Including Infant Formula Research Articles

Advances in our understanding and distribution of the Cronobacter genus in China.

This review considers how research in China has progressed our understanding and subsequent improved control of Cronobacter. This emergent bacterial pathogen is associated with neonatal infections through the ingestion of contaminated prepared feed. The review includes large-scale surveys of various sources of the organism, including infant formula production facilities. The analysis of over 20,000 samples is presented. Over 10,000 being from powdered infant formula and other infant foods as well as environmental sampling of production facilities, the remaining being from food, food ingredients, and human carriage. A major advance in China was adopting DNA-sequence-based methods (that is, multilocus sequence typing, clustered regularly interspaced short palindromic repeats-cas array profiling, and single-nucleotide polymorphism analysis) for the identification and genotyping of the organism. These methods have considerably advanced our understanding of the taxonomy, ecology, and virulence of this organism. In turn, this has improved source tracking of the organism both in infant formula production facilities and epidemiological investigations. Furthermore, whole-genome sequencing has revealed a range of virulence and persistence mechanisms as well as plasmid-borne multidrug resistance traits. China now has reliable and robust methods for accurate microbial source tracking of Cronobacter for use both in the food production environment and epidemiological analysis.

Assessment of intestinal absorption/metabolism of 3-chloro-1,2-propanediol (3-MCPD) and three 3-MCPD monoesters by Caco-2 cells

3-chloro-1,2-propanediol (3-MCPD) and 3-MCPD esters are contaminants present in a variety of processed foods, including infant formulas. Toxicological data are unavailable in humans, but rodent studies have demonstrated renal and testicular toxicity from 3-MCPD and 3-MCPD esters. There is evidence that 3-MCPD esters are hydrolyzed in the digestive system, releasing 3-MCPD that would be absorbed and induce damage. We assessed absorption and metabolism of 3-MCPD and three 3-MCPD monoesters, 1-oleoyl (1-Ol), 1-linoleoyl (1-Li) and 1-palmitoyl (1-Pa) commonly found in U.S. infant formula using differentiated Caco-2 cells. After 1-hour incubation, all three monoesters released free 3-MCPD and free fatty acids (FFA) into Caco-2 cell supernatants. Free 3-MCPD had a high apparent permeability (Papp = 30.36 ± 1.31 cm/s × 10−6) suggesting that it is freely diffusible and highly absorbed by intestinal epithelium. 1-Li released 3–4-fold more 3-MCPD than 1-Ol and 1-Pa over 1 h, suggesting that this variable release rates might contribute to the overall in vivo exposure to 3-MCPD. None of the monoesters or FFA were detected in basolateral supernatants, suggesting that these compounds do not cross the intestinal wall without further transformation. In summary, this study provides relevant data to advance knowledge of in vivo intestinal absorption and metabolism of 3-MCPD monoesters.

The Overview of Food Technology to Process Soy Protein Isolate and Its Application toward Food Industry

Soy protein isolate (SPI) is the purest form of protein from soybean with minimum protein content of 90%. Due to its high protein content, SPI is commonly used in food processing for improving the quality of food products, including infant formula. The use of SPI in infant formula is mainly designed for infant who cannot tolerate cow’s milk-based formula. This report reviews the benefit of using SPI in soy-based infant formula rather than soymilk from whole soybean itself. It will also review the technology of soy protein isolation which can result SPI for high quality infant formula, including the reducing of unfavourable ingredients which will ensure the safety of soy protein-based infant formula.

Chemical Sensor Based on Molecularly Imprinted Organic-Organic Nanocomposite Film for the Analysis of Melamine in Infant Formula

Melamine (MA) is non-edible synthetic chemical intentionally added to foodstuffs such as milk and milk products including infant formula as the fake source of protein. Many evidences confirmed that this molecule is primarily responsible for kidney failure by forming kidney stones and then blocking the renal tubes. Therefore, a simple, sensitive and rapid method for online quantification of MA can avoid the consumption of milk products contaminated with MA and possible health risks resulted both socially and economically. The present work reports the development of a chemical sensor from aniline (ANI) and acrylic acid (AA) copolymer films by using concurrent electrochemical polymerization method using MA as a template. After optimization of important parameters and template removal, the performance of the film formed was studied by cyclic and differential pulse voltammetry. The proposed sensor showed a linear current response with various concentrations of MA. The linear range, limit of quantification and limit of detection were 0.1-200 x10-9 M, 5.73x10-11M, 1.72 x10-11M respectively. The sensitivity and selectivity of the sensor achieved may attribute to the synergistic effect of different functional units found on the polymer composite. These functional units competitively bind the template with non-covalent interactions. The sensor was also applied to analyze milk samples and obtained a recovery of 95.87-105.63% with RSD 1.11-2.23%. The good stability and reusability as well as strong affinity to the imprint molecule make the imprinted copolymers the promising functional materials for future sensor development.

The relationship between joining a US free trade agreement and processed food sales, 2002-2016: a comparative interrupted time-series analysis.

To examine changes in sales of highly processed foods, including infant formulas, in countries joining free trade agreements (FTAs) with the US. Annual country-level data for food and beverage sales come from Euromonitor International. Analyses are conducted in a comparative interrupted time-series (CITS) framework using multivariate random-effects linear models, adjusted for key confounders: gross domestic product (GDP) per capita, percent of the population living in urban areas and female labor force participation rate. Memberships in other FTAs and investment treaties are also explored as possible confounders. Changes are assessed between 2002 and 2016. Ten countries joining US FTAs are compared with eleven countries without US FTAs in force; countries are matched on national income level, world region and World Trade Organization membership. After countries join a US FTA, sales are estimated to increase by: 0·89 (95 % CI 0·16, 1·6; P = 0·016) kg per capita per annum for ultra-processed products, 0·81 (95 % CI 0·47, 1·1; P < 0·001) kg per capita per annum for processed culinary ingredients and 0·17 (95 % CI 0·052, 0·29; P = 0·005) kg per capita under age 5 per annum for baby food. No significant change is estimated for minimally processed foods. In statistical models, large unexplained variations in country-specific trends suggest additional unmeasured country-level factors also impact sales trends following entry into US FTAs. These findings strongly support the conclusion that joining US FTAs can contribute to detrimental changes in national dietary consumption that increase population risk of non-communicable diseases.

Omics analysis reveals variations among commercial sources of bovine milk fat globule membrane

Milk fat globule membrane (MFGM) is a glycosylated, protein-embedded, phospholipid fraction that surrounds triglycerides in milk. Commercial bovine sources have recently come to the market as a novel food ingredient and have been added to various products, including infant formula. Considering that MFGM is a heterogeneous mixture of fat, protein, and carbohydrate, it can be expected that variations among MFGM products exist. For this reason, our aim was to characterize the composition of commercial MFGM samples through a combination of proteomic and lipidomic analyses. Six bovine milk fractions, represented as MFGM fractions or phospholipid fractions, were obtained from various commercial sources. Additionally, the MFGM samples were compared with 2 infant formulas, a standard formula as well as a premium formula containing MFGM. For proteomic analysis, bottom-up data-dependent liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on each MFGM fraction, and nearly a thousand proteins were identified across all samples, with 364 of them having different abundance across the samples tested. One hundred twelve proteins differed by a fold-change of 10 or greater, 14 by a fold-change of 50, and 2 by a fold-change of 100 in at least 1 pair, suggesting large differences in the proteins present in these fractions. Even though the classical MFGM proteins were enriched in the MFGM fractions, the relative protein composition varied considerably, and all contain an abundance of milk (casein and whey) proteins. Lipidomic analysis identified a total of 393 lipid species across both positive and negative ionization modes, with the major classes detected being triglycerides, sphingomyelins, and several phospholipids. Across all samples, triglycerides comprised at least 50% of total lipids, with phosphatidylcholine and sphingomyelin being the second and third most abundant lipid classes, respectively. These findings demonstrate the heterogeneous nature of various bovine commercial MFGM fractions. This variation must be considered when evaluating and describing potential functional benefits of these products shown in clinical trials.

Arsenic in baby foods: health effects and dietary exposure

Health effects of arsenic in adults have been studied and documented more extensively than in infants and children, despite the fact that the younger population is more sensitive and is more exposed. Although rice has the highest concentrations of inorganic arsenic among foods, it is widely consumed by infants and children. The present article discusses the latest global literature on the presence of inorganic arsenic and total arsenic in foods consumed by infants, focusing on the adverse chronic health effects as well as dietary exposure and risk assessment in this population. It also discusses international regulations related to arsenic in rice and other products consumed by infants. Finally, it reviews mitigation strategies to reduce arsenic intake or its effects. It was concluded that the limits regarding iAs in rice-based products need to be included in the Codex Alimentarius standards since these standards are currently used by countries that have not developed their own regional or national regulations. Early-life chronic exposure to arsenic has the potential to affect several systems of the human body, and while some of these effects are likely to remain latent, others become evident in the short term. Worldwide, numerous foods intended for infants and children, including infant formula, contain high levels of iAs, particularly rice and rice-based products.

Nutritional composition of processed baby foods targeted at infants from 0–12 months

In the last decade, there has been an increasing demand for ready-to-eat infant meals in Spain. However, the used food composition databases do not include processed products intended for infants under 12 months of age. Thus, we aimed at the creation of a food composition table including these products. Nutritional composition data were collected from the label of 568 products including infant formula, cereal products and complementary foods (mixed puree or snacks and desserts); additionally, breast-milk (11 preterm and term samples) nutritional composition were estimated from scientific publications available to date. Information was compiled into a table available in “open access” at the website http://ucc.uniovi.es/formacioncientifica/recursosinv including information about 53 dietary components of infant formula, 28 for breast milk, 30 for cereal products and 14 for complementary foods. The infant formula´s composition from 0 to 6 months were found similar to the values reported for mature breast milk (10–12 weeks) but different from breast milk from 1 to 4 weeks. One serving of cereals has been found to satisfy more than the 50% of the DRI for protein and carbohydrates. This information constitutes an essential step in order to understand the diet-health relationships in the early stages of life in the Spanish population.

Federal Regulation of Infant and Toddler Food and Drink Marketing and Labeling.

Foods and beverages marketed for infants, babies, and toddlers through 3 years of age is a $7 billion industry in the United States, incorporating a wide range of products, including infant formula and other types of drinks, foods, and snacks. The World Health Organization (“WHO”) found that mothers “are often inundated with incorrect and biased information” from direct advertising, health claims on products, information packs from sales representatives, and the distribution of samples of infant formula and “educational materials” by infant formula manufacturers. To address these problematic practices, in 1981, the WHO established the International Code of Marketing of Breast-milk Substitutes (the “Code”) to end the inappropriate marketing of infant formula and other food and drinks intended for children up to age two. In 2016, WHO expanded the definition of breastmilk substitutes to include milk and milk products specifically marketed for feeding infants and young children up to age three. However, the United States is one of a minority of countries that has not passed any legislation or regulation to implement the Code. Furthermore, U.S. regulation and enforcement actions have not kept pace with the introduction of new products and product categories and the profusion of labeling and marketing claims questionably implying nutritional and developmental benefits from these products.

Method development and validation for total haloxyfop analysis in infant formulas and related ingredient matrices using liquid chromatography-tandem mass spectrometry.

According to the European Commission directive 2006/141/EC, haloxyfop residue levels should not exceed 0.003mg/kg in ready-to-feed infant formula, and the residue definition includes sum of haloxyfop, its esters, salts, and conjugates expressed as haloxyfop. A simple method for total haloxyfop analysis in infant formula and related ingredient matrices was developed and validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The sample preparation consisted of an alkaline hydrolysis with methanolic sodium hydroxide to release haloxyfop (parent acid) from its bound forms prior to the extraction with acetonitrile. A mixture of magnesium sulfate (MgSO4) and sodium chloride (NaCl) (4:1, w/w) was added to the extract to induce phase separation and force the analyte into the upper acetonitrile-methanol layer and then a 1-mL aliquot was subsequently cleaned up by dispersive solid phase extraction with 150mg of MgSO4 and 50mg of octadecyl (C18) sorbent. The analytical procedure was developed and carefully optimized to enable low-level, total haloxyfop analysis in a variety of challenging matrices, including infant formulas and their important high-carbohydrate, high-protein, high-fat, and emulsifier ingredients. The final method was validated in two different laboratories by fortifying samples with haloxyfop and haloxyfop-methyl, which was used as a model compound simulating bound forms of the analyte. Mean recoveries of haloxyfop across all fortification levels and evaluated matrices ranged between 92.2 and 114% with repeatability, within-lab reproducibility, and reproducibility RSDs ≤14%. Based on the validation results, this method was capable to convert the haloxyfop ester into the parent acid in a wide range of sample types and to reliably identify and quantify total haloxyfop at the target 0.003mg/kg level in infant formulas (both powdered and ready-to-feed liquid forms). Graphical abstract LC-MS/MS-based workflow for the determination of the total haloxyfop in infant formula and related ingredients.

Structural strength analysis of amorphous trehalose-maltodextrin systems

Fundamental knowledge of physical state of materials gives practically important information for food, biological and pharmaceutical industry. Based on Williams-Landel-Ferry (WLF) equation, the strength concept was introduced. This concept provides a simple parameter, S, to express resistance of solids to flow above the glass transition temperature. To develop this approach, miscible trehalose-maltodextrin (0:100; 20:80; 40:60; 60:40; 80:20 and 100:0) systems with different ratios of components were used in the present study. Such systems represent various food products including infant formula and many nutritional formulations. Amorphous solids were prepared from 20% solids in water solutions by freeze-drying. Fractional water sorption analysis of trehalose-maltodextrin miscible systems allows control of water content at high water activities. Glass transition temperatures were measured by DSC. DMA and DEA in a multi-frequency mode allowed determination of corresponding α-relaxation temperatures at various structural relaxation times. Volume rheology gives structural relaxation time – temperature dependence for high water content systems. The strength showed linear dependence on maltodextrin concentration and its value decreased significantly with increasing water content in miscible systems.

Furan in commercial baby foods from the Spanish market: estimation of daily intake and risk assessment

ABSTRACTThe occurrence of furan in commercial baby food samples from the Spanish market was evaluated using an automated headspace solid-phase microextraction method coupled to gas chromatography-mass spectrometry (HS-SPME-GC-MS). A total of 76 baby food samples including infant formula, baby cereals, fruit in cans and/or jars, vegetables, meat, and fish, were surveyed for furan content. The lowest concentration of this compound was found in infant formula (<0.02–0.33 ng ml−1), and cereal-based food (0.15–2.1 ng g−1) while baby food containing fish showed the highest concentrations (19–84 ng g−1). Following recommendation of the European Food Safety Authority (EFSA), the effect on furan content was evaluated of consumer home preparation of foods, heating and handling. Furan concentrations were reduced by up to 35% when samples were heated in a dish using microwave oven and by up to 53% when a hot water bath was used. Finally, we estimated the furan intake from baby food consumption (0.002–1.18 µg kg−1 body weight day−1) and we calculated the margin of exposure (MOE) from samples as purchased and also after home preparation of the food. For infant formula and cereal baby foods, the MOEs (26,278–412,776) indicated no infant health concern or priority, while for meat and fish-based baby foods the values pointed to a potential public health risk, even considering the furan losses during preparation at home.

Quantitative analysis of cow whole milk and whey powder adulteration percentage in goat and sheep milk products by isotopic dilution-ultra-high performance liquid chromatography-tandem mass spectrometry.

The aim of the study was to develop a method for quantification of cow's whey and whole milk powder in goat or sheep milk products including infant formula. A ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for simultaneous quantification of four caseins and two major whey proteins by detecting their signature peptides, which were able to act as markers for differentiating goat or sheep from cow whey and whole milk powder in infant formulas. The signature peptides were screened based on the computational prediction by Biolynx software, and confirmed by database searching after analysis of liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (LC-Q-TOF-MS). The isotopic-labeled signature peptide was used as internal standard to compensate the matrix effect. The limits of quantification were 0.01-0.05g/100g for target proteins. The observed recovery rates ranged from 82.3 to 116.6% and the reproducibility was excellent (RSD <12%) at different spiking levels. The RSDs of intra- and inter-day precision were 2.8-6.2 and 3.3-9.8%, respectively. The multiple reaction monitoring method was successfully applied to milk powder with different composition, showing high specificity and accuracy in detection of species involved. The calculating formula was designed to assess the composition of adulteration in the actual detection of infant formulas. These results highlight applicability of this method for the detection of infant formulas with complicated matrix.

Isotope dilution-liquid chromatography/mass spectrometric method for the determination of riboflavin content in multivitamin tablets and infant formula

Isotope dilution liquid chromatography/mass spectrometry (ID-LC/MS) was developed to measure riboflavin contents in infant formula and multivitamin tablets. 13C4, 15N2-riboflavin was used as an internal standard. For infant formula, acid hydrolysis was conducted after spiking internal standard to release bound riboflavin. Free riboflavin in the multivitamin tablets was extracted with distilled water for 3h at 4°C. The mass spectrometer was operated in selected ion monitoring mode to observe the collisionally induced dissociation channels of m/z 377→243 for riboflavin and m/z 383→249for13C4,15N2-riboflavin. The developed method was validated based on tests of repeatability and reproducibility, evaluation of measurement uncertainty, and analysis of available certified reference materials. For infant formula and multivitamin samples, the repeatability was 0.1–0.4% and 0.3–1.2%, and the reproducibility was 1.4 and 1.0%, respectively. The realtive expanded uncertainty of analyzing homogenized sample was ranging 1–4%. Therefore, the developed method showed that it has as a high-order metrological quality. The developed ID-LC/MS method was also tested for the accurate analysis of riboflavin in commercial food products including infant formula, breakfast cereals, milk, and multivitamin tablets.

Promotion and consumption of commercially produced foods among children: situation analysis in an urban setting in Senegal.

This study assessed the promotion of commercially produced foods and consumption of these products by children less than 24 months of age in Dakar Department, Senegal. Interviews with 293 mothers of children attending child health clinics assessed maternal exposure to promotion and maternal recall of foods consumed by the child on the preceding day. Promotion of breastmilk substitutes and commercially produced complementary foods outside health facilities was common with 41.0% and 37.2% of mothers, respectively, reporting product promotions since the birth of their youngest child. Promotion of commercially produced snack food products was more prevalent, observed by 93.5% of mothers. While all mothers reported having breastfed their child, only 20.8% of mothers breastfed their newborn within the first hour after delivery, and 44.7% fed pre‐lacteal feeds in the first 3 days after delivery. Of children 6–23 months of age, 20.2% had consumed a breastmilk substitute; 49.1% ate a commercially produced complementary food, and 58.7% ate a commercially produced snack food product on the previous day. There is a need to stop the promotion of breastmilk substitutes, including infant formula, follow‐up formula, and growing‐up milks. More stringent regulations and enforcement could help to eliminate such promotion to the public through the media and in stores. Promotion of commercial snack foods is concerning, given the high rates of consumption of such foods by children under the age of 2 years. Efforts are needed to determine how best to reduce such promotion and encourage replacement of these products with more nutritious foods.

Fast Determination of Sodium Monofluoroacetate in Liquid Milk and Dairy Powder by Hydrophilic Interaction Liquid Chromatography-Triple Quadrupole Mass Spectrometry

Manufactured sodium monofluoroacetate is a rodenticide with high acute toxicity. A fast and sensitive analytical method in liquid milk and dairy powder was developed both for routine analysis and for fast tackling the terrorist threat. Monofluoroacetate was extracted from liquid milk using acetonitrile and from dairy powder with water and acetonitrile, cleaned using a solid phase extraction (SPE) cleanup procedure with polymeric anion exchange (PAX) cartridge, and measured by hydrophilic interaction liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS). The conditions for the monofluoroacetate extraction, SPE cleanup, and instrument injection solvent were optimized. A BEH amide chromatographic column with hydrophilic interaction was used to directly separate monofluoroacetate under basic conditions. The limits of detection in liquid milk and dairy powder were 1 and 2 μg kg-1, respectively. The recoveries at three spiking levels (10, 50, and 200 μg kg-1) were 72.5–97.6 % with relative standard deviations (RSDs) of 4.7–5.9 % for liquid milk and 70.1–92.8 % with RSDs of 4.7–6.8 % for dairy powder. The method has been applied to analyze sodium monofluoroacetate in dairy products including infant formula.

Food sources of energy and nutrients in the diets of infants and toddlers in urban areas of China, based on one 24-hour dietary recall

Food sources of nutrients in young children in China have been little investigated. The objective of this study was to understand feeding patterns of young children through an analysis of food sources of energy and nutrients in the diets of infants and toddlers living in urban areas in China. This study was part of the cross-sectional study, Maternal Infant Nutrition Growth (MING). One 24-h dietary recall was completed for a total of 1409 children (infants 6–11 months, younger toddlers 12–23 months, and older toddlers 24–36 months) via face-to-face interviews with the primary caregivers. All food, beverage and dietary supplements that the child consumed on the previous day were recorded and processed with a database including data from Chinese Food Composition tables. All reported foods and supplements were assigned to one of 83 food groups developed for the study. Percent contribution of each food group to nutrient intakes was calculated. Children in all 3 age groups received a majority of energy (52 to 69 %) from few foods including infant formula or growing-up (fortified) milk, rice, noodles, pork and eggs, with rice becoming the number one source of energy by 24 months. Rice and noodles were not only top sources of energy (17 to 26 %) and carbohydrate (27 to 40 %) but also top sources of protein (13 to 16 %), iron (13 to 18 %) and zinc (11 to 18 %). Supplements made substantial contributions to intakes of vitamin A, zinc, iron and calcium. Salt added during home-cooking was the main source of sodium (60 to 80 %). Few foods made up the core of the diets of infants and young toddlers living in urban areas in China. Low nutrient-dense rice and noodles were top sources of many nutrients, which could potentially lead to inadequate intakes of some key nutrients.

Persistence of furan-induced epigenetic aberrations in the livers of F344 rats.

Furan is a heterocyclic organic compound produced in the chemical manufacturing industry and also found in a broad range of food products, including infant formulas and baby foods. Previous reports have indicated that the adverse biological effects of furan, including its liver tumorigenicity, may be associated with epigenetic abnormalities. In the present study, we investigated the persistence of epigenetic alterations in rat liver. Male F344 rats were treated by gavage 5 days per week with 8 mg furan/kg body weight (bw)/day for 90 days. After the last treatment, rats were divided randomly into 4 groups; 1 group of rats was sacrificed 24 h after the last treatment, whereas other groups were maintained without further furan treatment for an additional 90, 180, or 360 days. Treatment with furan for 90 days resulted in alterations in histone lysine methylation and acetylation, induction of base-excision DNA repair genes, suggesting oxidative damage to DNA, and changes in the gene expression in the livers. A majority of these furan-induced molecular changes was transient and disappeared after the cessation of furan treatment. In contrast, histone H3 lysine 9 and H3 lysine 56 showed a sustained and time-depended decrease in acetylation, which was associated with formation of heterochromatin and altered gene expression. These results indicate that furan-induced adverse effects may be mechanistically related to sustained changes in histone lysine acetylation that compromise the ability of cells to maintain and control properly the expression of genetic information.

Determination of bovine lactoferrin in dairy products by ultra-high performance liquid chromatography–tandem mass spectrometry based on tryptic signature peptides employing an isotope-labeled winged peptide as internal standard

A new and sensitive determination method was developed for bovine lactoferrin in dairy products including infant formulas based on the signature peptide by ultra high-performance liquid chromatography and triple-quadrupole tandem mass spectrometry under the multiple reaction monitoring mode. The simple pretreatment procedures included the addition of a winged peptide containing the isotope-labeled signature peptide as internal standard, followed by an enzymatic digestion with trypsin. The signature peptide was chosen and identified from the tryptic hydrolyzates of bovine lactoferrin by ultra high-performance liquid chromatography and quadrupole-time-of-flight tandem mass spectrometry based on sequence database search. Analytes were separated on an ACQUITY UPLC BEH 300 C18 column and monitored by MS/MS in seven minutes. Quantitative result bias due to matrix effect and tryptic efficiency was corrected through the use of synthetic isotope-labeled standards. The limit of detection and limit of quantification were 0.3mg/100g and 1.0mg/100g, respectively. Bovine lactoferrin within the concentration range of 10–1000nmolL−1 showed a strong linear relationship with a linear correlation coefficient (r) of >0.998. The intra- and inter-day precision of the method were RSD<6.5% and RSD<7.1%, respectively. Excellent repeatability (RSD<6.4%) substantially supported the application of this method for the determination of bovine lactoferrin in dairy samples. The present method was successfully validated and applied to determination of bovine lactoferrin in dairy products including infant formulas.

Possible reservoir and routes of transmission of Cronobacter (Enterobacter sakazakii) via wheat flour

Contamination of food products is a major route of transmission for opportunistic foodborne pathogen. Cronobacter spp. (formerly Enterobacter sakazakii) are an important group of emerging opportunistic foodborne pathogens that may cause rare but severe systemic infection such as neonatal meningitis, septicemia, and enterocolitis in neonates. Here, we tried to investigate the possible natural reservoir and routes of transmission of this pathogen. Samples of food (N = 141) were screened for the presence of the bacterium, and 34 isolates of Cronobacter spp. were collected in total. Of the initial of 108 samples including infant formulas, dry milk powder, milk and milk related beverages, cereal (products), chocolate and candy, miscellaneous food products, 12% were positive for Cronobacter spp., with the highest percentage (69.3%) coming from cereal products. Further investigation indicated that 100% of 13 wheat flours, 5 of 8 (62.5%) wheat kernel samples, 100% of 5 different brands of dry noodles, and 3 of 7 (42.9%) deep-frozen dumpling samples contained Cronobacter spp. Biochemical analysis (ID32E strips) of the 34 isolates resulted in 11 different biogroups based on the utilization of 8 substrates. In addition, pulsed field gel electrophoresis (PFGE) demonstrated extremely high genetic diversity as 24 unique PFGE profiles were revealed among the 25 isolates acquired mainly from wheat flour or related products. We therefore conclude that wheat flour is one likely reservoir and/or transmission route for Cronobacter spp., and special care should be taken while preparing infant foods or formulas in order to avoid cross-contamination from this source.