Identification and characterization of potential autochthonous starter cultures from a Croatian "brand" product "Slavonski kulen".

The microbial population of a traditional Croatian fermented sausage "Slavonski kulen" was isolated, identified and subjected to technological and functional characterization in order to select potential autochthonous functional starter cultures. Dominant microflora were lactic acid bacteria (LAB), followed by staphylococci. Identification of isolated lactobacilli showed domination of Leuconostoc mesenteroides and Lactobacillus acidophilus while Staphylococcus xylosus and Staphylococcus warneri outnumbered the staphylococcal microbiota. Most of the isolated LAB and Staphylococcus species displayed good growth in the presence of 5% of NaCl and at 12, 18 and 22°C. All LAB and most of the staphylococci possess proteolytic activity and only Staphylococcus xylosus had lipolytic activity. All lactobacilli and staphylococci isolates produced significant concentrations of lactic acid (as determined by HPLC) and showed antimicrobial activity against pathogenic test microorganisms. Dominant LAB and Staphylococcus species displayed growth in the presence of 1% bile. Most of the staphylococci and all of lactobacilli showed sensitivity to all antibiotics tested.

Importance of S-layer proteins in probiotic activity of Lactobacillus acidophilus M92.

AIMS: To investigate the functional role of surface layer proteins (S-layer) in probiotic strain Lactobacillus acidophilus M92, especially its influence on adhesiveness to mouse ileal epithelial cells. METHODS AND RESULTS: Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S-layer) proteins, ca at 45 kDa in L. acidophilus M92. Southern blot with pBK1 plasmid, containing slpA gene, gave a positive signal, suggesting that L. acidophilus M92 has a slpA gene coding for the S-layer proteins. S-layer proteins of this strain are present during all phases of growth. The S-layer proteins appeared when cells treated with 5 mol l(-1) LiCl were allowed to grow again. Removal of the S-layer proteins reduced adhesion of L. acidophilus M92 to mouse ileal epithelial cells. Furthermore, the viability of cells without S-layer were reduced in simulated gastric juice at low pH range (2, 2.5, 3) and simulated pancreatic juice with bile salts (1.5 and 3 g l(-1)). S-layer proteins of L. acidophilus M92 were resistant to pepsin and pancreatin, in contrast, the treatment with proteinase K led to a significant proteolysis of the S-layer proteins. CONCLUSIONS: These results demonstrated functional role of S-layer; it is responsible for adhesiveness of Lactobacillus acidophilus M92 to mouse ileal epithelial cells and has a protective role for this strain. SIGNIFICANCE AND IMPACT OF THE STUDY: S-layer proteins have an important role in the establishment of probiotic strain Lactobacillus acidophilus M92 in the gastrointestinal tract.

Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92.

AIMS: To investigate aggregation and adhesiveness of Lactobacillus acidophilus M92 to porcine ileal epithelial cells in vitro, and the influence of cell surface proteins on autoaggregation and adhesiveness of this strain. METHODS AND RESULTS: Lactobacillus acidophilus M92 exhibits a strong autoaggregating phenotype and manifests a high degree of hydrophobicity determined by microbial adhesion to xylene. Aggregation and hydrophobicity were abolished upon exposure of the cells to pronase and pepsin. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S-layer) proteins, approximated at 45 kDa, in L. acidophilus M92. The relationship between autoaggregation and adhesiveness to intestinal tissue was investigated by observing the adhesiveness of L. acidophilus M92 to porcine ileal epithelial cells. Removal of the S-layer proteins by extraction with 5 mol l-1 LiCl reduced autoaggregation and in vitro adhesion of this strain. CONCLUSIONS: These results demonstrate that there is relationship between autoaggregation and adhesiveness ability of L. acidophilus M92, mediated by proteinaceous components on the cell surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation has shown that L. acidophilus M92 has the ability to establish in the human gastrointestinal tract, which is an important determinant in the choice of probiotic strains.

Abortion controversy hinders state reform effort.

PIP: A review of the progress and problems of the state of Maryland on health care reform is provided. The Standard Benefits Package Task Force completed and approved in a preliminary 7 to 2 vote a long list of benefits, including abortion services, to be covered in health plans that must be made available to small businesses by July 1, 1994. Small businesses are those employing at least 2 workers and no more than 50 workers. The value of benefits cannot exceed $3519, or policies, $3034. The new health care law passed in summer, 1993 stipulated that the value of basic policies for small companies cannot exceed 12% of wages. The development of a minimum package of benefits was the first phase of the health care reform law. Final decision on benefits will be made sometime in the first week of November and presented to the full 7-member Health Care Access and Cost Commission by November 4, 1993. Abortion opponents have opposed insurance coverage of abortions on the grounds that it is not health care and it forces employers opposed to abortion to accept this package, or deny coverage to employees. A public hearing is expected to hear from abortion foes about their notion to offer abortion coverage as an option or "rider" to the standard insurance policy. The task force has another option: to describe the benefit in terms that do not mention abortion per se, but refer to "family planning services and services for pregnant women," which is the wording in the Clinton health care plan. The Director of Public Affairs for Planned Parenthood of Maryland, Bebe Verdery, reported that the organization is strongly opposed to abortion as optional coverage. Kevin Appleby, Associate Director of Social Concerns for the Maryland Catholic Conference, finds that respect for rights of those with moral concerns against abortion should be respected. The task force is confronted with issues of cost, since its initial benefits package was too high, and, most troubling, the controversy over abortion rights. Unfortunately, the controversy portends to obscure other difficult decisions about extent of coverage of hospitalization, psychiatric services, home health care, laboratory work, and prescription drugs.^ieng