Heifers (n = 103) ranging in age from 1d to 2 yr were sampled to determine the coagulase-negative staphylococcal flora of haircoat, nares, vagina, teat skin, and streak canal. A total of 2706 staphylococal strains were identified from 3612 bacterial isolates. Other genera or groups identified included Bacillus, Micrococcus, Corynebacterium, and coliforms. Staphylococci were identified utilizing a simplified biochemical scheme. Staphylococcus xylosus, S. chromogenes, and S. warneri were the predominant species recovered from anatomic sites and streak canal. Staphylococcal strains identified from specific body sites (expressed as percentage of heifers harboring these species) were: nares 74% S. xylosus and 48% S. warneri; haircoat, 70% S. xylosus and 57% S. chromogenes; vagina, 60% S. chromogenes and 54% S. xylosus; teat skin 62% S. chromogenes and 61% S. warneri; streak canal 53% S. chromogenes, and 43% S. warneri. The prevalent staphylococcal strains identified differed from heifers in confined housing compared with heifers on pasture. Differences observed in distribution of Staphylococcus species among body sites, particularly those between teat skin and streak canal, suggest that establishment of staphylococcal microflora depends on the ability of a species to adapt to and colonize anatomic sites as well as on environmental conditions present.

The body sites of 163 heifers, bedding and feedstuff samples, flies, and hands and nares of the research personnel were sampled in order to determine the sources of Staphylococcus aureus in a dairy herd other than the lactating mammary gland. Lesions on the udder of lactating animals and the air in the milking parlor were also sampled. Staphylococci isolated from bedding samples were identified as to species. Staphylococcus aureus was isolated from all sources examined except flies. An enrichment procedure was necessary for isolating S. aureus from two bedding samples although other Staphylococcus species were present in high numbers. The designation "environmental staphylococci" is proposed for Staphylococcus species that were apparently free-living in the environment.

The ability of the pathogenic yeast Candida albicans to interconvert between budded and hyphal growth states, herein termed the budded-to-hyphal transition (BHT), is important for C. albicans development and virulence. The BHT is under the control of multiple cell signaling pathways that respond to external stimuli, including nutrient availability, high temperature, and pH. Previous studies identified 21 small molecules that could inhibit the C. albicans BHT in response to carbon limitation in Spider media. However, the studies herein show that the BHT inhibitors had varying efficacies in other hyphal-inducing media, reflecting their varying abilities to block signaling pathways associated with the different media. Chemical epistasis analyses suggest that most, but not all, of the BHT inhibitors were acting through either the Efg1 or Cph1 signaling pathways. Notably, the BHT inhibitor clozapine, a FDA-approved drug used to treat atypical schizophrenia by inhibiting G-protein-coupled dopamine receptors in the brain, and several of its functional analogs were shown to act at the level of the Gpr1 G-protein-coupled receptor. These studies are the first step in determining the target and mechanism of action of these BHT inhibitors, which may have therapeutic anti-fungal utility in the future.

Effects of intramammary infections in herds 1) either not teat dipping or postmilking teat dipping with either 2) linear dodecyl benzene sulfonic acid, 3) chlorhexidine, or 4) iodophor containing products were determined. Duplicate quarter foremilk samples were collected once from all lactating cows in 16 herds, four for each practice. Mean percentage of quarters infected with Staphylococcus species other than Staph. aureus was 11.0 in herds not teat dipping and 7.2 in herds teat dipping. Prevalence of Staphylococcus species intramammary infections in herds teat dipping with linear dodecyl benzene sulfonic acid was comparable to herds not teat dipping and greater than in herds using chlorhexidine and iodophor. The predominant Staphylococcus species in herds not teat dipping was Staph. epidermidis (37.1%). Staphylococcus hyicus constituted 48.5% of total Staphylococcus species intramammary infections in herds teat dipping and 12.4% in herds not teat dipping. Differences were not observed among SCC for quarters infected with different Staphylococcus species. Application of germicidal teat dips appeared to have selectively altered both prevalence and distribution of Staphylococcus species intramammary infections.

The efficacy of Baird-Parker agar, Baird-Parker agar with bacitracin (0.8 μg/ml), Baird-Parker agar supplemented with acriflavin (7 μg/ml), polymyxin (20 units/ml) and sulphonamides (55 μg/ml), KRANEP agar, mannitol salt agar, and Staphylococcus medium 110 agar for the isolation and enumeration of Staphylococcus species was investigated. Bovine blood agar was used as the control medium. Thirty-seven staphylococci strains representing 23 species and 19 non-Staphylococcus species were tested. None of the six selective media supported the growth of all 37 Staphylococcus strains. The number of Staphylococcus species that grew on a specific medium ranged from only the S. aureus strains on Baird-Parker agar supplemented with acriflavin, polymyxin, and sulphonamides, to all but S. warneri BG 647 on Staphylococcus medium 110 agar. Strains of Bacillus, Corynebacterium, and Micrococcus grew on all six selective media.

A fluorogenic assay for the detection of P-glucosidase was developed as part of a simplified conventional method to distinguish Staphylococcus warneri and Staphylococcus hominis isolated from bovine body sites. The assay is based on the fact that strains of S. warneri produce P-glucosidase, while strains of S. hominis do not.

Foodborne diseases are increasingly recognized as a significant global public health problem despite major advances and improvements in the quality of food , water, sanitation and hygiene. However, detection and characterization of foodborne pathogens during outbreak scenarios remains a laborious and time-consuming task. The aim of this work was to develop an oligonucleotide microarray for rapid detection and characterization of the most important infectious bacterial (Campylobacter, Salmonella and Yersinia) and viral (Noroviruses) pathogens found 1n swine and associated pork products. A total of 272 target regions and genes were identified that were specific for pathogen identification and characterization of specific antimicrobial resistance and virulence determinants. We designed multiple probes (up to three) per gene to increase the sensitivity and specificity of the microarray. After BLAST analysis, a total of 562 probes were finally selected to be printed on to glass slides. Appropriate control strains that were previously characterized in our laboratories by PCR were selected to test the developed arrays. Preliminary results indicated that the designed probes were highly specific and sensitive for identification of tested pathogens and known res1stance and virulence genes present in the selected control strains.