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Biomedical subjects

J S Hogan

Publications and source records attributed to J S Hogan.

At least 37 records · Page 2Linked to original sources

Evaluation of an acriflavine disk assay for differentiating Staphylococcus aureus from other staphylococci isolated from bovine milk.

OBJECTIVE: To develop an acriflavine disk assay for identification of Staphylococcus aureus and to test whether the acriflavine disk assay could be used to differentiate S aureus from other staphylococci isolated from bovine milk samples. DESIGN: Prospective study. SAMPLE POPULATION: 882 staphylococcal isolates from bovine milk samples and 3 S intermedius isolates from dogs. PROCEDURE: Paper disks saturated with various amounts of acriflavine were used in a growth inhibition assay to determine the amount of acriflavine that would most reliably differentiate S aureus from other staphylococci. For all isolates, hemolytic pattern, results of tube coagulase tests after 4 and 24 hours of incubation, growth on acriflavine-supplemented media, results of an acriflavine disk assay, and results of an automated identification system were determined. RESULTS: 10 micrograms of acriflavine/disk was determined to be the most appropriate concentration for use in the assay. All 112 isolates identified as S aureus by the automated identification system were resistant to this concentration of acriflavine, and only 1 of 236 isolates identified as non-S aureus staphylococci was resistant. There was substantial agreement between results of using the acriflavine disk assay as a diagnostic criterion for differntiating S aureus isolates from non-S aureus staphylococci and results of the automated identification system. Agreement between results of determining hemolytic pattern and results of other diagnostic tests was only moderate. CLINICAL IMPLICATIONS: The acriflavine disk assay, using 10 micrograms of acriflavine/disk, was a practical, accurate method for differentiating S aureus isolates from non-S aureus staphylococci.

Acriflavine↗

Inhibition of in vitro growth of coliform bacteria by a monoclonal antibody directed against ferric enterobactin receptor FepA.

The ability of a murine monoclonal antibody that blocks the enterobactin ligand-binding site of the ferric enterobactin receptor FepA to inhibit the growth of coliform bacteria derived from a bovine intramammary infection (IMI) was determined in an iron-restricted medium. Bacterial isolates from bovine IMI in five herds were tested by the chrome azurol sulfonate assay to detect siderophore production. Each of the isolates of Escherichia coli (n = 25) and Klebsiella pneumoniae (n = 25) were positive for siderophore production. Each isolate expressed iron-regulated outer membrane proteins when grown in trypticase soy broth plus the iron chelator alpha-alpha'-dipyridyl. Immunoblots revealed that the monoclonal antibody recognized FepA that was expressed by each of the E. coli isolates (n = 25). Only 4 of 25 K. pneumoniae isolates produced FepA that reacted with the monoclonal antibody. This result coincided with the results of an in vitro growth assay. Growth of all E. coli isolates was significantly inhibited by the addition of monoclonal antibody to synthetic medium containing apolactoferrin. Antigenic variation in the enterobactin-binding site resulted in a low percentage of K. pneumoniae isolates that were inhibited by the monoclonal antibody. Inhibition of bacterial growth by the monoclonal antibody was dose-dependent. As little as 50 micrograms/ml of purified antibody had an inhibitory effect on bacterial growth in the synthetic iron-restricted medium. Monoclonal antibody specific for the enterobactin ligand-binding site of FepA inhibited the growth of E. coli that was isolated from bovine IMI.

Animals↗

Immunization of cows with ferric enterobactin receptor from coliform bacteria.

The serum and milk immunoglobulin (Ig) G responses of lactating dairy cows were determined following immunization with ferric enterobactin receptor FepA. Escherichia coli 471 was cultured in iron-depleted medium, and outer membrane proteins were extracted by 2% N-lauroylsarcosine sodium salt and 2% Triton X-100. The FepA was isolated from the outer membrane proteins by ion-exchange chromatography. Twenty cows were assigned to four treatment groups of 5 cows blocked by breed and days in milk. Treatment groups were vaccinated with 100 micrograms of FepA, 500 micrograms of FepA, Escherichia coli J5 bacterin, or sterile phosphate-buffered saline. Primary immunization was at approximately 200 d in milk, and booster immunizations were given 14 and 28 d later. Serum and whey IgG titers to FepA in cows vaccinated with FepA were significantly higher than those from cows vaccinated with either E. coli J5 bacterin or phosphate-buffered saline. Serum and whey IgG titers to FepA were elevated by 14 d in cows vaccinated with FepA. Significant differences were not observed between doses of FepA. The degree of cross-reactivity of purified IgG from cows vaccinated with FepA to E. coli and Klebsiella pneumoniae isolates was significantly higher than that to a control isolate that lacked FepA production. Immunization with FepA elicited an immunological response in serum and milk.

Ammonium Sulfate↗

Dietary vitamin E and selenium affect mastitis and milk quality.

Vitamin E and selenium (SE) are essential nutrients that are integral components of the antioxidant defense of tissues and cells. Soils in many of the important dairy regions of the world are Se-deficient, and feedstuffs grown on these soils will not provide adequate dietary Se. Cattle consuming stored forages are likely to be low in vitamin E unless supplemented, and vitamin E deficiencies are frequently observed in peripartum dairy cows. Many new intramammary infections (IMI) occur in the 2 wk before and after calving. Deficiencies of either vitamin E or Se have been associated with increased incidence and severity of IMI, increased clinical mastitis cases, and higher somatic cell counts (SCC) in individual cows and bulk tank milk. Somatic cell counts are a primary indicator of mastitis and milk quality in dairy herds. The polymorphonuclear neutrophil (PMN) is a major defensive mechanism against infection in the bovine mammary gland. A know consequence of vitamin E and Se deficiency is impaired PMN activity and postpartum vitamin E deficiencies are frequently observed in dairy cows. Dietary supplementation of cows with Se and vitamin E results in a more rapid PMN influx into milk following intramammary bacterial challenge and increased intracellular kill of ingested bacteria by PMN. Subcutaneous injections of vitamin E approximately 10 and 5 d before calving successfully elevated PMN alpha-tocopherol concentrations during the periparturient period and negated the suppressed intracellular kill of bacteria by PMN that commonly is observed around calving.

Animals↗

Bacteria counts in sawdust bedding.

Bacteria counts in untreated sawdust bedding were compared with those in sawdust bedding after the addition of lime and after daily replacement of bedding in the back one-third of the stalls. Addition of 1 kg of lime to 10 kg of sawdust reduced Gram-negative bacteria, coliforms, Klebsiella spp., and streptococci prior to use as bedding. Sawdust treated with lime also showed decreased bacteria counts compared with bacteria counts for sawdust that was replaced daily and compared with bacteria counts for control bedding after 1 d in the stall. The decrease in bacterial populations was related to an increase in bedding pH. Mean pH in the sawdust treated with lime was greater prior to use and after 1 d in the stall than the pH of other treatments. After 2 and 6 d in stalls, however, bacteria counts and pH were similar among treatments. Dry matter content of bedding did not differ among bedding treatments. Bacteria counts in bedding were positively correlated with teat skin swabs. Gram-negative bacteria and Klebsiella spp. counts on teat swabs were lower for cows housed on bedding treated with lime on d 2 compared with those for cows housed on control bedding and bedding that was replaced daily. Addition of lime to sawdust in the back one-third of tie stalls caused a decrease in exposure of teats to environmental mastitis pathogens in bedding for 1 d. Daily replacement of bedding had a minimal effect on bacteria counts in bedding and on teat skin.

Animals↗

Effect of vitamin E supplementation in diets with a low concentration of selenium on mammary gland health of dairy cows.

Sixty-six cows and heifers (Holsteins and Jerseys) were assigned to one of three treatments at 60 d before anticipated calving. Treatment 1 consisted of 100 IU/d of supplemental vitamin E during the dry period and 100 IU/d during the first 30 d of lactation. Treatment 2 was 1000 IU/d of vitamin E during the dry period and 500 IU/d during lactation. Treatment 3 was 1000 IU/d of vitamin E during the first 46 d of the dry period, 4000 IU/d during the last 14 d of the dry period, and 2000 IU/d during lactation. Plasma concentrations of alpha-tocopherol decreased at calving for cows fed dietary treatments with low or intermediate concentrations of vitamin E, but not for cows fed the high vitamin E treatment. High dietary vitamin E increased concentrations of alpha-tocopherol in blood neutrophils at parturition, but no difference was found for the other two treatments. The percentage of quarters with new infections at calving was not different (32.0%) between cows receiving treatments that contained low and intermediate concentrations of vitamin E but was reduced (11.8%) in cows receiving the high vitamin E treatment. Clinical mastitis affected 25.0, 16.7, and 2.6% of quarters during the first 7 d of lactation for cows receiving the low, intermediate, and high vitamin E treatments, respectively. Cows with plasma concentrations of alpha-tocopherol < 3.0 micrograms/ml at calving were 9.4 times more likely to have clinical mastitis during the first 7 d of lactation than were cows with plasma concentrations of alpha-tocopherol > 3.0 micrograms/ml.

Animal Feed↗

Responses of antibody titers to intramammary immunization with Escherichia coli J5 bacterin.

The effect of an immunization schedule on responses of antibody titers was tested following vaccination with an Escherichia coli J5 bacterin. Eighteen cows were equally distributed among three immunization schedules: 1) subcutaneous injection at 14 d prior to the end of lactation, intramammary immunization at 7 d after drying off, and subcutaneous injection at 30 d into the dry period; 2) subcutaneous injections at drying off, at 30 d into the dry period, and within 12 h after calving; and 3) unimmunized controls. The E. coli J5 bacterin consisted of 5 ml of 10(9) boiled cells/ml of 0.9% NaCl plus 0.005% phenol emulsified with 5 ml of Freund's incomplete adjuvant. Subcutaneous injections were administered on the upper part of the rib cage, posterior to the scapula. Intramammary immunizations of 2.5 ml of bacterin were infused via the teat canal into each of the four mammary glands. Intramammary immunization increased rectal temperatures at 12 h after infusion, but subcutaneous injections did not induce febrile responses. Intramammary immunization enhanced immunoglobulin G titers in serum and whey on d 0 of lactation compared with subcutaneous immunizations. Immunoglobulin G titers in serum also were greater at d 30 of the dry period and at d 14 and 21 of lactation for cows that received intramammary immunization than for cows that were vaccinated by subcutaneous injections only. Immunoglobulin M titers in whey and serum on d 21 of lactation were greater for cows that received intramammary immunizations than for cows that were immunized by subcutaneous injections only.

Animals↗

Concentrations of alpha-Tocopherol after intramammary infusion of Escherichia coli or lipopolysaccharide.

Fifteen Holstein cows were used in a trial involving intramammary challenge to determine the effects of acute clinical mastitis on the concentrations of alpha-tocopherol in milk and plasma and the concentrations of neutrophils in milk and blood. Cows were assigned to one of three experimental groups challenged by intramammary infusion of lipopolysaccharide, Escherichia coli, or sterile phosphate-buffered saline. All quarters infused with lipopolysaccharide or E. coli were diagnosed with clinical mastitis on d 1 and 2 after challenge. Acute inflammation caused by intramammary infusion of lipopolysaccharide or E. coli resulted in increased concentrations of alpha-tocopherol in milk in challenged quarters but had no effect on concentrations of alpha-tocopherol in plasma. Concentrations of alpha-tocopherol in milk and blood neutrophils did not differ among treatment groups. Concentrations of alpha-tocopherol did not differ between milk and blood neutrophils. Approximately 25% of the alpha-tocopherol in milk from glands with clinical mastitis was associated with neutrophils, and < 10% of the alpha-tocopherol in milk from nonmastitic glands was associated with neutrophils. A shift toward sources of alpha-tocopherol other than synthesized milk fat occurred during acute inflammation in the mammary gland.

Animals↗

alpha-Tocopherol concentrations in milk and plasma during clinical Escherichia coli mastitis.

Eighteen cows were challenged by intramammary infusion with Escherichia coli 727 to determine the effects of acute clinical mastitis on alpha-tocopherol concentrations in plasma and milk. Cows were fed diets supplemented with 1000 IU of vitamin E/d from calving through the experimental period. At challenge, geometric mean DIM was 33 d. Each mammary quarter was diagnosed with an IMI and clinical mastitis at 24 and 48 h after challenge. The alpha-tocopherol concentrations in milk from challenged quarters were approximately 60% greater by 24 and 48 h after challenge than concentrations at prechallenge and 168 h postchallenge. Plasma alpha-tocopherol concentrations did not change after intramammary challenge. The alpha-tocopherol in plasma and milk was correlated at 48 and 168 h postchallenge but not at prechallenge or 24 h postchallenge. Milk alpha-tocopherol and SCC were correlated positively across all sample periods. Milk fat and milk alpha-tocopherol concentrations were correlated at each sample period except 24 h postchallenge. Increases in milk alpha-tocopherol during clinical mastitis were not correlated to milk production, DMI, or BSA concentration in milk. Changes in milk alpha-tocopherol concentration during clinical mastitis were similar to the dynamics of milk SCC.

Animals↗

Development of a PCR amplification assay as a screening test using bulk milk samples for identifying dairy herds infected with bovine viral diarrhea virus.

The approach of cDNA synthesis followed by polymerase chain reaction (PCR) amplification was used to develop a rapid screening test for the detection of bovine viral diarrhea virus (BVDV) in bulk tank milk samples. The initial development of this detection method was done using lactating Holstein cows; 1 acutely infected with BVDV following experimental inoculation and 2 persistently infected (PI) with BVDV. Viral RNA was extracted from somatic cells purified from whole milk using a guanidinium isothiocyanate and phenol/chloroform extraction method. Oligonucleotide primers were selected from the 5'untranslated region (5'UTR) and p80 region of BVDV genome. In the acutely infected cow, BVDV RNA was identified from days 6 to 10 postinoculation. Viral RNA extracted from somatic cells of milk from PI cows was detected by PCR using both 5'UTR and p80 primer sets. The sensitivity of PCR detection was determined by preparing dilutions of whole milk obtained from the BVDV persistently infected animals with milk from a BVDV-negative cow followed by purification of somatic cells and RNA extraction. BVDV was detected in milk serially diluted to 1:640 using PCR amplification. In addition, PCR amplification was 14.6 times more sensitive than virus isolation in detecting BVDV RNA in purified milk somatic cells. PCR detected BVDV RNA from a minimum of 580 somatic cells while the detection limit of virus isolation was 8500 cells. The sensitivity and specificity of BVDV amplification were confirmed by Southern hybridization analysis. BVDV RNA was detected using PCR in 33 out of 136 bulk milk samples collected from 124 individual herds using the 5'UTR primer set. These results indicate that PCR analysis of bulk tank milk samples may provide a rapid and sensitive method of screening herds for the presence of BVDV infections.

Animals↗

Effects of an Escherichia coli J5 vaccine on mild clinical coliform mastitis.

Efficacy of an Escherichia coli (O111:B4) J5 bacterin was tested in an experimental challenge trial. Nineteen cows were vaccinated with an E. coli J5 bacterin, and 10 cows were injected with a placebo containing adjuvant only. Vaccine and placebo were administered at drying off, 30 d after drying off, and within 48 h after calving. Cows were challenged approximately 30 d after calving by intramammary infusion with a smooth heterologous strain of E. coli previously shown to cause mild clinical mastitis. Vaccination with the J5 bacterin reduced duration of IMI and local signs of clinical mastitis. Concentrations of BSA in milk 24 h after challenge were greater in control cows than in cows vaccinated with J5. The SCC at 7 d postchallenge were greater for cows vaccinated with the placebo than for cows vaccinated with J5. Bacterial counts were lower for cows vaccinated with the placebo than for cows vaccinated with J5 at 3, 6, and 9 h postchallenge. In contrast, cows vaccinated with J5 had lower bacterial counts at 2, 3, and 4 d postchallenge than did cows vaccinated with placebo. Systemic signs of clinical mastitis were relatively mild and similar between treatment groups. Rectal temperature, DMI, and milk production did not differ between control and cows vaccinated with J5 following challenge.

Administration, Topical↗

Efficacy of recombinant bovine interleukin-2 as an adjunct to dry cow therapy.

Recombinant bovine interleukin-2 was tested as adjunct therapy to intramammary infusion of antibiotics at drying off. Cows were assigned randomly within three commercial herds to either recombinant bovine interleukin-2 or PBS treatment groups at drying off. Ninety-five cows received intramammary infusions of 1 mg of recombinant bovine interleukin-2 in each quarter immediately preceding intramammary infusion of a product for antibiotic therapy of dry cows. Ninety-four cows received intramammary infusion of 10 ml of endotoxin-free PBS in each quarter immediately preceding the antibiotic therapy. Cure rates for IMI present at drying off did not differ between cows treated with recombinant bovine interleukin-2 and those treated with PBS. Intramammary infusion of recombinant bovine interleukin-2 did not affect the rate of new IMI during the dry period. Milk production and SCC during the first 3 mo of lactation subsequent to therapy did not differ between treatment groups. Intramammary infusion of recombinant bovine interleukin-2 was not effective as an adjunct to antibiotic therapy for dry cows.

Animals↗

Effect of forage to concentrate ratio on disappearance of vitamins A and E during in vitro ruminal fermentation.

The effects of forage to concentrate ratio and the commercial form of vitamins A and E on in vitro ruminal disappearance of retinol and alpha-tocopherol were studied. Ruminally fistulated cows were fed diets with either 80 or 50% forage. In vitro substrates that were similar to those fed to the donor cows were incubated with buffered ruminal fluid for 24 h. Different commercial forms of vitamin E (spray-dried, silicic acid adsorbate, and lipid-encased forms) and vitamin A (gelatin beadlet and lipid-encased forms) were added to the flasks. The vitamin E was all-rac-alpha-tocopheryl acetate, and the vitamin A was all-trans-retinyl acetate. The amount of alpha-tocopherol in the flasks was not affected by diet or form of vitamin E and did not change over the 24-h incubation. Retinol disappearance was not affected by form of vitamin A but was substantially higher for the 50% forage diet than for the 80% forage diet (72 vs. 20% at 24 h). These data suggest that ruminal metabolism of vitamin E is minimal and not affected by forage to concentrate ratio. Additionally, vitamin A destruction in the rumen was much higher when cows were fed a typical lactation diet than when fed a typical dry cow diet.

Animal Feed↗

Immunization of dairy cows with an Escherichia coli J5 lipopolysaccharide vaccine.

Development of a lipopolysaccharide-protein conjugate vaccine and the immunological response to the vaccine were investigated. Lipopolysaccharide derived from Escherichia coli J5 was detoxified by mild alkaline hydrolysis. Detoxification reduced endotoxin activity 2500-fold compared with that of native J5 lipopolysaccharide. The conjugate vaccine was synthesized by covalently coupling detoxified lipopolysaccharide to chicken serum albumin by reductive amination. Dairy cows were immunized with 8.35 mg of conjugate (n = 3) or 5 x 10(9) heat-killed J5 bacterin (n = 5) at 215 DIM and received a secondary immunization 14 d later. Control cows were not immunized. Immunization enhanced serum antibody titer to J5 lipopolysaccharide antigens. Whey IgG and IgM titers to J5 lipopolysaccharide were not different among treatment groups. Serum and whey IgG titers to J5 whole-cell antigens were elevated in immunized cows within treatment groups. Immunization did not enhance whey IgM to J5 whole-cell antigens. Conjugate immunization elicited an immune response comparable with or greater than that of immunized cows with J5 bacterin.

Animals↗

Environmental streptococcal intramammary infections of the bovine mammary gland.

Characteristics of environmental streptococcal IMI were investigated over a 7-yr period for a herd in total confinement. A total of 374 new environmental streptococcal IMI was detected. Approximately 50.5% of IMI were new in the dry period, and 49.5% were new in lactation. The rate of new IMI was .00312 IMI/cow day during the dry period and .00054 IMI/cow day during lactation. The percentages of cows and quarters with an environmental streptococcal IMI present at calving were 10.6 and 3.2%, respectively. The percentage of heifers with an environmental streptococcal IMI at calving was similar to that for cows. The rate of new environmental streptococcal IMI was greater during the 1st mo of lactation than during the remainder of lactation. The rate of IMI during late lactation was higher for older cows than for either heifers or cows in second lactation. The rate of environmental streptococcal IMI during the dry period and during lactation was greatest during summer. The mean days of lactation that cows were infected for all IMI was 12.3 d. Approximately 41% of IMI had a duration of < 8 d. Stage of lactation, season of the year, and parity influenced the rate of new IMI.

Aging↗

Efficacy of a barrier teat dip containing .55% chlorhexidine for prevention of bovine mastitis.

A natural exposure trial was conducted for 12 mo in a commercial herd of 125 lactating cows to compare the efficacy of an experimental barrier teat dip containing .55% chlorhexidine gluconate with the efficacy of a 1% iodophor for preventing new IMI and clinical mastitis. Teats of half of the cows were dipped in the experimental barrier product, and teats of the remaining half of the herd were dipped in the 1% iodophor product. Quarters dipped with the experimental barrier product had fewer new IMI caused by Escherichia coli, coagulase-negative staphylococci, or Gram-positive bacilli than did quarters dipped in the 1% iodophor. Incidence of new IMI caused by Serratia spp. and Pseudomonas spp. was greater for quarters dipped in the experimental barrier product than for quarters dipped in the 1% iodophor. Efficacy of the two teat dips against new IMI caused by Staphylococcus aureus, environmental streptococci, and Klebsiella spp. did not differ. Incidence of bacteriologically negative clinical cases of mastitis was greater in quarters dipped in the 1% iodophor than in quarters dipped in the experimental barrier product. Incidence of clinical mastitis cases caused by Staph. aureus, environmental streptococci, E. coli, Klebsiella spp., Serratia spp., and Pseudomonas spp. did not differ between treatment groups.

Animals↗

Antigenic crossreactivity and lipopolysaccharide neutralization properties of bovine immunoglobulin G.

We investigated a possible mechanism by which immunization against core and lipid A determinants of lipopolysaccharide reduced clinical cases of mastitis and symptoms commonly associated with heterologous Gram-negative IMI. The IgG fraction of sera from cows immunized with either Escherichia coli J5 bacterin, E. coli J5 lipopolysaccharide conjugate vaccine, or unimmunized controls was purified by precipitation with caprylic acid and ammonium sulfate. The degree of IgG crossreactivity with Gram-negative bacteria that were isolated from clinical quarters was greater than that with Gram-positive isolates of Staphylococcus aureus. The highest magnitude of crossreactivity was against smooth strain E. coli isolates, followed by heterologous species of Enterobacter, Serratia, and Klebsiella isolates. Serum IgG from cows immunized with conjugate was highly crossreactive to E. coli J5, E. coli O111:B4, Serratia marcescens, Klebsiella pneumoniae, and Salmonella typhimurium lipopolysaccharides. The magnitude of antibody crossreactivity with lipopolysaccharides coincided with the ability of IgG to suppress the mitogenic effect of lipopolysaccharides on bovine lymphocytes.

Ammonium Sulfate↗

Therapy of experimentally induced coliform mastitis with a Propionibacterium acnes product.

The efficacy of a Propionibacterium acnes product for treatment of coliform mastitis was evaluated following intramammary infusion of Escherichia coli. Ten cows were injected intravenously with a product containing heat-killed P. acnes at 3 h, 3 d, and 7 d after bacterial challenge. Five cows served as untreated negative controls. Bacterial counts in milk, rectal temperatures, and milk SCC did not differ between treated cows and control cows. Intravenous infusion of P. acnes had no effect on milk production or DMI of treated cows compared with untreated controls. Therapy of experimentally induced coliform mastitis with a P. acnes product had no effect on severity or duration of clinical signs.

Adjuvants, Immunologic↗