Evaluation of a surfactant mixture C31G as a teat dip by a modified excised teat model

A mixture of amphoteric surfactants with antimicrobial properties, C31G, was evaluated as a teat dip by a modified excised teat model. The model was modified to avoid misinterpretation of results from indigenous bacterial microflora on teats. In three experiments, bacterial numbers recovered from teats dipped in .75, 1.5, and 3.0% solutions of C31G were compared with bacteria recovered from undipped teats (negative controls) and teats dipped with 1.0% iodophor (an effective teat dip). Mean log reductions of teat end concentrations of Staphylococcus, aureus, Streptococcus fecalis, and Escherichia coli were determined in each experiment. Efficacy was greatest for the 3.0% solution of C31G with mean log reductions (Log reduction = Log 10 of controls - Log 10 of dipped teats) 3.53 for Staphylococcus aureus, 2.89 for Streptococcus fecalis, and 3.16 for Escherichia coli. Iodophor (1%) yielded mean log reductions of 3.26 for Staphylococcus aureus, 2.16 for Streptococcus fecalis, and 2.83 for Escherichia coli. For all three organisms, number of viable organisms decreased with increasing concentrations of C31G. The efficacy of C31G should be evaluated in vivo.     

Germicidal teat dip in a herd with low prevalence of Streptococcus agalactiae and Staphylococcus aureus mastitis

Effectiveness of an iodophor teat dip in reducing new intramammary infection and clinical mastitis was tested in a herd free of Streptococcus agalactiae and with a low prevalence of Staphylococcus aureus infections. In this 6-mo study with an average of 152 lactating cows on trial on sampling dates, right rear and left front teats were dipped after each milking while right front and left rear teats were undipped controls. Teat dipping significantly reduced new infections by Staphylococcus aureus, streptococci (other than Streptococcus agalactiae), coagulase-negative staphylococci, and Corynebacterium bovis and reduced clinical cases caused by infections established during the trial. However, the dip did not reduce new infections or clinical mastitis caused by coliform bacteria. Therefore, use of a germicidal dip is recommended for herds with a low prevalence of Streptococcus agalactiae and Staphylococcus aureus, but the practice should not be expected to control coliform mastitis.     

Efficacy of chlorhexidine as a postmilking teat disinfectant for the prevention of bovine mastitis during lactation

A natural exposure trial was conducted for 12 mo in a herd of 150 lactating Jersey cows to determine efficacy of a .35% chlorhexidine teat dip containing a glycerine emollient for the prevention of bovine intramammary infections. Right teats of cows were dipped in the experimental teat dip after milking machine removal and left teats were not dipped. The herd was free of Streptococcus agalactiae and had a low prevalence of Staphylococcus aureus. Most new major pathogen intramammary infections resulted from Streptococcus species, primarily Streptococcus uberis and Streptococcus dysgalactiae. New infections by Streptococcus species were significantly lower in teats dipped in chlorhexidine than in undipped teats. Overall efficacy of the chlorhexidine teat dip against major mastitis pathogens was 50%. The experimental teat dip also reduced coagulase-negative Staphylococcus species infections 49.0% and Corynebacterium bovis infections 65.2%. Overall efficacy against minor mastitis pathogens was 54.0%. No irritation or chapping of teats dipped in the experimental teat dip was observed.     

Efficacies of chlorine dioxide and lodophor teat dips during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae

We tested two postmilking teat dips for efficacy against Staphylococcus aureus and Streptococcus agalactiae using experimental challenge procedures recommended by the National Mastitis Council. The chlorine dioxide teat dip that contained 0.7% sodium chlorite reduced the number of new intramammary infections (IMI) caused by Staph. aureus by 86.6% and reduced new IMI caused by Strep. agalactiae by 88.4%. The 0.5% iodophor teat dip reduced the number of new IMI caused by Staph. aureus by 92.9% and reduced the number of new IMI caused by Strep. agalactiae by 43.4%. Teat skin and teat end conditions were evaluated before and after the study, and no deleterious effects were noted among dipped quarters compared with undipped control quarters for either teat dip.     

Hygiene in the prevention of udder infections. IV. Evaluation of teat dips with excised cows' teats.

Suspensions of Staphylococcus aureus, Streptococcus agalactiae, Escherichia coli, and Pseudomonas aeruginosa were used to contaminate teats excised from cows. Commercially available teat dips were applied for evaluation of comparative germicidal activity. Iodophors, sodium hypochlorite, and sodium dichloro-s-triazenetrione were highly effective against all four test organisms. Quarternary ammonium, chlorhexidine, and cetylpyridinium chloride were effective against Staphylococcus aureus, and Streptococcus agalactiae but not Escherichia coli and Pseudomonas aeruginosa. Bronopol and 8-hydroxyquinoline sulfate were marginally effective on the two Gram positive organisms and ineffective on the two Gram negatives. Three of the dips were ineffective against all four pathogens; two were in nonaqueous bases, and the other was a blend of pine oil and antibodies with udder origin. In other trials, iodophor at .3% titratable iodine was highly effective against Staphylococcus aureus and Streptococcus agalactiae. Products containing .5 and 1% iodophor maintained effectiveness after each was used to dip 140 teats under conditions of commercial dairying. Logarithmic reduction in the geometric mean number of organisms recovered from teats was more meaningful for evaluating data than percent reduction.     

Effect of postmilking teat antisepsis on teat canal infections in lactating dairy cows

Effects of a .18% iodophor teat dip and a fatty acid plus lactic acid teat dip on teat canal infections were studied in two separate trials under experimental challenge procedures using Staphylococcus aureus and Streptococcus agalactiae. The iodophor teat dip was 90% effective in preventing new S. aureus teat canal infections and 95.6% effective in reducing progression of S. aureus teat canal infections to intramammary infections. The dip also reduced persistence of S. aureus teat canal colonizations from week to week by 94.4%. The fatty acid plus lactic acid dip had no effect on preventing S. aureus teat canal infections but reduced progression of S. aureus teat canal infections to intramammary infections by 43.8%. The fatty acid plus lactic acid dip also reduced the persistence of S. aureus teat canal infections by 39%. An insufficient number of Strep. agalactiae teat canal infections was detected in both trials for valid analyses. Neither dip had any effect on prevalence of naturally occurring, coagulase-negative staphylococcal teat canal infections or on teat canal infections by other organisms.     

Determination of teat dip germicidal activity using the excised teat model

Fifteen postmilking teat antiseptics were evaluated in seven trials using the excised teat model. All products were tested against Staphylococcus aureus Newbould 305 and Streptococcus agalactiae Cornell 48. Eight of nine iodophor products provided log reductions greater than 3 against Staph. aureus and Strep. agalactiae. A .045% iodophor product was ineffective. A .5% chlorhexidine product yielded a log reduction of 3.22 against Staph. aureus but only 2.92 against Strep. agalactiae. Two products containing fermented milk emulsified in detergent were tested upon receipt and after 1 yr storage. Fresh product provided log reductions greater than 3 against both pathogens. Storage for 1 yr resulted in minimal loss of germicidal activity. Three products containing 1, .5, and .2% dodecylaminoalkyl glycine provided log reductions greater than 3.5 against Staph. aureus and greater than 4 log reduction against Strep. agalactiae.     

Reduction of mastitis caused by experimental challenge with Staphylococcus aureus and Streptococcus agalactiae by use of a quaternary ammonium and halogen-mixture teat dip

A teat-dip formulation containing sodium dichloro isocyanuric acid, bronopol, and quaternary ammonium was tested for efficacy against Staphylococcus aureus and Streptococcus agalactiae intramammary infections (IMI) using an experimental challenge model. Sixty-two Jersey cows from the Hill Farm Research Station (Homer, LA) were used in an 8-wk controlled infection trial to evaluate the teat dip. During the afternoon milking, Monday through Friday for 8 wk, all teats of each cow were immersed to a depth of approximately 25 mm in a challenge suspension containing approximately 5 x 10(7) cfu of Staphylococcus aureus and approximately 5 x 10(7) cfu of Streptococcus agalactiae immediately after milking machines were removed. Immediately after challenge, the distal 25 mm of two contralateral teats were dipped with the experimental teat dip; the remaining two teats served as undipped controls. The experimental teat dip reduced the number of new Staph. aureus IMI by 70.9% and reduced the number of new Strep. agalactiae IMI by 60.0%. Teat end and teat skin condition were characterized as normal and without irritation at the completion of the study. The combination of the three germicides in this experimental teat dip is unique and an effective formulation without adverse effects on condition of teat ends or teat skin.     

Efficacy evaluation of two new teat dip formulations under experimental challenge

Two new teat dip formulations were evaluated against Streptococcus agalactiae and Staphylococcus aureus in half-udder, experimental challenge studies. The first product contained a combination of 1.9% linear dodecyl benzene sulfonic acid - .55% iodophor. Incidence of infection was reduced 52.7 and 70.7% for Streptococcus agalactiae and Staphylococcus aureus in dipped quarters compared to undipped controls. The second formulation was a barrier-type product consisting of milk protein solubilized with lauryl sulfate, a surface active detergent and 4.8% glycerin. Rates of infection were reduced 60.3 and 67.9% for Streptococcus agalactiae and Staphylococcus aureus.     

Effects of an automatic postmilking teat dipping system on new intramammary infections and iodine in milk

A technology of automatically applying a postmilking teat dip via the milking machine prior to machine detachment was compared to manual postmilking teat dipping with a teat dip cup for effects on new IMI and iodine content in milk. One hundred twenty Holstein cows were experimentally challenged in a 22-wk trial with Streptococcus agalactiae and Staphylococcus aureus and 148 Holstein cows were experimentally challenged with Streptococcus uberis in another 22-wk trial. The bacterial suspensions were applied to teats of all of the cows after premilking udder preparation and immediately prior to milking machine attachment. In both trials, cows were divided among four treatments: no postmilking teat dipping; manual postmilking teat dipping with a proven efficacious iodophor teat dip; manual postmilking teat dipping with an iodophor teat dip formulated for an automatic postmilking teat dipping system; and automatically postmilking teat dipping via milking machines with an iodophor teat dip formulated for the automatic postmilking teat dipping system. The postmilking teat dipping treatments reduced new Staph. aureus IMI by 64.5, 76.5, and 88.2%; new Strep. agalactiae IMI by 61.5, 77.8, and 94.4%; and new Strep. uberis IMI by 63.5, 82.5, and 93.8%, respectively, against the treatment of no postmilking teat dipping. The treatment applying the postmilking teat dip automatically via milking machines had the lowest number of new IMI caused by the three pathogens. Teat end and teat skin condition were characterized as normal at the end of the study with no differences between treatments. There were no differences with regard to iodine content in milk between treatments.     

Efficacy of two acidified chlorite postmilking teat disinfectants with sodium dodecylbenzene sulfonic acid on prevention of contagious mastitis using an experimental challenge protocol

Two acidified sodium chlorite postmilking teat disinfectants were evaluated for efficacy against Staphylococcus aureus and Streptococcus agalactiae by using National Mastitis Council experimental challenge procedures. The effect of these teat dips on teat skin and teat end condition was also determined. Both dips contained 0.32% sodium chlorite, 1.32% lactic, and 2.5% glycerin. Dips differed in the amount of sodium dodecylbenzene sulfonic acid (0.53 or 0.27%) added as a surfactant. Both dips significantly reduced new intramammary infection (IMI) rates compared with undipped controls. The dip containing 0.53% dodecylbenzene sulfonic acid reduced new IMI by Staph. aureus by 72% and Strep. agalactiae by 75%. The dip containing 0.27% dodecylbenzene sulfonic acid reduced new IMI by Staph. aureus by 100% and by Strep. agalactiae by 88%. Changes in teat skin and teat end condition for treatment and control groups varied in parallel over time. Teats treated with either teat dip had higher mean teat skin and teat end scores than control teats at some weeks. However, teat skin and teat end condition did not tend to change from the start to the completion of the trial. Application of the two new postmilking teat dips was effective in reducing new IMI from contagious mastitis pathogens. (Key words: teat dip, contagious mastitis, chlorous acid)     

Evaluation of a nisin-based germicidal formulation on teat skin of live cows.

A purified preparation of the nontoxic antimicrobial peptide, nisin (AMBICIN N), was used in the formulation of a germicidal sanitizer suitable for use on cow teats. The germicidal activity of the formulation against mastitis pathogens was measured on teat skin of live cows. The nisin-based formulation gave a mean log reduction of 3.90 against Staphylococcus aureus and 4.22 log reduction against Escherichia coli after exposure for 1 min to the germicide. This activity was comparable with that exhibited by a 1% iodophor teat dip but was significantly greater than that exhibited by the .1 and .5% iodophors and by the .5% chlorhexidine digluconate teat dips. The nisin-based formulation showed little or no potential for skin irritation after multiple application to skin, but iodophor and chlorhexidine digluconate teat dips showed significant potential for skin irritation in comparable studies.     

Effects of a 1.94% sulfonic acid teat dip and a 1% iodophor teat dip on teat canal infections in lactating dairy cows

Sixty-three cows were used in a natural exposure trial for 1 yr to determine the effects of a 1.94% linear dodecyl benzene sulfonic acid and a 1% iodophor teat dip on teat canal and intramammary infections. Overall, total teat canal infections increased 28.9% and total intramammary infections increased 30% in quarters dipped with the linear dodecyl benzene sulfonic acid compared with a 14.3% increase in teat canal infections and a 17.4% decrease in intramammary infections for the iodophor group. Higher frequencies were observed for teat canal and intramammary infections caused by Staphylococcus epidermidis in the linear dodecyl benzene sulfonic acid group. Staphylococcus hyicus was the predominant organism isolated from teat canal and intramammary infections in the iodophor group. No difference was observed between the two teat dips in the persistence of Staphylococcus aureus teat canal infections. A greater number of S. hyicus teat canal infections persisted in the iodophor group, whereas teat canal infections produced by S. epidermidis and Staphylococcus xylosus persisted longer in the sulfonic acid group. Results of this study indicated that use of a linear dodecyl benzene sulfonic acid teat dip for 1 yr was not associated with increased S. aureus teat canal infections and intramammary infections. However, linear dodecyl benzene sulfonic acid appeared to affect the distribution of coagulase-negative staphylococci isolated from teat canal infections and intramammary infections.     

Field studies on linear dodecyl benzene sulfonic acid teat dip

A teat dip containing 1.94% linear dodecyl benzene sulfonic acid and 12% glycerine, an anionic detergent formulation, was evaluated in two field trials on a commercial dairy. In Trial 1, the teat dip was compared to an undipped control in a split-herd design. Incidence of infection with Streptococcus agalactiae was reduced 57.8%; infections with presumptively identified Staphylococcus aureus were not reduced. Milking machine function and milking procedures may have affected teat dip effectiveness and incidence of udder infection. Streptococcus agalactiae was eradicated prior to Trial 2 in which the detergent teat dip was compared to a 1% iodophor product of proven efficacy. Incidence of Staphylococcus aureus infection was 40.5% lower in the detergent than in the iodophor group. Taxonomic studies on Staphylococcus species indicated that presumptive microbial identification resulted in erroneous conclusions concerning efficacy of the teat dip products. Importance of speciation of staphylococcal isolates is discussed.     

Evaluation of a postmilking teat disinfectant containing a phenolic combination for the prevention of mastitis in lactating dairy cows

A trial was conducted for 12 months in a herd of 120 Holstein cows in order to determine the efficacy of a teat disinfectant, which contained a phenolic combination, for the prevention of bovine intramammary infections during lactation. Postmilking teat disinfection was compared to a negative control using a split-udder experimental design. The percentage of quarters newly infected by mastitis pathogens was 45% lower in mammary glands with teats that had been dipped in the experimental teat disinfectant after milking than it was in undipped controls. New infections caused by Streptococcus uberis, Staphylococcus aureus, coagulase-negative Staphylococcus species, and Corynebacterium bovis were significantly lower in mammary glands with teats that had been dipped in the experimental teat disinfectant than in undipped controls. No statistical differences in the incidence of clinical mastitis between treatment groups were observed. No irritation or chapping of teats dipped in the experimental teat disinfectant were observed. The results of this study suggest that the experimental teat disinfectant containing a phenolic combination is an effective postmilking teat disinfectant for use in the prevention of new intramammary infections by both contagious and environmental mastitis pathogens.     

Colonization by Staphylococcus aureus on chapped teat skin: effect of iodine and chlorhexidine postmilking disinfectants.

The abilities of common postmilking teat disinfectant solutions and a teat skin ointment to retard Staphylococcus aureus colonization and promote healing of chapped skin were studied. Ten Holstein cows, free of S. aureus intramammary infections and mammary epidermal colonization, were used. All teats were experimentally chapped with 1N sodium hydroxide and dipped in S. aureus broth culture of 5 x 10(6) cfu/ml after three consecutive milkings. Postmilking teat disinfectant treatments were applied, starting after the second application of S. aureus broth culture, and were continued for the next 19 milkings. Quarters of each cow received one of the following postmilking teat treatments: 1% iodine plus 10% glycerin solution; .5% chlorhexidine plus 4.87% glycerin solution; 1% chlorhexidine ointment; and no postmilking treatment. After milking and before treatment application, samples of skin swabbings of the teat end and teat side and samples of milk were collected. Daily teat skin chapping visual scores were made to assess chapping and healing. Teat skin chapping decreased with time, and the decline was similar among treatments. The concentration of S. aureus recovered from teat skin swabbing solutions decreased over time. Staphylococcus aureus were recovered at significantly lower concentrations from swabbing solutions of teats that were dipped with iodine and chlorhexidine solutions as compared with untreated teats. Staphylococcus aureus was in marginally higher concentrations on ointment-treated teats than on dipped teats. Results indicate the importance of using postmilking teat dips on chapped teats and suggest no advantage in applying a disinfectant ointment to chapped teats to reduce S. aureus colonization and improve skin healing.     

Prolonged in vitro exposure of Staphylococcus aureus to germicidal teat dips

Eight strains of Staphylococcus aureus were tested to determine if prolonged exposure to commercial teat dips could enhance bacterial tolerance to teat dips in vitro. All strains of S. aureus were serially plated 15 times on chemically defined agar medium containing sublethal concentrations of linear dodecyl benzene sulfonic acid, chlorhexidine, sodium hypochlorite, and iodophor teat dips. Growth responses of S. aureus to chlorhexidine, sodium hypochlorite, and iodophor were not affected by prolonged exposure to these teat dips. Isolates subcultured on agar containing .1% linear dodecyl benzene sulfonic acid teat dip subsequently had a greater mean growth response to .1% solution of the germicide than did controls subcultured on basal medium. Hemolytic patterns, tube coagulase, clumping factor, and protein A reactions of S. aureus were not altered by exposure to any of the teat dips tested. In general, prolonged exposure to commercial teat dips did not alter germicidal susceptibility of S. aureus.     

Prevention of bovine mastitis by a postmilking teat disinfectant containing chlorous acid and chlorine dioxide in a soluble polymer gel

A natural exposure study was conducted in a herd of 150 lactating dairy cows for 18 mo to determine the effectiveness of chlorous acid and chlorine dioxide in a soluble polymer gel as a postmilking teat disinfectant for the prevention of bovine mastitis. Right quarters of cows were dipped in the experimental teat dip after milking machine removal. Left quarters were not dipped and served as within-cow negative controls. The experimental teat dip reduced Staphylococcus aureus infections 67.4%, Streptococcus dysgalactiae infections 63.8%, and Streptococcus uberis infections 27.8%. Overall efficacy of the chlorous acid and chlorine dioxide teat dip against major mastitis pathogens was 52.2%. The experimental teat dip reduced Corynebacterium bovis infections and coagulase-negative staphylococcal infections also by 45.8 and 38.7%, respectively. Overall efficacy against minor mastitis pathogens was 43.4%. Under conditions of this trial, the experimental teat dip containing chlorous acid and chlorine dioxide was effective in preventing new intramammary infections against a variety of mastitis pathogens.     

Hygiene in the prevention of udder infections. V. Efficacy of teat dips under experimental exposure to mastitis pathogens.

Efficacy (reduction of new intramammary infections) was determined against Staphylococcus aureus in 14 trials and against Streptococcus agalactiae in 5 trials. Against Staphylococcus aureus seven 1% iodophor products had an average efficacy of 89.9% while three .5% iodophor products averaged 76.4%. A solution of sodium dichloro-s-triazenetrione reduced infections 79.0% and 2% dodecyl benzene sulfonic acid was 50.4% effective. Efficacies of two quaternary ammonium products at .18 and .5% were 84.8 and 60.9%. A placebo vehicle and .15 M saline as teat dips reduced infections 8.9 and 9.2%. Infection rates with Streptococcus agalactiae were low; efficacy ranged from 50.5 to 100.0%, average 65.6%. The model of experimental exposure provided a better estimate of effectiveness of a teat dip than would be possible under conditions of most commercial dairying.     

Germicidal persistence of teat dips by modified excised teat procedure

An excised teat protocol was modified to evaluate persistence of germicidal activity of teat dips over 8 h. Five teat dip formulations, iodophor (1%), chlorhexidine gluconate (.55%), linear dodecyl benzene sulfonic acid (1.94%), sodium chlorite-lactic acid in a water base, and sodium chlorite-lactic acid in a gel base were tested against Escherichia coli and Klebsiella oxytoca. Iodophor and chlorhexidine had high germicidal activity throughout 8 h, whereas dodecyl benzene sulfonic acid had little activity. Germicidal activity of both sodium chlorite-lactic acid teat dips was high initially but declined with time. The gel base dip, however, remained more germicidal than the water base dip. Results were similar for either organism for most teat dips. However, chlorhexidine was less effective and the gel base dip more effective against Klebsiella oxytoca than Escherichia coli. Standard errors often appeared higher for Klebsiella oxytoca than for Escherichia coli. These assays may prove useful for laboratory screening of teat dips to determine germicidal persistence over time.