Bovine subclinical mastitis caused by different types of coagulase-negative staphylococci

Subclinical mastitis caused by intramammary infections (IMI) with coagulase-negative staphylococci (CNS) is common in dairy cows and may cause herd problems. Control of CNS mastitis is complicated by the fact that CNS contain a large number of different species. The aim of the study was to investigate the epidemiology of different CNS species in dairy herds with problems caused by subclinical CNS mastitis. In 11 herds, udder quarter samples were taken twice 1 mo apart, and CNS isolates were identified to the species level by biochemical methods. The ability of different CNS species to induce a persistent infection, and their associations with milk production, cow milk somatic cell count, lactation number, and month of lactation in cows with subclinical mastitis were studied. Persistent IMI were common in quarters infected with Staphylococcus chromogenes, Staphylococcus epidermidis, and Staphylococcus simulans. The results did not indicate differences between these CNS species in their association with daily milk production, cow milk somatic cell count, and month of lactation in cows with subclinical mastitis. In cows with subclinical mastitis, S. epidermidis IMI were mainly found in multiparous cows, whereas S. chromogenes IMI were mainly found in primiparous cows.     

A partial budget model to estimate economic benefits of lactational treatment of subclinical Staphylococcus aureus mastitis

Subclinical Staphylococcus aureus mastitis is rarely treated during lactation because it is widely believed to be uneconomical, although there are no economic studies that support this view. Partial budgeting was used to develop a deterministic simulation model to estimate the net cost or benefit of antibiotic treatment of subclinical S. aureus mastitis during lactation. Direct and indirect effects of treatment were taken into account, including prevention of clinical flare-ups and contagious transmission. Input variables were based on literature and on 2003-2004 prices in the Netherlands. When contagious transmission of S. aureus was likely (reproductive ratio R = 5.3), 3- and 8-d treatments resulted in an average net profit of €95.62 and €142.42, respectively, compared with no treatment. When the probability of S. aureus transmission was low (R = 0.32), the average economic benefit of 3- or 8-d treatment was − €21.12 and − €57.70, respectively. On low-transmission farms, 3-d treatment was profitable when the appropriate cows were selected for treatment using known risk factors for cure. Sensitivity analysis showed that the 6 most influential input variables in the model were chance of bacteriological cure, R, probability of culling, retention pay-off, and cost of antibiotics and bacterial culture. Although the economic outcome of lactational treatment of subclinical S. aureus mastitis is highly herd-, cow-, and strain-dependent, treatment is economically justified in many situations.     

Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine mastitis in Argentina

A total of 123 isolates of coagulase-negative staphylococci isolated from bovine clinical and subclinical mastitis in Argentina from March 1998 to March 2000 was investigated for in vitro susceptibility to several antimicrobial agents. Minimum inhibitory concentrations that inhibit 90% of the isolates tested (reported in micrograms per milliliter) were: 4.40, 0.38, 4.00, 0.75, 0.75, 3.60, and 2.00 for penicillin, oxacillin, cephalothin, gentamicin, erythromycin, clindamycin, and ampicillin-sulbactam, respectively. Resistance was detected in 34 (27.6%), 4 (3.2%), 7 (5.7%), and 6 (4.8%) isolates for penicillin, oxacillin, erythromycin, and pirlimycin, respectively. No resistance was detected for gentamicin, cephalothin, or ampicillin-sulbactam. Results indicated that coagulase-negative staphylococci isolates in Argentina exhibited the highest degree of resistance to penicillin of all antimicrobial agents tested.     

Production loss due to new subclinical mastitis in Dutch dairy cows estimated with a test-day model

Milk, fat, and protein loss due to a new subclinical mastitis case may be economically important, and the objective of this study was to estimate this loss. The loss was estimated based on test-day (TD) cow records collected over a 1-yr period from 400 randomly selected Dutch dairy herds. After exclusion of records from cows with clinical mastitis, the data set comprised 251,647 TD records from 43,462 lactations of 39,512 cows. The analysis was carried out using a random regression test-day modeling approach that predicts the cow production at each TD based on the actual production at all previous TD. The definition of new subclinical mastitis was based on the literature and assumed a new subclinical case if somatic cell count (SCC) was >100,000 cells/mL after a TD with SCC <50,000 cells/mL. A second data set was created by applying an adjustment to correct low SCC for the dilution effect when determining if the previous test-day SCC was <50,000 cells/mL. Thereafter, the loss was estimated for records with SCC >100,000 cells/mL. The production (milk, fat, or protein) losses were modeled as the difference between the actual and predicted production (milk, fat, or protein) at the TD of new subclinical mastitis, for 4,382 cow records, and 2,545 cow records after dilution correction. Primiparous cows were predicted to lose 0.31 (0.25-0.37) and 0.28 (0.20-0.35) kg of milk/d at an SCC of 200,000 cells/mL, for unadjusted and adjusted low SCC, respectively. For the same SCC increase, multiparous cows were predicted to lose 0.58 (0.54-0.62) and 0.50 (0.44-0.56) kg of milk/d, respectively. Moreover, it was found that the greater the SCC increase above 100,000 cells/mL, the greater the production losses. The estimated production losses were more precise than previously reported estimates.     

Effect of naturally occurring coagulase-negative staphylococci infections on new infections by mastitis pathogens in the bovine

Microbiological data from 1123 uninfected quarters and 216 quarters with preexisting coagulase-negative staphylococci infections were analyzed to determine the influence of infection status on subsequent new infection rate. Overall, prevalence of new infections in uninfected quarters was approximately two times that in quarters already harboring a coagulase-negative Staphylococcus infection. New infections by coagulase-negative staphylococci were greater in uninfected quarters than in quarters with preexisting coagulase-negative staphylococci infections. However, no differences were observed between uninfected and infected quarters in number of new infections by major pathogens (Staphylococcus aureus, streptococci, and coliforms). No differences were observed in uninfected or coagulase-negative Staphylococcus-infected quarters in infections with minor pathogens compared with major pathogens. The influence of individual coagulase-negative Staphylococcus species on new infections was also analyzed. However, numbers of existing infections by Staphylococcus species other than Staphylococcus chromogenes were limited. Therefore, the protective capacity of each coagulase-negative Staphylococcus species was difficult to assess. Overall, a significant restriction of bacterial invasion was observed in quarters with a preexisting infection. These results suggest that quarters harboring a coagulase-negative Staphylococcus infection suppress colonization of the mammary gland by mastitis-causing pathogens.     

A method for assessing loss in milk yield due to subclinical mastitis

A method for assessing the loss in quarter milk yield due to subclinical mastitis is proposed. It is based upon two indices calculated from quarter milk yields and results from the assessment of mastitis obtained by measurement of the electrical conductivity of milk. The results were obtained from the cows through repeated quarter-milkings during several months of lactation. The grade of abnormality of a quarter with subclinical mastitis was expressed by the first index, the mean quarter difference of electrical conductivity of milk produced by the quarter (mQdEC, 25 degrees C). The reduction in milk yield of a quarter presumed to be due to subclinical mastitis was expressed by another index, the quarter yield ratio, R, which is that percentage of normal milk yield produced by a quarter with mastitis. Examination of the relationship between these two indices showed that subclinical mastitis caused a reduction in milk yield in the affected quarters. From the linear regression, it was found that approximately 3% reduction in quarter milk yield occurred for every 1 X 10(-4)S/cm increase in the mQdEC of the quarter with presumed subclinical mastitis.     

Antimicrobial resistance and genetic characterization of coagulase-negative staphylococci from bovine mastitis milk samples in Korea

Coagulase-negative staphylococci (CNS) are one of the most common bovine mastitis pathogens found worldwide. In this study, we investigated the prevalence and distribution of CNS species in mastitis milk samples and further characterized the methicillin-resistant (MR) CNS. A total of 311 CNS were isolated from 3,692 quarter milk samples from 1,373 dairy cattle at 81 farms between 2013 and 2017. Further evaluation of the CNS isolates revealed 14 CNS species among the samples and 3 predominant species—namely, Staphylococcus chromogenes, Staphylococcus simulans, and Staphylococcus epidermidis. Resistance was higher in S. epidermidis than in other CNS species except for resistance against oxacillin in Staphylococcus sciuri. Resistance to β-lactams was the most common in all CNS species (8.4% in ampicillin, 21.2% in oxacillin, and 13.5% in penicillin). Conversely, only minimal resistance to cephalothin, ceftiofur, and pirlimycin/novobiocin was found. Twenty-one isolates from 4 species were mecA-carrying MRCNS strains, including 18 S. epidermidis and 1 each of S. sciuri, Staphylococcus equorum, and Staphylococcus hominis. The majority of the mecA-carrying MRCNS isolates were produced in the biofilm. Furthermore, multidrug-resistant sequence type 179 isolate produced the strongest biofilm. Seven genotypes were detected in the 18 MR S. epidermidis strains, the most predominant of which persisted on a farm for 2 yr. Our findings for the antimicrobial susceptibility profiles and genotypic characterization of the MRCNS isolates could provide valuable information for controlling the spread of resistance and the selection of appropriate antimicrobial therapies for mastitis in the future. Further, strategic antibiotic use for mastitis treatment and hygienic management practices aimed at the prevention of the growth of resistant bacteria are urgently needed on dairy farms.     

Milk and serum IL-4, IL-6, IL-10, and amyloid A concentrations in cows with subclinical mastitis caused by coagulase-negative staphylococci

The aim of the study was to evaluate the concentrations of cytokines IL-4, IL-6, and IL-10 and acute phase protein amyloid A in milk and in serum from cows with subclinical mastitis caused by coagulase-negative staphylococci and from healthy cows. The blood and milk samples were obtained from 35 midlactation, multiparous (between parities 2 and 4) Holstein-Friesian cows. In the milk samples from 20 cows with subclinical mastitis, the following species of Staphylococcus were detected: Staphylococcus xylosus (8 samples), Staphylococcus chromogenes (6 samples), Staphylococcus haemolyticus (2 samples), Staphylococcus simulans (2 samples), and Staphylococcus sciuri (2 samples). The results of the present study indicate that the level of IL-6 in cows suffering from subclinical mastitis tended to be high in both serum and milk (432.09 and 254.32 pg/mL) compared with the level in healthy cows (164.47 and 13.02 pg/mL, respectively). Amyloid A value also was significantly higher in milk of unhealthy cows compared with cows without subclinical mastitis (790.2 and 360.5 ng/mL). No significant differences were found in levels of amyloid A in serum of both tested groups of cows (2,680.0 and 2,720.0 ng/mL). In contrast, concentration of IL-4 was significantly lower both in serum and in milk of cows with staphylococcal mastitis (86.1 and 123.17 pg/mL) compared with control animals (413.5 and 670.2 pg/mL). The level of IL-10 also was significantly higher in milk of healthy cows than in infected cows (39.78 and 22.5 pg/mL); however, differences in serum levels of this cytokine between tested groups were significantly less important (220.6 and 175.1 pg/mL).     

Susceptibility of coagulase-negative staphylococci to a kanamycin and cefalexin combination

A combination of kanamycin and cefalexin was licensed in Europe in 2008 to treat bovine clinical mastitis. Preliminary broth and disk clinical breakpoints for this antibiotic combination have been proposed for Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus uberis, and Escherichia coli. This study indicates that these proposed breakpoints also hold for coagulase-negative staphylococci (CNS), a group of bacteria frequently isolated in milk samples from cows with clinical mastitis. The data show that clinical bovine mastitis isolates of CNS from Europe have a high degree of susceptibility to the kanamycin/cefalexin combination, with minimal resistance to either agent alone. The use of the available kanamycin and cefalexin combination disk for testing the susceptibility of bovine mastitis isolates of Staph. aureus, Strep. uberis, Strep. dysgalactiae, and E. coli is also reliable for use in the testing of CNS, as disk results correlated with broth minimum inhibitory concentrations. The study reports, for the first time, the approved Clinical Laboratory Standards Institute quality control ranges for the kanamycin/cefalexin combination and wild-type cutoff values for major bacterial pathogens implicated in bovine mastitis.     

Systemic treatment of subclinical mastitis in lactating cows with penethamate hydriodide

A randomized controlled field trial was performed to evaluate the efficacy of a 3-d treatment regimen with i.m. penethamate hydriodide compared with no treatment in lactating cows with subclinical mastitis. To be included, a cow had to have 2 somatic cell counts (SCC) 300,000 cells/mL at the last 3 monthly controls, 1 or more quarters with SCC >250,000 cells/mL, and the same bacterial species isolated in 2 consecutive samples 2 to 4 d apart. A total of 151 quarters from 92 cows were monitored for 2 mo following treatment. Quarter milk samples were examined for bacteriological cure (BC) and SCC at 14, 28, and 60 d after treatment. Bacteriological cure was defined as not having the same bacterial species isolated from the quarter milk samples taken at 14 and 28 d posttreatment as in the samples taken before treatment. Systemic treatment with penethamate resulted in BC in 59.5% of quarters and 52.2% of cows, compared with 16.7 and 10.9% in the untreated cows. Somatic cell count decreased significantly in the penethamate-treated cows, steadily in the case of BC and transiently when the infections persisted. This study confirms that systemic treatment of subclinical mastitis with penethamate is effective and that BC of infected quarters has a sustained positive effect on milk SCC during the 2 mo following treatment.     

Intramammary antimicrobial treatment of subclinical mastitis and cow performance later in lactation

The aim of this study was to evaluate long-term therapeutic effects of antimicrobial treatment of recently acquired subclinical mastitis (RASCM) during lactation. Quarter-level clinical mastitis (CM) follow-up, composite somatic cell counts (SCC), and cow-level milk yield later in lactation were evaluated using follow-up data from 2 previously published linked randomized field trials. The first trial randomly assigned antimicrobial treatment with any intramammary product or negative control to culture-positive quarters of cows having a first elevated composite SCC after 2 consecutive low composite SCC measurements. Untreated cows that had a second elevated composite SCC at the next measurement and were staphylococci-positive (i.e., Staphylococcus aureus or non-aureus staphylococci) were randomly assigned to treatment or control. Quarter-level CM cases were reported by the participating herd personnel, and milk yield and composite SCC data were obtained from the regular test-day recording. Frailty survival models were used to evaluate the long-term therapeutic effects of antimicrobial treatment of RASCM on quarter-level CM follow-up. Mixed linear regression models were applied to quantify the effect on milk yield and composite SCC. Data of 638 quarters from 486 cows in 38 herds were available for statistical analyses, of which 229 quarters of 175 cows received antimicrobial treatment for RASCM. Antimicrobial treatment culminated in reduced composite SCC levels later in lactation but did not result in different milk yield levels or CM follow-up compared with control cows. Antimicrobial treatment of cows with RASCM should therefore only be considered in exceptional situations given the current focus on antimicrobial usage reduction in animal husbandry.     

Diagnosis and treatment of subclinical mastitis in early lactation in dairy goats

The objectives of the study were to define the sensitivity and specificity of the California Mastitis Test (CMT) in determining the presence of intramammary infection in postpartum dairy goats and to determine whether antibiotic therapy increased bacteriological cure rate and lowered somatic cell count (SCC) compared with untreated controls. A CMT was performed and milk samples were collected for bacteriology from 211 glands of 106 does between 0 and 10 d after kidding. From a population of 3,239 glands from goats in 4 commercial herds, goats with one or both glands with a CMT score of >1 and from which bacteria were isolated were either assigned to be treated with 3 intramammary infusions at 12-h intervals of 75 mg of sodium ampicillin and 250 mg of sodium cloxacillin (n = 57 glands) or left as untreated controls (n = 49 glands). Milk samples were collected again 14 ± 3 and 21 ± 3 d later for bacteriology and SCC determination. Composite milk yield, goat SCC, length of lactation, and survival data were collected. A partial budget was constructed to assess the cost effectiveness of treatment. At a cut point of greater than trace, the sensitivity, specificity, and positive and negative predictive values of the CMT were 0.74, 0.74, 0.42, and 0.92, respectively. Treatment increased the bacteriological cure rate compared with no treatment [30/57 (53%) vs. 6/49 (12%)], but there was a pathogen by treatment interaction whereby treatment increased cure proportion in glands infected with minor, but not major, pathogens. Treatment reduced the foremilk gland-level SCC [1,595 (95% CI = 1,106-2,300) vs. 3,028 (95% CI = 2,091-4,385) geometric mean (× 1,000) cells/mL] but not the SCC at goat level [1,596 (95% CI = 1,219-2,090) vs. 1,488 (95% CI = 1,132-1,955) geometric mean (× 1,000) cells/mL] compared with no treatment. Milk yield, risk of removal from the herd, and length of lactation were not altered by treatment. Treatment resulted in a loss of NZ$20.39/doe. It was concluded that use of the CMT as a screening test resulted in a higher likelihood of finding a gland that would be infected than selecting a gland at random. Treatment increased bacteriological cure rate and reduced SCC at gland level compared with no treatment. However, at goat level, milk yield, SCC, and survival were not altered, resulting in no economic benefit of treatment.     

Stochastic modelling to assess economic effects of treatment of chronic subclinical mastitis caused by Streptococcus uberis

Chronic subclinical mastitis is usually not treated during the lactation. However, some veterinarians regard treatment of some types of subclinical mastitis to be effective. The goal of this research was to develop a stochastic Monte Carlo simulation model to support decisions around treatment of chronic subclinical mastitis caused by Streptococcus uberis. Factors in the model included the probability of cure after treatment, probability of the cow becoming clinically diseased, transmission of infection to other cows, and physiological effects of the infection. Using basic input parameters for Dutch circumstances, the average economic costs per cow of an untreated chronic subclinical mastitis case caused by Str. uberis in a single quarter from day of diagnosis onwards was euro109. With treatment, the average costs were higher (euro120). Thus, for the average cow, treatment was not efficient economically. However, the risk of high costs was much higher when cows with chronic subclinical mastitis were not treated. A sensitivity analysis showed that profitability of treatment of chronic subclinical Str. uberis mastitis depended on farm-specific factors (such as economic value of discarded milk) and cow-specific factors (such as day of diagnosis, duration of infection, amount of transmission to other cows and cure rate). Therefore, herd level protocols are not sufficient and decision support should be cow specific. Given the importance of cow-specific factors, information from the current model could be applied to automatic decision support systems.     

Bioeconomic modeling of lactational antimicrobial treatment of new bovine subclinical intramammary infections caused by contagious pathogens

This study determined the direct and indirect epidemiologic and economic effects of lactational treatment of new bovine subclinical intramammary infections (IMI) caused by contagious pathogens using an existing bioeconomic model. The dynamic and stochastic model simulated the dynamics of Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, and Escherichia coli during lactation and the dry period in a 100-cow dairy herd during 1 quota year. Input parameters on cure were obtained from recent Dutch field data. The costs of clinical IMI, subclinical IMI, and intervention were calculated into the combined total annual net costs of IMI per herd. The cost effectiveness of 4 scenarios with lactational intervention was determined; scenarios included no intervention, treatment after 1 mo of infection, treatment after 2 mo of infection, and treatment after 1 mo of infection and culling of uncured cows after 2 mo of infection. Model behavior was observed for variation in parameter input values. Compared with no lactational intervention, lactational intervention of new subclinical IMI resulted in fewer clinical flare ups, less transmission within the herd, and much lower combined total annual net costs of IMI in dairy herds. Antimicrobial treatment of IMI after 1 mo of infection and culling of uncured cows after 2 mo of infection resulted in the lowest costs, whereas treatment after 2 mo of infection was associated with the highest costs between the scenarios with intervention. Changing the probability of cure resulted in a nonlinear change in the cumulative incidence of IMI cases and associated costs. Lactational treatment was able to prevent IMI epidemics in dairy herds at high transmission rates of Strep. uberis, Strep. dysgalactiae, and E. coli. Lactational treatment did not limit the spread of Staph. aureus at high transmission rates, although the associated costs were lower compared with no intervention. To improve udder health in a dairy herd, lactational treatment of contagious subclinical IMI must therefore be preceded by management measures that lower the transmission rate. Lactational treatment of environmental subclinical IMI seemed less cost effective. Detection of subclinical IMI needs improvement to be able to most effectively treat subclinical IMI caused by contagious pathogens during lactation.     

Binding to extracellular matrix proteins and formation of biogenic amines by food-associated coagulase-negative staphylococci

In connection with a study on the DNA microarray based detection of genes involved in safety and technologically relevant properties (Seitter (née Resch) et al., 2011), food-associated coagulase-negative staphylococci (CNS) were investigated phenotypically with regard to their ability to bind to the extracellular matrix proteins (ECM) and to produce biogenic amines. The properties have been shown to be involved in the colonization of injured tissue and invasion into host cells as well as in pharmacologic effects on humans, respectively. The CNS exhibited a low, but nevertheless clearly measurable ECM binding capacity, except for strains of Staphylococcus equorum and Staphylococcus succinus, which show a comparable or even higher binding to fibrinogen and fibronectin than that of the control strain Staphylococcus aureus Cowan. Formation of biogenic amines could be often detected in S. carnosus, S. condimenti and S. strains, but rarely in S. equorum and not in S. succinus and S. xylosus strains. Mostly, 2-phenylethylamine, tyramine and tryptamine were formed by resting cells in amounts < 25 mg/l, whereas growing cells formed high amounts (> 100 mg/l) of 2-phenylethylamine and putrescine. This study confirmed the need of consideration of ECM binding and biogenic amine formation in the safety assessment of CNS used in the production of fermented foods.     

Negatively controlled,randomized clinical trial to evaluate intramammary treatment of nonsevere,gram-negative clinical mastitis

The objective of this negatively controlled, randomized clinical trial was to examine clinical outcomes of 2-d or 8-d treatment using an approved intramammary (IMM) product containing ceftiofur hydrochloride compared with no antimicrobial treatment of nonsevere, gram-negative cases of clinical mastitis (CM). Additionally, we contrasted clinical outcomes of cases caused by Escherichia coli (n = 56) or Klebsiella pneumoniae (n = 54). Cases (n = 168) of nonsevere (abnormal milk or abnormal milk and udder) CM were randomly assigned to receive 2 d (n = 56) or 8 d (n = 56) of IMM ceftiofur or assigned to a negative control group (n = 56). At enrollment, quarter milk samples were collected and used for on-farm culture, somatic cell count (SCC), and confirmatory microbiological analysis. Quarter milk samples were collected weekly from 7 to 28 d after enrollment for microbiological and SCC analysis. Clinical outcomes were followed for 90 d or until the end of lactation (follow-up period, FUP). Overall, no significant differences in quarter-level recurrence of CM (32% for negative control, 34% for the 2-d treatment, and 32% for the 8-d treatment), culling (18% for negative control, 12% for 2-d treatment, and 11% for 8-d treatment), voluntary dry-off of affected quarters (20% for negative control, 30% for 2-d treatment, and 27% for 8-d treatment), days until return to normal milk (4.2 days for negative control, 4.8 days for 2-d treatment, 4.5 days for 8-d treatment), weekly quarter-SCC during the FUP (6.1, 6.3, and 6.0 log₁₀SCC for the negative control, 2-d, and 8-d treatments, respectively), or daily milk yield during the FUP (37.1, 36.3, and 37.6 kg/cow per day for the negative control, 2-d, and 8-d treatments, respectively) were observed among experimental groups. Days of discarded milk were greater for cows assigned to 8-d IMM ceftiofur (11.1 d) than for cows assigned to 2-d (6.9 d) or cows assigned to negative control (5.6 d). Bacteriological cure (BC) at 14 and 21 d after enrollment was greater in cows assigned to 8-d (89%) and 2-d (84%) treatment than in cows assigned to negative control (67%), but this outcome was confounded by pathogen. For CM caused by Kleb. pneumoniae, BC was greater for quarters assigned to receive treatment (combined 2-d and 8-d groups; 74% BC) than for quarters assigned to negative control (18%). In contrast, no differences in BC were observed for CM caused by E. coli (97–98%). Culling and voluntary dry-off of affected quarters were significantly greater for cows with quarters affected by Kleb. pneumoniae (22% culled, 39% voluntary dry-off of quarters) than for cows with quarters affected with E. coli (7% culled, 11% voluntary dry-off of quarters). Overall, use of IMM ceftiofur did not result in improvement of most clinical outcomes, but differences between E. coli and Kleb. pneumoniae were evident. In contrast to E. coli, Kleb. pneumoniae caused chronic intramammary infection and induced worse clinical outcomes. Intramammary antibiotic treatment of most mild and moderate cases of CM caused by E. coli is not necessary, but more research is needed to identify which quarters affected by Kleb. pneumoniae may benefit from antimicrobial therapy.     

Acute phase proteins in bovine milk in an experimental model of Staphylococcus aureus subclinical mastitis

The objectives were to establish the origin of 2 acute phase proteins in milk during subclinical bovine mastitis and to characterize the relationship between those proteins in milk and blood. Haptoglobin (Hp) and mammary-associated serum amyloid A (M-SAA3) appear in milk during mastitis, whereas Hp and serum amyloid A increase in serum during mastitis. The concentrations of these proteins were determined in an experimental model using a field strain of Staphylococcus aureus to induce subclinical mastitis in dairy cows. The expression of mRNA coding for these proteins was assessed and the presence of M-SAA3 in mammary tissues was determined using immunocytochemistry. Increases of M-SAA3 and Hp in milk occurred within 12 h of Staphylococcus aureus infusion, with peak concentrations occurring 3 d after infusion of the bacteria. The increase of acute phase proteins in milk (15 h) preceded the increase in serum concentrations of both proteins (24 h). Expression of mRNA for M-SAA3 and Hp increased in both mammary and hepatic tissues 48 h after infusion of the mammary glands. In mammary tissue, the increase of M-SAA3 mRNA was greater than the increase in Hp mRNA expression, whereas in hepatic tissue, the increase in M-SAA3 mRNA was less than that for Hp mRNA. Immunocytochemistry demonstrated that M-SAA3 protein was present within secretory epithelial cells at significantly higher levels in infected mammary glands than in control tissues. These proteins, which have host defense and antibacterial activities, may play a significant role in the early response to invasion of mammary tissues by pathogenic bacteria.     

Heifers infected with coagulase-negative staphylococci in early lactation have fewer cases of clinical mastitis and higher milk production in their first lactation than noninfected heifers

Intramammary infections (IMI) in recently calved dairy heifers are more common than was formerly believed but their relevance for future performance has been studied only rarely. In the present study, the association between the IMI status of fresh heifers and their subsequent udder health, milk production, and culling in first lactation was explored. Quarter milk samples were collected between 1 and 4 d in milk (DIM) and between 5 and 8 DIM from 191 dairy heifers in 20 dairy herds for bacteriological culturing and somatic cell count (SCC) analysis. Monthly milk recording data including composite milk SCC and test-day milk yield (MY) were obtained for the first 285 DIM or until culling. Farmer-recorded clinical mastitis cases were available. Data were analyzed using mixed models and survival analysis. Approximately 80% of the fresh heifers (79.8%) had at least one culture-positive quarter. Coagulase-negative staphylococci (CNS) were the most frequently isolated pathogens (72%), followed by esculin-positive streptococci (4.6%) and Staphylococcus aureus (3.5%). Overall geometric mean SCC at quarter level decreased between the first and second samplings from 348,000 to 116,000 cells/mL. Heifers infected with CNS had an intermediate average test-day SCC (84,000 cells/mL) during the first 285 DIM compared with noninfected heifers (53,000 cells/mL) and heifers infected with major pathogens (195,000 cells/mL). Heifers infected with major pathogens had a much lower average daily MY (18.3 kg) during first lactation compared with noninfected animals (21.3 kg). That CNS-infected heifers out-produced their noninfected counterparts could be at least partially explained by their significantly lower incidence of clinical mastitis (incidence risk 3.6 vs. 21.0%) during first lactation compared with noninfected heifers. We conclude that although CNS cause the majority of IMI in heifers around calving, they should not be a reason for serious concern.     

Efficacy of extended ceftiofur intramammary therapy for treatment of subclinical mastitis in lactating dairy cows

Little research has focused on treatment of cows with subclinical mastitis during lactation. Ceftiofur is a new broad-spectrum, third-generation cephalosporin antibiotic for veterinary use that inhibits bacterial cell wall synthesis by interfering with enzymes essential for peptidoglycan synthesis. Ceftiofur should be effective against a wide range of contagious and environmental mastitis pathogens. Objectives of the present study were to evaluate the efficacy of ceftiofur for treatment of subclinical mastitis in lactating dairy cows, and to determine if extended therapy regimens enhanced efficacy of ceftiofur. Holstein and Jersey dairy cows (n = 88) from 3 dairy research herds were used. Cows were enrolled in the study based on milk somatic cell counts >400,000/mL and isolation of the same mastitis pathogen in 2 samples obtained 1 wk apart. Cows with one or more intramammary infections (IMI) were blocked by parity and DIM and allocated randomly to 1 of 3 different ceftiofur treatment regimens: 2-d (n = 49 IMI), 5-d (n = 41 IMI), and 8-d (n = 38 IMI) treatment regimens. For all groups, 125 mg of ceftiofur hydrochloride was administered via intramammary infusion. Eighteen cows with 38 IMI were included as an untreated negative control group. A bacteriological cure was defined as a treated infected mammary quarter that was bacteriologically negative for the presence of previously identified bacteria at 14 and 28 d after the last treatment. Efficacy of ceftiofur therapy against all subclinical IMI was 38.8, 53.7, and 65.8% for the 2-, 5-, and 8-d ceftiofur treatment regimens, respectively. Four of 38 (10.5%) IMI in control cows were cured spontaneously without treatment. All 3 ceftiofur treatment regimens were significantly better than the negative control, and the 8-d extended ceftiofur treatment regimen treatment group was significantly better than the standard 2-d treatment group. Pathogen groups had significantly different cure rates from one another. The cure rate for the 8-d extended ceftiofur treatment regimen was 70% for Corynebacterium bovis, 86% for coagulase-negative Staphylococcus species, 36% for Staph. aureus, 80% for Streptococcus dysgalactiae ssp. dysgalactiae, and 67% for Strep. uberis.