60 rats were trained in a runway at 1 trial each day through 4 phases of 15 trials each. In each phase reward was either large (24 pellets) or small (2 pellets). Rats were shifted from one reward magnitude to the other in the absence of previous experience with the shift magnitude (nontransfer shift) and following experience with the shift magnitude (transfer shift). A positive contrast effect was not obtained under either condition. A negative contrast effect was obtained under nontransfer shift conditions but not under transfer shift conditions. Transfer upshifts (small reward to large) were faster than nontransfer upshifts and transfer downshifts were slower than nontransfer downshifts.
Glucan plays a central role in sucrose-dependent biofilm formation by the dental pathogen Streptococcus mutans. This organism synthesizes several proteins capable of binding glucan. These are divided into the glucosyltransferases that catalyze the synthesis of glucan and the nonglucosyltransferase glucan-binding proteins (Gbps). The biological significance of the Gbps has not been thoroughly defined, but studies suggest that these proteins influence virulence and play a role in maintaining biofilm architecture by linking bacteria and extracellular molecules of glucan. We engineered a panel of Gbp mutants, targeting GbpA, GbpC, and GbpD, in which each gene encoding a Gbp was deleted individually and in combination. These strains were then analyzed by confocal microscopy and the biofilm properties were quantified by the biofilm quantification software comstat. All biofilms produced by mutant strains lost significant depth, but the basis for the reduction in height depended on which particular Gbp was missing. The loss of the cell-bound GbpC appeared dominant as might be expected based on losing the principal receptor for glucan. The loss of an extracellular Gbp, either GbpA or GbpD, also profoundly changed the biofilm architecture, each in a unique manner.
The survival and growth of Bacillus stearothermophilus and Bacillus licheniformis, naturally present (30–300 colony forming units/ml) in late season skim milk, was monitored in a three effect evaporator during low heat skim milk powder manufacture. Substantial growth was shown to occur in the preheating stages prior to direct steam heating. A typical heat treatment (77°C, 15 s) used in the manufacture of low heat powder did not inactivate the bacteria, which continued to grow in the heater. The importance of preheaters in influencing thermophile growth in the evaporator is demonstrated by the finding that growth in the preheater stages was accompanied by growth in subsequent evaporator effects which significantly exceeded that observed when the final two preheaters were bypassed. A mid‐run mini‐clean procedure incorporating 0.2% hydrogen peroxide for decontaminating the evaporator was tested and may prove useful in extending evaporator run times
One hundred human-derived coagulase negative staphylococci (CoNS) were screened for antimicrobial activity using agar-based deferred antagonism assays with a range of indicator bacteria. Based on the findings of the screen and subsequent well assays with cell free supernatants and whole cell extracts, one strain, designated CIT060, was selected for further investigation. It was identified as Staphylococcus capitis and herein we describe the purification and characterisation of the novel bacteriocin that the strain produces. This bacteriocin which we have named capidermicin was extracted from the cell-free supernatant of S. capitis CIT060 and purified to homogeneity using reversed-phase high performance liquid chromatography (RP-HPLC). Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometric (MS) analysis revealed that the capidermicin peptide has a mass of 5,464 Da. Minimal inhibitory concentration (MIC) experiments showed that capidermicin was active in the micro-molar range against all the Gram-positive bacteria that were tested. Antimicrobial activity was retained over a range of pHs (2–11) and temperatures (10–121°C x 15 mins). The draft genome sequence of S. capitis CIT060 was determined and the genes predicted to be involved in the biosynthesis of capidermicin were identified. These genes included the predicted capidermicin precursor gene, and genes that are predicted to encode a membrane transporter, an immunity protein and a transcriptional regulator. Homology searches suggest that capidermicin is a novel member of the family of class II leaderless bacteriocins.
BackgroundSevere-early childhood caries (S-ECC) is one of the most common infectious diseases in children and is prevalent in lower socio-economic populations. American Indian children suffer from the highest levels of S-ECC in the United States. Members of the mutans streptococci, Streptococcus mutans, in particular, are key etiologic agents in the development of caries. Children typically acquire S. mutans from their mothers and early acquisition is often associated with higher levels of tooth decay.MethodsWe have conducted a 5-year birth cohort study with a Northern Plains Tribe to determine the temporality and fidelity of S. mutans transmission from mother to child in addition to the genotypic diversity of S. mutans in this community. Plaque samples were collected from 239 mother/child dyads at regular intervals from birth to 36 months and S. mutans were isolated and genotyped by arbitrarily primed-polymerase chain reaction (AP-PCR).ResultsHere we present preliminary findings from a subset of the cohort. The focus for this paper is on initial acquisition events in the children. We identified 17 unique genotypes in 711 S. mutans isolates in our subset of 40 children, 40 mothers and 14 primary caregivers. Twelve of these genotypes were identified in more than one individual. S. mutans colonization occurred by 16 months in 57.5% of the children and early colonization was associated with higher decayed, missing and filled surface (DMFS) scores (p=0.0007). Children colonized by S. mutans shared a common genotype with their mothers 47.8% of the time. While multiple genotypes were common in adults, only 10% of children harbored multiple genotypes.ConclusionThese children acquire S. mutans at an earlier age than the originally described ‘window of infectivity’ and often, but not exclusively, from their mothers. Early acquisition is associated with both the caries status of the children and the mothers.
Raw cow's milk inoculated with four laboratory strains (102−105 cfu/mL) of Mycobacterium avium ssp. paratuberculosis (Map) was pasteurized in a custom designed pilot plant pasteurizer having a maximum throughput of 580 L/h under turbulent flow conditions. Following 16 pasteurizer trials none of the Map strains survived high‐temperature short‐time conditions (72.5°C × 27 s) whether milk was homogenized or not. Two dairy herds containing animals which were faecal positive for Map were sourced and milk was collected for pasteurization studies. Milk collected from one herd on five occasions in the autumn did not contain any detectable Map organisms, and the second herd that was sampled on only one occasion in early winter was shown to contain Map at low concentration. Map was not detected in any of these milks following pasteurization at 72.5°C for 27 s. Two natural isolates of Map inoculated into milk were likewise inactivated on pasteurization.
Dextran-dependent aggregation (DDAG) of Streptococcus mutans is an in vitro phenomenon that is believed to represent a property of the organism that is beneficial for sucrose-dependent biofilm development. GbpC, a cell surface glucan-binding protein, is responsible for DDAG in S. mutans when cultured under defined stressful conditions. Recent reports have described a putative transcriptional regulator gene, irvA, located just upstream of gbpC, that is normally repressed by the product of an adjacent gene, irvR. When repression of irvA is relieved, there is a resulting increase in the expression of GbpC and decreases in competence and synthesis of the antibiotic mutacin I. This study examined the role of irvA in DDAG and biofilm formation by engineering strains that overexpressed irvA (IrvA؉) on an extrachromosomal plasmid. The IrvA؉ strain displayed large aggregation particles that did not require stressful growth conditions. A novel finding was that overexpression of irvA in a gbpC mutant background retained a measure of DDAG, albeit very small aggregation particles. Biofilms formed by the IrvA؉ strain in the parental background possessed larger-than-normal microcolonies. In a gbpC mutant background, the overexpression of irvA reversed the fragile biofilm phenotype normally associated with loss of GbpC. Real-time PCR and Northern blot analyses found that expression of gbpC did not change significantly in the IrvA؉ strain but expression of spaP, encoding the major surface adhesin P1, increased significantly. Inactivation of spaP eliminated the small-particle DDAG. The results suggest that IrvA promotes DDAG not only by GbpC, but also via an increase in P1.
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