Bacterial counts associated with sawdust and recycled manure bedding treated with commercial conditioners.

Bacteria counts associated with untreated organic bedding materials were compared with those of bedding treated with either an alkaline commercial bedding conditioner, acidic commercial bedding conditioner, or hydrated lime. Bedding materials were recycled manure and kiln-dried sawdust. The effects of bedding treatments on bacteria counts differed between bedding types. Each of the bedding treatments significantly reduced bacteria in recycled manure prior to use. The alkaline conditioner and hydrated lime effectively inhibited bacteria in recycled manure for 1 d. Bedding counts and teat swabs of cows housed on recycled manure treated with the alkaline conditioner were reduced on d 2. The use of the acid conditioner in recycled manure had little effect on bacteria in bedding. Sawdust differed from recycled manure in that bacteria in untreated sawdust prior to use were minimal, and populations increased rapidly during the first 2 d after use as bedding. The acid conditioner had a bacteriostatic effect in sawdust, evident by the reduction of bacteria on d 2. The alkaline conditioner and hydrated lime did not alter bacteria counts in sawdust compared with untreated sawdust. Antibacterial activity of each conditioner deteriorated between d 2 and d 6 in both beddings. The antibacterial activities of conditioners were related to the pH of bedding materials. The use of commercial bedding conditioners initially reduced bacterial counts; however, the antibacterial effects had diminished between d 2 and 6 after use in bedding.

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.

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.

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.

Efficacy of dry cow therapy and a Propionibacterium acnes product in herds with low somatic cell count.

Dry cow therapy and a Propionibaterium acnes product were evaluated in four commercial herds with low SCC. Cows were randomly assigned within herds to treatment groups of approximately 90 cows receiving dry cow therapy, P. acnes, dry cow therapy plus P. acnes, or no treatment in a factorial arrangement. Each lactating quarter of cows that received dry cow therapy was infused via the teat duct with 300 mg of cephaprin at drying off. Cows that received P. acnes were infused intravenously with .4 mg of killed P. acnes at drying off, 7 to 10 d prepartum, and within 7 d after calving. A second prepartum injection of P. acnes immunostimulator was administered to cows that did not calve within 10 d after the first prepartum injection. Dry cow therapy enhanced bacteriological cures of IMI by Staphylococcus aureus and Corynebacterium bovis at drying off. Dry cow therapy reduced incidence of new IMI by environmental streptococci and C. bovis that originated during the dry period. Cows treated with P. acnes alone had a greater incidence of new IMI by Gram-negative bacilli originating during the dry period than did cows in the other treatment groups. Incidence of clinical mastitis at calving was greater for cows receiving no treatment than for cows receiving dry cow therapy, P. acnes, or dry cow therapy plus P. acnes.

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.

Growth responses of coliform bacteria to recombinant bovine cytokines.

Growth responses of 10 coliform isolates to recombinant bovine cytokines were measured in vitro. Six Escherichia coli and four Klebsiella pneumoniae isolates obtained from bovine IMI were tested for growth responses to recombinant bovine interleukin-1 beta, interleukin-2, and interferon-gamma. Cytokines were tested at 10(4), 10(3), 10(2), and 10 U/ml of media. Media used were a synthetic tissue culture medium, a chemically defined synthetic bacterial growth medium, and UHT sterilized milk. Bacterial counts in the synthetic tissue culture medium and UHT milk increased slightly as concentration of interferon-gamma in the media increased. Recombinant bovine interferon-gamma increased bacterial populations during the log growth phase but did not affect the number of bacteria in stationary growth phase. Bacterial growth responses were not related to either interleukin-2 or interleukin-1 beta concentrations in any of the three media. Bacterial growth responses to cytokines were not related to differences in either serum susceptibility, growth of isolates in dry cow secretion, duration of IMI from which isolates were obtained, or bacterial species.

Vitamin E as an adjuvant in an Escherichia coli J5 vaccine.

Vitamin E was tested as an adjuvant in an Escherichia coli (O111:B4) J5 vaccine. Twenty cows were assigned to five groups of 4 cows. Cows in four groups were vaccinated with an E. coli J5 bacterin containing 5 ml of 10(9) boiled cells/ml. Vaccinations were at drying off, 30 d after drying off, and within 48 h after calving. Vaccine adjuvants differed among groups. The four treatment adjuvants were 5 ml of Freund's incomplete adjuvant, 5 ml of vitamin E, 2.5 ml of Freund's plus 2.5 ml of vitamin E, and 5 ml of PBS. Cows in the fifth group were unimmunized controls. A front mammary quarter of each cow was challenged by infusion of 10 micrograms of E. coli J5 lipopolysaccharide approximately 4 wk into lactation. Vitamin E alone enhanced serum IgM titers but had no effect on milk IgM or serum and milk IgG titers. The mixture of Freund's plus vitamin E resulted in peak IgG titers in serum and milk comparable with that of Freund's alone. Persistency of IgG titers in cows immunized with the Freund's plus vitamin E mixture was greater than the persistency of titers for cows immunized with the vaccine containing Freund's alone as the adjuvant. The mixture of Freund's plus vitamin E had a synergistic effect in reducing severity of systemic clinical signs.

Susceptibility of Escherichia coli isolated from intramammary infections to phagocytosis by bovine neutrophils.

Thirteen Escherichia coli isolated from naturally occurring IMI were tested for susceptibility to phagocytosis by bovine blood neutrophils. Isolates were opsonized in pooled serum collected from nine healthy lactating cows. Bacteria isolated from IMI first diagnosed within 3 d after calving were more resistant to phagocytosis than were isolates from IMI originating during either the first half of the dry period or later during lactation. Duration of the IMI was negatively correlated with both phagocytic index and percentage of neutrophils phagocytizing within bacterial isolates from IMI originating at calving and during lactation. Phagocytosis was independent of duration of IMI within isolates from IMI originating during the first half of the dry period. Susceptibility to in vitro phagocytosis by neutrophils was not related to O antigen serotype, encapsulation, or growth in dry cow secretion.

Bovine neutrophil responses to parenteral vitamin E.

Twenty-eight Holsteins were tested to determine effects of dietary and parenteral vitamin E supplementation during the dry period on plasma alpha-tocopherol and in vitro neutrophil functions at calving. Cows were assigned to one of four experimental groups receiving either supplemental dietary vitamin E, injections of vitamin E, both dietary and injections of vitamin E, or neither source of supplemental vitamin E during the dry period in a 2 x 2 factorial arrangement. Cows receiving parenteral vitamin E were injected subcutaneously with 3000 IU of vitamin E (dl-alpha-tocopherol) at 10 and 5 d prior to anticipated calving. Cows not receiving parenteral vitamin E were injected with a placebo. Experimental groups receiving dietary vitamin E during the dry period were supplemented with 1040 IU/d compared with none for controls. Cows injected with vitamin E had greater plasma alpha-tocopherol concentration 5 d after the first injection, at calving, and 1 wk after calving than did cows injected with placebo. Plasma alpha-tocopherol concentrations did not differ between dietary vitamin E treatment groups from calving through 4 wk postpartum. No interaction was found between dietary and parenteral supplementation of vitamin E on plasma alpha-tocopherol concentration. Neutrophils from cows injected with vitamin E had greater intracellular kill of bacteria at calving than did neutrophils from placebo-injected cows. Neither phagocytic index nor percentage of neutrophils phagocytizing differed between vitamin E-injected and placebo-injected cows. Dietary vitamin E during the dry period had no effect on neutrophil function at calving. Intracellular kill and plasma alpha-tocopherol were correlated at calving.

Efficacy of an Escherichia coli J5 mastitis vaccine in an experimental challenge trial.

An Escherichia coli (O111:B4) J5 bacterin was tested for efficacy in reducing IMI and severity of clinical coliform mastitis in an experimental challenge trial. Ten cows were immunized at drying off, 30 d after drying off, and at calving. Ten control cows were not immunized. Right front quarters of all cows were infused with a heterologous strain of E. coli approximately 30 d after calving. Vaccinated cows had lower bacterial counts in milk and lower rectal temperatures than unvaccinated controls following intramammary challenge. Milk production and DMI were more depressed in controls than in vaccinated cows. Milk SCC did not differ between experimental groups. Mean serum IgG titer to whole cell E. coli J5 was significantly greater in vaccinated than in unvaccinated cows at 30 d after drying off, day of challenge, and 7 d postchallenge. Milk IgG titer to E. coli J5 was higher at challenge in vaccinated than in control cows. Vaccination with the E. coli J5 bacterin did not prevent IMI but did reduce severity of clinical signs following intramammary experimental challenge with a heterologous E. coli strain.

Opsonic activity of bovine serum and mammary secretion after Escherichia coli J5 vaccination.

Six pairs of cows were used to determine the effects of immunization with an Escherichia coli (O111:B4) J5 bacterin on in vitro opsonization of a smooth heterologous strain of E. coli. One cow in each pair was either immunized with the vaccine or sham-immunized at drying off, 30 d after drying off, and at calving. Opsonizing bacteria with serum collected from vaccinated cows 21 d after calving resulted in higher mean number of intracellular bacteria per phagocytosing neutrophil than opsonizing bacteria with serum collected from control cows. Phagocytic parameters using serum collected at drying off and calving did not differ between treatment groups. A trend for enhanced opsonic activity of colostrum from vaccinates was noted. Enhanced opsonization by serum from vaccinated cows coincided with higher serum IgM titer to E. coli J5 whole cell antigen compared with controls. Serum IgG titers to E. coli J5 did not differ between groups. Colostrum IgG titers to E. coli J5 were greater at calving in vaccinated than in control cows. Colostrum and milk collected 21 d after calving from vaccinated cows had higher IgM titers to E. coli J5 than did mammary secretions from control cows. Numbers of intracellular bacteria per phagocytizing neutrophil were correlated positively with IgM titers to E. coli J5 in both serum and colostrum.

Field trial to determine efficacy of an Escherichia coli J5 mastitis vaccine.

Efficacy of an Escherichia coli (O111:B4) J5 bacterin for preventing naturally occurring IMI and clinical mastitis was tested in a 2.5-yr field trial in a 225-cow commercial herd. Cows with odd-numbered identification were vaccinated, and cows with even-numbered identification served as unvaccinated controls for each lactation during the study. Immunizations were subcutaneous on the upper part of the rib cage just posterior to the scapula at drying off, 30 d after drying off, and at calving. Percentage of quarters infected at calving with Gram-negative bacteria did not differ between treatment groups. A total of 67% of Gram-negative bacterial IMI present at calving in control cows became clinical during the first 90 d of lactation compared with 20% in vaccinated cows. Rate of Gram-negative bacterial clinical mastitis was higher in control cows than in vaccinated cows during the first 90 d of lactation. Immunization with the E. coli J5 bacterin did not reduce level of Gram-negative bacterial IMI at calving but did reduce incidence of clinical mastitis.

Costs associated with selected preventive practices and with episodes of clinical mastitis in nine herds with low somatic cell counts.

Nine dairy herds (mean size, 149 cows) with bulk-tank milk somatic cell counts of less than 300,000 cells/ml and greater than 80% of cows with Dairy Herd Improvement Association linear somatic cell counts less than or equal to 4 were selected for study. Each herd was monitored for 12 consecutive months. Duplicate quarter-milk specimens were collected from each cow for bacteriologic culturing at beginning of lactation, cessation of lactation, and at the time of each clinical episode of mastitis. Streptococcus agalactiae was never isolated and Staphylococcus aureus was isolated from less than 1% of all quarters. There were 554 episodes of clinical mastitis. During the year of study, the incidence rate of clinical mastitis varied from 15.6 to 63.7% of cows among the 9 herds. Mean costs per cow per year in herd for mastitis prevention were: $10 for paper towels, $3 for nonlactating cow treatment, and $10 for teat disinfectants. Mean cost associated with clinical mastitis was $107/episode. Approximately 84% ($90) of the costs attributed to a clinical episode were associated with decreased milk production and nonsalable milk. Costs of medication and professional veterinary fees per clinical episode varied significantly among the 9 herds. Three of the herds did not have a veterinarian treat a clinical episode of mastitis during the year of study even though 2 of these herds had the first and third highest incidence rates of clinical mastitis. When calculated on a per cow in herd basis, mean costs of $40/cow/year were attributed to clinical mastitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Gram-negative bacterial infections of the mammary gland in cows.

Naturally acquired gram-negative bacterial intramammary infections (n = 160) were studied in 99 cows over a 2-year period. Escherichia coli, Klebsiella spp, Serratia spp, Enterobacter spp, and unidentified gram-negative bacteria were isolated from 28.8, 39.4, 9.4, 5.0, and 11.2%, respectively, of infected mammary glands. A majority (61%) of intramammary infections were first detected during the nonlactating period. Gram-negative bacteria isolated during the first half of the nonlactating period were predominantly Klebsiella spp, Serratia spp, and Enterobacter spp. Onset of E coli intramammary infections was more prevalent during the second half of the nonlactating period and during the first 7 days of lactation. The majority (59%) of infections were less than 28 days in duration, but Klebsiella spp and Serratia spp infections were of significantly (P less than 0.05) greater duration than infections with E coli. The greatest percentage (47%) of gram-negative bacterial intramammary infections were first detected during the summer.

Hemagglutination and hemolysis by Escherichia coli isolated from bovine intramammary infections.

Seventy-six Escherichia coli isolated from bovine intramammary infections were tested for hemagglutination and hemolysis of erythrocytes. Fifty-seven percent of isolates were hemagglutination-positive for bovine erythrocytes compared with 46% that agglutinated guinea pig erythrocytes. Twenty-eight percent of isolates were hemagglutination-positive for erythrocytes from both species. Only 14.5 and 2.6% of isolates were mannose-resistant, hemagglutination-positive for bovine and guinea pig erythrocytes, respectively. Neither duration nor severity of infection from which isolates were obtained differed between isolates that were hemagglutination-positive and hemagglutination-negative. Percentage distribution of hemagglutination-positive isolates did not differ among isolates from infections that originated at calving, during lactation, or the first half of the dry period. Hemagglutination reactions were also not related to in vitro growth in cell-free dry cow secretion. Percent of isolates that caused hemolysis of washed bovine erythrocytes was 2.6% compared to 3.9% for sheep erythrocytes. Hemolysis was not related to hemagglutination. Hemagglutination and hemolysis of erythrocytes did not appear to be virulence factors for E. coli isolated from bovine intramammary infections.

Bacterial counts associated with recycled newspaper bedding.

Bacterial counts associated with recycled newspaper, wood shavings, and pelleted corn cobs used as bedding for lactating dairy cows were compared. Chopped newspaper and pelleted corn cobs had similar gram-negative bacterial, coliform, and streptococcal bedding counts. Staphylococcal counts in pelleted corn cobs were greater than in chopped newspaper. Conversely, gram-negative bacterial, coliform, and staphylococcal counts in chopped newspaper were greater than in wood shavings. Coliform and streptococcal counts did not differ between chopped newspaper and wood shavings bedding materials. Teat swab counts from cows bedded on pelleted corn cobs were greater than those from cows bedded on chopped newspaper for gram-negative bacterial, coliform, Klebsiella species, and staphylococci. Streptococcal teat swab counts did not differ between cows bedded on chopped newspaper and pelleted corn cobs. Cows bedded on chopped newspaper and wood shavings had similar gram-negative bacterial, coliform, and Klebsiella species teat swab counts. Streptococcal and staphylococcal teat swab counts were greater from cows bedded on chopped newspaper than those from cows bedded on wood shavings. Teat swab and bedding counts were correlated. In general, bacterial counts in bedding suggest no advantage in using chopped newspaper over pelleted corn cobs or wood shavings in reducing exposure of teats to environmental mastitis pathogens.

Collaborative Study to Determine Effects of Milk Composition on Somatic Cell Counts.

Quarter, composite, and bulk tank milk samples were analyzed in a three laboratory collaborative study to determine the relationship of milk fat and protein content with milk somatic cell counts. Milk somatic cell counts were determined by two Coulter counters, a Fossomatic counter, and by direct microscopic somatic cell counting. In general, variability among somatic cell counts measured by different procedures was not related to protein or fat content of milk. The greatest percentage of variation between counts that could be explained by fat content of milk was 20.2% between a Coulter and direct microscopic somatic cell counts. The greatest percentage of variation between counts that could be explained by protein content of samples was 12.9% between a Coulter and Fossomatic counts. Breed of cow from which samples were collected also had little influence on differences among milk somatic cell counts. Differences among milk somatic cell counts due to counting methods did vary among quarter, composite, and bulk tank milk samples.

Serum susceptibility of coliforms isolated from bovine intramammary infections.

A total of 35 coliforms isolated from bovine intramammary infections were tested for serum susceptibility. Growth responses of Klebsiella pneumoniae isolated from intramammary infections originating during the dry period were inhibited more than were growth responses of Klebsiella pneumoniae isolated from infections originating during lactation. Escherichia coli growth responses did not differ between isolates originating from dry period and lactational intramammary infections. Mean growth responses to serum did not differ between Klebsiella pneumoniae and Escherichia coli. Bactericidal properties of serum were due to complement activity. The classical pathway of complement appeared to be the principal source of bactericidal activity of serum toward Klebsiella pneumoniae. Alternative and classical pathways of complement were both important for bactericidal effects of serum on Escherichia coli. Iron saturation of serum did not affect growth responses of either Klebsiella pneumoniae or Escherichia coli. These data illustrate that coliforms isolated from intramammary infection originating during the dry period were not more resistant to the bactericidal activity of bovine serum than were coliforms isolated from intramammary infection occurring during lactation.

Field survey of clinical mastitis in low somatic cell count herds.

Nine commercial dairy herds, each with low herd milk somatic cell counts, were monitored for 1 yr to determine prevalence of intramammary infections and rates of clinical mastitis. Staphylococcus species was the bacterial group most frequently isolated from quarters at calving and at drying off. Environmental streptococci and coliform intramammary infections totaled less than 6% of quarters at both calving and at drying off. Staphylococcus aureus were isolated from less than 1% of quarters and Streptococcus agalactiae from 0% of quarters at both calving and drying off. A total of 646 clinical cases of mastitis were diagnosed in 548 quarters of 406 cows. Mean rate of clinical mastitis among herds was .457 clinical cases/305 cow-days. Rates of clinical mastitis ranged among herds from .273 to .748 clinical cases/305 cow-days. Coliforms and bacteriologically negative and environmental streptococci accounted for 82.3% of clinical cases. Rates of clinical mastitis and severity of clinical signs differed among herds, seasons of the year, parity groups, and stages of lactation. Rates of clinical mastitis were highest during summer, in first lactation cows, and during the first 7 d of lactation.