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L K Fox

Publications and source records attributed to L K Fox.

At least 37 records · Page 2Linked to original sources
PubMed2001

Identity of activation molecule 3 on superantigen-stimulated bovine cells is CD26.

Most CD8(+) T cells in cultures of bovine mononuclear cells stimulated with staphylococcal enterotoxin C1 develop an unusual phenotype characterized by expression of activation molecule 3 (ACT3). This superantigen-dependent phenotype may be relevant to immunopathogenesis mediated by certain microbial toxins. The size and N-terminal sequence of immunoprecipitated ACT3 indicate that ACT3 is the bovine orthologue of CD26.

Amino Acid Sequence↗
PubMed2001

Technical note: Therapeutic cessation of lactation of Staphylococcus aureus-infected mammary quarters.

The objective of the present study was to compare the ability of chlorhexidine and povidone-iodine to cause cessation of lactation in Staphylococcus aureus-infected mammary quarters, assess milk production in the treated quarter in the subsequent lactation, and evaluate whether microbiological cure was obtained. Fourteen mid- to late-lactation Holstein-Friesian dairy cattle from the Washington State University dairy herd with single mammary quarter S. aureus intramammary infections were studied. Cows were randomly assigned to one of two treatment groups, povidone-iodine or chlorhexidine. Cows in the povidone-iodine group were infused with 120 ml of 5% povidone-iodine solution (0.5% iodine) after complete milk-out. Chlorhexidine-treated cows were infused with a proprietary chlorhexidine suspension after two milkings 24 h apart. Treated mammary quarters were not milked for the rest of the lactation. Milk production from each mammary quarter (kg of milk/quarter) was measured using in-line volume flow meters for 5 consecutive days before treatment and again at the start of the subsequent lactation. Povidoneiodine caused permanent cessation of lactation in the treated quarter, whereas 71% of the chlorhexidine-treated mammary quarters returned to function in the subsequent lactation. Hence, if the primary objective is to eliminate the mammary quarter from lactation, and thereby presumably lower the risk of herdmates acquiring new S. aureus intramammary infection, then povidone-iodine appears to be the best of the two methods. No difference in total milk production between lactation one and two in either group was found, suggesting that permanent loss of a quarter was not detrimental to overall milk production.

Animals↗
PubMed2001

Bovine mammary immune response to an experimental intramammary infection with a Staphylococcus aureus strain containing a gene for staphylococcal enterotoxin C1.

Staphylococcal enterotoxin C (SEC), a superantigen, is the most frequently expressed enterotoxin by bovine strains of Staphylococcus aureus causing mastitis. To examine the possible impact of SEC on the immune response of the bovine mammary gland, we monitored changes in lymphocyte subpopulations in mammary glands of four lactating cows after intramammary instillation of S. aureus strain Rn4220 transformed with a plasmid containing a gene coding for SEC1. Four other lactating cows received the same strain transformed with the plasmid without the SEC1 gene (positive control), and four cows were untreated (negative control). Mammary quarter milk samples for somatic cell count (SCC) analysis and determination of N-acetyl-beta-D-glucosimindase (NAGase) activity levels were collected daily for 21 d postinstillation. Flow cytometry utilizing three-color analysis was used to phenotype lymphocyte subpopulations isolated from milk samples collected on d 0, 4, 7, 11, 14, 18, and 21 postinstillation from all the cows. Milk from mammary gland halves (positive control and experimental) or all mammary quarters (negative control) was collected for flow cytometric analysis. Increased NAGase activity, SCC, and isolated S. aureus demonstrated that infection was established in mammary quarters intrammarily instilled with bacteria. There were no significant differences (P > 0.05) in the proportions of BoCD4 helper T lymphocytes or BoCD8 cytotoxic T lymphocytes between the two infected treatment groups. There was a significant day x treatment difference of the proportion of a gammadelta T cell subpopulation that did not express BoCD2, but did express the ACT2 activation molecule and a significant treatment difference of a gammadelta T cell subpopulation that expressed BoCD2, but not the ACT2 activation molecule (P < 0.05). Results do not support the hypothesis that the presence of the gene for SEC1 alters the mammary BoCD4 or BoCD8 T lymphocyte response to infection.

Acetylglucosaminidase↗
PubMed2000

Isolation of bovine mammary lymphocytes for fluorescent activated flow cytometry.

A detailed methodology is described for the isolation of T lymphocytes from bovine mammary gland secretions for flow cytometry. Mammary gland secretions are collected and centrifuged to separate the leukocytes from the mammary supernatant. The leukocytes are purified using a density gradient and diluted for fluorescent staining. Using tri-color fluorescent staining, the T lymphocytes are identified by monoclonal antibodies and stained with secondary antibodies. Flow cytometry is used to quantify the cellular subpopulations of the T lymphocytes. Proportional and statistical analysis of the flow cytometric data is conducted with computer software, CELLQuest and SAS.

Animals↗
PubMed2000

The ability of the enzyme-linked immunosorbent assay to detect antibody against Staphylococcus aureus in milk following experimental intramammary infection.

Changes in the milk antibody levels against Staphylococcus aureus were measured at the start of an experimental intramammary instillation of either S. aureus (Study I) or Staphylococcus hyicus (Study II). A commercial enzyme-linked immunosorbent assay system was used. Twenty-one Holstein cows were enrolled in Study I and 15 Holstein cows were used in Study II. Pathogen instillation began 21 days before the start of the non-lactating period. Cows received intramammary antibiotic treatment in all quarters immediately after the last milking, the start of the non-lactating period. Lacteal secretions were collected before the start of the non-lactating period, and during the immediate postpartum period in both studies, and during the non-lactating period in Study I. Milk was cultured for mastitis pathogens and S. aureus antibody levels and somatic cell counts were determined from all samples. There was an approximate 2-week delay in the elevation in antibody levels in response to the instillation of S. aureus. Antibody levels remained elevated in cows with S. aureus intramammary infections postpartum, but were below threshold in cows where intramammary infections were cured during the non-lactating period. Antibody levels were elevated by S. hyicus intramammary infections, remained elevated for the first 12 days postpartum, but were below threshold by day 21 postpartum. Cows with incipient intramammary S. aureus infections might be misclassified as false negatives by the antibody test. However, results suggest that cows with S. hyicus intramammary infections that were not cured would not be misclassified if milk is withheld from test for the first 30 days postpartum, as recommended by the manufacturer of the test.

Animals↗
PubMed1999

Comparison of tilmicosin and cephapirin as therapeutics for Staphylococcus aureus mastitis at dry-off.

Forty-four cows (26 Jerseys and 18 Holsteins) that had at least 1 mammary quarter that was naturally (n = 12) or experimentally (n = 84) infected with Staphylococcus aureus were allotted to three treatment groups of approximately equal number at the end of lactation. Cows were dried off by abrupt cessation of milking, and dry cow therapy was administered as an intramammary infusion of cephapirin benzathine at 10 ml per quarter, an intramammary infusion of tilmicosin (solution containing 300 mg/ml) at 5 ml per quarter, or a subcutaneous injection of tilmicosin at 5 mg/kg of body weight on the day of drying off and another injection 4 d later. Mammary secretions were monitored during the dry period and postpartum for antimicrobial residues, intramammary infection (IMI) status, and somatic cell counts. Results demonstrated the following percentage cures for IMI caused by Staph. aureus at 28 d postcalving based on individual mammary quarters: cephapirin benzathine, 78.1%; tilmicosin infused, 74.2%; and tilmicosin injected, 9.1%. During the first 4 wk after drying off, the mean concentration of tilmicosin in mammary secretions from cows infused with the antibiotic remained approximately 10-fold higher than that in secretions from cows injected with the antibiotic (3.43 vs. 0.32 ppm), and, by the time of calving, concentrations for cows treated with both methods were below the dilution limit of the assay (< 0.1 ppm). Results demonstrated that intramammary infusion of tilmicosin was equally as effective as cephapirin benzathine in curing IMI caused by Staph. aureus at drying off; however, the subcutaneous injection of tilmicosin at the dose used was not effective as a dry cow therapeutic against Staph. aureus.

Animals↗
PubMed1998

Outbreak of mastitis caused by one strain of Staphylococcus aureus in a closed dairy herd.

The Washington State University dairy experienced an outbreak of intramammary infections (IMI) caused by Staphylococcus aureus during autumn 1993 through summer 1995. The outbreak was believed to be a result of transmission of 1 strain of S aureus in a herd that historically had excellent control of contagious mastitis. Control practices included strict hygiene at time of milking and preferential culling of cows infected with S aureus. Mastitis caused by Streptococcus agalactiae was not found in this herd. Despite excellent control practices, the strain of S aureus caused a new infection rate of approximately 3% of the herd per month. Moreover, a second strain of S aureus, isolated from a cow with mastitis, was introduced into the herd experimentally, and it failed to transmit disease. The outbreak of S aureus mastitis in this herd was eventually controlled by maintaining a program of strict milking time hygiene, by intensifying the program of preferentially culling infected cows, and by segregating cows with S aureus IMI in a separate pen and milking these infected cows last.

Animals↗
PubMed1998

Factors affecting retirement migration to Idaho: an adaptation of the amenity retirement migration model.

The purpose of this study was to determine factors affecting retirement migration of older persons, using Haas and Serow's (1993) model as the framework. Data were based on 462 persons aged 50 and older who had migrated to Idaho and surrendered their out-of-state drivers' licenses during 1992 and 1993. Most moved from another state in the west and were motivated to move to Idaho by such factors as outdoor recreation and quality and pace of life. They had visited their new community at least five times before moving and developed social ties quickly upon arriving. Businesses and community planners can more effectively anticipate changes necessary to accommodate older migrants to their community.

Aged↗
PubMed1998

Intracellular Staphylococcus aureus escapes the endosome and induces apoptosis in epithelial cells.

We examined the invasion of an established bovine mammary epithelial cell line (MAC-T) by a Staphylococcus aureus mastitis isolate to study the potential role of intracellular survival in the persistence of staphylococcal infections. S. aureus cells displayed dose-dependent invasion of MAC-T cells and intracellular survival. An electron microscopic examination of infected cells indicated that the bacteria induced internalization via a mechanism involving membrane pseudopod formation and then escaped into the cytoplasm following lysis of the endosomal membrane. Two hours after the internalization of S. aureus, MAC-T cells exhibited detachment from the matrix, rounding, a mottled cell membrane, and vacuolization of the cytoplasm, all of which are indicative of cells undergoing programmed cell death (apoptosis). By 18 h, the majority of the MAC-T cell population exhibited an apoptotic morphology. Other evidence for apoptosis was the generation of MAC-T cell DNA fragments differing in size by increments of approximately 180 bp and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling of the fragmented nuclear DNA of the infected host cells. These results demonstrate that after internalization S. aureus escapes the endosome and induces apoptosis in nonprofessional phagocytes.

Animals↗
PubMed1998

Sources of intramammary infections from Staphylococcus aureus in dairy heifers at first parturition.

The study objective was to identify probable sources and modes of transmission of 91 Staphylococcus aureus isolates obtained from the colostrum of 76 heifers at parturition. Sources cultured were milk (including colostrum), heifer body sites (teats, muzzle, rectum, vagina, and lacteal secretions), and environmental sites (bedding, insects, housing, water, feedstuffs, humans, nonbovine animals, air, and equipment). Staphylococcus aureus isolates were characterized by 63 phenotypic traits. A similarity coefficient was calculated by herd to identify the S. aureus that most closely resembled the S. aureus obtained from heifer colostrum. Staphylococcus aureus from a heifer's colostrum was compared with all preexisting S. aureus isolates from that heifer's herd. Isolates that were > or = 90% similar were considered to be identical. Because 30 (of the 91) S. aureus isolates from heifer colostrum were collected prior to environmental sampling, only 61 S. aureus isolates from heifer colostrum were available for comparison among all three sources. Possible sources of S. aureus from heifer colostrum at parturition were milk (70%, 43 of 61 isolates), heifer body sites (39%, 24 of 61), environmental sites (28%, 17 of 61), or no identified source (16%, 10 of 61). Three heifers with intramammary infection (IMI) from S. aureus at parturition had the same S. aureus on their teats prior to parturition. Milk was the only source identified for 41% (25 of 61) of isolates from heifer colostrum. Isolates from heifer body sites were the only source identified for 5% (3 of 61) of heifer colostrum isolates. Staphylococcus aureus from the environment was never the sole possible source for S. aureus from heifer colostrum. Data suggest that the major sources of S. aureus IMI in heifers at parturition are milk and heifer body sites. Contact among heifers may be an important mode of transmission of S. aureus leading to IMI in heifers at parturition.

Animals↗
PubMed1998

A comparison of two methods of evaluation of teat skin pathology.

A split-plot design with repeated measures was used to test the relationship between visual teat skin condition score, the degree of transepidermal water loss from the skin, and the colonization by Staphylococcus aureus on experimentally chapped and inoculated teats of 20 lactating Holstein cows. Visual teat skin chapping score and the number of S. aureus colonies obtained from a teat skin swab were correlated (r = 0.53). Transepidermal water loss and S. aureus count were not correlated (r = 0.02). Results indicated that visual teat skin evaluation is superior to measurements of transepidermal water loss in the prediction of the susceptibility of teat skin to colonization by S. aureus.

Animals↗
PubMed1998

Effects of premilking and postmilking teat disinfectants on teat skin condition.

An incomplete block split-plot design with repeated measures was used to test the effects of eight combinations of premilking and postmilking teat disinfectants on teat skin and teat end condition over a 6-wk period. These combinations included iodine, sodium chlorite, lactic acid, and orthophosphoric acid. Visual assessments of teat skin and teat ends were made on the teats of 101 cows. Teat skin thickness, teat end thickness, and transepidermal water loss were assessed for the teats of 50 cows. The interaction of predip and postdip had no effect on teat condition. Postdip had a significant effect on transepidermal water loss, visual teat skin condition, and visual teat end condition. The 1% iodine postdip was associated with the poorest teat skin condition.

Animals↗
PubMed1998

Characterization of staphylococcal bovine mastitis isolates using the polymerase chain reaction.

A polymerase chain reaction (PCR) assay was adapted to detect toxin genes of staphylococcal isolates from cases of bovine mastitis. Samples were obtained from three geographical areas: Korea and Idaho and Washington in the northwest United States. Samples from Korea and Washington were randomly chosen. Idaho samples were from a prospective study of mastitis etiology. Forty-one milk samples from 25 commercial farms in south-central Idaho were collected from cows with symptoms of mastitis. Although Staphylococcus aureus constituted 37.5% of mastitis isolates, these isolates lacked genes for staphylococcal enterotoxins (SEs), toxic shock syndrome toxin, and exfoliative toxins. In contrast, 4 of 13 isolates from Washington and 6 of 20 isolates from South Korea expressed SEs. These results suggest that PCR may be an effective means of screening bovine isolates for toxins. They also emphasize the potential for significant geographic differences in mastitis etiology.

Animals↗
PubMed1998

Induction of type 2 cytokines by a staphylococcal enterotoxin superantigen.

Persistent intramammary infections of dairy cows with Staphylococcus aureus may involve immunosuppression mediated by bacterial toxins such as enterotoxins and other super-antigens (SAgs). Previously we found that stimulation of bovine PBMC with staphylococcal enterotoxin C (SEC) induced a unique phenotype of activated CD8+ T cells expressing a newly identified activation molecule, ACT3. In the present study we found that SEC induced the expression of interleukin (IL)-4 and IL-10 mRNAs, two cytokines associated with type 2 responses. Elevated levels of IL-4 and IL-10, observed between day 0 and day 4 of culture, were associated with temporary inhibition of proliferative responses of T cells, evidenced by a decrease in numbers of CD4+ T cells and a small increase in numbers of CD8+ T cells. Vigorous proliferation of T cells occurred between days 4 and 7 of culture and with a bias towards CD8+ T cells. Acquisition of the ACT3+ phenotype by CD8+ T cells was preceded by induction of IL-4 mRNA. Thus, in the bovine system, SAgs may hinder protective responses by inducing type 2 cytokines, which interfere with immune clearance of many microbial pathogens. The results of the study are consistent with the hypothesis that SAgs are involved in immunosuppression, and suggest possible immunomodulatory mechanisms.

Animals↗
PubMed1997

Genetic association between parameters of inmate immunity and measures of mastitis in periparturient Holstein cattle.

Relationships between genetic measures of mastitis (somatic cell score, score for clinical mastitis, and scores for IMI with major or minor pathogens) and immunological parameters (physiological and molecular markers) were examined for periparturient Holstein cows. Physiological markers included 11 in vitro immunological assays. Molecular markers included the second exon of the DRB3 locus of the bovine major histocompatibility complex, the IgG2 isotype genotype, and the CD18 genotype (the locus responsible for bovine leukocyte adhesion deficiency). A gene substitution model was used to estimate the additive genetic effects of alleles of the three molecular markers on estimated breeding value (EBV) for mastitis measures. Pearson correlation coefficients between EBV for immunological assays and EBV for mastitis measures were computed. Molecular markers explained up to 40% of the variation in EBV for measures of mastitis. The presence of allele DRB3.2*16 was associated with higher EBV for SCS. Allele DRB3.2*8 was associated with increased EBV for clinical mastitis, as was the IgG2b allele and the normal CD18 allele. Alleles DRB3.2*11, *23, IgG2a, and the recessive allele for bovine leukocyte adhesion deficiency were associated with decreased clinical mastitis. A positive genetic association was found between allele DRB3.2*24 and EBV for IMI by major pathogens and between DRB3.2*3 and IMI by minor pathogens. Several correlations between EBV for immunological assays and EBV for mastitis measures were significantly different from 0. Cows with low EBV for SCS tended to have neutrophils that had greater functional ability at maximal immunosuppression, low serum IgG1, and high numbers of circulating mononuclear cells. Immunological parameters, including physiological and molecular markers, are useful aids to understand the genetics of resistance to mastitis.

Alleles↗
PubMed1996

Evaluation of a coagulase-negative variant of Staphylococcus aureus as a cause of intramammary infections in a herd of dairy cattle.

A coagulase-negative variant of Staphylococcus aureus was identified in a herd of 250 lactating dairy cows. During testing of the entire herd, this strain of S aureus was isolated from aseptically collected milk samples of 25 cows. Cows with intramammary infections attributable to coagulase-negative S aureus had an increased somatic cell count in their milk, which was indicative of mastitis infection. Speciation of the Staphylococcus organisms was made, using a series of biochemical tests. A strain of a coagulase-positive S aureus also caused intramammary infections in the herd and shared identical biochemical characteristics with the coagulase-negative strain. Moreover, both strains could not be typed by the use of the International Set of Bovine Phages. Analysis of these findings indicated that a coagulase-negative variant of S aureus can cause intramammary infections in cattle, coagulase-negative variants of S aureus that cause mastitis can be more prevalent in herds than coagulase-positive variants, and clinicians should avoid misclassifying coagulase-negative S aureus as organisms that are clinically unimportant.

Animals↗
PubMed1996

Environmental reservoirs for Serratia marcescens intramammary infections in dairy cows.

Via special media, Serratia marcescens isolates were found in 3 bedding pack samples and in 2 milking parlor floor samples, and in milk samples from 19 cows during an episode of mastitis in a dairy cow herd. Chromosomal digest patterns of isolated S marcescens were indistinguishable for 18 of the milk samples and all bedding pack samples. Our findings provide strong evidence that the bedding pack was the reservoir of S marcescens associated with the outbreak of intramammary infections. Additionally, our ability to match digest patterns of isolates in the bedding pack and milk confirms the theory that S marcescens is an environmental pathogen capable of causing mastitis.

Animals↗