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S P Oliver

Publications and source records attributed to S P Oliver.

At least 19 recordsLinked to original sources

Clinical, epidemiological and molecular characteristics of Streptococcus uberis infections in dairy herds.

A longitudinal observational study (18 months) was carried out in two Dutch dairy herds to explore clinical, epidemiological and molecular characteristics of Streptococcus uberis mastitis. Infections (n = 84) were detected in 70 quarters of 46 cows. Bacterial isolates were characterized at strain level by random amplified polymorphic DNA (RAPD) fingerprinting. Persistent infections were usually attributable to one strain, while recurrent infections could be caused by different strains. When multiple quarters of a cow were infected, infections were mostly caused by one strain. In each herd, multiple strains were identified yet one strain predominated. The majority of all infections were subclinical, and infections attributed to predominant strains were more chronic than infections attributed to other strains. Epidemiological and molecular data suggest infection from environmental sources with a variety of S. uberis strains as well as within-cow and between-cow transmission of a limited number of S. uberis strains, with possible transfer of bacteria via the milking machine.

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Prepartum antibiotic treatment of heifers: milk production, milk quality and economic benefit.

Prepartum intramammary antibiotic infusion of heifer mammary glands at 7 or 14 d before expected parturition is an effective procedure for eliminating many infections in heifers during late gestation and for reducing the prevalence of mastitis in heifers during early lactation and throughout lactation. Mastitis pathogens were isolated from 76% of samples obtained from untreated control quarters 7 d before expected calving, from 47% of samples obtained 3 d after calving, and from 29% of samples obtained 10 d postpartum. Mastitis pathogens were isolated from about 30% of control quarters through 240 d of lactation. A similar percentage of samples (70%) was positive for mastitis pathogens at C-7 before antibiotic treatment. However, only 8% of samples obtained at 3 d after calving and 4% of samples obtained at 10 d postpartum from quarters of antibiotic-treated heifers contained mastitis pathogens. Throughout the remainder of lactation, mastitis pathogens were isolated from an average of about 11% of quarters. The percentage of samples with mastitis pathogens was higher in untreated controls than in antibiotic-treated quarters at all sampling intervals during lactation. A similar response was observed in heifers that were treated with antibiotics at 14 d before expected parturition. Prepartum antibiotic-treated heifers produced significantly more milk than control heifers and had significantly lower somatic cell count scores than untreated control heifers. These observations are likely associated with or due to the lower prevalence of mastitis pathogen isolation in prepartum antibiotic-treated heifers throughout lactation. Prepartum antibiotic-treated heifers produced 531 kg more milk than heifers in the untreated control group. Multiplying this increase by a milk price of 0.407 dollars/kg yielded a 216.24 dollars per-heifer increase in gross revenue. The cost of treatment, including the cost of testing for antibiotic residues, was estimated at 15.60 dollars for a net revenue of 200.64 dollars per heifer. Prepartum antibiotic treatment to reduce the rate of mastitis in heifers during lactation was highly effective and economically beneficial.

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Binding of bovine lactoferrin to Streptococcus dysgalactiae subsp. dysgalactiae isolated from cows with mastitis.

Three strains of Streptococcus dysgalactiae subsp. dysgalactiae (UT516, UT519, ATCC 27957) were used to determine if bovine lactoferrin (Lf) binds to bacterial cells by biotin avidin binding assay (BABA), enzyme-linked immunosorbent assay (ELISA), and binding inhibition assay. Binding assays revealed that all strains of S. dysgalactiae subsp. dysgalactiae (S. dysgalactiae) evaluated in this study bound to Lf. However, differences in Lf binding capability among strains and between methods used were detected. Binding of Lf was not inhibited by transferrin (Tf) and Lf moiety molecules (mannose, galactose, and lactose) but by Lf. This study demonstrates that S. dysgalactiae bound to bovine Lf in a specific manner.

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Prevalence and molecular characterization of Escherichia coli O157:H7 in bulk tank milk and fecal samples from cull cows: a 12-month survey of dairy farms in east Tennessee.

A study on the prevalence of Escherichia coli O157:H7 was conducted on 30 dairy farms in east Tennessee between May 2000 and April 2001. This pathogen was isolated from 8 of 30 (26.7%) dairy farms at various sampling times. A total of 415 fecal samples from cull dairy cows and 268 bulk tank milk samples were analyzed. Overall, 10 of 683 (1.46%) samples (2 of 268 [0.75%] milk samples and 8 of 415 [1.93%] fecal samples) tested positive for E. coli O157:H7. Food and Drug Administration Bacteriological Analytical Manual protocols were used for the conventional isolation and confirmation of E. coli O157:H7. Samples were shake cultured (150 rpm) at 42 degrees C for 24 h in tryptic soy broth containing 2 mg of novobiocin per liter. White colonies isolated on cefixime-tellurite sorbitol MacConkey agar plates were evaluated for fluorescence on sorbitol MacConkey agar supplemented with 0.025 g of methylumbelliferyl-beta-D-glucuronide per liter. Nonfluorescing white colonies were biochemically typed and serologically confirmed. Multiplex polymerase chain reaction profiles of E. coli O157:H7 isolates indicated the presence of common virulence factors (Shiga toxin, enterohemolysin, and intimin) of Shiga toxin-producing E. coli, suggesting the potential human pathogenicity of bacterial isolates. Pulsed-field gel electrophoresis profiles of SpeI and XbaI restriction enzyme-digested genomic DNA were used to establish relatedness among bacterial isolates. Data from this study indicate that both cull dairy cows and bulk tank milk pose a potential hazard with regard to human foodborne illness. It is therefore imperative to develop on-farm and preharvest pathogen reduction programs to control the carriage of E. coli O157:H7 pathogens.

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Molecular characterization of Salmonella spp. isolated from bulk tank milk and cull dairy cow fecal samples.

The consumption of meat from cull dairy cows and of raw milk has been associated with foodborne salmonellosis. This survey was conducted to establish the prevalence of Salmonella in cull dairy cow fecal samples and bulk tank milk and to determine the proportion of Salmonella-positive dairy farms (n = 30) in east Tennessee. Food and Drug Administration bacteriological analytical protocols were generally used for Salmonella isolation. Primary enrichment was performed with lactose broth, and secondary enrichment was conducted with tetrathionate broth. Eosin methylene blue, hektoen enteric, xylose lysine desoxycholate, bismuth sulfite, and brilliant green (BG) were used as isolation agars. BG agars supplemented with individual antibiotics and/or sulfur compounds were also evaluated. Six of 268 (2.24%) bulk tank milk samples and 9 of 415 (2.17%) fecal samples from 7 of 30 (25.3%) dairy farms were Salmonella-positive. Most isolates (11 of 15) were obtained between September and December. Salmonella isolates were further characterized using polyvalent somatic O Salmonella antiserum, o-nitrophenyl-beta-D-galactopyranoside (ONPG), and Analytical Profile Index (API) 20E strips for Enterobacteriaceae. Serological evaluation of presumptive positive Salmonella isolates resulted in substantial numbers of false positives (41.2%). ONPG and API 20E tests enabled further biochemical distinction of the majority of Salmonella spp. from Salmonella Arizonae and closely related members of Enterobacteriaceae like Citrobacter youngae. Pulsed-field gel electrophoresis of SpeI-digested Salmonella DNA was used to subtype isolates. The isolates grouped into four clusters. The baseline information generated in this survey is being used to develop preharvest pathogen reduction programs on selected farms.

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Interaction of coagulase-negative Staphylococcus species with bovine mammary epithelial cells.

Three coagulase-negative Staphylococcus species (CNS) (Staphylococcus epidermidis, Staphylococcus xylosus and Staphylococcus hyicus), from the milk of cows with mastitis, were used to evaluate adherence to and internalization by bovine mammary epithelial cells, and to investigate involvement of host cell signal transduction and host cell cytoskeleton rearrangement on internalization of CNS. S. xylosus showed highest adherence and internalization values of the species evaluated. Host cell cytoskeleton polymerization and protein kinase (PK) phosphorylation were required for internalization of CNS. Both protein kinase C (PKC) and tyrosine kinase (TPK) pathways were involved, but internalization of S. xylosus occurred preferentially through epidermal growth factor TPK activity. S. epidermidis and S. hyicus seemed to exploit other TPK pathways. Results of this study showed that S. xylosus, S. hyicus and S. epidermidis were able to adhere and internalize bovine mammary cells in a process that appeared to be receptor(s) mediated and exploited host signal transduction and cytoskeleton to induce an uptake signal.

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Role of collagen in adherence of Streptococcus uberis to bovine mammary epithelial cells.

We reported previously that pre-incubation of Streptococcus uberis with collagen induced expression of S. uberis surface proteins. In a subsequent study, we showed that incubation of S. uberis with extracellular matrix proteins, particularly collagen, increased adherence and internalization of S. uberis to mammary epithelial cells. In the present report, the potential mechanism by which S. uberis exploits the presence of collagen to enhance adherence to bovine mammary epithelial cells was evaluated. Adherence assays were conducted with S. uberis pre-treated with and without collagen and co-cultured in medium supplemented with or without collagen. Pre-incubation with collagen followed by co-culture in medium containing collagen up-regulated ligands that enhanced adherence of S. uberis to mammary epithelial cells. Collagen-up-regulated ligand(s) also increased adherence of S. uberis to mammary epithelial cells in the absence of collagen, but adherence was lower than when collagen was present during the adherence assay. Chloramphenicol was added to the culture medium to inhibit bacterial protein synthesis. Adherence decreased significantly in chloramphenicol-treated S. uberis pre-treated or co-cultured in the presence of collagen. These results suggest that S. uberis expresses ligands with affinity for collagen that are up-regulated by collagen. We hypothesize that these ligands increase adherence by using collagen as a bridge between the bacterium and host cell and/or by direct interaction with host cell receptor(s).

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Influence of subclinical mastitis during early lactation on reproductive parameters.

Our objective was to determine the effects of mastitis during early lactation on the reproductive performance of Jersey cows. From 1986 to 1997, quarter foremilk samples were collected every 4 to 8 wk during lactation, at drying off, near calving, and when clinical mastitis was diagnosed and were evaluated microbiologically to identify causative bacteria. Services per conception, days open, and days to first service were obtained from DHIA records on 752 cows. Cows were separated by mastitis type (clinical, n = 186; subclinical, n = 240; control, uninfected or infected after confirmed pregnancy, n = 326). Cows were reclassified based on the time of clinical or subclinical mastitis as follows: period 1, before first service (n = 374); period 2, between first service and pregnancy (n = 52); and period 3, after confirmed pregnancy or uninfected (control; n = 326). Milk production did not differ for any group separations. Reproductive performance did not differ between gram-negative or gram-positive mastitis pathogens. Cows with clinical or subclinical mastitis before first service had increased days to first service (77.3+/-2.7 and 74.8+/-2.7 d), days open (110.0+/-6.9 and 107.7+/-6.9 d), and services per conception (2.1+/-0.2 and 2.1+/-0.2) compared with controls (67.8+/-2.2 d, 85.4+/-5.8 d, 1.6+/-0.2; P < 0.05). Days to first service were not increased in cows with clinical or subclinical mastitis during period 2 (70.6+/-3.3 and 61.2+/-7.8 d). However, days open (143.6+/-8.5 d) and services per conception (3.0+/-0.2) were increased (P < 0.05) in cows with clinical mastitis during period 2, but not in cows with subclinical mastitis (90.9+/-20.2 d and 2.1+/-0.5). Cows initially diagnosed subclinical that became clinical during period 2 exhibited increased days to first service (93.9+/-10.1 d), days open (196.0+/-26.2 d), and services per conception (4.3+/-0.7) compared with control animals (P < 0.05). In conclusion, subclinical mastitis reduced reproductive performance of lactating cows similar to clinical mastitis. Subclinical mastitis followed by clinical mastitis resulted in the most severe loss in reproductive performance.

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Efficacy of a new premilking teat disinfectant containing a phenolic combination for the prevention of mastitis.

A teat disinfectant containing a phenolic combination was evaluated in a natural exposure study in two dairy research herds. Premilking teat disinfection was compared with a negative control using a split-udder experimental design. In both herds, premilking and postmilking teat disinfections with the phenolic combination were significantly more effective in preventing new intramammary infection (IMI) than was postmilking teat disinfection only. Clinical mastitis and new IMI by Streptococcus uberis, Streptococcus dysgalactiae, Gram-negative pathogens, and coagulase-negative Staphylococcus species were significantly lower in quarters of cows with teats predipped and postdipped than in quarters with teats postdipped only. No chapping or teat skin irritation was observed. Premilking teat disinfection with the phenolic combination in association with good udder preparation and postmilking teat disinfection can further reduce the occurrence of new IMI by numerous mastitis pathogens during lactation.

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Adhesion and invasion of Escherichia coli from single and recurrent clinical cases of bovine mastitis in vitro.

Seven strains of Escherichia coli, originating from clinical cases of bovine mastitis, and one Salmonella typhimurium control strain were tested for their ability to adhere to, and invade, bovine mammary epithelial cells (MAC-T cells) in vitro. Four of the seven strains were isolated from cows with chronic intramammary infections with recurrent episodes of clinical mastitis and three strains were isolated from single cases of clinical mastitis. Both adhesion and invasion of all strains were dose and time dependent. The four E. coli strains isolated from recurrent cases of clinical mastitis invaded twice as frequently as and three times faster than the strains isolated from single cases of clinical mastitis. By contrast, there was no difference in the amount or speed of adhesion between the two types of strains of E. coli. Adhesion and invasion curves of E. coli resembled a two-step chain reaction, where invasion was the rate-limiting step. Although adhesion and invasion of E. coli has not been demonstrated in vivo yet, the results of the present study may contribute to an understanding of the pathogenesis of chronic intramammary infections caused by E. coli.

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Endocrine profiles of dairy cows following experimentally induced clinical mastitis during early lactation.

Concentrations of LH, cortisol, estradiol-17beta (E(2)), prolactin and 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM) were determined in cows with experimentally induced clinical mastitis during early lactation. Cows free of intramammary infection (IMI) and in the luteal phase of the estrous cycle were balanced by lactation number and days in milk and assigned to either control (n=5) or treatment (n=5) groups. Treated cows were infected experimentally (day 0), in two mammary quarters, with Streptococcus uberis and developed clinical mastitis within 60 h after inoculation as evidenced by increased mastitis scores, elevated rectal temperatures, mammary swelling and isolation of S. uberis pathogen. Four days following bacterial challenge, blood samples were collected every 20 min for 8 h for determination of PGFM and LH following administration of oxytocin and GnRH, respectively. Blood samples were also collected on days 0, 4 and 7 of the experiment to determine concentrations of E(2), prolactin and cortisol. Four days after bacterial challenge, concentrations of cortisol were higher (P=0.04) in experimentally infected cows than controls. Experimentally challenged cows had increased (P=0.02) concentrations of cortisol on days 4 and 7 compared with day 0. Control cows had no significant increase in blood cortisol during the experimental period. Baseline concentrations of PGFM did not differ between groups; however, peak concentrations of PGFM following oxytocin challenge were elevated (P=0.006) in cows with clinical mastitis compared with control animals. Prolactin, E(2) and LH did not differ between cows with clinical mastitis or controls. Experimentally induced mastitis during early lactation elevated concentrations of cortisol during the luteal phase of the estrous cycle. Furthermore, mastitic cows demonstrated an increased PGFM response following oxytocin administration. Altered reproductive efficiency in cows with clinical mastitis caused by Gram-positive pathogens may be the result of increased uterine sensitivity to prostaglandin F(2alpha) (PGF(2alpha)).

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Influence of protein kinase inhibitors on Streptococcus uberis internalization into bovine mammary epithelial cells.

Previous reports indicated that bovine mammary epithelial cells internalized Streptococcus uberis, a bovine mastitis pathogen, and that inhibitors of F-actin microfilament polymerization inhibited bacterial internalization into mammary epithelial cells. In the present report, we show that inhibitors of eukaryotic cell tyrosine protein kinase (TPK) and protein kinase C (PKC), staurosporine, genistein and tyrphostin, significantly reduced internalization of S. uberis into mammary epithelial cells. Short-term treatment (15 min) of mammary epithelial cells with 12- O -tetradecanoylphorbol-13-acetate (TPA), shown previously to up-regulate activity of PKC, significantly increased internalization of S. uberis. Conversely, long-term incubation (24 h) of epithelial cells with TPA, which down-regulates PKC activity, significantly reduced the number of internalized S. uberis. These results suggest that protein kinases (TPK and PKC) are involved in internalization of S. uberis into bovine mammary epithelial cells. Identification of host cell surface receptor(s) and ligands that trigger the uptake signal by S. uberis need to be delineated.

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Determination of the lowest dilution of aluminium acetate solution able to inhibit in vitro growth of organisms commonly found in chronic suppurative otitis media.

Burow's solution has been found to inhibit the in vitro growth of most commonly occurring bacteria found in the discharging ear. These organisms were inoculated onto appropriate agar plates that contained serial dilutions of aluminium acetate. Results show that the lowest dilution able to inhibit the growth of these organisms lies between a 1:80 and a 1:160 dilution of Burow's solution.

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Effects of lactoferrin and milk on adherence of Streptococcus uberis to bovine mammary epithelial cells.

OBJECTIVE: To determine whether lactoferrin (LF) or milk influenced adherence of Streptococcus uberis to bovine mammary epithelial cells. SAMPLE POPULATION: Three strains of S uberis from cows with mastitis, pooled milk samples from 3 clinically healthy Jersey cows early in the lactation period, and bovine mammary epithelial cells from a clonal cell line. PROCEDURES: Adherence of S uberis to bovine mammary epithelial cells in the presence of various concentrations of LF or milk and after pretreatment of bacteria with LF or milk was tested. Bacteria were cultured with mammary epithelial cell monolayers for 1 hour. The culture supernatant was removed, and the epithelial cells were lysed. Adherence index was calculated as number of colony-forming units (CFU) in the cell lysate divided by number of CFU in the supernatant times 10,000. RESULTS: All 3 strains of S uberis were found to bind to purified LF and LF in milk. Addition of LF to the culture medium enhanced adherence of all 3 strains to mammary epithelial cells, whereas addition of milk enhanced adherence of 2 strains and decreased adherence of the third. Pretreatment of bacteria with LF or milk increased adherence of 1 of the strains but decreased adherence of the other 2. Increased adherence was antagonized by rabbit antibovine LF antibody. CONCLUSIONS: Results suggest that LF may function as a bridging molecule between S uberis and bovine mammary epithelial cells, facilitating adherence of the bacteria to the cells.

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Incubation of Streptococcus uberis with extracellular matrix proteins enhances adherence to and internalization into bovine mammary epithelial cells.

Two strains of Streptococcus uberis (UT 888 and UT 366) isolated from cows with clinical mastitis were co-cultured with bovine mammary epithelial cells (MAC-T) with and without laminin, fibrinogen, fibronectin or collagen. Incubation of S. uberis with extracellular matrix proteins (ECMPs) increased adherence to and internalization into MAC-T cells. Both strains of S. uberis exhibited greater adherence when co-cultured in the presence of collagen than with any other ECMP. However, adherence was always higher when strains were co-cultured with ECMP than in medium alone. S. uberis UT 888 adhered better to MAC-T cells than S. uberis UT 366. The influence of ECMPs on bacterial internalization into MAC-T cells was similar to adherence, however, differences among ECMPs were less noticeable. S. uberis UT 888 had a higher internalization index than S. uberis UT 366. It is possible that ECMPs induce or up-regulate proteins that selectively adhere to ECMPs which could serve as a bridge between the eukaryotic cell and the bacterial pathogen that leads to internalization of the ECMP-bound pathogen into the mammary epithelial cell.

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Adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells are mediated by host cell proteoglycans.

The role of cell glycosaminoglycans (GAG) on adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells was evaluated by adherence/internalization competition assays, by removal of GAG from the host cell surface and by inhibition of GAG glycosylation in the host cell. Heparin (HEP), heparan sulfate (HSA), chondroitin sulfate A (CSA) and chondroitin sulfate B (CSB) inhibited adherence and internalization of S. uberis in a dose-dependent manner. However, inhibition was lower with CSA and CSB than that observed with HEP and HSA. Adherence and internalization were also inhibited upon treatment of mammary epithelial cells with GAG lyases. The greatest inhibition was observed with heparinase I. Tunicamycin, an inhibitor of mammalian cell glycosylation of cell surface glycoproteins, markedly inhibited internalization of S. uberis into mammary epithelial cells. Differences between strains were observed. These results suggest that a HSA proteoglycan receptor on the host cell surface may mediate S. uberis adherence to and internalization of bovine mammary epithelial cells.

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Identification of lactoferrin-binding proteins in bovine mastitis-causing Streptococcus uberis.

All strains of Streptococcus uberis evaluated bound to lactoferrin (Lf) in milk as detected by polyacrylamide gel electrophoresis and Western blotting. A biotin-avidin-based microplate binding assay and ELISA also revealed that these bacterial strains bound to purified Lf. Binding of bacteria of Lf was not inhibited by mannose and galactose, indicating that glycosidic domains of the Lf molecule were not involved in binding. Lf binding was also unaffected by bovine transferrin. Western blot analysis demonstrated that there were at least two bacterial proteins involved in Lf-binding. Lf binding by S. uberis could enable this bacterium to acquire iron necessary for its growth.

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