Influence of Staphylococcus aureus strain-type on mammary quarter milk somatic cell count and N-acetyl-beta-D-glucosaminidase activity in cattle from eight dairies

The hypothesis tested was that there are differences in pathogenicity between strains of Staphylococcus aureus that cause bovine mastitis. Mammary quarter milk somatic cell count (SCC) and N-acetyl-beta-D-glucosaminidase (NAGase) activity were used as indicators of the pathogenicity of different strains of S. aureus that infect the bovine udder. Eight commercial dairy herds with a history of S. aureus in bulk tank milk cultures were studied. Initially, composite foremilk samples were collected from all lactating cattle in each herd and cultured for staphylococci. Subsequently, all cows with a coagulase-positive staphylococcal intramammary infection (IMI) at the initial sampling that were still present in the herd of origin had individual mammary quarter foremilk samples collected. Coagulase-positive staphylococcal isolates were confirmed as S. aureus using a commercial biotyping system. Staphylococcus aureus isolates were strain-typed using pulsed-field gel electrophoresis. Mammary quarter milk SCC and N-acetyl-beta-D-glucosaminidase activity were determined for each cow. The difference in mean somatic cell count and mean NAGase activity for mammary quarters infected with the same strain of S. aureus and for uninfected quarters on the same cow was calculated. One-way analysis of variance was used to assess differences between strains within a herd. Overall, no significant differences were found between strains, suggesting that the degree of udder parenchymal injury induced by S. aureus IMI is in general significantly affected by factors other than strain type.     

Comparison of probiotic and antibiotic intramammary therapy of cattle with elevated somatic cell counts.

The effects of treating subclinical mastitis with intramammary infusions of either a Lactobacillus or an antibiotic preparation on intramammary infection cure rate and on milk SCC were compared. Cows with two consecutive monthly DHIA composite SCC greater than 300,000 cells/ml (5.4771 log10/ml) were defined as high SCC cows. Twenty-six subclinical cows were randomly assigned to one of two treatments. Quarter foremilk samples were obtained from all quarters at d 0, 7, and 14 following infusion to determine the microbiological status and SCC. Composite milk SCC were determined monthly by DHIA and at d 0, 7, and 14 of the study. Coagulase-negative staphylococci were the predominantly isolated pathogens. Treatment of cows with Lactobacillus cured 21.7% of infected quarters, whereas 73.7% of infections treated with antibiotic were eliminated. Treatment of quarters with antibiotic did not reduce quarter SCC unless infected quarters were cured. Intramammary infusion of quarters with Lactobacillus increased quarter SCC, mainly because of an increase in SCC of initially uninfected, low SCC quarters. Monthly composite SCC were similar between treatments. The results indicate that administering Lactobacillus or antibiotic treatment to all quarters based on elevated composite SCC should not be adopted. Lactobacillus treatment increased SCC with no effect on infection rate.     

Experimental Staphylococcus aureus intramammary challenge in late lactation dairy cows: quarter and cow effects determining the probability of infection

The purpose of this study was to identify factors at the quarter and cow level that determine whether a quarter remains infected after an intramammary challenge with Staphylococcus aureus Newbould 305. A total of 135 cows were studied. Information on animal characteristics, cow-conformation, cow somatic cell count (SCC), and bacteriology, blood vitamin E levels, serology for retro-viral infections, bovine leukocyte adhesion deficiency-carrier status, and the presence of bovine lymphocyte antigens class I alleles was collected on each animal. All quarters of all cows were then challenged with Staphylococcus aureus Newbould 305. The challenge with S. aureus Newbould 305 resulted in 28 cows (20.7%) that did not establish infection in any of the quarters, 21 (15.6%) cows had 1 quarter infected, 35 (25.9%) had 2 quarters infected, 24 (17.8%) had 3 quarters infected, and 27 (20.0%) had all quarters infected. A higher prechallenge SCC decreased the risk of infection. An infection with Corynebacterium bovis prior to challenge decreased the risk of S.aureus infection. Of the bovine lymphocyte antigen alleles, the presence of the W20A allele proved to be significantly associated with a decreased risk of infection. No other factors proved to be significant.     

Intramammary challenge with Escherichia coli following immunization with a curli-producing Escherichia coli

Holstein and Jersey cattle were immunized with a curli-producing strain of Escherichia coli (pCRL65/A012) or a noncurli-producing strain (pUC18/HB101) to determine differences in resistance to establishment of experimental intramammary infection. Cows (n = 6 per group) were immunized at 14 d prior to drying off, 7 d of involution, and at calving with 3 x 10(10) E. coli in Freund's Incomplete Adjuvant. At 30 d of lactation, one mammary quarter of each cow was infused with a wild strain of E. coli (727). Escherichia coli 727 was isolated from a naturally occurring intramammary infection and produced curli. All challenged quarters became infected, and all cows developed acute clinical mastitis. Geometric mean duration of intramammary infections was 6 d for both immunization groups. All infections were spontaneously eliminated within 10 d. No differences occurred between immunization groups in blood selenium and glutathione peroxidase activity, plasma selenium, number of E. coli 727 isolated from secretion after challenge, rectal temperature and SCC response, clinical status of mammary quarters, or DMI. Reduction in milk production after challenge was greater for cows immunized with E. coli pCRL65/A012. Immunization of dairy cattle with a curli-producing strain of E. coli did not protect against experimental intramammary challenge during lactation.     

Determination of milk and blood concentrations of lipopolysaccharide-binding protein in cows with naturally acquired subclinical and clinical mastitis

Blood and milk concentrations of the acute phase protein lipopolysaccharide-binding protein (LBP) were evaluated in cows with naturally occurring mastitis. Blood and milk samples were collected from 101 clinically healthy dairy cows and 17 dairy cows diagnosed with clinical mastitis, and the LBP concentrations of the samples were measured by an ELISA. Concentrations of LBP were greater in the blood and milk of cows with clinical mastitis than in those with healthy quarters. Concentrations of LBP also differed between uninfected and subclinically infected quarters with low somatic cell count. Blood concentrations of LBP in cows with subclinical intramammary infections could not be differentiated from those of cows with all healthy quarters. Together, these data demonstrate that increased blood and milk concentrations of LBP can be detected in dairy cows with naturally acquired intramammary infections that cause clinical mastitis.     

A study of the incidence and significance of intramammary enterobacterial infections acquired during the dry period

We assessed the incidence of enterobacterial infection of the mammary glands of 629 cows, from six commercial herds in Somerset, during the nonlactating period; samples were collected from all clinical quarters of these cows during the subsequent lactation. A rise in the incidence of intramammary enterobacterial infection was detected between drying off and before calving. Quarters infected with an enterobacterial organism during the dry period were more likely to develop mastitis due to that pathogen than were uninfected quarters. Of all enterobacterial mastitis occurring in the first 100 d of lactation, 52.6% arose in quarters previously infected, during the dry period, with the same strain of bacteria, as identified by DNA fingerprinting using enterobacterial repetitive intergenic consensus primers. When compared with unsampled controls, quarters sampled during the dry period did not show a higher incidence of infection at calving or of subsequent clinical mastitis. These findings suggest that chronic infections are important in the epidemiology of enterobacterial mastitis and that environmental management during the dry period may greatly impact the incidence of enterobacterial mastitis in the subsequent lactation.     

Experimentally induced intramammary infection with multiple strains of Streptococcus uberis

The effect of infusing a mixture of 5 Streptococcus uberis strains into mammary quarters of 10 lactating cows was investigated. All 5 strains, which included 2 originally isolated from the dairy environment and 3 from clinical cases of mastitis, were capable of establishing an intramammary infection when infused individually. However, when the 5 strains were infused together, a single strain predominated in 7 out of 10 quarters. One strain in particular prevailed in 4 mammary quarters and was also found to inhibit the growth of the other 4 strains with deferred antagonism on esculin blood agar. The genes required for the production of bacteriocins nisin U and uberolysin were identified in this strain, whereas the other 4 strains contained only uberolysin genes. Direct competition may have occurred between strains within the mammary gland but competition was not apparent when cultured together in UHT milk, where no strain predominated. Although the mechanism is unknown, these results imply that a selection process can occur within the mammary gland, leading to a single strain that is detected upon diagnosis of mastitis.     

Bovine intramammary Escherichia coli challenge infections in late gestation demonstrate a dominant antiinflammatory immunological response

Coliform mastitis that presents itself at parturition or in the early weeks of bovine lactation is often characterized by severe inflammation and impaired milk production and can lead to death of the animal. Chronic intramammary infections caused by persistent strains of Escherichia coli may result in high production losses. The aim of this study was to determine the inflammatory response to a teat-canal challenge of bovine mammary glands with a persistent strain of E. coli during late gestation (dry period) and into early lactation. Two weeks before parturition, animals were challenged in 2 quarters with 30 cfu of a persistent strain of E. coli; control quarters were vehicle-infused and not infused, respectively. Samples of dry cow secretions were taken from all quarters before challenge and at 6, 12, 18, 24, 48, 72, 96, and 120 h following challenge. Colostrum samples and milk samples were taken from all quarters at parturition and 6, 12, 18, 24, 48, 72, 96 and 120 h postpartum. Bacterial culture, combined with random amplified polymorphic DNA genetic strain-typing analysis, indicated recovery of the bacterial challenge strain until 48 to 96 h postchallenge, and again at parturition and up to 6 and 12h postpartum. One animal exhibited clinical mastitis and the bacterial challenge strain was evident to at least 12 d postpartum. During twice-daily milkings, production levels were lower in bacteria-challenged quarters compared with controls. Somatic cell counts decreased to normal levels at a slower rate in challenged quarters compared with control quarters. Cytokine analysis indicated a minimal proinflammatory cytokine response, including interleukin-1β and tumor necrosis factor-α in challenged-quarter dry cow samples up to 120 h postchallenge. Interleukin-10 levels were significantly increased by 12h postchallenge in secretions from challenged and control quarters. These preliminary results in 2 cows indicate that proinflammatory signaling after intramammary bacterial infection may be actively suppressed during late gestation. We hypothesize that this immune-inhibitory response allows intramammary infections to become persistent in the dry period and cause clinical signs immediately after parturition.     

Study of the efficacy of a Streptococcus uberis mastitis vaccine against an experimental intramammary infection with a heterologous strain in dairy cows

Streptococcus uberis is a worldwide pathogen that causes intramammary infections in dairy cattle. Nevertheless, commercial vaccines are currently not available and measures to control S. uberis mastitis are limited to the implementation of good management practices. The aim of the present study was to evaluate the efficacy of an S. uberis subunit vaccine against bovine mastitis (Laboratorios Hipra S.A., Amer, Spain) administered precalving against an experimental intramammary challenge with a heterologous S. uberis strain in dairy cows postcalving. With this objective, 25 gestating Holstein-Friesian heifers were randomly assigned to 1 of 2 groups: group 1 (n = 13), vaccinated by intramuscular route with the vaccine, and group 2 (n = 12), vaccinated by intramuscular route with phosphate-buffered saline as a control group. Both groups were immunized 60 and 21 d before the expected parturition date (75 and 36 d before challenge). Fourteen days after calving all cows were challenged by intramammary infusion of 100 colony-forming units of a heterologous S. uberis strain in 2 quarters per cow. Then, challenged quarters were monitored for clinical signs of mastitis, bacterial count, and somatic cell count for the following 21 d. Rectal temperature and daily milk yield per cow were also assessed. Results showed that all challenged quarters developed clinical mastitis. Nevertheless, vaccination significantly reduced the clinical signs of mastitis, bacterial count, rectal temperature, and daily milk yield losses after the intramammary infection and significantly increased the number of quarters with no bacterial isolation and somatic cell count <200,000 cells/mL at the end of the study (d 19, 20, and 21 after challenge). To confirm the efficacy of this vaccine, further studies under field conditions are needed.     

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.     

Evaluation of protein A and a commercial bacterin as vaccines against Staphylococcus aureus mastitis by experimental challenge

Protein A and a commercial staphylococcal bacterin were evaluated by experimental challenge with Staphylococcus aureus (ATCC 29740). Thirty cows in first lactation were in three treatment groups, protein A, bacterin, and nonvaccinated controls. Studies were through three lactations and included bacteriological and cytological analyses of quarter milk samples. Rate of intramammary infection with Staphylococcus aureus was similar for vaccinated and unvaccinated cows. Rates of spontaneous cure within each lactation were significantly higher for vaccinated cows. For all three lactations, spontaneous cure rates were 83, 73, and 47% for protein A, bacterin, and control cows. Somatic cell counts were significantly lower for vaccinated cows for quarters infected with Staphylococcus aureus, but no differences were demonstrated for milk production by lactation. Incidence of clinical mastitis was higher in unvaccinated cows, but too few developed for a valid comparison.     

Determination of milk and mammary tissue concentrations of ceftiofur after intramammary and intramuscular therapy

Twenty-five Staphylococcus aureus strains isolated from bovine mastitis were tested for their susceptibility to ceftiofur. Zone diameter for 30 micrograms disks averaged 39 mm, and minimum inhibitory concentrations ranged from .5 to 1 microgram/ml. Tissue and milk concentrations were determined from biopsy and quarter milk samples collected from eight cows treated with either intramammary infusion of 100 or 200 mg of ceftiofur, one or two intramuscular injections of 500 mg of ceftiofur, or combination therapy of intramammary infusion coupled with intramuscular injection. Three additional cows received two intramammary infusions of 200 mg of cephapirin at 24-h intervals. Intramuscular injections of ceftiofur resulted in tissue and milk concentrations below detectable limits. Staphylococcus aureus was not eliminated from infected mammary glands by infusion of 100 mg of ceftiofur or by injection of 500 mg of ceftiofur by 48 h after treatment. Combination therapy of 100 mg of ceftiofur infused and 500 mg injected reduced S. aureus numbers in milk and tissue markedly, as did infusion of 200 mg of ceftiofur. Cows receiving intramammary infusion of 200 mg of ceftiofur (two doses at 24-h intervals) had highest concentrations in milk (450 micrograms/ml at 4 and 6 h) and in tissue (.08 microgram/mg at 30 h). These concentrations are similar to those obtained with two 200-mg doses of cephapirin at 24-h intervals. Histologic analysis of mammary parenchymal tissues showed that combination therapy resulted in higher percentages of alveolar luminal area and lower percentages of interalveolar stroma compared with infusion or injection alone. Histology of quarters receiving combination therapy was not different from that of quarters receiving cephapirin infusion alone.     

Bovine mammary immune response to an experimental intramammary infection with a Staphylococcus aureus strain containing a gene for staphylococcal enterotoxin C1

Staphylococcal enterotoxin C (SEC), a superantigen, is the most frequently expressed enterotoxin by bovine strains of Staphylococcus aureus causing mastitis. To examine the possible impact of SEC on the immune response of the bovine mammary gland, we monitored changes in lymphocyte subpopulations in mammary glands of four lactating cows after intramammary instillation of S. aureus strain Rn4220 transformed with a plasmid containing a gene coding for SEC1. Four other lactating cows received the same strain transformed with the plasmid without the SEC1 gene (positive control), and four cows were untreated (negative control). Mammary quarter milk samples for somatic cell count (SCC) analysis and determination of N-acetyl-beta-D-glucosimindase (NAGase) activity levels were collected daily for 21 d postinstillation. Flow cytometry utilizing three-color analysis was used to phenotype lymphocyte subpopulations isolated from milk samples collected on d 0, 4, 7, 11, 14, 18, and 21 postinstillation from all the cows. Milk from mammary gland halves (positive control and experimental) or all mammary quarters (negative control) was collected for flow cytometric analysis. Increased NAGase activity, SCC, and isolated S. aureus demonstrated that infection was established in mammary quarters intrammarily instilled with bacteria. There were no significant differences (P > 0.05) in the proportions of BoCD4 helper T lymphocytes or BoCD8 cytotoxic T lymphocytes between the two infected treatment groups. There was a significant day x treatment difference of the proportion of a gammadelta T cell subpopulation that did not express BoCD2, but did express the ACT2 activation molecule and a significant treatment difference of a gammadelta T cell subpopulation that expressed BoCD2, but not the ACT2 activation molecule (P < 0.05). Results do not support the hypothesis that the presence of the gene for SEC1 alters the mammary BoCD4 or BoCD8 T lymphocyte response to infection.     

Protection against Staphylococcus aureus mastitis in dairy cows using a bismuth-based teat seal containing the bacteriocin, lacticin 3147

We assessed the effectiveness of a novel dry cow treatment containing lacticin 3147 using deliberate challenge studies in lactating cows. Infection-free quarters of lactating cows were infused with Teat seal (Cross Vetpharm Group, Ltd., Dublin, Ireland) combined with the food-grade bacteriocin, lacticin 3147. Natural infection of the teat was simulated by deliberately introducing Staphylococcus aureus into the teat duct and teat sinus. Relative to control quarters, teat seal plus lacticin 3147 reduced the number of teats shedding viable cells when an inoculum of either approximately 1.7 x 10(3) or approximately 6.8 x 10(3) cfu per teat was used. In addition, the numbers of challenge organisms in those teats from which S. aureus was subsequently recovered were also reduced. However, when the concentration of bacteriocin in the teat seal formulation was reduced by approximately 50%, the number of teats shedding S. aureus cells was not reduced. These data indicate the potential for lacticin 3147 to prevent staphylococcal mastitis infections when a sufficient concentration of the bacteriocin is present. This study also highlights the application of a lactating-cow model to assess the effectiveness of antimicrobial intramammary products on mastitic cell populations.     

Effects of a selective regimen of dry cow therapy on intramammary infection and on antibiotic sensitivity of surviving pathogens

Selective dry cow therapy with 1 million U of procaine penicillin G and 1 g of dihydrostreptomycin in a slow-release base was administered to all quarters of cows currently infected or treated for mastitis in the previous lactation. All other cows in the herd that were dried off during the experimental period were left untreated. Milk samples were cultured for detection of intramammary infection at drying off, 1 wk later (at time of treatment), and after next calving from 227 treated and 143 untreated cows. Isolants were tested for sensitivity to 12 antibiotics including penicillin and streptomycin. Although treatment resulted in clearance of Staphylococcus aureus, Staphylococcus epidermidis, streptococci other than agalactiae, and coliforms from 78, 82, 88, and 87% of infected glands, the net effect on udder health of the herd was a gain of 1.4% in infected quarters. Incidences of new infection in the early dry period (1 wk) were 18.1 and 12.3% among treated and untreated cows. Therapy failed to control new infection in the late dry and peripartum periods; incidence was about 18% of quarters among both long and short dry periods. Among 46 instances of apparent survival of an intramammary pathogen through a treated dry period, 7 of 10 strains previously sensitive to streptomycin became resistant and 8 of 10 strains previously sensitive to penicillin became resistant. The surviving pathogen population was not large enough to be considered a threat to herd udder health in the next lactation.     

Efficacy of intramammary tilmicosin and risk factors for cure of Staphylococcus aureus infection in the dry period

The objective ofthis study was to evaluate the efficacy of intramammary tilmicosin, administered at drying-off, for eliminating Staphylococcus aureus infection, and to identify risk factors for S. aureus cure during the dry period. A total of 219 naturally infected cows, representing 308 quarters, were randomized to receive either one of two treatments at drying-off. Cows received either an intramammary infusion of 500 mg of benzathine cloxacillin, or a sterile solution containing 1,500 mg of tilmicosin. All cows had quarter milk samples taken aseptically three times before dry-off, and at wk 1, 2, and 4 of the subsequent lactation. Overall, 62% of cows and 67.5% of quarters infected with S. aureus cured during the dry period. The cure following administraton of tilmicosin was 67.3 and 72.5% for cows and quarters, respectively. By comparison, the cure achieved with cloxacillin was 56.9 and 62.9% of cows and quarters. Cows receiving tilmicosin were 2.1 times more likely to cure. The cure rate for cows decreased as the linear score on the last DHI test increased, and as the amount of S. aureus being shed increased. Quarters that cultured positive multiple times before drying-off were less likely to cure. Staphylococcus aureus infections located in front quarters of the udder were 2 times more likely to cure than those in hind quarters. Results of this study demonstrate that intramammary tilmicosin at drying-off is efficacious in curing existing S. aureus during the dry period. Risk factors associated with the cure of S. aureus were identified.     

Functional activity of neutrophils from bovine mammary glands infected with Staphylococcus aureus

To test the effect of Staphylococcus aureus infection on mammary neutrophil function, intramammary neutrophils from S. aureus-infected quarters (n = 8), from adjacent uninfected quarters (n = 8) of S. aureus-infected cows, and from quarters (n = 8) of uninfected cows were collected and incubated with S. aureus in vitro. Mean percent neutrophils phagocytizing, number S. aureus per neutrophil, and log10 viable phagocytized S. aureus/ml were: 53.2, 6.4, and 4.72. Differences in function of neutrophils collected from infected and uninfected quarters were not statistically significant. Results indicate that function of neutrophils from S. aureus-infected quarters is similar to function of neutrophils from uninfected quarters.     

Environmental gram-positive mastitis treatment: in vitro sensitivity and bacteriologic cure

A clinical trial was conducted in a large dairy herd to determine the efficacy of intramammary pirlimycin hydrochloride administration during lactation for bacteriologic clearance of gram-positive environmental clinical and subclinical mastitis infections. Quarters infected with environmental streptococci that received pirlimycin therapy (13/28) were 1.8 times more likely to resolve infection than untreated quarters (5/14). The small numbers of quarters infected with coagulase-negative staphylococci resulted in inadequate power to assess treatment differences in cure rate. Although the association was not statistically significant, quarters from cows with sensitive environmental streptococci isolates from composite samples (8/13) resolved infection with treatment at approximately twice the rate of treated quarters with resistant isolates (3/10).     

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.     

Systemic dry cow therapy--a preliminary report

Efficacy of three different treatment regimens in the elimination and prevention of Staphylococcus aureus intramammary infection was studied in 106 dry cow periods. At drying off, norfloxacin nicotinate was given subcutaneously to 44 cows at 10 mg/kg, oxytetracycline-HCl was administered intramuscularly to 18 cows at 20 mg/kg, 500 mg cephapirin benzathine were infused into each udder quarter of 21 cows, and a group of 23 cows served as an untreated control. Number of existing Staphylococcus aureus intramammary infections was reduced only in the norfloxacin nicotinate treatment group. New infection rate appeared lower in the two systemic treatment groups. The percentage of infected quarters remained the same throughout the dry period in the norfloxacin treatment group but number of infected quarters increased by 33 to 85% (significant in the cephapirin group) in the other groups. Minimal inhibitory concentration of the drugs for 57 S. aureus isolates was determined. Isolates were sensitive to norfloxacin and cephapirin and moderately sensitive to oxytetracycline. Results suggest that systemic dry cow therapy using norfloxacin nicotinate, which possesses large distribution volume, long half-life, and is highly active against the pathogen involved, was more effective than the other treatments.