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.     

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.     

In vitro germicidal activity of teat dips against Nocardia asteroides and other udder pathogens.

Nine commercial teat dip formulations containing 1.94% linear dodecylbenzene sulfonic acid, or 1% available iodine from nonylphenoxypoly (ethyleneoxy) ethanol-iodine complex, or .5% chlorhexidine acetate were tested for contamination with aerobic and anaerobic bacteria and their in vitro germicidal activity against Staphylococcus aureus, Streptococcus agalactiae, Escherichia coli, and Nocardia asteroides. All products were free of bacteria when neutralized samples were tested on blood agar or liquid thioglycollate media. To test for in vitro efficacy, each teat dip preparation was mixed with a suspension of one of the pathogenic test organisms containing 10(8) bacteria/ml (final concentration) for .5 to 15 min. Viable bacteria were evaluated by direct plating of neutralized aliquots and by filtration techniques. All products were effective against E. coli, Staph. aureus, and Strep. agalactiae. With N. asteroides, the direct plating method gave equivocal results. The filtration experiments indicated that all teat dips containing dodecylbenzene sulfonic acid and nonylphenoxypoly (ethyleneoxy) ethanol-iodine complex were effective against all four pathogens. Three of the teat dips containing chlorhexidine acetate were ineffective against N. asteroides. The fourth teat dip, containing chlorhexidine acetate and an emollient, was partially effective.     

Evaluation of postmilking teat germicides containing Lauricidin, saturated fatty acids, and lactic acid.

The efficacies of postmilking teat germicides containing Lauricidin (glyceryl monolaurate), saturated fatty acids, lactic acid, and lauric acid were determined against new IMI caused by Staphylococcus aureus and Streptococcus agalactiae in three controlled infection trials. In trial 1, a germicide was evaluated containing 1% Lauricidin, 5% caprylic and capric acids, 6% lactic acid, and .85% lauric acid. New IMI with Staph. aureus and Strep. agalactiae were reduced 81.3 and 49.6%, respectively. Trial 2 germicide involved an artificially aged sample of the formulation evaluated in trial 1. The germicide was aged at 40 degrees C for 5 mo, which was approximately equal to 2 yr at room temperature (24 degrees C). Reductions in new IMI were 81.2 and 27.5% for Staph. aureus and Strep. agalactiae, respectively. In trial 3, a teat germicide aged at ambient temperature for 33 mo, which was originally formulated to contain 1% Lauricidin, 5% caprylic and capric acids, and 6% lactic acid, was evaluated. Reductions in new IMI were 75.5 and 40.4% for Staph. aureus and Strep. agalactiae, respectively. The formulation evaluated in trial 1 was superior to other formulations in reducing new IMI by the two test organisms.     

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.     

Evaluation of .5% and 1% iodophor teat dips on commercial dairies

Two postmilking teat dips containing .5 and 1% iodine were evaluated for approximately 8 mo under conditions of natural exposure to mastitis pathogens on four commercial dairy farms. In the two herds using .5% iodophor, incidence of intramammary infection with Staphylococcus aureus and Streptococcus agalactiae was reduced 68.3 and 46.2%, respectively. When numbers of new infections with both pathogens were combined, efficacy for the .5% iodophor was 62.3%. In the two herds using 1% iodophor, incidence of intramammary infection with Staph. aureus and Strep. agalactiae was reduced 52.4 and 70.7%, respectively. When numbers of new infections with both pathogens were combined, efficacy for the 1% iodophor was 64.3%. Efficacy of teat dips was variable against coagulase-negative staphylococci and Corynebacterium bovis.     

Efficacy of two iodine teat dips during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae

An experimental challenge trial was performed against Staphylococcus aureus and Streptococcus agalactiae following the procedures recommended by the National Mastitis Council. The efficacy of two teat dips, product 1 (Bovadine with I-Tech II) and product 2 (Bovadine with I-Tech, used as a positive control), was determined. Both teat dips contain 1% iodine and 10% glycerin. Product 1 established an 89.7% reduction in infections against Staph. aureus and 73.1% reduction in infections against Strep. agalactiae. Product 2 demonstrated an 86.2% reduction in infections against Staph. aureus and 78.4% reduction in infections against Strep. agalactiae. Teat skin and teat ends were evaluated before and after the trial. No significant change in teat condition was observed for either product.     

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.     

Evaluation of powdered 0.5% chlorhexidine acetate-based postmilking teat dip compared with a foamed 1% iodine-based postmilking teat dip under cold weather conditions in northern New York

The objective of this trial was to compare a powdered 0.5% chlorhexidine acetate-based postmilking teat dip with a foamed 1% iodine-based postmilk teat dip during winter on clinical mastitis, subclinical mastitis (somatic cell count ≥200,000 cells/mL), linear score, teat skin condition, teat end score for hyperkeratosis, and risk of developing a new intramammary infection (IMI). Holstein cows (n = 331) housed in freestall and tiestall barns on one farm were blocked by pen, parity, lactation stage, and lactation performance. They were assigned randomly to a powdered chlorhexidine postmilking teat dip (PD; Derma Soft n' Dry, IBA Inc., Millbury, MA) or a foamed iodine-based postmilking teat dip (ID; FS-103, IBA Inc.). Treatments were applied for 6 wk starting January 4, 2016, for 3 milkings per day. Milk samples were collected from each quarter at the beginning and end of the trial and analyzed for aerobic culture and somatic cell count. Cows that had a clinical mastitis event during the trial were quarter sampled for aerobic culture at the time of clinical event. Teat skin condition and teat end score for hyperkeratosis were evaluated at the beginning, middle, and end of the trial based on a 3- and 5-point scale, respectively. No treatment difference was observed for linear score or teat skin condition. Teat end score was greater for ID cows compared with PD cows (2.72 vs. 2.77) at the conclusion of the trial. At the beginning of trial 102 PD quarters and 129 ID quarters had an IMI identified on aerobic culture, 402 PD and 457 ID quarters cultured negative, and 109 PD and 125 ID samples were classified as “no significant growth.” At the conclusion of the trial, 129 PD and 101 ID quarters had an IMI. Use of PD resulted in a greater risk for developing a new IMI, based on bacteriological culture, at the conclusion of the trial as compared with ID (relative risk = 1.51; confidence interval: 1.10–2.07). Additionally, use of PD resulted in a greater risk as compared with ID of coagulase-negative staphylococci (relative risk = 1.5; confidence interval: 1.10–2.25) and Staphylococccus aureus (relative risk = 2.30; confidence interval: 1.04–5.07) to be present at the conclusion of the trial. In conclusion, use of PD led to a lower teat end score, an increase in new IMI, and an increased risk of coagulase-negative staphylococci and Staph. aureus compared with ID after 6 wk of product use.     

Evaluation of 1.6% phenol as a premilking and postmilking teat dip in preventing new bovine intramammary infections

We evaluated the effectiveness of a 1.6% phenol-based teat dip using both a teat skin assay and natural exposure field trial. A teat skin assay was conducted to ascertain the concentration of phenol + phenate to be used in the field study. One percent and 0.5% iodine, and 0.5, 1.1, and 1.6% phenol + phenate were compared using Escherichia coli and Staphylococcus aureus. Logarithmic reductions for S. aureus were 2.2 and 2.8 for 0.5 and 1% iodine, and 1.3, 2.1, and 2.8 for 0.5, 1.1, and 1.6% phenol + phenate, respectively. Logarithmic reductions for E. coli were 3.3 and 3.8 for 0.5 and 1% iodine, and 1.2, 1.9, and 2.6 for 0.5, 1.1, and 1.6% phenol + phenate, respectively. A concentration of 1.6% phenol + phenate was chosen as experimental teat dip, and 0.5% iodine served as control. The field study was conducted at Beltsville (n = 185) and Clarksville (n = 100) dairy herds using a split herd design. Teat dips were used premilking and postmilking for 12 mo. The number of new intramammary infections (IMI) for the Beltsville herd in iodine and phenol + phenate teat dipped cows were: 29 and 35 for major pathogens, and 81 and 72, for minor pathogens. For the Clarksville herd, number of new intramammary infections in iodine and phenol + phenate teat dipped cows was 9 and 10 for major pathogens, and 50 and 60 for minor pathogens. Rates of IMI per quarter day per lactation were not different for either herd or when herd data were combined. The number of clinical mastitis cases per 100 cows per month were similar in both treatments. The incidences of new IMI and clinical mastitis were similar using both dips.     

Efficacy of .18% iodine teat dip against Staphylococcus aureus and Streptococcus agalactiae

Effective postmilking teat dip products with lower iodine concentrations are being formulated as concern increases about iodine residues in milk. Increased free iodine concentration with greater germicidal activity in teat dip products is also possible with special formulation procedures. Low iodine concentration dips are cheaper and have reduced teat irritation. A concentrated iodine teat dip containing .18% iodine and 8 ppm free iodine upon dilution was evaluated under experimental bacterial challenge to determine efficacy for prevention of new intramammary infections. The undiluted product also contained 15% collagen protein emollient as a teat skin conditioner. Efficacy of the teat dip was 93.6 and 51. 7% for Staphylococcus aureus (Newbould 305) and Streptococcus agalactiae (McDonald 44). No adverse effects of the dip on teat skin were noted.     

Evaluation of experimental teat dip containing sodium chlorite and lactic acid by excised teat assay

An experimental teat dip containing sodium chlorite and lactic acid, diluted in water, was evaluated by excised teat protocol. The teat dip was tested against 21 microorganisms. Included were: Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Numerous strains were tested for strain differences. Environmental bacteria were included because of their increasing importance as a cause of bovine mastitis. All excised teats were dipped in a bacterial suspension containing about 1 X 10(8) cfu/ml. Negative control teats were not dipped in a germicidal compound. Positive controls were dipped in 1% iodophor. Effectiveness of the experimental teat dip was expressed as the percent reduction in mean log of bacteria recovered from dipped teats as compared to numbers recovered from control teats. The sodium chlorite - lactic acid dip caused a greater percent log reduction than iodophor for 14 of 21 strains tested. However, differences were generally slight. The experimental teat dip appeared effective against Gram-negative bacteria. Some differences in percent log reduction were observed between strains of the same species. Lowest effectiveness and greatest strain variation were observed with Staphylococcus aureus for both dips tested.     

Short communication: disinfectant containing a complex of skin conditioners

The efficacies of 2 new teat dip formulations were tested against experimental challenge by contagious mastitis pathogens Staphylococcus aureus and Streptococcus agalactiae over a 12-wk period. Formulations contained an iodine complex (0.5 or 1.0% iodine) and skin conditioning agents (propylene glycol, polyvinylpyrridone, glycerine, lanolin, allantoin, and aloe). Percentage reduction (dipped vs. control mammary quarters) in new contagious mastitis pathogen intramammary infections for the 0.5 and 1.0% iodine dips was 65.4 and 84.5, respectively. Both dips were significantly effective in reducing new contagious intramammary infections. Teat skin scores and teat end scores varied over time but were virtually identical for both treated and control teats, for both treatments. Thus, both dips were effective in reducing new contagious mastitis infections without untoward effects on teat skin condition.     

Evaluation of nine teat dip formulations under experimental challenge to staphylococcus aureus and streptococcus agalactiae

Nine postmilking teat dips were evaluated by an experimental challenge model against either Staphylococcus aureus, Streptococcus agalactiae, or both. Formulations containing .9 and .6% sodium hypochlorite, 1% sodium dichloro-s-triazene-trione, .55% chlorhexidine gluconate, and .35% povidone iodine reduced incidence of Staphylococcus aureus infections 56.8, 28.3, 75.9, 92.5, and 77.9%. Incidence of infections with Streptococcus agalactiae was reduced 48.1 and 63.2% by 1.7 and 1% sodium dichloro-s-triazene-trione formulations. The 1% chlorhexidine gluconate and .35% povidone iodine products reduced Streptococcus agalactiae infections 71.0 and 67.0%. Three experimental 1% iodophor formulations reduced Streptococcus agalactiae infections 28.9, 44.8, and 50.7%. The experimental challenge model was refined further and provided an efficient method to determine efficacy of postmilking teat dips.     

Evaluation of a chlorous acid-chlorine dioxide teat dip under experimental and natural exposure conditions

A postmilking teat dip containing chlorous acid-chlorine dioxide was evaluated by experimental challenge and in two herds under natural exposure. The test product had an efficacy of 78.9% against Staphylococcus aureus and 52.5% against Streptococcus agalactiae in the experimental challenge trial. The product was compared with a 1% iodine product in a 15-mo natural exposure study. Post-dipping with chlorous acid-chlorine dioxide reduced incidence of udder infection by major mastitis pathogens 36.1% when data were combined from the two herds. The 1% iodine and the chlorous acid-chlorine dioxide products were not equivalent for major mastitis pathogens; the test product was more effective. Incidence of udder infection by environmental mastitis pathogens was reduced 36.8% in both herds combined. Efficacy of the two teat dips was equivalent for environmental pathogens.     

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.     

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.     

Effects of teat dipping on intramammary infections by staphylococci other than Staphylococcus aureus

Effects of intramammary infections in herds 1) either not teat dipping or postmilking teat dipping with either 2) linear dodecyl benzene sulfonic acid, 3) chlorhexidine, or 4) iodophor containing products were determined. Duplicate quarter foremilk samples were collected once from all lactating cows in 16 herds, four for each practice. Mean percentage of quarters infected with Staphylococcus species other than Staph. aureus was 11.0 in herds not teat dipping and 7.2 in herds teat dipping. Prevalence of Staphylococcus species intramammary infections in herds teat dipping with linear dodecyl benzene sulfonic acid was comparable to herds not teat dipping and greater than in herds using chlorhexidine and iodophor. The predominant Staphylococcus species in herds not teat dipping was Staph. epidermidis (37.1%). Staphylococcus hyicus constituted 48.5% of total Staphylococcus species intramammary infections in herds teat dipping and 12.4% in herds not teat dipping. Differences were not observed among SCC for quarters infected with different Staphylococcus species. Application of germicidal teat dips appeared to have selectively altered both prevalence and distribution of Staphylococcus species intramammary infections.     

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.     

Efficacy of a new premilking teat disinfectant containing a phenolic combination for the prevention of mastitis.

A teat disinfectant containing a phenolic combination was evaluated in a natural exposure study in two dairy research herds. Premilking teat disinfection was compared with a negative control using a split-udder experimental design. In both herds, premilking and postmilking teat disinfections with the phenolic combination were significantly more effective in preventing new intramammary infection (IMI) than was postmilking teat disinfection only. Clinical mastitis and new IMI by Streptococcus uberis, Streptococcus dysgalactiae, Gram-negative pathogens, and coagulase-negative Staphylococcus species were significantly lower in quarters of cows with teats predipped and postdipped than in quarters with teats postdipped only. No chapping or teat skin irritation was observed. Premilking teat disinfection with the phenolic combination in association with good udder preparation and postmilking teat disinfection can further reduce the occurrence of new IMI by numerous mastitis pathogens during lactation.