Enterobacter sakazakii found rarely in infant formula

The Centre for Food Safety (CFS) recently conducted a targeted food surveillance to assess the situation of Enterobacter sakazakii in powdered infant formula. "Enterobacter sakazakii can be found in the environment, but it generally causes disease only in people with weakened immune systems. Enterobacter sakazakii can cause invasive infection such as bacteria in blood or brain infection.

Pre-term infants, neonates (28 days old or below), infants less than 2 months of age, low-birth-weight infants (below 2.5 kg) and infants with weakened immunity, are at greater risk," a CFS spokesman said today (December 21).  The centre collected 100 samples of powdered infant formula from the local market for testing of Enterobacter sakazakii. The samples, which involve 17 different brands, are from different countries.

All results were satisfactory. "The World Health Organisation recommends that infants should be exclusively breastfed for the first six months of life to achieve optimal growth, development and health. For those who are not breastfed, risk of infection can be greatly reduced by correctly preparing and storing the powdered infant formula.

Reconstitution of powdered infant formula with water that is no less than 70 degrees Celsius can significantly inactivate Enterobacter sakazakii. Feeds prepared in advance should be cooled immediately after preparation, stored in a refrigerator and used within 24 hours of preparation. Reconstituted feed should be re-warmed, no more than 15 minutes, immediately before feeding," the spokesman said.

"For high-risk infants who cannot be breastfed, caregivers should use commercially sterile liquid formula," he added.  The spokesman also urged the trade to implement preventive measures (such as good manufacturing practice) as well as monitoring and environmental management programmes to reduce the risk of product contamination.

The Naming Of Cronobacter Sakazakii

"Classifications are theories about the basis of natural order, not dull catalogues compiled only to avoid chaos." Stephen Jay Gould, Wonderful Life (1989), 98.

Enterobacter sakazakii, a gram-negative bacillus, is a rare cause of bloodstream and central nervous system infections. In 2007, following extensive study, it was proposed that the original taxonomy of Enterobacter sakazakii be revised, to consist of five new species moved to a new genus, identified as "Cronobacter". (1) A review of the what, the how, and the why the change was first proposed, and why it was eventually approved, provides an insight into the related scientific process of taxonomy at work, involving this notorious neonatal pathogen.

Initially, taxonomy is the science of classifying organisms, identifying and naming species, and organizing them into systems of classification. At least 1.7 million species of living organisms have been discovered, and the list grows longer every year. Ideally, classification should be based on homology, i.e., the shared characteristics that have been inherited from a common ancestor. Until recent decades, the study of homologies was limited to anatomical structures and pattern of embryonic development. However, since the birth of molecular biology, homologies can now also be studied at the level of proteins and DNA. (2)

More specifically, E. sakazakii is a rare, but life-threatening cause of neonatal meningitis, sepsis, and necrotizing enterocolitis. In general, E. sakazaii kills 40-80 % of infected newborns diagnosed with this type of severe infection. (5) E. sakazakii meningitis may lead to cerebral abscess or infarction with cyst formation and severe neurologic impairment. E. sakazakii can cause a variety of infections, though central nervous system infection has been most commonly described. (6) For infants, infection typically manifests through signs of sepsis in the first week of life: irritability or lethargy, temperature instability, and feeding intolerance. Meningitis often produces overwhelming infection that rapidly moves through cerebral hemorrhage, infarct, necrosis, liquefaction, and eventually, cyst formation. (7)

E. sakazakii invasive infections occur more frequently in infants than in older children. (9) The neonate's immature immune system may increase the risk of acquiring an E. sakazakii infection. (10) In a study of E. sakazakii cases over a 47 year period, investigators found that the median age at infection onset was two days and 94% of cases were less than 28 days old. (11)

While the reservoir for E. sakazakii is unknown in many cases, a growing number of reports have established powdered infant formula as the source and vehicle of infection. In several investigations of outbreaks of E. sakazakii infection that occurred among neonates in neonatal intensive care units, investigators were able to show both statistical and microbiological association between infection and powdered infant formula consumption. These investigations included cohort studies which implicated infant formula consumed by the infected infants. In addition, there was no evidence of infant-to-infant or environmental transmission; all cases had consumed the implicated formula. The stomach of newborns, especially of premature babies, is less acidic than that of adults: a possible important factor contributing to the survival of an infection with E. sakazakii in infants. (13)

The first cases attributed to this organism occurred in 1958 in England (Urmenyi and Franklin, 1961). Since then, up to July 2008, around 120 documented cases of E. sakazakii infection, and at least 27 deaths, have been identified from all parts of the world in the published literature and in reports submitted by public health organizations and laboratories. (12)

E. sakazakii used to be previously known as a "yellow pigmented Enterobacter cloacae", until 1980. E. sakazakii was first defined as a novel species in 1980, when it was introduced as a new species based on differences in DNA-DNA hybridization, biochemical reactions, and antibiotic susceptibility. The bacteria was named sakazakii in honour of the Japanese microbiologist, Riichi Sakazaki, when the species was first designated in 1980. (15) Enterobacter sakazakii (E. sakazakii) then became identified as one of sixteen distinct species in the genus Enterobacter, within the Enterobacteriaceae family. (14)

From the beginning, however, many different biogroups were defined as E. sakazakii, with the existence of these divergent geno- and biogroups suggesting that E. sakazakii could in fact represent multiple species. (14) Accordingly, in 2007, a research group clarified the taxonomic relationship among the various E. sakazakii strains, by using sophisticated new means of viewing and analyzing the bacteria. Iverson et al were thus able to distinguish numerous separate species. Their work resulted in the proposal of an alternative classification of E. sakazakii into a new genus, Cronobacter. (16)

The new techniques used by the research group provide clear proof of the substantial advances made in molecular biology, and included f-AFLP, automated ribotyping, full-length 16S rRNA gene sequencing and DNA-DNA hybridization. (14) F-AFLP (fluorescent amplified fragment length polymorphism) is a means to genotype bacteria, by selecting pre-adapted fragments of DNA and amplifying them to easily detectable and accurately sizeable concentrations. Automated ribotyping is a genotyping method that can be used to generate genetic fingerprints of bacterial isolates. Full-length 16S rRNA gene sequencing provides a means to compare a stable part of the genetic code (the 16S rRNA gene) amongst different bacteria. The technique of DNA-DNA hybridization provides genetic comparisons amongst the total genome of two species.

E. sakazakii has thus now been reclassified as 6 separate species in the new genus, Cronobacter, gen. nov., within the Enterobacteriaceae family. The new species are presently Cronobacter sakazakii; C. turicensis; C. malonaticus; C. muytjensii and C. dublinensis; the sixth species is identified simply as genomospecies I, as currently it includes only two representative strains. (19)

The name Cronobacter was appropriately derived from Greek mythology. E. sakazakii constitutes a microbiological hazard in the infant food chain, with historic high mortality in neonates. Accordingly, it was named after the Greek mythological god, Cronos. (17). Cronos was the son of Uranus (Heaven) and Gaea (Earth), being the youngest of the 12 Titans. He eventually became the king of the Titans, and took for his consort his sister Rhea. Rhea bore him a number of children, including Hestia, Demeter, Hera, Hades, and Poseidon. Cronos, however, had been previously warned by his parents that he would be overthrown by his own child. Accordingly, he swallowed all those children. When Zeus was born, however, Rhea hid him in Crete, and tricked Cronus into swallowing a stone instead. Zeus grew up, forced Cronus to disgorge his brothers and sisters, waged war on Cronus, and was victorious. (18)

It was proposed that these species be moved to the new genus, "Cronobacter", in order to facilitate their identification for the diagnosis of infection and the microbiological monitoring of food products. (20) All these species have been linked retrospectively to clinical cases of infection in either infants or adults, and therefore all these species should be considered pathogenic. (21) The correct and more detailed identification of these organisms will improve the understanding of the broader epidemiology of the members of the new genus. 


It is also important, however, that this reclassification of species not be detrimental to health protection measures already in place, and that all these risk organisms continue to be recognized. (22) As the genus Cronobacter is synonymous with Enterobacter sakazakii, current identification schemes developed for E. sakazakii remain applicable for the Cronobacter genus. The reclassification of E. sakazakii to the new genus Cronobacter will not require the modification of dedicated culture-based laboratory isolation and detection protocols. All currently valid laboratory methods will continue to facilitate the recognition of all of the organisms defined within the new taxonomy. (23) Furthermore, the reclassification does not require any change to the regulatory framework currently in place. (24)

REFERENCES:

Continue Reading...

Cronobacter sakazakii advice, policy and research in Canada

Editor's Note: We return to one of the presentations made at last January's Dublin conference on Cronobacter sakazakii.  This one by Dr. Jeff Farber of Health Canada.

Biography: Dr Jeffrey M. Farber gained an MSc and PhD from McGill University, Montreal, Canada, in Medical Microbiology & Immunology and Food Microbiology respectively. He worked as a research scientist and as Division Chief in the Microbiology Research Division before becoming Associate-Director of the Bureau of Microbial Hazards Food Directorate, Health Products and Food Branch, Health Canada where he is currently the Director. Dr Farber`s major interests are Listeria monocytogenes, Enterobacter sakazakii in foods, produce safety, molecular typing, food safety risk assessment and policy. He is a member of the Editorial Board of the International Journal of Food Microbiology; a reviewer for a number of other Journals; and Content Editor for IAFP Report. He is also an Adjunct Professor, at the University of Ottawa where he co-supervise undergraduate and graduate students; sits on a number of graduate student advisory committees; and is a member of thesis committees. Dr Farber is the treasurer of the International Commission on the Microbiological Specifications for Foods (ICMSF), and is a Past-President of the International Association for Food Protection.

Cronobacter sakazakii advice, policy and research in Canada

Although Canada has not had many reported cases of Cronobacter sakazakii, Health Canada has been actively studying this organism since 1998. In 2002, as a result of an outbreak in Tennessee  in the USA, Health Canada issued an advisory to inform Health Professionals in Canada what  measures they could take to reduce the risk to infants, of consuming powdered-infant formula (PIF).

After reviewing the situation at the national level and due to health concerns with powdered formulae and its international trade, in 2003, Health Canada raised this issue at the international level by proposing to revise the Code of Practice for Powdered Formulae for Infants and Young Children at the Codex Alimentarius Committee of Food Hygiene.

Canada volunteered to chair the Working Group that would be developing the Code. Because of the high level of interest in this issue, the Code was completed in four years, which is a relatively short time considering the complexity and politics behind this issue. The Code has contributed to a big improvement in the hygienic conditions in plants manufacturing PIF, resulting in a lower level of product contamination with C. sakazakii. Canada has produced a document detailing Good Manufacturing Practices (GMPs) for Infant Formula in Canada.

The purpose of this text is to establish and document the current GMPs for the production and quality control of infant formula products made for distribution in Canada. Health Canada uses the GMPs as a basis on which to assess the manufacturing information received in pre-market notifications for new or changed infant formulas.

Health Canada does have microbiological criteria for C. sakazakii in PIF; however, they are currently being revised to be more in line with recent Codex thinking. At present, unfortunately, there are no active or passive surveillance systems for C. sakazakii, in Canada, although this has been discussed.

Health Canada has recently adapted and condensed FAO/WHO guidelines to develop a draft guidance document on the preparation and handling of PIF in home and hospitals/care settings, which outline requirements for parents, caregivers, and staff in hospitals and day-care centers. The guidance document can be used to educate parents, caregivers and staff in hospitals and day-care centres, on the potential hazards associated with PIF.

Health Canada’s Bureau of Microbial Hazards conducts research focussed at examining the ecology, biology and pathogenesis of the organism. Some of the research projects include specific aspects of molecular typing, virulence studies involving animal models, as well as in-vitro tissue culture work to examine adhesion and invasion. Collaborative research is also being done with the National Research Council, using NMR and mass spectroscopy to reveal the structure of the O- polysaccharide of the various Cronobacter species.  For more, see his Powerpoint.

Codex Addresses Powdered Formula For Infants Out Of Concern About E. Sakazakii

The Codex Alimentarius Commission (CAC) adopted 30 new international standards and guidelines to improve food safety and protect consumers, including some that impact powdered infant formula.  The Codex changes came during a just ended week-long meeting.

The Commission adopted criteria for salmonella and other bacteria in powdered follow-up formulae for children six months of age or older and for special medical purposes for young children.

A bacterium of special concern is E. sakazakii, for which Codex adopted specific criteria for powdered formula for infants (0 to 6 months) in 2008. The Commission decided that in countries with particular risk for E. sakazakii from consumption of follow-up formulae (i.e. countries with substantial populations of immunocompromised babies) similar criteria for E. sakazakii could be introduced for follow-up formula as for powdered formula for infants.

Follow-up formulae should only be used for the intended target population. Unfortunately, they are often consumed by babies younger than six months of age. The standard stresses the need to address such product misuse issues through education campaigns and training. 

Codex, established in 1963, has 181 member states plus the European Community.  It is an offshoot of the United Nations.

Nestle Role In Alberta Conference Called Into Question

1st Annual Canadian Perinatal & Pediatric Nutrition Conference

Date & Time: September 25 - 26, 2009, 8:15 a.m. - 4:30 p.m.

Location: Bernard Snell Hall, University of Alberta Hospital

City/Town: Edmonton, AB

Description: This conference features two full days on obesity, allergies and other nutritional "hot topics" with expert faculty from across North America. It will be of interest to all those working with infants, children and nutrition including physicians, nurses, dietitians and pharmacists.

Early Bird Registration: $350
After August 15th: $400
One-day and student rates are also available

More Information: Call 780-735-1359 or e-mail RNFSEduc@cha.ab.ca

 

OUR COMMENT: This is the conference that the Infant Feeding Action Coalition (INFACT Canada) is out with an alert on, saying that  organizers are allowing "unethical marketing" by Nestle.

Here is what INFACT Canada says:

INFACT Canada has learned that a pediatric nutrition conference slated for September has listed Nestlé Nutrition as a major sponsor. The Alberta Health Services First Annual Canadian Perinatal and Pediatric Nutrition Conference being held in Edmonton on September 25-26 is not only being sponsored by Nestlé, but one of its speakers will be a Nestlé employee.

It is a serious conflict of interest for a public health agency such as Alberta Health Services (AHS) to stage a pediatric nutrition conference in partnership with an infant formula manufacturer like Nestlé. Nestlé is notorious for its aggressive marketing of infant formula and obstinate rejection of the WHO International Code of Marketing of Breastmilk Substitutes. The company recently launched a deceptive marketing campaign claiming that probiotics added to its formula will mimic the bifidus factor of human milk. It is no coincidence that one of the conference’s themes will be probiotics in infant nutrition.

As reported in a previous INFACT email, Nestlé’s new formula contains so-called “natural cultures” of bifidus bacteria, which are found in breastmilk. The company is marketing the new brand as equivalent to breastmilk. “There are only two places your baby can get natural cultures,” reads the advertising tagline, “The first is you. The other is from Nestlé Good Start Natural Cultures.” No scientific study is cited as proof that the bacterial cultures in this formula have the same effect on infants as breast milk.

We've seen no response to this from Alberta Health Services, the conference organizer.

Dr. Keith Lampel From U.S. Food & Drug Administration (FDA) Addresses Standard Method Of Detection

Editor's Note:  We continue presentations from the international Cronobacter conference held earlier this year in Ireland.  In this segment, we hear from the USA's Dr. Keith Lampel from FDA.

Biography: Dr. Keith Lampel is director of the Division of Microbiology at the U.S. Food & Drug Administration (FDA). He joined the FDA as a research microbiologist in 1987 after five years as a senior staff fellow at the National Institutes of Health, National Institute of Neurological Disorders and Stroke.
Dr. Lampel received his PhD in Microbiology from the University of Miami and was a postdoctoral fellow at the State University of New York at Stony Brook.
At FDA today, he is responsible for developing bacteriological and molecular-based methods to isolate, detect, and subtype foodborne pathogens. Dr Lampel is also the Editor of FDA’s Bacteriological Analytical Manual and serves as the FDA’s expert on the detection and isolation of the foodborne bacterial pathogen Shigella.
Other professional activities include serving on several editorial boards; NIH and USDA study panels, and ad’hoc review panels for several journals and extramural grant programs. He also serves as an adjunct professor at the University of Maryland and Uniformed Services University of the Health Sciences, and is a member of several PhD dissertation committees.

Summary - Development of an FDA/AOAC standard method for detection of Cronobacter

Although the number of incidences of illnesses caused by the ingestion of the bacterial pathogen Cronobacter (Enterobacter sakazakii) has not been as dramatic as other foodborne pathogens, a need remains for a robust isolation method to recover this microbe from powdered infant formula (PIF).
The current method described on the FDA website was developed in response to one such incident. Although C. sakazakii was a rather novel pathogen in an unusual food matrix, a method was devised quickly and applied to PIF samples. Unfortunately, this method requires multiple steps and at least 3-4 days for complete analysis of PIF for isolation and confirmation of C. sakazakii from the formula sample.
The revised method, however, includes a bacteriological enrichment and isolation protocol as well as the integration of a PCR-based assay. As for the bacteriological application, plating follows one-step enrichment on chromogenic agar(s) for presumptive identification of C. sakazakii. Suspected colonies are confirmed by either biochemical analysis or a real-time PCR-based assay. Therefore, isolation and identification of E. sakazakii from PIF is markedly improved and can be accomplished in 24-28 hrs.

Dr. Lampel's POWERPOINT can be found here

Development of a CEN-ISO horizontal standard method

Editor's Note: Another of the presentations follows from last January's 1st International Conference on Cronobacter (Enterobacter Sakazakii) held at University College Dublin.  Dr. Han Joosten from the Nestle Research Center in Switzerland addresses the standard method for detection of Cronobacter.

Biography: Dr. Han Joosten is a Senior Scientist at the Nestle Research Center in Lausanne, Switzerland. At Nestle since 1996, he is responsible for providing scientific guidance on various research projects and early identification of emerging microbiological safety issues. He also provides advice to the business and quality management on analytical methods, hurdle technology, safety assessments and HACCP.
After finishing his studies at the University of Nijmegen in the Netherlands in 1983 Joosten worked five years at NIZO Food Research on the formation of biogenic amines in cheese, obtaining his PhD degree from the University of Wageningen on this subject.
From 1989 to 1991 he worked as a postdoc at the Autonomous University of Madrid on molecular characterization of African Swine Fever Virus. After this he headed the microbiological laboratory of Coberco Research in Deventer, the Netherlands and moved back to Spain in 1994 where he worked for two years at the National Instititute for Agricultural and Food Research (CIT-INIA) in Madrid on a bacteriocin-producing Enterococcus strain.


Summary - Development of a CEN-ISO horizontal standard method for detection of Cronobacter
The availability of a reliable and internationally accepted reference method for detection of Cronobacter in powdered infant formula is an essential tool to verify compliance with regulatory requirements by public health authorities and manufacturers. ISO-TS 22964:2006 was developed as a temporary solution for this purpose, but shortly after being issued it was decided to prepare a full-fledged horizontal CEN-ISO standard.
A summary will be given of the work done thus far, in particular with respect to the modifications that are envisaged to address the main shortcomings of TS 22964:

  • The scope will be extended to all types of powdered infant formula (incl. soy- based) and infant formula ingredients.
  • It will take into account the latest taxonomical revisions (e.g. definition of the genus Cronobacter and phenotypically related species)
  • It will no longer use yellow pigment production as a confirmation criterion
  • The enrichment broth (mLST) and chromogenic isolation agar (ESIATM) are too selective and need to be replaced by media that will also allow the detection of strains that are very susceptible to commonly used inhibitors of gram positive microorganisms.
  • The main performance characteristics of the new standard will be determined

Based on the results obtained during an extensive comparative/collaborative trial a method based on the utilization of Cronobacter Screening Broth (CSB) in combination with modified DFI agar appears to be the most suitable procedure to be adopted in the new standard.

His POWERPOINT can be found here.

Oxoid Thermofisher Scientist Patrick Duggan Address Culture Media For Isolation and Detection of Cronobacter Species

 Editor's Note: This is another report on the presentations that were made in Dublin earlier this year at the 1st International Meeting on Cronobacter (Enterobacter Sakazakii).  In this segment, we hear from Dr. Patrick Druggan, Oxoid Ltd., Thermo Fisher Scientific, Basingstoke, Hampshire RG24 8PW, United Kingdom.

Biography: Patrick Druggan  is Principle Scientist, Oxoid Thermofisher, Basingstoke, UK.

He received an Honours BSc in Food Science from the University of Strathclyde, Glasgow, UK. He has worked in the diagnostics industry for 22 years. He designed his first chromogenic medium in 1989 while working at Gibco.

Patrick studied part-time for his Ph.D. at the Pharmacy Department of University of Brighton, UK. His thesis was on improvements in the resuscitation of heat-injured Salmonella species from processed food samples.

He synthesized a number of autocytotoxic compounds that could be used during pre-enrichment to inhibit competitive microflora while allowing injured Salmonella spp. to resuscitate and grow.

This invention lead to the development of Inhibigens.™ His skills in chemistry and microbiology have allowed him to design a number of successful rapid biochemical tests and chromogenic culture media, including Druggan-Forsythe-Iversen Agar for the isolation of Cronobacter spp. 

Summary – - Culture media for isolation and detection of Cronobacter species

In 2001 a pre-term infant died of meningitis caused by Enterobacter sakazakii (Cronobacter spp.).  Infant formula milk (IFM) was implicated as a potential source of the infection.

The Food and Drug Administration (FDA) independently develop a method for enumeration of this emerging  pathogen in IFM using culture collections from national bodies that have later been shown to be poorly defined.

This method was introduced in 2002 and has regulatory standing for the import of IFM and skimmed milk powder in to the USA and a number of other countries. The FDA method is a modification of the procedure for the detection of Enterobacteriaceae, with the addition of yellow pigmentation of colonies for presumptive identification of Cronobacter spp.

It should be remembered that the FDA method was developed in a short time due to a public health concern, and this would have put a time constraint and significant pressure on those working on Enterobacter sakazakii (Cronobacter) to get a working method in the field as soon as possible. The FDA method has been shown to have a sensitivity of around 50 percent and a specificity of around 70 percent. Only 75 percent of Cronobacter strains phenotypically express yellow pigmentation, and the low specificity of the method coupled with the recommendation that only five presumptive Enterobacteriaceae colonies are tested from Violet Red Bile Glucose Agar (VRBGA) may explain the poor sensitivity of the method.

Assuming the prevalence of Cronobacter spp. in IFM is around 2 percent, the FDA method will fail to detect around 50 percent of batches contaminated with Cronobacter, while around 95 percent of rejected batches will not contain this organism.

This high rate of failure has lead many stakeholders to question the usefulness of the FDA method. This presentation reviews developments in culture media since the release of the FDA method in 2002, with specific emphasis on media that have improved the specificity of methods for Cronobacter spp. The unique phenotypic trait of this emerging pathogen that aid and hinder design of methods is discussed.  

POWERPOINT is here.

Dublin's Professor Fanning Speaks About Molecular Identification Methods for Cronobacter spp.

Editor’s Note: We next are going to hear from Professor Seamus Fanning of University College Dublin, addressing Molecular identification methods for Cronobacter. He also spoke at the Dublin conference on Cronobacter.


Biography:  Seamus Fanning is the Professor of Food Safety & Zoonoses and the Director of the Centre for Food Safety, University College Dublin. Professor Fanning is an editor of Research in Microbiology and a member of the editorial board of the Journal of Food Protection.

Professor Fanning received an Honours BSc in Biochemistry from NUI, Cork, where he also completed his PhD in Microbiology and Molecular Genetics. Current research interests include the application of molecular methods to Food Safety to control zoonotic microorganisms associated with human disease. A significant part of this work relates to the characterization of the genetic mechanisms contributing to the emergence of multiple drug resistance (MDR); the role of membrane bound efflux pumps in MDR and virulence and how these phenotypes are regulated at a local and global level.
Also, in the past few years the UCD Centre for Food Safety has published several papers describing the detection and characterization of Cronobacter. Professor Fanning is a member of the Microbiology Sub-Committee of the Food Safety Authority of Ireland (FSAI), the Scientific Advisory Committee of safe food and was recently appointed by the European Food Safety Authority (ESFA) to a working group to provide expert opinion on the emergence of antibiotic resistance in food. He also served as a member on the FAO/WHO expert panel on Enterobacter sakazakii in follow-up formula.


Summary: Molecular identification methods for Cronobacter spp.


Historically the ancestry of the genus Enterobacter can best be described as nebulus and confusing. In the 1970’s and 1980’s considerable movement of species, originally assigned to this genus occurred, and these re-designations arose because of initial misplacements, based on older phenotypic and morphological approaches to describing taxonomy.
Currently the genus Enterobacter comprises a large and heterogenous group of organisms within the Enterobacteriaceae family being accounted for by 16 distinct species. Enterobacter sakazakii (E. sakazakii) is one of these species and the only member of the genus recognised as a food-borne pathogen. Following a revision of Enterobacter taxonomy, a new genus Cronobacter was devised which is synonymous with E. sakazakii. Cronobacter consists of a least five distinct species and an additional genomospecies, Cronobacter sakazakii (C. sakazakii), C. dublinensis, C. malonaticus, C. muytjensii, C. turicensis and C. genomospecies I. A further three sub-species of C. dublinensis are also recognised. Correct identification of these organisms is important in order to improve our understanding of the broader epidemiology of the members of this new genus.
In recent years there have been rapid improvements in the provision of microbiologically-based culture approaches to isolate and identify these organisms. A number of molecular identification methods have also been proposed, however the recent recognition of multiple species that share less than 70 percent DNA-DNA similarity has important implications for the sensitivity and specificity of these methods. In this paper, three examples of the application of molecular-based detection strategies for the identification of Cronobacter will be presented.
These will include strategies to identify the genus, specific targets that are thought to be related to pathogenicity and the development of a molecular-based approach to begin to define the O-serotypes of C. sakazakii. Although by no means complete, these examples will illustrate some of the current and future challenges to enable a more refined and reliable molecular-based approach to the identification of all Cronobacter spp.
The development of appropriate molecular methods will facilitate not only a rapid identification of an isolate, but in addition complement the more traditional microbiological-based methods.


POWERPOINT is here.

It's All In The Enterobacteriaceae Family For University of Bern's Dr. Kuhnert

Editor's Note: This is another report on the presentations that were made in Dublin earlier this year at the 1st International Meeting on Cronobacter (Enterobacter Sakazakii). Below we hear from Professor Peter Kuhnert of the Institute of Veterinary Bacteriology, University of Bern, Switzerland.

Biography:  Peter Kuhnert is working at the University of Bern, Switzerland as an associate professor in bacteriology. He completed a PhD in molecular biology studying gene regulation of the porcine TNF-locus.
As a postdoc at the Weizmann Institute of Science in Israel he focused on the regulation of the human TNF-receptor genes and its role in the immune response. After his return 1994 he switched topics and is since then working at the Institute of Veterinary Bacteriology where he focuses on bacterial virulence, phylogeny and taxonomy.  His work includes pathotyping of E. coli, genotyping of foodborne pathogens with an emphasis on Campylobacter, pathogenesis and virulence mechanisms of Pasteurellaceae as well as work on Mycoplasma hyopneumoniae.

Summary - Multilocus sequence analysis (MLSA) of Cronobacter and related taxa Genetic similarity as determined by DNA-DNA hybridization is still considered the ‘gold standard’ method to determine relatedness between bacterial species. Nevertheless, it is very time consuming and cumbersome to perform and requires cross-hybridization between representatives of a species and related taxa. Moreover, variation between experiments, techniques and laboratories make exchange and comparison of data difficult. Whole genome sequence comparisons could be an alternative to DNA-DNA hybridization however, data handling and the
open question as to what genes should be used for defining genome similarity cannot be neglected. Therefore, for taxonomic purposes investigating as many isolates of a species as possible in order to respect the biodiversity of taxa, a few representative genes indicative for genetic similarity between isolates is the optimal way to go. Recently, we showed that the three genes recN, rpoA, and thdF can be used to estimate whole genome similarity of representatives of the family Pasteurellaceae [Kuhnert & Korczak (2006) Int.J.Syst.Evol.Microbiol 152: 2537-2548]
In the presented work multilocus sequence analysis (MLSA) based on recN, rpoA and thdF genes was done on more than 30 species of the family Enterobacteriaceae with a focus on Cronobacter and the related genus Enterobacter. The sequences provided valuable data for phylogenetic, taxonomic and diagnostic purposes. Phylogenetic analysis showed that the genus Cronobacter forms a homogenous cluster related to recently described species of Enterobacter, but distant to
other species of this genus. Combining sequence information on all three genes is highly representative for the species’ percentage of GC-content used as taxonomic marker. Sequence similarity of the three genes and even of recN alone can also be used to extrapolate genetic similarities between species of Enterobacteriaceae, being an alternative to DNA-DNA hybridization. Finally, the rpoA gene sequence, which is the easiest one to determine, provides a powerful diagnostic tool to identify and differentiate pathogens of this family. The comparative analysis gives important
insights into the phylogeny and genetic relatedness of the family Enterobacteriaceae and will serve as a basis for further studies and clarifications on the taxonomy of this large and heterogeneous family.

Powerpoint: Find it here.

Dr. Iversen Introduces The Updated Version Of E. Sakazakii; Now Call It Cronobacter

Editor's Note:  In January, the 1st International Meeting on Cronobacter (Enterobacter sakazakii) was held in Dublin. As we can get to it, we are featuring summaries from some of the main speakers along with links to their power point presentations.

We begin with Ireland's own Dr. Carol Iversen.

Biography:  After completing a PhD at Nottingham Trent University, Carol Iversen worked for Nestlé Research Centre, Switzerland, where she led an international collaboration to define the taxonomy of Enterobacter sakazakii and propose a new genus, Cronobacter.
Dr. Iversen then joined the research team at the Institute for Food Safety and Hygiene, University of Zürich, to continue work on isolation methods and virulence characterisation of these organisms before moving to Ireland under an IRCSET postdoctoral fellowship in 2008.
Dr. Iversen is currently engaged in research on foodborne pathogens at the Centre for Food Safety, UCD. She is also a representative for the National Standards Authority of
Ireland on the ISO and CEN committees for Microbiology and Microbial Contamination, and is co-project leader of the ad’hoc group for development of a horizontal EN ISO
standard for the detection of Cronobacter (E. sakazakii).

Summary: What is (and isn’t) Cronobacter?  

Enterobacter sakazakii is an opportunistic pathogen that can cause meningitis, necrotising enterocolitis, and bacteraemia infants. It was first designated as a species in 1980 by Farmer et al. and several outbreaks in NICUs have been linked to contaminated powdered infant formula.
The organism is therefore of concern to infant food manufacturers as well as clinical microbiologists and food safety regulators. In 2008 the taxonomy of E. sakazakii was updated using a polyphasic approach based on extensive geno- and phenotypic evaluations. This resulted in the description of five novel species and the proposal that these be incorporated into a new genus, Cronobacter, which is contaxic with E. sakazakii. The isolation of Cronobacter is complicated by the existence of closely related species, Enterobacter pulveris, E. helveticus and E. turicensis. These species share
similar characteristics to Cronobacter and occur in the same ecological niches including infant foods. However, no health risk has been attributed to these organisms.

Her Powerpoint is Here.

50 Year Old Species Gets New Name: Cronobacter

In January some of the world's most distinguished health scientists gathered in Dublin for what they called "1st International Conference on Cronobacter (Enterobacter sakazakii)"

We've been reviewing the conference proceedings.   It seems that one of the things they did was change the name of Enterobacter sakazakii to Cronobacter.

Being diplomatic about it, they also recognized both the memory of Dr. Riichi Sakazaki (August 21, 1920 - January 11, 2002) for whom the organism is named (see picture) and Dr. Don J. Brenner,  Frances Brenner, Richard Fanning,  Arnold J. Steigerwalt, and the late Mary Alyce Fife-Asbury who were all involved in the original Centers on Disease Control(CDC) studies that led to it being recognized as a separate species.  They also gave a salute to Dr. Harry Muytjens for "his pioneering work in uncovering its ecology and epidemiology in cases of neonatal meningitis and the important role of powdered infant formula."

It's been 50 years since the first case of neonatal meningitis due to the organism now known as Cronobacter (Enterobacter sakazakii) was documented.  That case occurred in 1958, and was reported in 1961 by Urmenyi and White-Franklin.

Obviously, this name-changing is going to eventually be recognized in the name of this blog.  In the meantime, and probably for a long time, we will do what every scientist in Dublin did, which is to use the old and new names interchangeably. 

Iowa Family Sues Maker Of Enfamil In Federal Court: Seeks Damages, Requests Jury Trial

 Daniel Korte was one of the twin boys born prematurely on April 23, 2007 at Mercy Medical Center in Des Moines, Iowa.  Because of the pre-mature birth, Daniel remained in Mercy’s NICU unit where he was fed Enfamil Powdered Human Milk Fortifier for the first time on May 5, 2007.

This week, Patrick and Michelle L. Korte, Daniel’s parents, sued Mead Johnson Nutritionals, the spin-off of Bristol Myers Squibb Co. that makes the Enfamil Powdered Human Milk Fortifier.

The family alleges that within hours of consuming the Enfamil product, Daniel was in tachycardia and blood cultures taken revealed the presence of the Enterobacter Sakazakii bacteria.

“The E. Sakazakii bacteria that infected Daniel Korte came from the powdered human milk fortifier he received out of the Enfamil container,” says the lawsuit filed in U.S. District Court in Iowa.

The two year old continues to suffer from severe brain and other physical damages caused by the E. Sakazakii, which the Korte family blames the manufacturing defect in the Enfamil product, which they says was produced in a defective and unreasonably dangerous condition of contamination.

Evansville, IN-based Mead Johnson just raised $724 million in its initial public offering.  Symbol for its now publicly traded stock is MJN.  Its IPO was priced at $24 a share.

The Korte family is represented by attorneys William R. King of the Davis, Brown, Koehn, Shors & Roberts law firm in Des Moines, and Andrew Weisbecker and Bruce Clark of Marler Clark in Seattle.  In seeking damages for the Korte family, the lawyers have asked for a jury trial.

E Sakazakii Discussion From A Settlement Demand Letter

Editor's Note:  A Settlement Demand Letter (SDL) is a communication that is sent to defense attorneys on behalf of the plaintiff.  In food-borne illness litigation, an SDL usually requires some academic discussion like the one that follows on Enterobacter Sakazakii.  Our hat-tip goes to Andy Weisbecker, one of the talented attorneys at Marler Clark, for sharing his work with us.

THE ENTEROBACTER SAKAZAKII BACTERIA

Characteristics, Epidemiology, and Virulence

Enterobacter sakazakii is a gram-negative , non-spore-forming bacterium belonging to the Enterobacteriaceae family.

Enterobacteriaceae have increasingly been identified as pathogens in the past 30 years. E. sakazakii is a relatively newly identified pathogenic bacteria and used to be known as a "yellow pigmented Enterobacter cloacae" until 1980, when it was introduced as a new species based on differences in DNA-DNA hybridization, biochemical reactions, and antibiotic susceptibility.

E. sakazakii is a rare, but life-threatening cause of neonatal meningitis, sepsis, and necrotizing enterocolitis. In general, E. sakazaii kills 40-80 percent of infected newborns diagnosed with this type of severe infection. E. sakazakii meningitis may lead to cerebral abscess or infarction with cyst formation and severe neurologic impairment.

E. sakazakii can cause a variety of infections, though central nervous system infection has been most commonly described. For infants, infection typically manifests through signs of sepsis in the first week of life: irritability or lethargy, temperature instability, and feeding intolerance. Meningitis often produces overwhelming infection that rapidly moves through cerebral hemorrhage, infarct, necrosis, liquefaction, and eventually, cyst formation.

With a single reported exception, E. sakazakii meningitis has been reported exclusively among infants. In a study of E. sakazakii cases over a 47 year period, investigators found that the median age at infection onset was two days and 94 percent of cases were less than 28 days old. Premature infants are thought to be at greater risk for E. sakazakii infections than term babies, older children and adults.

Enterobacter species are opportunistic and generally infect those with underlying illness or are immunosuppressed. E. sakazakii invasive infections occur more frequently in infants than in older children. The neonate’s immature immune system may increase the risk of acquiring an E. sakazakii infection. Among adults E. sakazakii infection appears uncommon.

Continue Reading...

The Food and Drink Innovation Network Nottingham Trent University Award for Research

The Food and Drink iNet Innovation Awards were recently announced at the UK's National Space Centre in Leicester.  

This one caught our eye:

Nottingham Trent University School of Science & Technology for creating an easy-to-use, colour-changing agar that detects a specific bacterium which can be dangerous in food.

The Enterobacter sakazakii bacterium detected by the chromogenic agar is particularly risky for newborn babies.

The agar can be used right across the food industry, by virtually anyone in a workforce…making it a simple but effective food safety tool. 

An Irish Priest Speaks Out On Dangers of Enterobactyer Sakazakii

Father Shay Cullen is an Irish Catholic  priest  working  mostly in the  Philippines with kids who live on the streets and jails.   His work has often been recognized in the U.K., Canada, and the United States. Father Cullen weighed in last Sunday with advice to mothers in the developing world in light of the recent melamine scandal.   His comments ran in the Manila Sunday Times, and we want to share some of what he said.

It’s very disturbing to learn that children face the risk of infection and even death because of the brainwashing of mothers that milk formula is better than mother’s milk for their babies. It just goes to show the power of persuasion, the ability of the subliminal advertising of the pharmaceutical and food companies to manipulate mothers. It has overturned the age-old proven practice that has assured the survival of the human race. Breast-feeding has been the way from the beginning of human existence and in this short past forty years it has been undone.

Children are now more in danger of sickness and death than ever before because of this one change caused by the greed and irresponsibility of the money-driven corporations. The World Heath Organization (WHO) has campaigned tirelessly, with government health departments and non-government organizations, to reverse the all-powerful influence of these economic behemoths that care more for profits than for people.

The China melamine catastrophe of recent weeks is just one more example of the corrupt practices that food corporations will do to profit at the expense of children. They deliberately contaminated milk powder with a toxic chemical to increase the protein reading and make it appear acceptable to the other food companies producing powder milk formula...

He goes on to say:  The Food and Agriculture Organization (FAO) and WHO in February 2004 convened a panel of experts who concluded that infant formula has inherent contamination of Enterobacter Sakazakii and Salmonella that cause serious illness leading to developmental sequelae and even death. The evidence is overwhelming that these milk formulas are bad for babies and here have been dozens of product recall around the world.

For the complete column, go here.

 

Jakarta Judges Want to Name Brands Contaminated with Enterobacter Sakazakii

We've found a whole panel of judges in Jakarta who are our kind of guys!

They've accepted David M.L Tobing’s lawsuit against the Bogor Institute of Agriculture (IPB), the Food and Drug Monitoring Agency (BPOM) and the Health Minister. In the verdict, the panel ordered the defendants to announce the brands of formula milk and baby food products contaminated by the Enterobacter sakazakii bacteria, as indicated in their research and
reported in the newspapers and websites.

The panel of judges led by Reno Listowo said if the brands are not published, people will worry and be afraid to buy baby milk and food products. The judge added that the public has the right to know which products are safe and which are contaminated.

David’s plea was triggered by an IPB study that indicated some formula milk and baby foods were Enterobacter sakazakii-contaminated.

This report comes to us from TEMPO Interactive.

For more, go here.

 

 

 

Health Canada Warns Against Enterobacter Sakazakii In Health Trek Products

Do some Wild Vineyard products, made by Health Trek  in Manitoba poise a health  risk,  including  Enterobacter  Sakazakii,   as  Health  Canada  has twice warned  or  not?


We  raise  the question  because  the Selkirk,  Manitoba  Journal gave  a lot of ink  last Friday to  Health Trek's owner , who  claims to be outraged by  twice being on the  wrong  end of  a  Health  Canada  warnings to the public  not to  consume his company's products.  The  Journal reports:

A Selkirk business owner is outraged Health Canada has issued its second warning in seven months, saying some of his product may pose a health risk to consumers.
Health Canada issued the warning July 22, saying 11 herbal Wild Vineyard products, distributed by Health Trek company owner Terry Bell, located at 371 Eveline St., may be unhealthy.
Paul Duchesne, a Health Canada spokesperson, said the newest release gives additional information on Wild Vineyard products that pose a health risk.

He said different bacteria have been detected in different products, adding some of the bacteria found that pose a potential health risk include E. Coli, Enterobacter sakazakii, Klebsiella pneumoniae and excessive levels of aerobic bacteria, including species of bacillus.
On the Health Canada website, a news release listed products which have been found to contain “unacceptable levels of bacterial contamination and/or heavy metal contamination, such as lead.”
Symptoms of exposure to heavy metals include nausea, abdominal pain, vomiting, muscle cramps, diarrhea, heart abnormalities, anemia, as well as bone, liver, kidney and nervous system problems.
Bacterial contamination of health products, meanwhile, can pose a risk to human health and may cause a variety of adverse effects, such as gastrointestinal upset and bacterial infection.

For more on the dispute, we suggest reading the entire story here.

Dr. Hook Uses Humor To Get At Enterobacter Sakazakii and Other Bacteria

We first took a look at what Dr. John Hong, MD, was writing because he was mentioning Enterobacter Sakazakii.   We kept reading his column at The Hook because in writing as Dr. Hook, Dr. Hong is a really funny guy.

"Eat up! But be sure food is bacteria-free" is Dr. Hong's walk through today's dangerous food environment.   It's a hoot and well worth the read.

The Hook says: Dr. Hook cracks a joke or two, but he's a renowned physician with a local practice. They invite Emails to the Doctor with questions.

From its local weather report, we think The Hook and Dr. Hong are based in Charlottesville, Virginia

Limits Set on Enterobacter Sakazakii Bacteria In Powdered Milk

The Codex Alimentarius Commission, created in 1963 by the United Nations and the World Health Organization to develop food standards, guidelines and related texts such as codes of practice under the Joint UN/WHO Food Standards Programme, has agreed to a new benchmark for powdered milk.

The new standard was adopted to reduce the risk of Enterobacter sakazakii, a bacteria that causes severe illness in babies and small children. The code sets maximum limits for the bacteria and provides guidelines on how to produce and handle prepared powdered formula.

All members of the Commission accepted the need for strict measures on this product, but some developing countries, led by Indonesia, inquired about possible alternative testing methods at lower cost.

Cuba pressed "for more scientific analysis of safe levels of bacteria, as well as a possible funding source for testing." The countries raised the possibility that the Codex Commission could provide more support for developing countries in meeting Codex standards, with the the UN and the WHO playing a greater role in capacity building.

As shown by their reservations with regard to standards that would require using costly production techniques, developing countries were concerned over potential implications for their export capacity and competitiveness.

Throughout discussion about standard adoption on June 30th, Commission members from various developing countries stressed the need for consumer safety - not only for exported goods, but also for products used domestically - while also voicing concerns with regard to resource constraints and difficulties they face in standard implementation.

Go here for a pretty good summary of the 31st annual meeting of the Commission.


China Picks System For Detecting Enterobacter Sakazakii And Other Pathogens

Dupont Qualicon's  BAX (R) system is the choice of the General Administration of Quality Supervision, Inspection and Quarantine (AQSIQ) of the People's Republic of China as the official method for pathogen detection in food imports and exports.

The decision by AQSIQ names the automated BAX(R) system as an approved PCR method for detecting Salmonella, E. coli O157:H7, Listeria monocytogenes, Campylobacter jejuni and E. sakazakii in food.
PRC officials are quoted in a statement from Dupont. In part, it said:

"When monitoring food imports and exports, efficiency is critical," said
Qin Zhenkui, president of the Chinese Academy of Inspection and Quarantine (CAIQ), AQSIQ. "PCR technology can provide the speed, sensitivity and accuracy in routine food testing that allows our agency to make faster release decisions with a high degree of confidence."

Another food safety monitoring authority in China -- the Beijing MunicipalCenter for Food Safety Monitoring (BFSM) -- is already using the BAX(R)
detection system to provide technical support for food safety management of the Beijing municipal government and the 2008 athletic competition.


"This is good news from AQSIQ," said Kevin Huttman, president - DuPont Qualicon. "Protecting the food supply is a major global concern, and we understand that China is also facing the increasing demands and emerging challenges on food safety. We're pleased that the BAX(R) system is helping government agencies in China and around the world with cost-effective and highly accurate food safety testing."

For the complete statement from Dupont, go here.


France's bioMerieux Offers Product To Detect E. sakazakii

Laboratorytalk, which bills itself as the lab equipment news source, reports that a company called bioMerieux is launching an “innovative chromogenic culture media—ChromID Sakazakii—for “precise, reliable controls” of Enterobacter sakazakii.

Here are the money quotes from the Laboratorytalk report:




Enterobacter sakazakii is considered to be a neonatal pathogen Its presence in infant food products, including powdered milk, can cause meningitis, sepsis and other serious infections, primarily in newborn or premature infants.
40 to 80 percent of children who become infected die, and those who survive can suffer severe and long-lasting complications.
Its innovative formula reveals two Enterobacter sakazakii-specific enzyme activities that offer a high level of detection sensitivity in just 24 hours of incubation.
Enterobacter sakazakii is an enterobacterium that is naturally present in the environment and can be found in various foods, most notably dehydrated products. Considered a neonatal pathogen, when present in infant food products, it can cause serious infections such as neonatal meningitis or necrotizing enterocolitis, as well as the deaths of 40 to 80 percent of children infected.
A French company involved vitro diagnostics for 45 years, bioMérieux is present in more than 150 countries through 38 subsidiaries and a large network of distributors. In 2007, revenues reached €1.063 billion with 84 percent of sales outside of France.

For story, Innovative ways to detect food-borne pathogens, go here.

Malaysia Ministry of Health Calls Anti-Mom's Milk Study Reckless

Dr. Safurah Jaafar, Director of the Family Health Division of Malaysia's Ministry of Health, has come down hard on the New Straits Times for a report it carried in early May on the potential for "toxins" in mother's milk.

In response, Dr. Jaafar writes:

The Health Ministry is concerned over the recklessness and insensitivity of the report, especially with regard to certain statements which seem to belittle breast-feeding and favour infant formula.

This clearly goes against the ministry's breast-feeding policy whereby mothers are encouraged to breast-feed their infants exclusively with breast milk for the first six months of the infant's life and thereafter to continue breast-feeding along with appropriate complementary foods up to the age of two years.

Your report has generalised the findings of a small-scale study on a group of mothers. It is certainly inappropriate to assume that all mothers are exposed to environmental contaminants on a similar scale, when, in fact, studies have shown that factors such as age, diet, cigarette smoking, occupation and geographic location of the mother influence the body burden of persistent organic pollutants (POPs) on varying scales.

Dr. Jaafar said the benefits of mother's milk far outweigh the risks.   The doctor then turned to the risks of the alternatives, saying:

As an example, formula-fed infants are at a high risk of exposure to life-threatening bacterial contamination.

Bacterial contamination of powdered infant formula, notably Enterobacter sakazakii, can cause fatal outcomes in newborns.

The doctor's complete article can be found here.

Most Infant Milk Supplements Get Contaminated with E. Sakazakii, Says Doctor

Most infant milk supplements get contaminated with Enterobacter sakazakii, which causes meningitis or enteritis, especially in infants, and that the World Health Organisation has issued strict guidelines about these products.

So says Dr. Arun Gupta, a paediatrician who is national coordinator of the Breastfeeding Promotion Network of India and regional coordinator of IBFAN Asia.

Dr. Gupta's comments were carried in Newindpress on Sunday in an articile titled "How Safe is your baby?" by C. Shivakumar.   Dr. Gupta  says multi-billion dollar baby-food industry, which  includes the manufacture of baby bottles,  isn't sufficiently regulated.

The online India news service reports that:

As infant formula is widely used, the presence of Enterobacter sakazakii and its potential effects could well be a significant public health problem in most countries, feels Gupta. He adds that most health workers in India are unaware of the dangers posed by these milk supplements.

Check here  for the rest of the story.



Is Ozone Technology Solution To E Sakazakii Illness?

Is there a penny stock company out there with a silver bullit to elimate food-borne illnesses?  We're often amused by reports that reach us from sources like "The World Stock Wire" and we sometimes wish we had access to those "Pink Sheet," which somehow have yet to make it to the internet in this 21st Century.

Nevertheless, sometimes we get something that gives us one of those Hmmmm moments.  Information we recieved this morning from Irving, TX-based Produce Safety & Security prompted one of those moments.    PDSC on those Pink Sheets "is promoting their patented ozone systems, equipment and machinery and other proprietary sanitizing product lines on a worldwide basis to commercial, government and major industrial users."

We're in no position to judge the claims the company makes about its ozone systems, but we did think it has an understanding of food borne illness.  For example, here's what the company said about Enterobacter Sakazakii:

One example of a newer food-borne pathogen is enterobacter sakazakii, which can cause serious illness such as sepsis (blood infection) and meningitis (inflammation of the membrane surrounding the brain and spinal cord).

In 2002, FDA, working with CDC, discovered and subsequently alerted health care professionals to clusters of E. sakazakii infections reported in a variety of locations among hospitalized newborns, particularly premature or other immuno-compromised infants who were fed powdered infant formulas. The emergence of new food-borne pathogens requires updated technologies that can detect the presence of new agents in a variety of foods. Addressing these emerging hazards requires cooperation among industry, academia, and government to share information and establish testing protocols.

Like we said, it is interesting, but cannot say more.   You can check it out here and here.



 

 

Enterobacter sakazakii

Irish Offer Ten Step Plan to Nail E. sakazakii

Vhi, the Irish private health insurer, and Safefood, Ireland's food safety promotion board, are promoting a new 10 step procedure to keep baby bottles safe from Enterobacter sakazakii.

Dr Cliodhna Foley-Nolan, Director, Human Health and Nutrition at Safefood warned that powdered infant formula is not a sterile product and has the potential to cause illness if not prepared properly. In rare circumstances, powdered infant formula can contain the bacterium Enterobacter sakazakii (E.sakazakii) and other harmful bacteria. These bacteria can cause illness in infants. Babies under 2 months are most at risk. However, making up the formula using water that is above 70°C will kill E.sakazakii and any other bacteria like salmonella that may be present, said Dr Foley-Nolan

Vhi and Safefood say if parents and carers follow the 30-minute rule any harmful bacteria will be killed. The 30 minute rule means that after boiling fresh water, the water is left to cool for 30 minutes before adding the powdered infant formula.

Here are the 10-steps on making baby’s bottles safely are:

  1. Boil water
  2. Leave to cool for 30 minutes but no longer
  3. Clean surfaces, wash hands
  4. Read the instructions on the formula’s label carefully
  5. Pour the boiled water into sterile bottle
  6. Add formula using scoop provided
  7. Shake well
  8. Cool quickly
  9. Check temperature is cool enough for baby
  10. Throw away any unused feed after 2 hours.

For more, go here.

Enterobacter sakazakii: Infections Associated with Powdered Infant Formula

Enterobacter sakazakii is a gram-negative rod-shaped bacterium within the family Enterobacteriaceae that is a rare cause of bloodstream and central nervous system infections. The majority of infections reported in the peer-reviewed literature have described neonates—newborn infants, including premature infants, post-mature infants, and full-term newborns—with sepsis, meningitis, or necrotizing enterocolitis as a consequence of the infection. (1)

Reported outcomes are often severe: seizures; brain abscess; hydrocephalus; developmental delay; and death in as many as 40%–80% of cases.  Premature infants are thought to be at greater risk than more mature infants, other children, or adults, and outbreaks have occurred in hospital units for newborns. (2)

Although E. sakazakii can cause illness in all age groups, infants are believed to be at greatest risk of infection and E. sakazakii was first implicated in a case of neonatal meningitis in 1958. Since that time, around 70 cases of E. sakazakii infection have been reported, but it is likely that is the number of cases is significantly under-reported in all countries and the incidence is believed to be more common. (3)

Experts from the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) met in 2004 to summarize information, and develop international guidelines and educational messages regarding E. Sakazakii. The meeting confirmed that there is very little known about virulence factors and pathogenicity of this bacterium.  Like other bacteria in the family Enterobacteriaceae, such as Escherichia coli, differences in virulence among E. sakazakiistrains appear to exist, and some strains may be non-pathogenic. Work done by Pagotto et al. (2003) was the first describing putative virulence factors for E. sakazakii.  Enterotoxin-like compounds were produced by some strains, but not all.  Using tissue cultures, some strains produced a cytotoxic effect. Two strains (out of 18 isolates) were capable of causing death in suckling mice by the peroral route. (4)

Mortality rates from E. sakazakii infection were once reported to be 50 percent, but this figure has declined to under 20 percent in recent years. Significant morbidity in the form of neurological deficits can result from infection, especially among those with bacterial meningitis and cerebritis. While the disease is usually responsive to antibiotic therapy, a number of authors have reported increasing antibiotic resistance to drugs commonly used for initial treatment of suspected Enterobacter infection. Long-term neurologic sequelae are well recognized. (4)

Enterobacter sakazakii kills 40%–80% of infected infants. In 2007, the CDC reviewed literature for reports of E. sakazakii and analyzed 46 cases of invasive infant infection to define risk factors and guide prevention and treatment.  The data available showed that of the 46 cases analyzed, twelve infants had bacteremia, 33 had meningitis, and 1 had a urinary tract infection. Compared with infants with isolated bacteremia, infants with meningitis had higher birth weight (2,454 g vs. 850 g, p = 0.002) and longer gestational age (37 weeks vs. 27.8 weeks, p = 0.02), and infection developed at a younger age (6 days vs. 35 days, p<0.001). Among meningitis patients, 11 (33%) had seizures, 7 (21%) had brain abscess, and 14 (42%) died. (2) 

Although E. sakazakii can cause illness in all age groups, infants (children <1 year) are at most risk. The groups of infants at greatest risk includes pre-term infants, low-birth-weight (<2.5 kg) infants or immunocompromised infants. However, infants who are compromised for any other reason may also be at greater risk of E. sakazakii infection. Infants of HIV-positive mothers are at risk because they may be immunocompromised, and may specifically require powdered infant formula. (3)

In 2006, an FAO/WHO expert working group concluded that neonates and infants under two months are at greatest risk and research has shown that premature infants who develop bacteremia after one month of age and term infants who develop meningitis during the neonatal period are still more at risk. (3)

In the United States, an incidence rate of 1 per 100,000 infants for E. sakazakii infection has been reported. This incidence rate increases to 9.4 per 100,000 in infants of very low birth weight, i.e. <1.5 kg. (3)

While the reservoir for E. sakazakii is unknown in many cases, a growing number of reports have established powdered infant formula as the source and vehicle of infection.  In several investigations of E. sakazakii outbreaks that occurred among neonates in neonatal intensive care units, investigators were able to show both statistical and microbiological association between infection and powdered infant formula consumption. These investigations included cohort studies which implicated infant formula as the source of the outbreaks. In addition, there was no evidence of infant-to-infant or environmental transmission; all cases had consumed the implicated formula. The stomach of newborns, especially of premature babies, is less acidic than that of adults: a possible important factor contributing to the survival of an infection with E. sakazakii in infants. (4)

Limited information was available on the numbers of E. sakazakii organisms that ill patients were exposed to in any of the various outbreaks and it is therefore not possible to develop a dose-response curve for E. sakazakii. However, it is possible that a small number of cells present in powdered infant formula could cause illness. (3)

The frequency of intrinsic E. sakazakii contamination in powdered infant formula is of concern, even though intrinsic concentration levels of bacteria appear to be typically very low. In a study of the prevalence of E. sakazakii contamination in 141 powdered infant formulas, 20 were found culture-positive, yet all met the microbiological specifications of the current Codex code for coliform counts in powdered infant formula (<3 cfu/g).  Such formula has been linked to outbreaks. (4)

Furthermore, outbreaks have occurred in which the investigators have failed to identify lapses in formula preparation procedures. Thus, it seems that neither high levels of contamination nor lapses in preparation hygiene are necessary to cause infection from E. sakazakii in powdered infant formula. While it can be assumed that lapses in preparation hygiene or extended holding at non-refrigerated temperatures could lead to increases in the levels of contamination at the time of consumption, it is not possible to assess the contribution that these factors have on the cases of infection that have been associated with powdered infant formula that contained low levels of E. sakazakii. Thus it must be currently assumed that low levels of E. sakazakii in infant formula (<3 cfu/100 g) can lead to infections. (4)

In the April 12th 2002 issue of Morbidity and Mortality Weekly Report, the Centers for Disease Control and Prevention (CDC) reported on a fatal case of meningitis in an intensive care nursery in Tennessee. The infecting organism was E. sakazakii - an unusual but often fatal invasive pathogen. In the fatal Tennessee case, the infection was traced to contaminated powdered infant formula. Other infants in the same nursery were screened for E sakazakii. Of 49 screened infants, 10 events were discovered (1 proven infection, 2 assumed infections, and 7 colonizations). This report detailed a direct link to an unopened product for the first time. The manufacturer voluntarily recalled the contaminated batch of powdered formula identified as the source. (5)

In 2004, powdered infant formula was microbiologically linked to two E. sakazakii outbreaks, in New Zealand and in France. The French outbreak involved nine cases, and resulted in the death of two infants. While eight of the cases were in premature infants of low birth weight (<2 kg), one case was in an infant born at 37 weeks and weighing 3.25 kg. The outbreak involved five hospitals, and a review of practices in the hospitals revealed that one hospital was not following recommended procedures for the preparation, handling, and storage of feeding bottles, and four were storing reconstituted formula for >24 hours in domestic-type refrigerators, with no temperature control or traceability. (3)

The FDA points out that powdered infant formulas are not commercially sterile products. Powdered milk-based infant formulas are heat-treated during processing, but unlike liquid formula products they are not subjected to high temperatures for sufficient time to make the final packaged product commercially sterile. FDA has noted that infant formulas nutritionally designed for consumption by premature or low birth weight infants are available only in commercially sterile liquid form. However, so-called "transition" infant formulas that are generally used for premature or low birth weight infants after hospital discharge are available in both non-commercially sterile powder form and commercially sterile liquid form. Some other specialty infant formulas are only available in powder form. (1)

The FDA has become increasingly aware that a substantial percentage of premature neonates in neonatal intensive care units are being fed non-commercially sterile dry infant formula. In light of the epidemiological findings and the fact that powdered infant formulas are not commercially sterile products, FDA recommends that powdered infant formulas not be used in neonatal intensive care settings unless there is no alternative available. If the only option available to address the nutritional needs of a particular infant is a powdered formula, risks of infection can be reduced by:

  • Preparing only a small amount of reconstituted formula for each feeding to reduce the quantity and time that formula is held at room temperature for consumption;
  • Recognizing differences in infant formula preparation among hospitals—individual facilities should identify and follow procedures appropriate for that institution to minimize microbial growth in infant formulas;
  • Minimizing the holding time, whether at room temperature or while under refrigeration, before a reconstituted formula is fed; and
  • Minimizing the "hang-time" (i.e., the amount of time a formula is at room temperature in the feeding bag and accompanying lines during enteral tube feeding), with no "hang-time" exceeding 4 hours. Longer times should be avoided because of the potential for significant microbial growth in reconstituted infant formula. (1)

WHO recommends that infants should be exclusively breastfed for the first six months of life to achieve optimal growth, development, and health. Thereafter, to meet their evolving nutritional requirements, infants should receive nutritionally adequate and safe complementary foods while breastfeeding continues for up to two years of age or beyond (WHO/UNICEF, 2003). It is important to support breastfeeding and promote its benefits to infants and young children.  (3)

There are, however, instances where breast milk is not available, where the mother is unable to breastfeed, where they have made an informed decision not to breastfeed, or where breastfeeding is not appropriate, e.g. where the mother is taking medication that is contraindicated for breastfeeding or the mother is HIV-positive.  Similarly, some very low-birth-weight babies may not be able to breastfeed directly, and in some cases expressed breast milk may not be available at all or available in insufficient quantities. Infants who are not breastfed require a suitable breast-milk substitute, for example, an infant formula prepared in accordance with the present guidelines. WHO Guidelines for infant formula preparation, storage, and handling (2007), in both care settings and at home, are specified at http://www.who.int/foodsafety/publications/micro/powdered infant formula (3)

In January 2006, a second meeting (after the initial one in 2004) of experts from the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) took place to summarize information, and develop international guidelines and educational messages. The meeting participants first re-endorsed the recommendations made by the 2004 FAO/WHO meeting on this issue. The additional recommendations made by the expert meeting to member countries included the following:

  • Develop prevention strategies for E. sakazakii infections caused by contaminated powdered infant formula that address the different stages of production and preparation and use of powdered infant formula, taking into consideration the risk to infants ?both within and beyond the neonatal period and of any immune status.
  • Develop educational messages on the safe handling, storage and use of powdered infant formula, including the health hazards of inappropriate preparation and use; target healthcare workers, parents and other caregivers, in both hospitals and the community, since E. sakazakii infections have occurred in hospital and home settings.
  • Review and revise product labels, as appropriate, to enable caregivers to handle, store and use the product safely, and to make clear the health hazards of inappropriate preparation.
  • Encourage member countries to establish surveillance and rapid response networks, and facilitate coordinated investigation by clinicians, laboratorians, and public health and regulatory officials, to enable the timely recognition and cessation of outbreaks of illness associated with E. sakazakii and the identification of contaminated powdered infant formula. (6)

REFERENCES:

(1)        “Health Professionals Letter on Enterobacter sakazakii Infections Associated With use of Powdered (Dry) Infant Formulas in Neonatal Intensive Care Units”, U. S. Department of Health and Human Services, U. S. Food and Drug Administration, April 11, 2002; Revised October 10, 2002.

(2)        “Invasive Enterobacter sakazakii Disease in Infants”, Emerging Infectious Diseases, Volume 12, Number 8–August 2006.  

(3)        “Safe Preparation, Storage and Handling of Powdered Infant Formula Guidelines” (2007), World Health Organization, in collaboration with Food and Agriculture Organization of the United Nations.

(4)        “Enterobacter sakazakii and other microorganisms in powdered infant formula” Microbiological Risk Assessment Series 6, World Health Organization (2004).

(5)        “Enterobacter sakazakii Infections Associated With the Use of Powdered Infant Formula—Tennessee, 2001”, JAMA. 2002; 287:2204-2205, Vol. 287 No. 17, May 1, 2002.

(6)        “Enterobacter sakazakii and Salmonella in powdered infant formula: Meeting report, MRA Series 10”, Microbiological Risk Assessment Series 10, World Health Organization (2006).