaining strain was C universalis. Multilocus sequence typing showed the strains were different sequence types.These results demonstrate the presence of different Cronobacter species in food ingredients used in the preparation of infant formula, and also the need for molecular identification and profiling methods to be revised according to taxonomic revisions.

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“…However 16S rRNA gene sequence analysis revealed that it was E. cloacae and had been misidentified. Several methods for the identification of E. sakazakii via PCR have been described and were used in this study (19,20,22). PCR products of the indicative size were obtained when genomic DNAs from E. sakazakii strains NCTC 11467 T and ATCC 12868 were probed as previously described.…”

Section: Patientsmentioning

confidence: 99%

Genotypic and Phenotypic Analysis of Enterobacter sakazakii Strains from an Outbreak Resulting in Fatalities in a Neonatal Intensive Care Unit in France

et al. 2007

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In 1994, an outbreak of Enterobacter sakazakii infections occurred in a neonatal intensive care unit in France from 5 May to 11 July. During the outbreak, 13 neonates were infected with E. sakazakii, resulting in 3 deaths. In addition, four symptomless neonates were colonized by E. sakazakii. The strains were subjected to 16S rRNA gene sequence analysis, genotyped using pulsed-field gel electrophoresis, and phenotyped for a range of enzyme activities. E. sakazakii was isolated from various anatomical sites, reconstituted formula, and an unopened can of powdered infant formula. A fourth neonate died from septic shock, attributed to E. sakazakii infection, during this period. However, 16S rRNA gene sequence analysis revealed that the organism was Enterobacter cloacae. There were three pulsotypes of E. sakazakii associated with infected neonates, and three neonates were infected by more than one genotype. One genotype matched isolates from unused prepared formula and unfinished formula. However, no pulsotypes matched the E. sakazakii strain recovered from an unopened can of powdered infant formula. One pulsotype was associated with the three fatal cases, and two of these isolates had extended-spectrum ␤-lactamase activity. It is possible that E. sakazakii strains differ in their pathogenicities, as shown by the range of symptoms associated with each pulsotype.Enterobacter sakazakii is an opportunistic pathogen associated with the ingestion of reconstituted infant formula and is a rare cause of neonatal meningitis, necrotizing enterocolitis (NEC), and sepsis (9, 10, 11, 23). Such cases often occur among low-birth-weight preterm neonates, who are generally more susceptible to gram-negative bacterial sepsis and endotoxemia associated with NEC (1, 26). The International Commission on Microbiological Specifications for Foods (14) has ranked E. sakazakii as a "severe hazard for restricted populations, life-threatening or substantial chronic sequelae or long duration." A number of reported E. sakazakii outbreaks have been attributed to contaminated reconstituted infant formula (4,7,13,18,31). Bowen and Braden (4) reviewed 46 cases of invasive E. sakazakii infections and showed a link between symptoms and birth weight but did not consider cases of NEC.The virulence of E. sakazakii has been studied by Pagotto et al. (23) and Mange et al. (21), who showed the presence of enterotoxins and adhesion to brain cells, respectively. Townsend et al. demonstrated the translocation of E. sakazakii and other intestinal bacteria across the rat intestinal wall in response to the presence of lipopolysaccharide (28). They also demonstrated that E. sakazakii causes chronic-patterned inflammation in the neonatal rat brain, invades capillary endothelial brain cells, is taken up by macrophages, and induces anti-inflammatory cytokine (interleukin-10) expression in vitro and in vivo at various levels according to strain (29). However, these publications did not report the individual case details associated with the isolates under study. Therefor...

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Treatment of winery effluent with upflow anaerobic sludge blanket (UASB) – granular sludges enriched withEnterobacter sakazakii

Cited by 70 publications

References 11 publications

Study of Cronobacter sakazakii Strains Isolated from Powdered Milk Infant Formula by Phenotypic and Molecular Methods in Iran

Study of Cronobacter sakazakii Strains Isolated from Powdered Milk Infant Formula by Phenotypic and Molecular Methods in Iran

Comparison of methods for the microbiological identification and profiling of Cronobacter species from ingredients used in the preparation of infant formula

Genotypic and Phenotypic Analysis of Enterobacter sakazakii Strains from an Outbreak Resulting in Fatalities in a Neonatal Intensive Care Unit in France

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