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

Diversity of O Antigens within the Genus Cronobacter: from Disorder to Order

Cronobacter species are Gram-negative opportunistic pathogens that can cause serious infections in neonates. The lipopolysaccharides (LPSs) that form part of the outer membrane of such bacteria are possibly related to the virulence of particular bacterial strains. However, currently there is no clear overview of O-antigen diversity within the various Cronobacter strains and links with virulence. In this study, we tested a total of 82 strains, covering each of the Cronobacter species. The nucleotide variability of the O-antigen gene cluster was determined by restriction fragment length polymorphism (RFLP) analysis. As a result, the 82 strains were distributed into 11 previously published serotypes and 6 new serotypes, each defined by its characteristic restriction profile. These new serotypes were confirmed using genomic analysis of strains available in public databases: GenBank and PubMLST Cronobacter. Laboratory strains were then tested using the current serotype-specific PCR probes. The results show that the current PCR probes did not always correspond to genomic O-antigen gene cluster variation. In addition, we analyzed the LPS phenotype of the reference strains of all distinguishable serotypes. The identified serotypes were compared with data from the literature and the MLST database (www.pubmlst.org/cronobacter/). Based on the findings, we systematically classified a total of 24 serotypes for the Cronobacter genus. Moreover, we evaluated the clinical history of these strains and show that Cronobacter sakazakii O2, O1, and O4, C. turicensis O1, and C. malonaticus O2 serotypes are particularly predominant in clinical cases

Protein sequences insight into heavy metal tolerance in Cronobacter sakazakii BAA-894 encoded by plasmid pESA3

The recently annotated genome of the bacterium Cronobacter sakazakii BAA-894 suggests the organism has the ability to bind heavy metals. This study demonstrates heavy metal tolerance in Cronobacter sakazakii, in which proteins with the heavy metal interaction were recognized by computational and experimental study. As the result, approximately one fourth of proteins encoded on the plasmid pESA3 are proposed to have potential interaction with heavy metals. Interaction between heavy metals and predicted proteins was further corroborated using protein crystal structures from protein data bank database and comparison of metal-binding ligands. In addition with, a phylogenetic study was undertaken for most toxic heavy metals, like arsenic, cadmium, lead and mercury and obtained related tree pattern for lead, cadmium and arsenic. Laboratory studies confirmed the organism's tolerance to tellurite, copper and silver. These experimental and computational study data extend our understanding of the genes encoding for proteins of this important neonatal pathogen and provides further insights into the genotypes associated with features that can contribute to its persistence in the environment. The information will be of value for future environmental protection from heavy toxic metals

Sustainability of water resources management in the Indus Basin under changing climatic and socio economic conditions

Pakistan is highly dependent on water resources originating in the mountain sources of the upper Indus for irrigated agriculture which is the mainstay of its economy. Hence any change in available resources through climate change or socio-economic factors could have a serious impact on food security and the environment. In terms of both ratio of withdrawals to runoff and per-capita water availability, Pakistan's water resources are already highly stressed and will become increasingly so with projected population changes. Potential changes to supply through declining reservoir storage, the impact of waterlogging and salinity or over-abstraction of groundwater, or reallocations for environmental remediation of the Indus Delta or to meet domestic demands, will reduce water availability for irrigation. <br><br> The impact of climate change on resources in the Upper Indus is considered in terms of three hydrological regimes – a nival regime dependent on melting of winter snow, a glacial regime, and a rainfall regime dependent on concurrent rainfall. On the basis of historic trends in climate, most notably the decline in summer temperatures, there is no strong evidence in favour of marked reductions in water resources from any of the three regimes. Evidence for changes in trans-Himalayan glacier mass balance is mixed. Sustainability of water resources appears more threatened by socio-economic changes than by climatic trends. Nevertheless, analysis and the understanding of the linkage of climate, glaciology and runoff is still far from complete; recent past climate experience may not provide a reliable guide to the future

The speciation and genotyping of Cronobacter isolates from hospitalised patients

The World Health Organization (WHO) has recognised all Cronobacter species as human pathogens. Among premature neonates and immunocompromised infants, these infections can be life-threatening, with clinical presentations of septicaemia, meningitis and necrotising enterocolitis. The neurological sequelae can be permanent and the mortality rate as high as 40 – 80 %. Despite the highlighted issues of neonatal infections, the majority of Cronobacter infections are in the elderly population suffering from serious underlying disease or malignancy and include wound and urinary tract infections, osteomyelitis, bacteraemia and septicaemia. However, no age profiling studies have speciated or genotyped the Cronobacter isolates. A clinical collection of 51 Cronobacter strains from two hospitals were speciated and genotyped using 7-loci multilocus sequence typing (MLST), rpoB gene sequence analysis, O-antigen typing and pulsed- field gel electrophoresis (PFGE). The isolates were predominated by C. sakazakii sequence type 4 (63 %, 32/51) and C. malonaticus sequence type 7 (33 %, 17/51). These had been isolated from throat and sputum samples of all age groups, as well as recal and faecal swabs. There was no apparent relatedness between the age of the patient and the Cronobacter species isolated. Despite the high clonality of Cronobacter , PFGE profiles differentiated strains across the sequence types into 15 pulsotypes. There was almost complete agreement between O-antigen typing and rpoB gene sequence analysis and MLST profiling. This study shows the value of applying MLST to bacterial population studies with strains from two patient cohorts, combined with PFGE for further discrimination of strains

Comparative Outer Membrane Protein Analysis of High and Low-Invasive Strains of Cronobacter malonaticus

are an important group of foodborne pathogens that has been linked to life-threatening infections in both infants and adults. The major infections associated with species are neonatal meningitis, necrotizing enterocolitis, and septicaemia. There are seven species in the genus, of which only three are of clinical importance; , and . To date most studies have focussed on as it is the major species associated with neonatal infections. However, recently , in particular sequence type 7 (ST7), has been noted as being prevalent in adult infections and therefore warranting further investigation. In this study, eight strains of ST7, that had been isolated from a wide range of sources and varied in their virulence, were chosen for proteomic analysis of their outer membrane proteins (OMPs). One-dimensional gel analysis revealed a ~29 kDa size band that was only present in the highly invasive strains. Subsequent mass spectrometric analysis identified several peptides that matched the flagellin protein. The presence of flagellin protein was confirmed in 2D gel spot. Mass spectrometry analysis of total OMPs revealed that the four highly invasive strains expressed the main flagellum proteins that were absent from the four low invasive strains. These were the flagellar hook protein FlgE, flagellar hook-associated protein 1, flagellar hook-associated protein, flagellin, and flagellar hook-filament junction protein FlgL. This data indicates that flagellar proteins may have an important role in the organism's invasion properties

Surface grafting of electrospun fibers using ATRP and RAFT for the control of biointerfacial interactions

BACKGROUND The ability to present signalling molecules within a low fouling 3D environment that mimics the extracellular matrix is an important goal for a range of biomedical applications, both in vitro and in vivo. Cell responses can be triggered by non-specific protein interactions occurring on the surface of a biomaterial, which is an undesirable process when studying specific receptor-ligand interactions. It is therefore useful to present specific ligands of interest to cell surface receptors in a 3D environment that minimizes non-specific interactions with biomolecules, such as proteins. METHOD In this study, surface-initiated atom transfer radical polymerization (SI-ATRP) of poly(ethylene glycol)-based monomers was carried out from the surface of electrospun fibers composed of a styrene/vinylbenzyl chloride copolymer. Surface initiated radical addition-fragmentation chain transfer (SI-RAFT) polymerisation was also carried out to generate bottle brush copolymer coatings consisting of poly(acrylic acid) and poly(acrylamide). These were grown from surface trithiocarbonate groups generated from the chloromethyl styrene moieties existing in the original synthesised polymer. XPS was used to characterise the surface composition of the fibers after grafting and after coupling with fluorine functional XPS labels. RESULTS Bottle brush type coatings were able to be produced by ATRP which consisted of poly(ethylene glycol) methacrylate and a terminal alkyne-functionalised monomer. The ATRP coatings showed reduced non-specific protein adsorption, as a result of effective PEG incorporation and pendant alkynes groups existing as part of the brushes allowed for further conjugation of via azide-alkyne Huisgen 1,3-dipolar cycloaddition. In the case of RAFT, carboxylic acid moieties were effectively coupled to an amine label via amide bond formation. In each case XPS analysis demonstrated that covalent immobilisation had effectively taken place. CONCLUSION Overall, the studies presented an effective platform for the preparation of 3D scaffolds which contain effective conjugation sites for attachment of specific bioactive signals of interest, as well as actively reducing non-specific protein interactions.This research was supported by the Cooperative Research Centre for Polymers (CRCP)

Solving the inhomogeneous Bethe-Salpeter equation

We develop an advanced method of solving homogeneous and inhomogeneous Bethe-Salpeter equations by using the expansion over the complete set of 4-dimensional spherical harmonics. We solve Bethe-Salpeter equations for bound and scattering states of scalar and spinor particles for the case of one meson exchange kernels. Phase shifts calculated for the scalar model are in agreement with the previously published results. We discuss possible manifestations of separability for one meson exchange interaction kernels.Comment: 9 pages, 11 eps-figures. Talk presented by S. S. Semikh at XVII International Baldin Seminar on High Energy Physics Problems "Relativistic Nuclear Physics and Quantum Chromodynamics", September 27 - October 2, 2004, Dubna, Russia; to appear in the proceedings of this conferenc

'Reclaiming the criminal' : the role and training of prison officers in England, 1877-1914

This article examines the role and training of prison officers in England, between 1877 and 1914. It is concerned with the changing penal philosophies and practices of this period and how these were implemented in local prisons, and the duties of the prison officer. More broadly, this article argues that the role of the prison officer and their training (from 1896) reflect wider ambiguities in prison policy and practice during this period