Effect of abraded intramammary device on outcome in lactating cows after challenge exposure with Streptococcus uberis

Intramammary devices (IMD) were abraded with medium-grade emery cloth or were left smooth. One IMD of each type was inserted into a mammary quarter of each of 5 lactating cows. The remaining 2 quarters served as controls. Quarter foremilk, bucket milk, and stripping milk samples were collected for 3 consecutive days at 2 weeks after IMD insertion, and milk somatic cell counts (SCC) were determined. Milk samples also were collected immediately after and 0.5, 1, 2, 4, 6, 8, and 11 hours after milking. All quarters were challenge exposed with 250 colony-forming units of Streptococcus uberis at 2 months after IMD insertion. Foremilk and stripping milk samples were collected for bacteriologic culture and SCC at the next 10 milkings. Mean foremilk, bucket milk, and stripping milk SCC (X 10(6) cells/ml) were 0.18, 0.07, and 0.91, respectively, for quarters with abraded IMD; 0.06, 0.05, and 0.43, respectively, for quarters with smooth IMD; and 0.03, 0.03, and 0.15, respectively, for control quarters. Mean SCC after milking (X 10(6) cells/ml) for the various intervals were 0.70, 1.29, 0.70, 0.97, 1.15, 1.17, 0.77, and 0.85 for quarters with abraded IMD; 0.43, 0.62, 0.61, 0.45, 0.64, 0.60, 0.31, and 0.26 for quarters with smooth IMD; and 0.15, 0.24, 0.15, 0.19, 0.15, 0.15, 0.14 and 0.06 for control quarters. After challenge exposure, 2 of 5 of the quarters with abraded IMD, 4 of 5 of the quarters with smooth IMD, and 8 of 9 control quarters became infected. Results indicated that abraded IMD increased SCC in stripping milk to concentrations that provided 60% protection against challenge exposure with S uberis.     

Effect on outcome of intramammary challenge exposure with Staphylococcus aureus of somatic cell concentration and presence of an intramammary device

Results of experimental Staphylococcus aureus intramammary challenge of all quarters of 6 cows, each fitted with an intramammary device (IMD) in 2 quarters, and of 10 quarters of 3 cows not fitted with an IMD were reported. Infection was established in all 34 quarters, regardless of presence or absence of an IMD. Neither the course nor severity of early S aureus intramammary infection were influenced by the presence of an IMD or by differences in milk somatic cell (MSC) concentration in the gland at the time of bacterial challenge infusion, up to a MSC concentration of nearly 1 million/ml. Cumulative success of experimental infection in this and a previous study from our laboratory was nearly 100% in glands in which the MSC concentration was less than 1 million/ml and about 17% when the MSC concentration exceeded 1 million/ml.     

Persistent Experimental Salmonella dublin Intramammary Infection in Dairy Cows

Experimental intramammary infections were induced in five post-parturient Holstein cows by inoculation of low numbers (5000 colony forming units) of virulent Salmonella dublin via the teat canal of mammary gland quarters. Rectal temperature, pulse and respiratory rates, milk yield, and milk quality as assessed by the California Mastitis Test (CMT) and somatic cell counts (SCC) were recorded every 12 hours at milking. Bacteriologic cultures of foremilk quarter samples and feces were obtained daily, as were complete blood counts. ELISA titers for IgG and IgM recognizing S. dublin lipopolysaccharide (LPS) were obtained weekly on serum and quarter milk samples. All cows excreted S. dublin intermittently from infected quarters, but no changes were detected in rectal temperature, appearance of the mammary gland or secretions, CBC, milk yield, and pulse and respiratory rates. Somatic cell counts were modestly increased in infected quarters as compared with uninfected quarters (P = .015, paired t test); however, CMT scores after infection remained low, and were not significantly different from pre-infection scores (P greater than .10, sign test). After infection, administration of dexamethasone resulted in signs of clinical mastitis and increased excretion of S. dublin from mammary quarters (P = .0004, paired t test). One cow had necrotizing mastitis and S. dublin septicemia and was euthanatized. In the four surviving cows, clinical improvement was observed after systemic gentamicin therapy and intramammary infusion with polymyxin B, but all cows continued to excrete S. dublin intermittently from one or more quarters and occasionally from feces for the remaining period of observation. All infected cows demonstrated a rise in IgG and IgM ELISA titers recognizing S. dublin LPS in serum and milk. At necropsy (13-25 weeks postinfection), S. dublin was recovered only from the mammary tissue or supramammary lymph nodes in three of four cows. In one cow, mammary gland and lymph-node samples were negative for S. dublin despite positive milk cultures. In all cows, histopathologic examination revealed multifocal areas of chronic active mastitis. These lesions were similar to histopathologic findings from mammary gland carriers with naturally acquired S. dublin infection.     

Efficacy of intramuscular treatment of beef cows with oxytetracycline to reduce mastitis and to increase calf growth

Spring-calving multiparous Angus x Hereford cows were used to determine the efficacy of intramuscular treatment with oxytetracycline to reduce the incidence of mastitis-causing bacteria, decrease milk somatic cell counts (SCC), and increase calf growth. During 2 yr, milk samples were collected from each quarter from a total of 319 cows at 8 to 14 d after calving and at weaning, to determine the presence of bacteria and SCC. A California mastitis test (CMT) was performed on milk from each quarter of each cow at the initial sample collection. Cows with a CMT score of 1, 2, or 3 in at least one quarter, were randomly assigned to receive either an intramuscular injection of oxytetracycline (n = 63) or the control vehicle (n = 60), and cows with a CMT score of 0 or trace in all four quarters were not treated (n = 196). Calf weights were determined at birth, early lactation, and weaning. The number of somatic cells in milk and the percentage of quarters that were infected increased as CMT score increased (P < 0.01). The presence of mastitis-causing bacteria at calving increased (P < 0.05) the incidence of infection at weaning. The presence of mastitis-causing bacteria at weaning was associated with increased SCC for quarters and average SCC for cows (P < 0.01). Average SCC per cow at weaning increased (P < 0.05) as the number of infected quarters per cow increased. Treatment did not alter (P > 0.10) the percentage of cows or quarters infected with mastitis-causing bacteria or SCC of cows or quarters at weaning. Average SCC per cow was negatively correlated (P < 0.05) with calf weights at early lactation, but not with weaning weights of calves. Treatment did not influence (P > 0.10) calf weights at early lactation or at weaning. Cows with one or more dry quarters after calving had calves that weighed less at early lactation and weaning than cows with four functional quarters (P < 0.01). Intramuscular oxytetracycline treatment of beef cows that had CMT scores of 1 or greater after calving did not reduce intramammary infection rates or increase calf weights at weaning.     

Endotoxin-induced bovine mastitis: arachidonic acid metabolites in milk and plasma and effect of flunixin meglumine

Arachidonic acid metabolites (AAM) were measured in milk and plasma during the course of acute endotoxin-induced mastitis in 12 lactating cows. Mastitis was induced by intramammary challenge exposure with 10 micrograms of Escherichia coli (026:B6) endotoxin. Endotoxin was injected into the teat cistern via the teat canal of a single randomly selected rear quarter of each cow. Concentrations of prostaglandin (PG) F2 alpha and thromboxane (Tx) B2 in fat-free unextracted milk and of 15-keto-13,14-dihydro-PGF2 alpha in plasma were measured by radioimmunoassay. Total production of AAM in milk was determined by measuring quarter milk production. The AAM were compared in 6 cows administered flunixin meglumine (1.1 mg/kg of body weight) and in 6 cows administered saline solution. Concentrations of TxB2 in milk were significantly (P less than 0.001) increased during the early course of acute mastitis in endotoxin-treated quarters of cows not administered flunixin meglumine. Peak concentrations of TxB2 in milk occurred at 8 hours after endotoxin inoculation. Flunixin meglumine treatment produced significant (P less than 0.05) reductions in milk TxB2 and plasma 15-keto-13,14-dihydro-PGF2 alpha concentrations. Concentrations of PGF2 alpha in milk and total PGF2 alpha and TxB2 production per quarter per milking were not significantly influenced by endotoxin challenge or by flunixin meglumine treatment.     

Intensification of milk somatic cell response to intramammary device

Treatments consisting of copper-impregnated polyethylene intramammary device (PIMD-Cu), PIMD-Cu which had been abraded to ensure exposure of surface copper (APIMD-Cu), PIMD which had been abraded in an identical manner (APIMD), and an untreated control were established in mammary quarters of 2 cows. Quarters selected were bacteria free and had milk somatic cell counts (MSCC) in strippings of less than 240 X 10(3) ml. Milk somatic cell counts were determined from strippings immediately after milking and 6 hours later. Cows were monitored for 4 weeks after devices were inserted. Milk samples (250 ml) were collected and analyzed for free fatty acids, a measure of hydrolytic rancidity, at 14 days. The devices were removed from mammary quarters after 4 weeks and examined with a scanning electron microscope. Surfaces of the devices were assessed for plaque buildup and presence of leukocytes. Immediately after and 6 hours later, geometric mean MSCC for APIMD, APIMD-Cu, PIMD-Cu, and control quarters average 946/1,556, 1,479/2,882, 512/1,148, and 161/190 X 10(3) ml, respectively. There was no difference in free fatty acid values of samples from treated and control quarters. Plaque formation was observed over the entire surface of APIMD. Numerous leukocytes were found associated with plaque and appeared to initiate development of plaque. Smaller amounts of plaque were found on APIMD-Cu, and minimal amounts were found on PIMD-Cu. Results indicate that modification of PIMD by abrading or addition of copper will increase MSCC to concentrations (greater than 900 X 10(3) ml) that should be protective against establishment of infection by mastitis pathogens.     

Elimination kinetics of ceftiofur hydrochloride after intramammary administration in lactating dairy cows

OBJECTIVE: To determine the elimination kinetics of ceftiofur hydrochloride in milk after intramammary administration in lactating dairy cows. DESIGN: Prospective study. ANIMALS: 5 lactating dairy cows. PROCEDURE: After collection of baseline milk samples, 300 mg (6 mL) of ceftiofur was infused into the left front and right rear mammary gland quarters of each cow. Approximately 12 hours later, an additional 300 mg of ceftiofur was administered into the same mammary gland quarters after milking. Milk samples were collected from each mammary gland quarter every 12 hours for 10 days. Concentrations of ceftiofur and its metabolites in each milk sample were determined to assess the rate of ceftiofur elimination. RESULTS: Although there were considerable variations among mammary gland quarters and individual cows, ceftiofur concentrations in milk from all treated mammary gland quarters were less than the tolerance (0.1 microg/mL) set by the FDA by 168 hours (7 days) after the last intramammary administration of ceftiofur. No drug concentrations were detected in milk samples beyond this period. Ceftiofur was not detected in any milk samples from nontreated mammary gland quarters throughout the study. CONCLUSIONS AND CLINICAL RELEVANCE: Ceftiofur administered by the intramammary route as an extra-label treatment for mastitis in dairy cows reaches concentrations in milk greater than the tolerance set by the FDA. Results indicated that milk from treated mammary gland quarters should be discarded for a minimum of 7 days after intramammary administration of ceftiofur. Elimination of ceftiofur may be correlated with milk production, and cows producing smaller volumes of milk may have prolonged withdrawal times.     

Effects of intramammary infection and parity on calf weaning weight and milk quality in beef cows

The objectives of this study were to determine 1) the effect of intramammary infection on calf weaning weight, milk somatic cell count, and milk composition, and 2) the effect of parity on percentages of infected cows, infected quarters, and blind quarters. The number of infected quarters, milk somatic cell counts, milk components, and intramammary infection were studied at weaning in 164 beef cows. The percentage of infected cows ranged from 61.9% at first parity to 66.7% at fifth to ninth parities. Cows with three or four infected quarters had higher (P < .01) milk somatic cell counts than cows with zero, one, or two infected quarters. Among bacterial isolates, Staphylococcus aureus-infected quarters had the highest (P < .01) milk somatic cell count. Percentages of butterfat and lactose were lower (P < .01) in milk from infected quarters than from uninfected quarters. Infections by S. aureus and coagulase-negative staphylococci were the most common and accounted for 67 to 78% of the infections. Percentages of infected quarters and infections caused by S. aureus increased with parity (P < .01). Intramammary infections did not affect (P > .10) calf weaning weight. In conclusion, intramammary infection had no effect on calf weaning weight but increased milk somatic cell count and decreased the percentage of protein, lactose, solids-not-fat, and butterfat. The number of infected and blind mammary quarters increased with parity.     

The prophylactic effect of a dry-cow antibiotic against Streptococcus uberis

The prophylactic use of a dry-cow antibiotic for reducing the incidence of mastitis due to Streptococcus uberis was studied in four seasonally calving dairy herds involving 378 cows. The treatment was a long-acting dry-cow antibiotic preparation administered immediately after the last milking of lactation. New intramammary infections were identified by comparing the bacteriological status of quarters at drying off with that after calving, or through manual udder palpation during the dry period. The administration of dry-cow antibiotic to uninfected quarters at drying off reduced the overall incidence of new infections with Streptococcus uberis from 12.3% for untreated quarters to 1.2% of quarters (p<0.01). The reduction was significant (p<0.01) for both dry-period and post-calving infections. The susceptibility of uninfected quarters to new infection by Streptococcus uberis appeared to be unrelated to the infection status of a cow at drying off. Clinical infections during the dry period were most prevalent (97%) in quarters identified as having open teat canals. Fewer open teat canals (p<0.05) were observed among antibiotic treated quarters over the first 4 weeks of the dry period. Treated quarters had a lower (p<0.05) incidence of new clinical infection during the ensuing lactation and lower somatic cell counts. This did not affect production levels of milk, milk fat or protein. The results clearly indicated a prophylactic benefit for the dry cow antibiotic treatment against new Streptococcus uberis infections during the dry period.     

Antibiotic Treatment of Experimental Staphylococcus aureus Infections of the Bovine Mammary Gland

Experimental infections were produced in 78 quarters of 17 cows by the infusion of small numbers of a single strain of Staphylococcus aureus. In each single experiment three quarters in a cow were infected, with the fourth left as a control. At times varying from three to 60 days after the infusion of organisms, a standard intramammary antibiotic treatment was administered on a single occasion. A cure was arbitrarily defined as the absence of the organism in foremilk, from direct plating and replated incubated milk, together with return to normal somatic cell count levels as determined by an electronic counter.

With these standardized conditions the effects of a number of cow associated factors on the outcome of the therapy were determined.

Forty-three of the 78 quarters (55%) were cured by the standard treatment. Significant differences in percentages of quarters cured were found to be associated with the duration of infection before therapy, the lactation age of the cow, the length of time in lactation, somatic cell count in milk at time of treatment, the location of the quarter in the udder and individual cows. No significant effects on the outcome of the standard treatment were found associated with the number of bacteria in the secretion at the time of treatment, previous infection and cure in a quarter nor the season of the year in which treatment was given. Of the 35 quarters in which infection recurred following treatment, organisms were reisolated from 12 within four days, 18 between five and nine days, four between ten and 17 days and one after 28 days.

From these data it is apparent that if, as has been suggested, models such as described are to be used for efficacy trials, standardization of some parameters is essential.

     

The prophylactic effect of a dry-cow antibiotic against Streptococcus uberis

The prophylactic use of a dry-cow antibiotic for reducing the incidence of mastitis due to Streptococcus uberis was studied in four seasonally calving dairy herds involving 378 cows. The treatment was a long-acting dry-cow antibiotic preparation administered immediately after the last milking of lactation. New intramammary infections were identified by comparing the bacteriological status of quarters at drying off with that after calving, or through manual udder palpation during the dry period. The administration of dry-cow antibiotic to uninfected quarters at drying off reduced the overall incidence of new infections with Streptococcus uberis from 12.3% for untreated quarters to 1.2% of quarters (p<0.01). The reduction was significant (p<0.01) for both dry-period and post-calving infections. The susceptibility of uninfected quarters to new infection by Streptococcus uberis appeared to be unrelated to the infection status of a cow at drying off. Clinical infections during the dry period were most prevalent (97%) in quarters identified as having open teat canals. Fewer open teat canals (p<0.05) were observed among antibiotic treated quarters over the first 4 weeks of the dry period. Treated quarters had a lower (p<0.05) incidence of new clinical infection during the ensuing lactation and lower somatic cell counts. This did not affect production levels of milk, milk fat or protein. The results clearly indicated a prophylactic benefit for the dry cow antibiotic treatment against new Streptococcus uberis infections during the dry period.     

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.     

Quarter,cow, and farm risk factors for intramammary infections with major pathogens relative to minor pathogens in Thai dairy cows

A cross-sectional study was carried out from May to September 2011 on 35 smallholder dairy farms in Chiang Mai, Thailand, to identify the quarter, cow, and farm factors that relate to intramammary infections (IMI) from major specified pathogens, compared to infections from minor pathogens. Data on general farm management, milking management, and dry cow management were recorded for each herd. Quarter milk samples were collected from either clinical or subclinical mastitis quarters. Dependent variables were binary data defining the specified major pathogens, including Streptococcus agalactiae (7.1 %), Streptococcus uberis (9.4 %), Streptococcus dysgalactiae (4.0 %), and other streptococci (16.7 %), as a case, and all minor pathogens as a control, in each dependent variable. The occurrence of S. agalactiae IMI was lower in first-parity cows and cows with short milking time. Cows with body condition score (BCS) <2.5 had higher occurrence of S. agalactiae IMI. The occurrence of S. uberis IMI was higher in quarters with California mastitis test (CMT) score 2, score 3, and having clinical mastitis and in farms with increasing age of vacuum system. Quarters with CMT score 3, having clinical mastitis, cow with manual milking after detaching milking cluster, and farms with high bulk milk somatic cell counts (BMSCC >500,000 cells/ml) had higher occurrence of S. dysgalactiae IMI. For other streptococci, quarters having clinical mastitis, BCS <2.5, and pulling down of milking cluster while milking increased occurrence of other streptococci IMI relative to minor pathogen IMI. These results highlight the importance of individual cow factors, milking characteristics, and BMSCC in determining the risk of IMI from major pathogens.     

Interleukin-1β infusion in bovine mammary glands prior to challenge with <Emphasis Type="Italic">Streptococcus uberis</Emphasis> reduces bacterial growth but causes sterile mastitis

Dairy cows are especially vulnerable to intramammary infection by the bacterial pathogen Streptococcus uberis in the dry period. Use of immunotherapeutic agents at drying off could increase cellular defences in the gland and prevent establishment of new S. uberis infections. This study investigated the potential of infusing recombinant bovine interleukin-1 beta (rbIL-1beta) in the mammary glands as a prophylactic agent against subsequent intramammary challenge with S. uberis in the early dry period. Immediately after the last milking at commencement of the dry period, one cow from each of 10 monozygous twinsets was infused with 10 microg of rbIL-1beta in two quarters and the other twin was infused with the carrier agent, sterile phosphate buffered saline. Twenty-four hours later, the quarters were infused with 10(3) colony-forming units (CFU) of S. uberis. Bacteriology, somatic cell count (SCC), concentrations of specific cytokines and antibody responses were monitored in mammary gland secretions and sera for the next 21 days. Infusion of rbIL-1beta into mammary glands at commencement of the dry period was associated with less new S. uberis intramammary infections, as determined by the number of quarters with bacterial growth. However, high SCC in quarters following infusion of rbIL-1beta masked the full beneficial effect of this procedure.     

The kinetics of inflammation and phagocytosis during bovine mastitis induced by Streptococcus agalactiae bearing the protein X

The protein X of Streptococcus agalactiae is a surface antigen borne by a high proportion of strains isolated from bovine mastitis. We have tested the capacity of two strains of X-bearing Streptococcus agalactiae to induce mastitis in dairy cows. The reference X-strain (411.07) produced an intramammary infection with local clinical signs in the three inoculated quarters. Another X-bearing strain (443.31) of bovine origin produced infection in all 11 quarters inoculated with only 25 or 85 colony-forming units. In naive cows, strain 433.31 induced less exudation of plasma into the milk, shedding of bacteria, macroscopic alteration, and a lower somatic cell count (SCC) than did the reference strain. Only one quarter spontaneously eliminated the infection before antibiotic treatment 9 days after inoculation.The serum of all the cows contained naturally acquired or induced antibodies to the challenge strain (443.31) and possessed opsonic activity. Before inflammation occurred, the milk was almost devoid of antibody or opsonic activities. The early phase of infection was characterized by rapid multiplication of streptococci in the milk, followed by a sharp drop in bacterial counts concomitant with the onset of inflammation.Three cows immunized with protein X displayed higher SCC and bactericidal activity in milk from the inoculated quarter at the onset of inflammation than non-immunized cows. Two of the three immunized cows underwent an early and transient febrile episode and eliminated the infection.     

Intramammary treatment of clinical mastitis of dairy cows with a combination of lincomycin and neomycin, or penicillin and dihydrostreptomycin

AIM: To compare clinical and bacteriological cure rates of clinical mastitis following treatment with intramammary preparations containing either lincomycin and neomycin or penicillin and dihydrostreptomycin. METHODS: Cases of clinical mastitis were sourced from four seasonal-calving dairy herds in the central Waikato region of New Zealand during the first 120 days of lactation. Affected quarters were infused three times at 12 h intervals with either 333 mg lincomycin plus 100 mg neomycin (lin/neo; 197 glands),or 1,000 mg penicillin plus 500 mg dihydrostreptomycin (pen/DHS; 207 glands). Milk samples were collected for bacteriology from each quarter immediately before and approximately 21 days after initiation of treatment. Additionally, a composite milk sample from each cow was collected, on average, 54 days after enrolment for assessment of milk yield, composition and somatic cell count (SCC). The probability of bacterial cure was initially analysed using Chi-squared analysis, and factors that were associated (p<0.2) were offered to a reverse stepwise logistic regression model. Continuous variables (e.g. milk solids production and log10 SCC) were analysed using general linear models. RESULTS: A total of 404 quarters diagnosed with clinical mastitis, from 282 cows in the first 120 days of lactation, were included. Streptococcus uberis, coagulase-negative staphylococci and Staphylococcus aureus were isolated from 56.5%, 18.8% and 10.0% of the bacteriologically positive quarters. There was no difference in the bacteriological cure rate (76.7% vs 76.7%, OR=0.94; p>0.8), the log10 SCC (2.1, SE 0.1, vs 2.0, SE 0.1; p>0.3) or milk production (1.2, SE 0.1, vs 1.2, SE 0.1, kg milksolids/cow/day; p>0.7) between lin/neo vs pen/DHS treatments, respectively. However, the proportion of cows re-treated following initial treatment was higher for the lin/neo compared to pen/DHS-treated group (16.3% vs 5.2%, OR=3.46; p<0.05). CONCLUSIONS: No difference in bacteriological cure rate, milk production or SCC was evident between lin/neo and pen/DHS intramammary treatments for clinical mastitis in dairy cows during the first 120 days of lactation. KEYWORDS: Dairy cow, mastitis, intramammary, antibiotic, treatment, somatic cell count.     

Efficacy of flunixin meglumine for the treatment of endotoxin-induced bovine mastitis

The clinical effect of flunixin meglumine administration was determined in cows with acute mastitis induced by intramammary administration of endotoxin. In 12 lactating cows, 10 micrograms of Escherichia coli 026:B6 endotoxin were administered via a teat cannula into the teat cistern of single randomly selected rear quarters. Cows were challenge exposed as pairs. One cow in each pair was administered parenteral flunixin meglumine (6 cows) and 1 cow per pair was administered saline solution (6 cows). Multiple doses (7) of 1.1 mg of flunixin meglumine/kg of body weight or saline solution were administered at 8-hour intervals beginning 2 hours after endotoxin. Cow and quarter clinical signs as well as milk somatic cell concentrations, bovine serum albumin, electrical conductivity, and milk production were determined before and for 14 days after endotoxin inoculation. Intramammary endotoxin produced signs characteristic of acute coliform mastitis. Quarter and systemic abnormalities occurred and milk production was reduced by approximately 50% at 12 hours after endotoxin. Flunixin meglumine therapy significantly (P less than or equal to 0.05) reduced rectal temperatures and quarter signs of inflammation and improved clinically graded depression when compared with these signs in saline solution-treated controls. Milk production and laboratory indicators of inflammation in milk were not significantly (P greater than 0.05) different for flunixin meglumine vs saline solution controls. The clinical response observed was consistent with the antipyretic, analgesic, and anti-inflammatory properties of flunixin meglumine.     

Bovine mycoplasma mastitis from intramammary inoculation of small numbers of Mycoplasma bovis. I. Microbiology and pathology

Intrammary inoculation of 70 colony forming units (cfu) of Mycoplasma bovis into one quarter of four previously non-infected cows resulted in severe mastitis in inoculated and uninoculated quarters. Hematogenous spread of the infection was most likely, as mycoplasma was isolated from the peripheral blood of three of four cows and the milking machine was designed to prevent quarter to quarter communication of milk and air. The presence of large numbers of mycoplasma exceeding 10⁶ cfu/ml of milk preceded the onset of overt sero-purulent mastitis by 1–3 days. In general, the severity, duration of the infection and within cow spread of mastitis to adjacent quarters after the inoculation of 70 cfu was indistinguishable from naturally occurring mycoplasma mastitis.The pathology of the chronically infected quarters consisted of alveolar involution and moderate to severe mononuclear infiltration and an increase in interalveolar and loose connective tissue. The quarters of one cow resolving the infections at the time of slaughter were not as severely affected and contained numerous milk-producing alveoli and many alveoli with hyperplasia of the alveolar cells.     

Experimental infection of lactating bovine mammary glands with Streptococcus uberis in quarters colonized by Corynebacterium bovis

Twenty-seven quarters of 18 lactating dairy cows were inoculated intramammarily with 3.6 X 10(4) colony-forming units (CFU) of a strain of Streptococcus uberis isolated from a cow with clinical mastitis. Before quarters were inoculated, 22 were considered as naturally colonized with Corynebacterium bovis, and 5 were considered bacteriologically negative. Streptococcus uberis was isolated from all quarters within 2 days after inoculation, and all quarters developed clinical mastitis by 3 days after inoculation. Mastitis was acute, and most cows had increased rectal temperatures. The number of somatic cells increased significantly (P less than 0.05), and milk production decreased significantly. In many cows, rectal temperatures remained increased, and Str uberis was isolated from infected glands after intramammary and systemic antimicrobial treatments were given. A decreased number (110 CFU) of the same strain of Str uberis caused equally severe mastitis in 3 quarters colonized with C bovis and in 1 bacteriologically negative quarter in 2 cows. Streptococcus uberis was isolated from all inoculated quarters, and all quarters developed clinical mastitis by 2 days after inoculation. Two quarters colonized with C bovis and 2 bacteriologically negative quarters were inoculated once with 25 CFU and once with 240 CFU of a different strain of Str uberis (ATCC 27958). Streptococcus uberis was never isolated from inoculated quarters, and changes in milk yield or number of somatic cells were not observed.     

Host response in bovine mastitis experimentally induced with Staphylococcus chromogenes

An experimental infection model was developed to study host response to intramammary infection in cows caused by Staphylococcus chromogenes. CNS intramammary infections have become very common in modern dairy herds, and they can remain persistent in the mammary gland. More information would be needed about the pathophysiology of CNS mastitis, and an experimental mastitis model is a means for this research. Six primiparous Holstein-Friesian cows were challenged with S. chromogenes 4 weeks after calving. One udder quarter of each cow was inoculated with 2.1 x 10(6)cfu of S. chromogenes. All cows became infected and clinical signs were mild. Milk production of the challenged quarter decreased on average by 16.3% during 7 days post-challenge. Cows eliminated bacteria in a few days, except for one cow which developed persistent mastitis. Milk indicators of inflammation, SCC and N-acetyl-beta-D-glucosaminidase (NAGase) returned to normal within a week. Milk NAGase activity increased moderately, which reflects minor tissue damage in the udder. Concentrations of serum amyloid A (SAA) and milk amyloid A (MAA) were both elevated at 12h PC. MAA was affected by the milking times, and was at its highest before the morning milking. In our experimental model, systemic acute phase protein response with SAA occurred as an on-off type reaction. In conclusion, this experimental model could be used to study host response in CNS mastitis caused by the main CNS species and also for comparison of the host response in a mild intramammary infection and in more severe mastitis models.