Exploitation of the diverse insertion sequence element content of dairy Lactobacillus helveticus starters as a rapid method to identify different strains.

The species Lactobacillus helveticus is a commonly used thermophilic starter and/or adjunct culture for Swiss and Cheddar cheese manufacture. Its use is normally associated with flavour improvement which is known to be associated with culture traits such as rapid autolysis and high proteolytic activity. The genome of the commercial strain, DPC4571, was recently sequenced and found to have an abundance of IS sequences in terms of both abundance (213 intact) and diversity (21 types).

The presence of pMRC01 promotes greater cell permeability and autolysis in lactococcal starter cultures.

Conjugative transfer of plasmid-associated properties is routinely used to generate food-grade derivatives of lactococcal starter strains with improved technological traits. However, the introduction of one or more plasmids in a single strain is likely to impose a burden on regular cell metabolism and may affect the growth characteristics of the transconjugant culture. The aim of this study was to evaluate the impact of the 60.

Administration of a live culture of Lactococcus lactis DPC 3147 into the bovine mammary gland stimulates the local host immune response, particularly IL-1beta and IL-8 gene expression.

Mastitis is one of the most costly diseases to the dairy farming industry. Conventional antibiotic therapy is often unsatisfactory for successful treatment of mastitis and alternative treatments are continually under investigation. We have previously demonstrated, in two separate field trials, that a probiotic culture, Lactococcus lactis DPC 3147, was comparable to antibiotic therapy to treat bovine mastitis.

Bacteriophage and their lysins for elimination of infectious bacteria.

When phages were originally identified, the possibility of using them as antibacterial agents against pathogens was immediately recognized and put into practise based on the knowledge available at the time. However, with the advent of antibiotics a decline in the use of phage as therapeutics followed. Phages did, however, become more useful in the study of fundamental aspects of molecular biology and in the diagnostic laboratory for the identification of pathogenic bacteria.

Evaluation of an antimicrobial ingredient prepared from a Lactobacillus acidophilus casein fermentate against Enterobacter sakazakii.

Previously two antimicrobial peptides, IKHQGLPQE (caseicin A) and VLNENLLR (caseicin B), were identified following the fermentation of sodium caseinate with the proteolytic strain Lactobacillus acidophilus DPC 6026. This study evaluated the ability of these peptides to kill Enterobacter sakazakii ATCC 12868 spiked in reconstituted infant formula. The survival of E.

The vexed relationship between Clostridium difficile and inflammatory bowel disease: an assessment of carriage in an outpatient setting among patients in remission.

Objectives: Comorbidity with Clostridium difficile may cause diagnostic delay in newly presenting inflammatory bowel disease (IBD) patients, trigger relapse in established disease, confound therapies, and serve as an indicator of an underlying defect in innate immunity. Retrospective analyses have suggested community acquisition; to address this we conducted a prospective analysis of C. difficile carriage in IBD patients using molecular methods specifically in an outpatient setting.

Evaluation of two food grade proliposomes to encapsulate an extract of a commercial enzyme preparation by microfluidization.

The entrapment by microfluidization of a commercial enzyme extract (Debitrase DBP20) in liposomes using two food grade proliposome (C and S) preparations was studied. Liposomes obtained at a low microfluidization pressure (4000 psi) were distributed in a bimodal population of small (30-40 nm) and large vesicles (300-700 nm). The composition of the proliposome influenced entrapment efficiency and the repartition of the enzyme between the core and the surface of the liposome.

Comparative genomics of lactic acid bacteria reveals a niche-specific gene set.

Background: The recently sequenced genome of Lactobacillus helveticus DPC4571 revealed a dairy organism with significant homology (75% of genes are homologous) to a probiotic bacteria Lb. acidophilus NCFM. This led us to hypothesise that a group of genes could be determined which could define an organism's niche.

Optimization of a reconstituted skim milk based medium for enhanced CLA production by bifidobacteria.

Aims: To determine the effect of a range of supplements on the bioconversion of linoleic acid to conjugated linoleic acid (CLA) by Bifidobacterium breve NCIMB 702258 in reconstituted skim milk (RSM).

Results: Seven supplements (yeast extract, casein hydrolysate, tryptone, l-cysteine hydrochloride, sodium acetate, sodium butyrate and sodium propionate) were identified as increasing the bioconversion of linoleic acid to c9, t11 CLA. Using these supplements, the percentage bioconversion of linoleic acid (0.

Role of gut microbiota in early infant development.

Early colonization of the infant gastrointestinal tract is crucial for the overall health of the infant, and establishment and maintenance of non-pathogenic intestinal microbiota may reduce several neonatal inflammatory conditions. Much effort has therefore been devoted to manipulation of the composition of the microbiota through 1) the role of early infant nutrition, particularly breast milk, and supplementation of infant formula with prebiotics that positively influence the enteric microbiota by selectively promoting growth of beneficial bacteria and 2) oral administration of probiotic bacteria which when administered in adequate amounts confer a health benefit on the host. While the complex microbiota of the adult is difficult to change in the long-term, there is greater impact of the diet on infant microbiota as this is not as stable as in adults.

Characterization of enterocin- and salivaricin-producing lactic acid bacteria from the mammalian gastrointestinal tract.

Bacteriocin production may be a factor contributing to bacterial dominance within complex microbial populations and may therefore be a common trait within the gut microbiota. However, of 278 antimicrobial-producing culturable lactic acid bacteria (LAB) isolated from a range of mammalian intestinal sources in this study, characterization revealed just 23 distinct strains producing bacteriocin-like inhibitory substances and one Streptococcus hyointestinalis strain producing a potentially novel protease-insensitive antimicrobial. Three class II bacteriocins previously isolated from intestinal-derived LAB were identified as enterocin A and two salivaricin P-like bacteriocins.

Antimicrobial activity of two peptides casecidin 15 and 17, found naturally in bovine colostrum.

Aims: To isolate and characterize peptides from bovine colostrum with antimicrobial activity.

Methods And Results: Three peptides were purified from fresh colostrum by a range of chromatographic methods using antimicrobial activity against Escherichia coli DH5alpha to screen for the most active fractions. Two peptides, with antimicrobial activity, casecidin 17 and casecidin 15, were identical to sequences in the C-terminal of bovine beta-casein (YQEPVLGPVRGPFPIIV and YQEPVLGPVRGPFPI) and had corresponding molecular masses of 1881.

Inhibitory effect of conjugated alpha-linolenic acid from bifidobacteria of intestinal origin on SW480 cancer cells.

In this study, we assessed the ability of six strains of bifidobacteria (previously shown by us to possess the ability to convert linoleic acid to c9, t11-conjugated linoleic acid (CLA) to grow in the presence of alpha-linolenic acid and to generate conjugated isomers of the fatty acid substrate during fermentation for 42 h. The six strains of bifidobacteria were grown in modified MRS (mMRS) containing alpha-linolenic acid for 42 h at 37 degrees C, after which the fatty acid composition of the growth medium was assessed by gas liquid chromatography (GLC). Indeed, following fermentation of one of the strains, namely Bifidobacterium breve NCIMB 702258, in the presence of 0.

Phage lysin LysK can be truncated to its CHAP domain and retain lytic activity against live antibiotic-resistant staphylococci.

A truncated derivative of the phage endolysin LysK containing only the CHAP (cysteine- and histidine-dependent amidohydrolase/peptidase) domain exhibited lytic activity against live clinical staphylococcal isolates, including methicillin-resistant Staphylococcus aureus. This is the first known report of a truncated phage lysin which retains high lytic activity against live staphylococcal cells.

Intramammary infusion of a live culture for treatment of bovine mastitis: effect of live lactococci on the mammary immune response.

In the accompanying article, we demonstrated that a live culture of Lactococcus lactis compares favourably with antibiotics for treatment of bovine mastitis in two initial field trials. In an effort to explain the mechanism involved, this study investigated the effect of culture administration on the local immune response. In this respect we initially observed that infusion of the live culture Lactococcus lactis stimulated substantial recruitment of polymorphonucleocytes (PMN) and lymphocytes to the udder.

Intramammary infusion of a live culture of Lactococcus lactis for treatment of bovine mastitis: comparison with antibiotic treatment in field trials.

A treatment containing a live food-grade organism, Lactococcus lactis DPC3147, was compared with conventional antibiotic therapy for its potential to treat bovine chronic subclinical or clinical mastitis in two separate field trials. Effects on disease symptoms and bacteriology were monitored in response to infusion with the culture in each trial. In the first trial, the live culture treatment was compared with an intramammary antibiotic (n=11 quarters for each treatment).

Effects of cutting intensity and stirring speed on syneresis and curd losses during cheese manufacture.

Recombined whole milk was renneted under constant conditions of pH, temperature, and added calcium, and the gel was cut at a constant firmness. The effects of cutting and stirring on syneresis and curd losses to whey were investigated during cheese making using a factorial design with 3 cutting modes designed to provide 3 different cutting intensity levels (i.e.

Characterisation of single nucleotide polymorphisms identified in the bovine lactoferrin gene sequences across a range of dairy cow breeds.

The lactoferrin gene sequences of 70 unrelated dairy cows representing six different dairy breeds were investigated for single nucleotide polymorphisms to establish a baseline of polymorphisms that exist within the Irish bovine population. Twenty-nine polymorphisms were identified within a 2.2kb regulatory region.

Tryptophan-mediated denaturation of beta-lactoglobulin A by UV irradiation.

beta-Lactoglobulin A, a genetic variant of one of the main whey proteins, was irradiated at 295 nm for 24 h. After irradiation, 18% of the protein was denatured (determined by reverse-phase chromatography). The fluorescence spectrum of the irradiated protein was red-shifted compared to that of the native protein, indicating a change in protein folding.

Towards a one-step Enterobacter sakazakii enrichment.

Aims: The current international standard method for detection of Enterobacter sakazakii from milk products is by the International Organization for Standardization and the International Dairy Federation documented method, a procedure involving two-step enrichment. This study aimed to assess enrichment of E. sakazakii using a one-step enrichment.

Presence of two Lactobacillus and Bifidobacterium probiotic strains in the neonatal ileum.

The overall purpose of this study was to examine the lactobacilli and bifidobacteria microbiota in the human ileum at a very early stage of life. Ileostomy effluents from two infants, taken at different time points, were plated on Lactobacillus selective agar and cys-MRS containing mupirocin to select for bifidobacteria. In one case, a stool sample following ileostomy reversal was subsequently analyzed microbiologically.

Genome sequence of Lactobacillus helveticus, an organism distinguished by selective gene loss and insertion sequence element expansion.

Mobile genetic elements are major contributing factors to the generation of genetic diversity in prokaryotic organisms. For example, insertion sequence (IS) elements have been shown to specifically contribute to niche adaptation by promoting a variety of genetic rearrangements. The complete genome sequence of the cheese culture Lactobacillus helveticus DPC 4571 was determined and revealed significant conservation compared to three nondairy gut lactobacilli.

Contamination of milk by enterococci and coliforms from bovine faeces.

Aim: To determine the contribution of enterococci and coliforms from bovine faeces and teats to contamination of raw milk.

Methods: Putative enterococci (n = 301) and coliforms (n = 365) were isolated from bovine faeces (n = 20), cows' teats (n = 20), the raw milk (n = 1) and the milking environment (n = 4) on one farm. The clonal relationships of each bacterial group were investigated using Pulsed-Field Gel Electrophoresis of genomic macrorestriction fragments.

Growth characteristics of Brevibacterium, Corynebacterium, Microbacterium, and Staphylococcus spp. isolated from surface-ripened cheese.

The growth characteristics of five bacteria, Brevibacterium aurantiacum 1-16-58, Corynebacterium casei DPC 5298(T), Corynebacterium variabile DPC 5310, Microbacterium gubbeenense DPC 5286(T), and Staphylococcus saprophyticus 4E61, all of which were isolated from the surface of smear cheese, were studied in complex and chemically defined media. All of the coryneforms, except M. gubbeenense, grew in 12% salt, while B.

Salmonella enterica phage-resistant mutant colonies display an unusual phenotype in the presence of phage Felix 01.

Aims: To investigate irregular colony morphology formation in Salmonella enterica serovar Typhimurium DPC6046 in the presence of a lytic phage, Felix 01.

Methods And Results: Phage-resistant derivatives of the parent strain DPC6046 were isolated which exhibited an irregular colony morphology. These were subjected to viability studies by using confocal scanning laser microscopy and live/dead BacLight stain to evaluate the cell viability within the colony.