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Biomedical subjects

R J Eberhart

Publications and source records attributed to R J Eberhart.

At least 19 recordsLinked to original sources

Comparison of microbiologic culture, an enzyme-linked immunosorbent assay, and determination of somatic cell count for diagnosing Staphylococcus aureus mastitis in dairy cows.

Results of using microbiologic culture of a single milk sample, determination of somatic cell count (SCC), an ELISA, and a combination of determination of SCC and ELISA to diagnose Staphylococcus aureus mastitis in dairy cattle were compared. Cows were considered to have S aureus intramammary infections if microbiologic culture of at least 2 of 3 consecutive sets of milk samples yielded growth of the organism. Data were analyzed from milk samples collected over a 4-month period from 185 cows in 5 herds. Sensitivity, specificity, and likelihood ratio of a positive test result for microbiologic culture of a single milk sample were 93%, 99%, and 93.0, respectively. Sensitivity, specificity, and likelihood ratio of a positive test result for ELISA were 69%, 61%, and 1.8, respectively, and for determination of SCC, they were 79%, 72%, and 2.9, respectively. Combination of determination of SCC and ELISA had sensitivity, specificity, and likelihood ratio of a positive test result of 80%, 62%, and 3.4, respectively. Results from microbiologic culture of consecutive milk samples were more consistent than results of ELISA performed on consecutive samples. These data suggest that microbiologic culture of a single milk sample is the best of the 3 tests studied for diagnosing S aureus intramammary infection.

Animals↗

The effect of milking machine liner slip on new intramammary infections.

New IMI of cows milked with high and low slip rate milking machine liners were compared. High slip liners used in this study averaged 6.1 initial slips and 7.6 major vacuum fluctuations per cow milking, and low slip liners averaged 3.6 initial slips and 3.1 major vacuum fluctuations per cow milking. Least squares means for new IMI rates per 100 cow days were .49 in the high slip group and .27 in the low slip group. Rates of clinical cases per 100 cow days were .35 in the high slip group and .20 in the low slip group. New IMI rates for cows entering the trial with an existing infection were .81 in the high slip group and .39 in the low slip group. More new IMI tended to occur with cows milked with the high slip liners than with low slip liners.

Animals↗

Post-milking teat dip use in dairy herds with high or low somatic cell counts.

Milk samples for bacteriologic culture were submitted from 71 dairy herds, 24 with low somatic cell count (SCC) and 47 with high SCC and high prevalence of subclinical mastitis. At the time of sample submission to the Mastitis Diagnostic Laboratory of Pennsylvania State University, information regarding the herd mastitis control practices was collected. A combined program of post-milking teat dipping (PMTD) and antibiotic treatment of all cows at the start of the nonlactating period was practiced more frequently for herds with low SCC, (P less than 0.001) than for herds with high SCC. Among all herds for which PMTD was practiced, a higher proportion (P less than 0.001) of those for which chlorhexidine-based products were used had low SCC than high SCC. Conversely, a higher proportion of herds for which a dip with an acrylic latex barrier was used had high SCC rather than low SCC (P = 0.002). For herds with high prevalence of subclinical mastitis, and despite a program of PMTD and treatment of all cows at the start of the nonlactating period, a change to a different germicidal teat dip product may be indicated to help reduce prevalence of infection.

Animals↗

Dietary selenium effects on milk eicosanoid concentration in dairy cows during coliform mastitis.

The effect of selenium deficiency on the product profile of arachidonic acid oxidation by enzymatic pathways in Holstein cows with experimentally-induced coliform mastitis was investigated. The animals were fed dairy rations containing 0.05 mg Se/kg dry matter, with the supplemented group receiving additional Se to increase the dietary concentration to approximately 0.35 mg Se/kg dry matter. Cows were inoculated intracisternally with 30 colony-forming-units of Escherichia coli at 14-16 weeks of lactation. Eicosanoids and bacteria numbers were recorded at various intervals of time for 60 h postinoculation. Milk from cows fed the Se-depleted diet had significantly higher (p less than 0.05) concentrations of TXB2 between 24 and 48 h and 6-keto-PGF1 alpha between 24 and 60 h postinoculation. Milk PGE2 concentration was significantly higher in the Se-deficient group at 24 h, whereas LTB4 was higher between 36 and 60 h postinoculation in the Se-deficient cows (p less than 0.05). Milk bacteria numbers were significantly higher between 16 and 24 h postinoculation in the Se-deficient group and three of the four cows in this group required euthanasia, whereas all four cows in the Se-supplemented group recovered without therapeutic intervention. These data indicate marked effects of dietary Se on milk eicosanoid concentrations in response to an E. coli infection. The changes in eicosanoid concentrations may be associated with the altered pathogenesis and outcome of mastitis in a Se-deficient state.

6-Ketoprostaglandin F1 alpha↗

Interactions between Pseudomonas aeruginosa and iodophor germicides in milking parlor udder wash water systems.

In a field study of 29 dairy farms, Pseudomonas aeruginosa was isolated more frequently (P = 0.05) from milking parlor udder wash water systems containing iodophor germicides than from those with no germicide. Most available iodine (AI2) concentrations were below the recommended level of 25 ppm (25 microgram/ml). Rubber and polyvinyl chloride hoses caused rapid decreases in the AI2 concentrations of 25 ppm iodophor solutions. AI2 dropped from 25 ppm to 6 ppm or less in 240 min for solutions contained in either polyvinyl chloride or rubber, compared with solutions in glass, which were unchanged in 240 min. Addition of inactivated iodophor solution to aqueous cultures resulted in significantly higher (P less than 0.05) numbers of P. aeruginosa at 10 and 24 h postinoculation. P. aeruginosa was grown in polyvinyl chloride tubing and exposed twice daily to 0, 10, or 25 ppm of AI2. None of the exposure concentrations eliminated the bacteria from the hoses, and bacterial numbers were not significantly different in hoses exposed to 0 and 10 ppm by the eighth treatment day. Bacteria taken from the water in these hoses were exposed to different concentrations of iodophor solution. Iodophor concentrations which will kill 50% of P. aeruginosa cultures previously exposed to 0, 10, and 25 ppm of AI2 were predicted to be 3.0, 11.8, and 20.8 ppm, respectively.

Animals↗

Mastitis in beef cows and its effects on calf weight gain.

Quarter milk samples from 51 purebred (Angus, Polled Hereford, and Simmental) and 69 crossbred (Angus x Simmental x Charolais three-way cross) beef cows were collected aseptically at three times during lactation to determine the prevalence of intramammary infection, milk somatic cell counts (SCC), and effects of infection on calf weight gain. Quarter infection prevalence was 13.1, 14.9, and 27.5% in early, mid, and late lactation; corresponding cow infection prevalence was 25.8, 29.2, and 54.4%. Staphylococcus aureus was isolated from 2.9, 2.7, and 3.2% of quarters in early, mid, and late lactation, respectively. Corynebacterium bovis, generally regarded as a minor pathogen, was isolated from 4.0, 7.6, and 18.2% of quarters at the three respective times. Geometric SCC means (10(3) cells/ml) were 1,522, 344, and 509 for S. aureus-infected quarters; 344, 899, and 221 for Staphylococcus hyicus-infected quarters; 65, 36, and 86 for C. bovis-infected quarters; and 20, 17, and 18 for uninfected quarters in early, mid, and late lactation, respectively. Adjusted 205-d weight gain for calves with S. aureus-infected dams was 9.6 kg less (P less than .05) than for calves with uninfected dams. Adjusted 205-d weight gain for calves with dams infected with any mastitis pathogen did not differ significantly from that of calves with uninfected dams. At weaning half of the infected cows and half of the uninfected cows were given an intramammary infusion product containing 300 mg of cephapirin benzathine in each quarter; the remaining cows were untreated controls. Quarter samples were collected aseptically from all cows 14 to 28 d after subsequent calving. Quarter prevalence of infection after calving was lower (P less than .05) in treated (8.2%) than in control (22.4%) cows. Significantly more infections present at weaning were eliminated in treated than in control cows, but the new infection rate during the dry period and early lactation did not differ between the two groups.

Age Factors↗

Effects of supplemental vitamin A or beta-carotene during the dry period and early lactation on udder health.

Effects of vitamin A or beta-carotene supplementation during the dry period and early lactation on the frequency of new intramammary infection and clinical mastitis and on SCC and milk yield were examined. Eighty-two Holstein cows were randomly assigned to one of three groups: 1) 50,000 IU/d of vitamin A per cow (approximately equivalent to 1978 NRC recommended daily intake for dairy cows); 2) 170,000 IU/d of vitamin A per cow; or 3) 50,000 IU/d of vitamin A plus 300 mg of beta-carotene per cow. Cows were supplemented during the 2 wk before drying off, throughout the dry period, and for the first 6 wk of lactation. Concentrations of serum vitamin A did not differ among treatment groups but tended to decrease for all treatment groups from 14 d before drying off to calving. After calving, serum vitamin A tended to increase in all groups through wk 6 of lactation. Serum beta-carotene tended to be higher in beta-carotene-supplemented cows at dry-off, in the early dry period, and again during lactation. Serum beta-carotene decreased sharply in all groups during the prepartum period. The frequency of clinical mastitis and of new intramammary infection during the dry period, near parturition, and for the first 6 wk of lactation did not differ among treatment groups. The percentage of quarters newly infected over the entire trial was 26.8 in the control, 25.0 in the high vitamin A, and 30.6 in the beta-carotene group. Pathogens isolated most frequently were coagulase-negative staphylococci, streptococci other than Streptococcus agalactiae, and coliforms.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

Changes in the bovine teat canal during the nonlactating period and early lactation, as measured by teat canal impressions.

Changes in dimensions of impressions of the lumen of the teat canals of 13 cows were examined at 17 intervals during the nonlactating period and early lactation. Impressions were made of teats of 2 diagonally opposed quarters of each cow, using dental impression material. Impression length was measured and cross sections of the impressions at the proximal (distal to Furstenburg rosette), distal (proximal to the teat orifice), and middle (midway between the 2), portions of the teat canal were prepared. Cross sections were photographed and enlarged, and circumference and area were determined by use of planimetry. Effects of making repeated impressions during the nonlactating period and early lactation on new infection rates and somatic cell counts were also assessed. Mean length of teat canal impressions decreased between days 0 and 3 of involution and during the prepartum periods. Depending on the level from which they were taken, cross-sectional areas of impressions tended to increase or increased significantly during the period of involution and again in the prepartum period. Significant changes in cross-sectional area were not observed during early lactation. Changes in circumference of proximal, middle, and distal cross sections followed trends similar to area measurements, but were more variable and differences were less statistically significant. On the basis of our findings, we suggest that heightened susceptibility to new infection during mammary involution and the prepartum period may be attributable, in part, to changes in the patency of the teat canal. Making impressions repeatedly throughout the nonlactating period and early lactation did not affect the number of new intramammary infections.

Animals↗

Herd benefit-to-cost ratio and effects of a bovine mastitis control program that includes blitz treatment of Streptococcus agalactiae.

Twelve dairy herds that had participated in the Pennsylvania Dairy Herd Improvement Association (DHIA) program for at least 12 months, that had a 12-month mean DHIA somatic cell count greater than 700,000 cells/ml, and that had greater than 25% of lactating cows infected with Streptococcus agalactiae participated in a herd blitz treatment program. Initially, quarter milk samples for bacteriologic culturing were collected from all lactating cows. Subsequently, all cows identified as infected with Str agalactiae were treated, using a commercial penicillin-novobiocin intramammary infusion product. In addition, a herd mastitis management program of postmilking teat dipping and treatment of all cows at the start of the nonlactating period was instituted. Thirty days after the initial herd visit, samples from all lactating cows were again cultured, and cows infected at that time were treated. Twelve months after the initial herd visit, samples from all lactating cows were again cultured. Mean prevalence of infection with Str agalactiae decreased (P less than 0.05) from 23.0% of quarters and 41.6% of cows initially to 3.4% of quarters and 9.3% of cows at 30 days and 1.6% of quarters and 4.2% of cows at 1 year. Mean herd DHIA somatic cell count decreased (P less than 0.05) from 918,000 cells/ml initially to 439,000 cells/ml at 30 days and 268,000 cells/ml at 1 year.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Performance studies of an enzyme-linked immunosorbent assay for detecting Staphylococcus aureus antibody in bovine milk.

An enzyme-linked immunosorbent assay (ELISA) for detecting Staphylococcus aureus antibody in bovine milk samples was examined for repeatability. A set of 51 bovine milk samples from 4 universities with confirmed culture results was assembled, and a panel of 30 milk samples was randomly selected. When the selected panel was tested at the collection laboratory, there was 97% agreement between the ELISA and the culture test. The panel was tested with the ELISA by the 4 university laboratories. Results were scored by both visual and optical density reader methods. When compared to reference ELISA results, the university laboratory ELISA results showed an agreement of 99.8% for negative samples, 98% for positive samples, and 99% for all samples. Additional studies on 19 milk samples that cultured positive for bacteria other than S. aureus showed 100% specificity. Overall comparison of ELISA and culture results showed high agreement between the 2 techniques. Disagreement appeared to result from explainable differences in antibody and bacterial levels and not from errors in either of the 2 techniques.

Animals↗

Effects of bovine mammary secretion during the early nonlactating period and antibiotics on polymorphonuclear neutrophil function and morphology.

The effect of bovine mammary secretion during the early nonlactating period and of antibiotic preparations on bovine polymorphonuclear neutrophil (PMN) phagocytic function and morphology were evaluated in a series of in vitro multifactorial experiments. Benzathine cloxacillin (CL), benzathine cephapirin (CE), sodium novobiocin (NO), and a combination of dihydrostreptomycin with procaine penicillin G (DP) were prepared in the presence and absence of a peanut oil aluminum monostearate vehicle. The PMN were isolated from bovine blood, and the effect of each antibiotic preparation on PMN function and morphology was evaluated in a buffer, fat, skin, and a combination of fat with skim from bovine mammary secretion during the nonlactating period. The fat and skim were diluted with buffer to approximate their concentration in mammary secretion. Phagocytic functions of PMN were monitored by fluorescent microscopy, which made it possible to estimate both ingestion and intracellular killing of bacteria by PMN. Changes in PMN morphology were monitored by transmission electron microscopy. The ability of PMN to ingest and kill Staphylococcus aureus ATCC 25923 was significantly decreased by fat, skim, CL, CE, NO, and DP. Effects of some antibiotics on ingestion and killing of bacteria by PMN were influenced by the addition of vehicle and by interactions with mammary secretion. Neutrophil morphology was altered by fat, skim, CL, CE, NO, and DP. The detrimental effects of CL, CE, NO, and DP on PMN morphology were influenced (some significantly) by the presence of vehicle and interactions with mammary secretion. There were significant correlations among secretion- and antibiotic-induced changes in PMN ingestion of bacteria, PMN killing of bacteria, and PMN morphology.

Animals↗

Effects of antibiotics on phagocyte recruitment, function, and morphology in the bovine mammary gland during the early nonlactating period.

The effects of 2 antibiotic preparations administered intramammarily on phagocyte recruitment, function, and morphology were evaluated at the beginning of the nonlactating period. Twelve cows with no clinical or microbiologic evidence of mastitis were assigned to 1 of 2 treatment groups. At the end of lactation, 1 of the antibiotic preparations was infused in a fore- and hind quarter of each cow; the remaining quarters were untreated controls. One group was given benzathine cephapirin; the second group was given sodium novobiocin. Secretion samples were collected from 1 treated and 1 control quarter at 16 hours, and from the remaining 2 quarters at 64 hours after treatment. Total and differential somatic cell counts were determined, and morphology of mammary polymorphonuclear neutrophils (PMN) and macrophages was observed by transmission electron microscopy. In vitro ingestion and killing of Staphylococcus aureus by mammary PMN and macrophages were assessed by fluorescent microscopy, using acridine orange stain. Cells resident in a fixed volume of secretion were incubated with a known concentration of S aureus. Total cell and PMN concentrations were higher in treated than in control quarters. Neutrophils were the predominant cell type in both treated and control quarters over the sampling period. As measured in this study, in vitro ingestion and killing of S aureus by individual PMN from treated quarters was reduced. Antibiotic treatment also increased the proportion of morphologically abnormal phagocytes. There were significant correlations among PMN ingestion, killing, and morphology. However, increased PMN concentrations tended to compensate for the reduced phagocytic function of individual cells. Therefore, efficacy of antibiotic treatment of nonlactating cows may depend, at least in part, on increased PMN concentration, which may tend to compensate for reduced phagocytic function. Compared with PMN, macrophages appeared to have only a minor role in phagocytosis of bacteria.

Animals↗

Experimentally induced Staphylococcus aureus mastitis in selenium-deficient and selenium-supplemented dairy cows.

Ten Holstein cows were fed a selenium-deficient (SeD) diet containing 0.04 mg of Se/kg of dry matter for 3 months before and throughout their first lactation. A selenium-supplemented (SeS) group of 10 cows was fed an additional 2 mg of Se/head/d to increase dietary Se concentration of the dry matter to approximately 0.14 mg/kg of body weight. An intracisternal challenge exposure of 40 to 60 colony-forming units (CFU) of Staphylococcus aureus was administered into 1 or 2 quarters of the udder of each trial cow at about the twenty-second week of lactation. Blood Se concentration (micrograms/ml +/- SEM) at the time of challenge exposure was 0.035 +/- 0.002 in SeD and 0.139 +/- 0.006 in SeS cows. Infections were established in 14/16 of the challenge-exposed quarters in SeD and 16/19 of the challenge-exposed quarters in SeS cows. The infection in 1 quarter of each Se group cleared without treatment by the end of the 8-week trial period. Log10 peak bacterial concentrations in milk from infected SeD quarters (5.04 +/- 0.25 CFU/ml) were higher (P less than 0.05) than those of infected SeS quarters (4.40 +/- 0.12 CFU/ml). Log10 peak somatic cell count (SCC) in milk from infected SeD quarters (7.18 +/- 0.08 cells/ml) did not differ from that of SeS quarters (7.17 +/- 0.05 cells/ml). Peak bacterial concentrations were attained sooner (P less than 0.05) in SeD quarters (9.5 +/- 4.0 days) than in SeS quarters (20.7 +/- 3.1 days). Similarly, peak SCC were reached earlier (P less than 0.05) in SeD (4.3 +/- 1.1 days) than in SeS quarters (13.3 +/- 3.8 days).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Phagocytosis, bactericidal activity, and oxidative metabolism of milk neutrophils from dairy cows fed selenium-supplemented and selenium-deficient diets.

Six primiparous Holstein cows were fed a Se-deficient diet, beginning at least 90 days before their first calving, and 6 other primiparous cows were given the same diet plus a supplement of 2 mg of Se/cow/d as sodium selenite. All cows were fed their diets for the duration of the experimental period. One uninfected quarter of each cow was injected with 25 micrograms of Escherichia coli endotoxin at postpartum week 5. Leukocytes were isolated by centrifugation from milk collected at postinjection hour 16. Isolated cells were 92 +/- 3% neutrophils and were incubated with Staphylococcus aureus or E coli in a 1:300 ratio. Phagocytosis and intracellular killing by neutrophils were assessed after 0, 30, 60, and 90 minutes by a fluorochrome assay, using acridine orange. Viability of neutrophils was assessed by use of trypan blue. Superoxide anion production and hydrogen peroxide production by neutrophils also were determined. Cows fed Se-deficient diets had significantly (P less than 0.05) lower blood Se concentration and blood glutathione peroxidase activity than cows fed Se-supplemented diets. Selenium status had no effect on the phagocytic capacity of neutrophils. Neutrophils obtained from cows fed Se-supplemented diets killed a significantly (P less than 0.05) higher percentage of ingested bacteria than did neutrophils from cows fed the Se-deficient diet. Viability was significantly (P less than 0.05) reduced by incubation with S aureus in neutrophils from both groups of cows, with neutrophils from Se-deficient cows having lower viability.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Induction of Escherichia coli mastitis in cows fed selenium-deficient or selenium-supplemented diets.

Ten Holstein heifers were fed a selenium-deficient (SeD) diet (0.04 mg of Se/kg on a total ration dry-matter basis) 3 months before calving and throughout their first lactation. A selenium-supplemented (SeS) diet (2 mg of Se/head/d) was fed to a group of 10 heifers. In about the 14th week of lactation, the cows were challenge-exposed to Escherichia coli by administering 15 to 40 colony-forming units (CFU) into 1 mammary gland. Selenium concentration (microgram/ml) in blood around the time of challenge exposure was 0.033 +/- 0.002 (mean +/- SEM) in SeD and 0.132 +/- 0.006 in SeS cows. Infections were established in all challenge-exposed quarters. The frequency of quarter atrophy and agalactia, and reduction in whole-udder milk yield in the first 4 days after challenge exposure, were greater (P less than 0.05) in the SeD cows. Log10 peak bacterial concentrations in milk were higher (P less than 0.05) in SeD (7.63 +/- 0.34 CFU/ml) than in SeS cows (5.57 +/- 0.66 CFU/ml). Mean log bacterial concentration was significantly higher (P less than 0.05) from 12 to 20 hours after challenge exposure in SeD than in SeS cows. Duration of infection was significantly greater (P less than 0.05) in SeD (162.0 +/- 12.0) than in SeS cows (114.4 +/- 18.0 hours). Milk somatic cell counts increased significantly more slowly (P less than 0.05) in SeD than in SeS cows from 8 to 16 hours after challenge exposure. Ratios of milk somatic cells to bacteria in milk were significantly lower (P less than 0.05) in SeD than in SeS cows at 12 and 16 hours after challenge exposure.

Animals↗

Incidence and types of clinical mastitis in dairy herds with high and low somatic cell counts.

Eighteen dairy herds were studied, 12 with a 12-month Dairy Herd Improvement Association herd mean somatic cell count (SCC) less than or equal to 150,000 cells/ml (low SCC) and 6 with a 12-month mean SCC greater than 700,000 cells/ml (high SCC). At the outset of the study, quarter samples for bacteriologic culture were collected (in duplicate) from all quarters of all lactating cows (whole herd culture). Subsequently, quarter milk samples for culture from all cows with clinical mastitis were collected for a period of 6 months. In the herds with low SCC, results of whole herd culture revealed low prevalence of intramammary infection attributable to all major pathogens (less than 4% of all quarters). Prevalence of infection with Streptococcus agalactiae (22.2% of all quarters) and Staphylococcus aureus (6.6% of all quarters) was significantly (P less than 0.05) higher in the herds with high SCC. Mean incidence of clinical mastitis in the herds with low SCC was 4.23 infections/100 cows/month (range, 0.42 to 10.25 infections). In the herds with high SCC, mean incidence was 2.91 infections/100 cows/month (range, 1.33 to 3.92 infections). In the herds with low SCC, infection type, as mean percentage of total clinically infected quarters sampled for culture/herd, was 0.0%, 2.2%, 12.3%, 43.5%, and 28.6% for Str agalactiae, S aureus, streptococci other than Str agalactiae, coliforms, and organisms not isolated, respectively. Respective percentages for the herds with high SCC were 41.5%, 18.3%, 12.6%, 8.0%, and 8.8%. During the study period (from April through January), incidence of clinical mastitis and clinical mastitis caused by coliform bacteria were highest in July and August for herds with low SCC.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Comparison of duplicate and single quarter milk samples for the identification of intramammary infections.

Results of bacteriological culture of 5426 pairs of duplicate quarter milk samples collected were analyzed for agreement. Overall, the percentage of agreeing pairs was 98.1%. The percentage agreement by infection type was greater for the contagious pathogens, Streptococcus agalactiae (96.4%) and Staphylococcus aureus (94.2%), than for other Streptococcus spp. (81.6%) and coliform organisms (55.6%). Single quarter samples may be adequate for determining status of quarter infection with Strep. agalactiae and Staph. aureus.

Animals↗

Pseudomonas mastitis: difficulties in detection and elimination from contaminated wash-water systems.

Histories of 4 dairy herds with increased incidence of Pseudomonas mastitis associated with contaminated wash hoses in milking parlors are described. Problems of detection and elimination of the organism from contaminated water sources and the inadequacy of iodide germicides in eliminating Pseudomonas are discussed. In problem herds, greater numbers of organisms often are found in the water left standing in the wash hoses between milkings. Thus, flushing of hoses before their use in the milking process may be beneficial in reducing exposure of the cows to the organism.

Animals↗