Sialic acid utilization by Cronobacter sakazakii

Background: The Cronobacter genus is composed of seven species, and can cause infections in all age groups. Of particular concern is C. sakazakii, as this species is strongly associated with severe and often fatal cases of necrotizing enterocolitis and meningitis in neonates and infants. Whole genome sequencing has revealed that the nanAKT gene cluster required for the utilisation of exogenous sialic acid is unique to the C. sakazakii species (ESA_03609-13). Sialic acid is found in breast milk, infant formula, intestinal mucin, and gangliosides in the brain, hence its metabolism by C. sakazakii is of particular interest. Therefore its metabolism could be an important virulence factor. To date, no laboratory studies demonstrating the growth of C. sakazakii on sialic acid have been published nor have there been reports of sialidase activity. The phylogenetic analysis of the nan genes is of interest to determine whether the genes have been acquired by horizontal gene transfer

Genotypic and phenotypic characteristics of Cronobacter species, with particular attention to the newly reclassified species C. helveticus, C. pulveris, and C. zurichensis [forthcoming]

In 2013, Enterobacter helveticus, E. pulveris and E. turicensis, were reclassified as Cronobacter helveticus, C. pulveris and C. zurichensis, respectively. Previously these species had been used as negative controls for some Cronobacter detection assays. This study examined cultural, biochemical and molecular Cronobacter detection and identification assays, with emphasis on the new species. Additionally, 32 Cronobacter genomes were examined for the presence of PCR target genes using the BLAST function of the online Cronobacter BIGSdb facility. The results of the cultural methods varied and no single medium was able to correctly detect all Cronobacter spp. Since the supporting databases have not been updated to include the Cronobacter genus, Enterobacter sakazakii was returned for four strains of the newly reclassified species with ID32E and none with API 20E. PCR probes targeting rpoB and ompA could not correctly identify the new Cronobacter spp., due to primer specificity or absent target genes. As neonates have been identified as a high-risk group for infection, international standards require the absence of all Cronobacter species in powdered infant formula. However, many conventional detection methods cannot correctly identify the newly recognized species. Conversely, DNA sequence-based methods can adapt to taxonomic revisions and will likely become more common

God in trinity, love in creation

This thesis attempts to display vital connections between themes not generally connected. On the one hand, it considers the notion of love and relation in connection with God and the idea of Trinity. On the other, it endeavours to show how the notion of a Triune God of Love may be used to safeguard an idea of human beings in which their dependence upon love and relationship is stressed. Fundamental to this is the demand of freedom in terms of choice. This is shown to be a direct consequence of the nature of love, and to necessitate considerable revision of conventional doctrines of grace and providence. Throughout the thesis, great stress is laid on an eclectic use of sources to demonstrate the argument. The thesis comprises two chapters of ontology, and three chapters of human concern. In the first two, the nature of Trinity and Creation are examined from a standpoint of the philosophy of loving relation. Three negative forces are identified in the shape of Cartesian Solipsism, the Platonic philosophy of Love, with its lack of relation and its abstract view of the Good, and thirdly Nygren's theology of agape, which it is held destroys man’s integrity. Coupled with his non-use of the Trinity, it also reduces God to an abstract monad. Positive thought is taken from Vanstone's Love's Endeavour Love’s' Expense which develops a doctrine of God from a theology of Love, and this is taken into the third chapter which considers the consequences for grace and providence. A revised portrait of providence coupled to a God who is neither impassible nor omniscient is provided. The remaining two chapters examine the consequences of such theology for man's own expectations concerning self-fulfillment and his obligations to others

Innovative construction and the role of boundary objects: a Gehry case study

© 2016 Informa UK Limited, trading as Taylor & Francis Group. Physical objects have long been used in addressing the challenges involved in constructing innovative buildings, yet their significance for collaborative problem solving in inter-organizational projects is rarely acknowledged. The aim of this research is to investigate what happens when a project team has to collaboratively innovate to address radical design challenges in a construction setting. We focus on the role of a full-scale mock-up of a façade in transforming the design intent for a building by Frank Gehry into design realization. The concept of boundary objects is used as an analytical lens via a case study methodology utilizing non-participant observation of weekly meetings and workshops over a period of 10 months covering client, consultant and contractor involvement. The research shows the role of mock-ups in radical construction settings is in tension along three delivery dimensions: performance, aesthetic and technical construction. Task completion competed with the requirements for experimentation around innovative problem solving with the how to construct it problem left unresolved. The findings suggest that co-location and synchronicity are critical conditions for collaborative and innovative problem solving in radical construction contexts. Project teams need to create open-ended ‘moments’ for iterating critical objects and the interactions that take place around them

Genomic dissection of the 1994 Cronobacter sakazakii outbreak in a French neonatal intensive care unit

Background: Cronobacter sakazakii is a member of the genus Cronobacter that has frequently been isolated from powdered infant formula (PIF) and linked with rare but fatal neonatal infections such as meningitis and necrotising enterocolitis. The Cronobacter MLST scheme has reported over 400 sequence types and 42 clonal complexes; however C. sakazakii clonal complex 4 (CC4) has been linked strongly with neonatal infections, especially meningitis. There have been a number of reported Cronobacter outbreaks over the last three decades. The largest outbreak of C. sakazakii was in a neonatal intensive care unit (NICU) in France (1994) that lasted over 3 months and claimed the lives of three neonates. The present study used whole genome sequencing data of 26 isolates obtained from this outbreak to reveal their relatedness. This study is first of its kind to use whole genome sequencing data to analyse a Cronobacter outbreak. Methods: Whole genome sequencing data was generated for 26 C. sakazakii isolates on the Illumina MiSeq platform. The whole genome phylogeny was determined using Mugsy and RaxML. SNP calls were determined using SMALT and SAMtools, and filtered using VCFtools. Results: The whole genome phylogeny suggested 3 distant clusters of C. sakazakii isolates were associated with the outbreak. SNP typing and phylogeny indicate the source of the C. sakazakii could have been from extrinsic contamination of reconstituted infant formula from the NICU environment and personnel. This pool of strains would have contributed to the prolonged duration of the outbreak, which was up to 3 months. Furthermore 3 neonates were co-infected with C. sakazakii from two different genotype clusters. Conclusion: The genomic investigation revealed the outbreak consisted of an heterogeneous population of C. sakazakii isolates. The source of the outbreak was not identified, but probably was due to environmental and personnel reservoirs resulting in extrinsic contamination of the neonatal feeds. It also indicated that C. sakazakii isolates from different genotype clusters have the ability to co-infect neonates

Screening forCronobacterSpecies in Powdered and Reconstituted Infant Formulas and from Equipment Used in Formula Preparation in Maternity Hospitals

Background/Aims: Cronobacter spp. have been identified as being of considerable risk to neonates. The occurrence of organism in infant formulas is therefore of considerable interest. Methods: The occurrence of Cronobacter spp. in infant feeds (formulas and fortified cow’s milk) was determined using most probable number (MPN) analysis, and from formula preparation utensils. Ninety nine samples were analyzed, of which 42 were unopened cans of powdered infant formula (PIF), 25 reconstituted infant formulas in feeding bottles, 27 utensils used from the preparation of infant formula, and 5 samples of fortified cow’s milk. Presumptive Cronobacter spp. isolates were identified using the 7 allele multilocus sequence typing (MLST) scheme. Results: C. sakazakii, C. malonaticus and C. muytjensii were recovered from PIF. Although the incidence of Cronobacter in PIF was 29% (12/42), the level was low with an average of 0.54 MPN/100g. According to MLST profiling, C. sakazakii was the most frequently isolated Cronobacter species, and C. sakazakii ST4 (associated with neonatal meningitis) was recovered from 2/42 PIF samples at 0.51 and 0.92 MPN/100g. Conclusions: Cronobacter spp. can be isolated from PIF and therefore strict hygienic practices during PIF preparation are important to minimize neonate exposure and reduce the risk of severe infections

Gender in agricultural mechanization: Key guiding questions

This tool describes case studies of gender implications in mechanization in RTB crops, illustrating both positive and negative outcomes. GENNOVATE research has shown that the beneficiaries of mechanization tend to be the wealthier rather than the poorer farmers and more often men than women. However, results also reveal that women have strong interest in mechanization as a way to improve their own circumstances

Draft genome sequence of "Candidatus Cronobacter colletis" NCTC 14934T, a new species in the genus Cronobacter

Members of the Cronobacter genus are associated with serious infections in neonates. This is the first report of the draft genome sequence for the newly proposed species Cronobacter colletis