A simple and sensitive aptasensor with rolling circle amplification for viable Cronobacter sakazakii detection in powdered infant formula
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Cronobacter sakazakii
Necrotizing Enterocolitis
Cronobacter sakazakii
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Cronobacter sakazakii
Necrotizing Enterocolitis
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Clinical Question
For preterm infants, is hydrolyzed formula associated with lower rates of feeding intolerance, lower rates of necrotizing enterocolitis, and faster rates of growth compared with standard cow's milk formula?Bottom Line
Compared with standard formula, feeding preterm infants hydrolyzed formula is not associated with a lower rate of feeding intolerance, a lower rate of necrotizing enterocolitis, or with faster growth.Necrotizing Enterocolitis
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Introduction: Certain strains of Cronobacter sakazakii can cause serious invasive infections in children, mainly those less than 2 months old and fed with powdered infant formula (PIF). The infectious dose of C. sakazakii is unknown but evidence suggests that it is approximately 1000 CFU. Powdered infant formula is currently considered safe if its end-product C. sakazakii level is <1 CFU/g. In this study, we determined the lag time, generation time, and growth rate of five pooled C. sakazakii isolates to evaluate the factors affecting contamination levels in reconstituted PIF. Methods: 1.71 log CFU/ml of C. sakazakii were inoculated into 100 ml and 3000 ml of reconstituted PIF and incubated at 22 °C and 35 °C. Growth was evaluated over a 24-h period. ComBase was used for modeling. Results: In 3000 ml, the growth rate was 0.45 ± 0.02 log CFU/h with a lag phase of 3±0.05 h and generation time of 0.67 h at 22 °C, while the growth rate was 0.73 ± 0.01 log CFU/h with a lag phase of 0.45±0.03 h and generation time of 0.41 h at 35 °C. Conclusions: C. sakazakii grows rapidly in reconstituted PIF, especially at 35 °C.
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Contamination of baby foods with Cronobacter spp. is still a persisting challenge to the baby food industry. The antimicrobial activity of chitosan and lactic acid (LA) against C. sakazakii in reconstituted infant formula (RIF) and wheat-based infant cereal (WBIC) was studied. The RIF and WBIC were spiked with a cocktail of three strains of C. sakazakii (ca. 5.5 × 105 cfu/mL or g of RIF or WBIC). Chitosan concentrations of 1.5 and 2.0% resulted in 2–3 log reductions in the C. sakazakii count after 6 h handling at room temperature. LA at 0.4% concentration elicited the highest inhibitory activity, which resulted in approximately 3–4 log reductions for WBIC and RIF, respectively. Our results demonstrate that chitosan and LA are natural nontoxic substances that may have the potential to be used to inhibit C. sakazakii in RIF and baby foods. Practical Applications Chitosan and lactic acid may be used as effective antimicrobial agents to inhibit Cronobacter sakazakii growth in infant formula and baby foods.
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Milk and whey powders are commonly used ingredients in powdered infant formula (PIF) and follow-up formula (FUF). In this study, Cronobacter sakazakii and Cronobacter dublinensis both of dairy origin and a reference strain, Cronobacter muytjensii, ATCC 51329 were investigated for thermal inactivation (D and z values). Heat resistance of the pathogen was studied between 52 and 60 °C in trypic soy broth. Among the strains, C. muytjensii ATCC 51329 was the most heat-resistant strain at lower temperatures (52 and 54 °C). The D-values at 52 and 54 °C of C. muytjensii ATCC 51329 were 33.30 (±1.17) and 6.79 (±1.05) min, respectively. At higher temperatures, e.g., 56, 58, and 60 °C, C. sakazakii strains 131 and 807 exhibited the highest D values. The D-values at 56 and 58 °C were 4.73 (±0.40) min for C. sakazakii 131 and 2.30 (±0.26) min for C. sakazakii 807. The D-values at 60 °C for C. sakazakii 131 and 807 were 1.17 (±0.03) and 1.14 (±0.02) min, respectively. The z-value of the reference strain (C. muytjensii ATCC 51329) was lower than the other strains. The findings of this study will be very helpful in understanding the heat resistance of C. sakazakii and guarding against this pathogen in PIF and FUF. Practical applications The results presented will play a significant role in eliminating Cronobacter from powdered infant formula and follow-up formula and ensuring safety against this pathogen.
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Necrotizing Enterocolitis
Cronobacter sakazakii
Enterocolitis
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Background: Cronobacter species is the most significant foodborne pathogens in infant milk formula (IMF). These pathogens have been incriminated in sever form of neonatal meningitis, sepsis and necrotizing enterocolitis with high mortality rate. Aim: The current study was performed to demonstrate the effect of heat stress on Cronobacter spp. (C. sakazakii and C. pulveris) in reconstituted IMF (RIMF). Methods: The reconstituted formula was inoculated with five C. sakazakii isolates and four C. pulveris isolates separately. The nine isolates of Cronobacter spp. were heated in RIMF at 48, 52, 56, 60, 64 and 66°C. The D- and z-values were determined by using linear regression analysis. Results: The D˗values of all isolates of C. sakazakii (CS1, CS3, CS4, CS5 and CS6) at 48°C, 52°C, 56°C, 60°C, 64°C, 66°C ranged from 7.29 ˗ 23.47, 2.77˗15.50, 0.62˗1.04, 0.62˗1.02, 0.62˗1.00, 0.62˗1.00 min, respectively, while the z˗ values extended from 2.50˗4.28°C. The D˗ values of C. pulveris isolates (CP1, CP2, CP3, CP4) ranged from 7.60 ˗ 22.32, 1.42 ˗ 8.45, 0.62 ˗ 1.08, 0.62 ˗ 0.78, 0.62 ˗ 0.78, 0.62 ˗ 0.79 min. at 48°C, 52°C, 56°C, 60°C, 64°C, 66°C, respectively and the calculated z-values ranged from 3.33 ˗ 4.89°C. Conclusion: This study may contribute to improve the understanding of behavior of C. sakazakii and C. pulveris isolates in RIMF at various heat stress temperature and may participate in effective control of this pathogens in infant’s foods production.
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Necrotizing Enterocolitis
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Enterobacter sakazakii may be related to outbreaks of meningitis, septicemia, and necrotizing enterocolitis, mainly in neonates. To reduce the risk of E. sakazakii in baby foods, thermal characteristics for Korean E. sakazakii isolates were determined at 52, 56, and 60 degrees C in saline solution, rehydrated powdered infant formula, and dried baby food. In saline solution, their D-values were 12-16, 3-5, and 0.9-1 min for each temperature. D-values increased to 16-20, 4-5, and 2-4 min in rehydrated infant formula and 14-17, 5-6, and 2-3 min in dried baby food. The overall calculated z-value was 6-8 for saline, 8-10 for powdered infant formula, and 9-11 for dried baby food. Thermal inactivation of E. sakazakii during rehydration of powdered infant formula was investigated by viable counts. Inactivation of cultured E. sakazakii in infant formula milk did not occur for 20 min at room temperature after rehydration with the water at 50 degrees C and their counts were reduced by about 1-2 log CFU/g at 60 degrees C and 4-6 log CFU/ml with the water at 65 and 70 degrees C. However, the thermostability of adapted E. sakazakii to the powdered infant formula increased more than two times. Considering that the levels of E. sakazakii observed in powdered infant formula have generally been 1 CFU/100 g of dry formula or less, contamination with E. sakazakii can be reduced or eliminated by rehydrating water with at least 10 degrees C higher temperature than the manufacturer-recommended 50 degrees C.
Cronobacter sakazakii
Necrotizing Enterocolitis
Food microbiology
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