Prevention of Bovine Mastitis by a Premilking Teat Disinfectant by tfo34034

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									                                OUR INDUSTRY TODAY
Prevention of Bovine Mastitis by a Premilking Teat Disinfectant
Containing Chlorous Acid and Chlorine Dioxide
                                       S. P. OUVER,1 M. J. LEWIS, 1 L. INGLE, B. E. GILLESPIE,
                                                                   .
                                                                         and K. R. MAITHEWS
                                                                           Institute of Agriculture
                                                                   Department of Animal Science
                                                                     The University of Tennessee
                                                                             Knoxville 37901-1071

                   ABSTRACT                                       INTRODUCTION

      The objective of this study was to             Remilking teat disinfection is a relatively
  evaluate the efficacy of a premilking teat      new concept that was developed as a potential
  disinfectant for the prevention of mastitis     method to control environmental mastitis by
  in dairy cows under natural exposure            reducing bacterial populations on teat skin
  conditions. Predipping was compared             prior to milking. Premilking and postmilking
  with a negative control using a split ud-       teat disinfection, in association with good ud-
  der experimental design. All teats were         der preparation, was significantly more effec-
  dipped after milking with the same teat         tive in preventing environmental pathogen IMI
  dip. Percentage of quarters newly in-           than good udder preparation and postmilking
  fected by major mastitis pathogens was          teat disinfection (IO).
  34% lower in quarters with teats                   Many dairy producers have incorporated
  predipped and postdipped than in quar-          premilking teat disinfection into their mastitis
  ters with teats postdipped only. New IMI        control strategy. Most of the teat disinfectants
  by Streptococcus uberis and Staphy-             now used for such purposes likely have not
  Zococcus aureus were significantly lower        been evaluated for safety and efficacy. To date,
  in quarters with teats predipped and post-      the effectiveness of premilking teat disinfec-
  dipped than in quarters with teats post-        tion has been described with low concentra-
  dipped only. Differences in incidence of        tion, iodine-based teat disinfectants only (10).
  clinical mastitis between treatment             The objective of the present study was to
  groups approached significance. Predip-         evaluate a teat dip containing chlorous acid
  ping and postdipping were no more ef-           and chlorine dioxide as a premilking teat disin-
  fective against Gram-negative bacteria,         fectant for the prevention of environmental
  coagulase-negative Staphylococcus spe-          mastitis.
  cies, and Corynebacterium bovis than
  postdipping only. No chapping or irrita-                  MATERIALS AND METHODS
  tion of teats was observed, and no ad-             A natural exposure study was conducted
  verse effects were noted using the test         following procedures described by the Na-
  product as a premilking and postmilking         tional Mastitis Council (4). The teat dip con-
                                   hs
  teat disinfectant. Results of ti study          sisted of separate solutions containing, as ac-
  suggest that predipping and then postdip-       tive ingredients, sodium chlorite (.64%) and
  ping with the test product was a more           lactic acid (2.64%) (4XLA@; Alcide Corp.,
  effective procedure against major masti-        Norwalk, CT) that were mixed daily in equal
  tis pathogens than postdipping only.            volumes. The antimicrobial activity of this teat
  (Key words: premilking, teat disinfec-          disinfectant is thought to be due to the trans-
  tion, postmilking, environmental mastitis       formation of the chlorite ion to chlorous acid,
  pathogens)                                      which, in concert with its conversion product,
                                                  chlorine dioxide, readily penetrates cell-wall
                                                  structures of bacteria and oxidatively destroys
  Received June 22, 1992.                         them (1).
  Accepted August 31, 1992.                          This study was conducted at The University
  'Reprint requests.                              of Tennessee in the Knoxville Experiment Sta-
1993 J Dairy Sci 76:287-292                     287
288                                             OLIVER ET AL.

tion dairy research herd. An average of 175            lytic characteristics, Gram stain reaction, and
cows (approximately 80% Holsteins and 20%              catalase test. Isolates identified presumptively
Jerseys) were lactating at each herd survey. A         as staphylococci were tested for coagulase
total of 423 cows were included in the                 production by the tube coagulase method. Iso-
15-mo trial. Cows that lactated at least 30 d          lates identified presumptively as streptococci
were included in data analysis. Cows were              were evaluated initially for growth in 6.5%
milked twice daily in a 12-stall trigon milking        NaCI, esculin hydrolysis, and CAMP reaction.
parlor. Milking equipment was maintained ac-           Streptococcal organisms were identified as to
cording to the manufacturer’s recomrnenda-             species by the API Rapid Strep System@
tions. Cows were housed in free stalls bedded          (Analytab Inc., Plainview, NY) and serotyped
with sawdust.                                          as described previously (5). Gram-negative
    Premilking teat disinfection was compared          isolates were characterized by the following
with a negative control using a split udder            tests: triple sugar iron, urea, motility, indole,
experimental design. Both left teats (controls)        and ornithine decarboxylase. The Vitek Gram-
were forestripped, washed with a minimum               Negative Identification System (Vitek Sys-
amount of water if necessary, and dried with           tems, Inc., Hazelwood, MO) was used to
single-service paper towels in preparation for         differentiate Serratia and Enterobacter species.
milking. Both right teats were forestripped,           Corynebacterium bovis was identified as a
washed with a minimum amount of water if               Gram-positive rod, catalase-positive, urea-
necessary, dried with a single-service paper           positive, and oxidase-positive organism that
towel, treated with experimental premilking            exhibited enhanced growth on brain-heart infu-
teat disinfectant applied with a minimum con-          sion agar supplemented with .l% Tween 80
tact time of 30 s, and dried thoroughly with a         (Fisher Scientific Co., Fair Lawn, NJ).
single-service paper towel. All teats of cows              At the onset of the study, a quarter was
were dipped full length with the same teat             considered to be infected when the same bac-
disinfectant after milking machine removal.            terial species was isolated in duplicate fore-
Thus, milking routine for all cows was consis-         milk samples. Diagnosis of IMI in mammary
tent except for application of premilking teat         glands of cows after parturition was based on
disinfectant to right teats and subsequent dry-        the isolation of the same bacterial species from
ing of teats after predip application.                 single foremilk samples obtained on two occa-
    Duplicate samples of foremilk were col-            sions during the first 10 d of lactation.
lected aseptically from all quarters of lactating          A quarter was diagnosed as newly infected
cows at the onset of the study, when clinical          when the same bacterial species was isolated
mastitis was observed, and from cows leaving           in consecutive monthly samples or was from
the herd. Single samples of foremilk were col-         mammary glands of cows with clinical masti-
lected aseptically from all quarters of lactating      tis. Abnormalities of the udder, such as swell-
cows monthly for 15 mo and from cows on                ing or hardness, and changes in the appearance
two occasions during the first 10 d after partu-       of the milk, such as flakes, clots, or watery
rition. Before sample collection, teats of cows        secretion, were criteria used by milkers to de-
were cleaned thoroughly and dried with single-         tect clinical mastitis. A quarter was eligible for
service paper towels, and teat ends were sani-         only one IMI per bacterial species during a
tized with swabs containing 70% isopropyl              lactation. Teats of cows were examined for
alcohol. All samples were collected prior to           irritation, chapping, or other abnormalities
premilking teat disinfection.                          throughout the study. Differences between
    Foremilk (10 pl) from each quarter was             treatment groups were determined by, Student’s
plated onto one quadrant of a brain-heart infu-        t test.
sion agar plate (Gibco Laboratories, Madison,
WI) supplemented with 5 % defibrinated sheep                               RESULTS
blood. Plates were incubated at 37’C, and bac-
terial growth was observed and recorded at                 Efficacy of chlorous acid and chlorine diox-
24-h intervals for 3 d. Bacteria on primary             ide as a premilking teat disinfectant is shown
culture medium were ideRtified tentatively ac-          in Table 1. New IMI by Staphylococcus aureus
cording to colony morphologic features, hemo-           (P e .OS) and Streptococcus species (P e .025)
Journal of Dairy Science Vol. 76, No. 1, 1993
                                             OUR INDUSTRY TODAY                                                    289
TABLE 1. Efficacy of a teat dip containing chlorous acid and chlorine dioxide as a premilking teat disinfectant for the
prevention of new IMI.

                                           Treatment                                   New IMI
Organism                                   group                            Quarters                        Reduction
                                                                (n)                                 (%)
Staphylococcus aureus                      Control'              11                     1.3                 69.2
                                           Redip2                 3                       .4a
Streprococcus species                      Control               38                     4.6                 44.6
                                           Predip                21                     2.5b
Gram-negative bacteria                     Control               37                     4.4
                                           Predip                21                     3.2
Other                                      Control               11                     1.3
                                           Predip                13                     1.6
All major pathogens                        Control               91                    11.6                 34.0
                                           Predip                64                     7.T
Staphylococcus species                     Control               43                     5.1
                                           Predip                33                     4.0
Corynebacterium bovis                      Control               42                     5.0
                                           Predip                33                     4.0
All minor pathogens                        Control               85                    10.2
                                           Predip                 66                    7.9
All major and minor                        Control               182                   21.8                 28.6
 pathogens                                 Redip                 130                   15.6c
   %fferent from controls (P e .05).
   bDifferent from controls (P < .025).
   CDifferent from controls (P e .01).
   In = 835 quarters available for IMI.
   2n = 834 quarters available for IMI.




and percentage of quarters newly infected by                    Mastitis pathogens isolated from mammary
major mastitis pathogens (P < .01) were lower                glands of cows with clinical mastitis are
in quarters with teats predipped and postdipped              presented in Table 4. A total of 125 cases of
than in quarters with teats postdipped only.                 clinical mastitis were observed: 72 in post-
Redipping and postdipping treatment was no                   dipped and 53 in the predipped and postdipped
more effective against Gram-negative bacteria,               groups. Differences in the incidence of clinical
coagulase-negative Staphylococcus species,                   mastitis between the two treatment groups ap-
and C. bovis than postdipping only.                          proached significance (P < .lo). Clinical
   A heterogeneous mixture of Sfreptococcus                  mastitis was caused by several different masti-
species was isolated from mammary glands of                  tis pathogens. Most clinical mastitis was
cows with subclinical and clinical mastitis (Ta-             caused by E. coli, Klebsiella species, and
ble 2). Most Streptococcus species IMI were                  Strep. uberis. About 84% of E. coli, 91% of
caused by Streptococcus uberis (46%) and                     Klebsiella species, and 56% of Strep. uberis
Streptococcus dysgalactiae (3 1 %). New IMI                  IMI detected during lactation were of clinical
by Strep. uberis were lower (P < .025) in                    origin. Mastitis pathogens were not isolated in
quarters with teats predipped and postdipped                 23.2% of samples obtained from cows with
than in quarters with teats postdipped only.                 clinical mastitis.
   Most new Gram-negative IMI were caused
by Escherichia coli (39%) and Klebsiella spe-                                     DISCUSSION
cies (45%) (Table 3). Redipping and postdip-
ping treatment was no more effective against                    Remilking and postmilking teat disinfec-
Gram-negative mastitis pathogens than post-                  tion with the chlorous acid-chlorine dioxide
dipping only.                                                teat disinfectant, in association with good
                                                                       Journal of Dairy Science Vol. 76, No. 1, 1993
290                                             OLIVER ET AL.
                                                        M
TABLE 2. Distribution of Sfreprococcus species causing I I
                                                    ~




                                                   Treatment
Organism                                           group                             M
                                                                                New I I in quarters
                                                                          (n)                         (96)
Srreprococcus uberis                               Control]               20                          2.4
                                                   Predip2                 7                           .Sa
Srreprococcus dysgalactiae                         Control                11                          1.3
                                                   Predip                  7                           .8
Srreprococcus equinus                              Control                 1                           .I
                                                   Predip                  2                           .2
Other Streprococcus species                         oto
                                                   Cnrl                    2                           .2
                                                   Pttdip                  1                           .1
Enterococcus species                               Control                 4                           .5
                                                   Predip                  4                           .5
All Srreprococcus species                          Control                38                          4.6
                                                   Predip                 21                          2.5a
   aDifferent from controls (P < .05).
   ln = 835 quarters available for IMI.
   2n = 834 quarters available for IMI.




udder preparation, was significantly more ef-              To date, effectiveness of premilking teat
fective against IMI caused by major mastitis            disinfection has been described (10) using low
pathogens than good udder preparation and               concentration, iodine-based teat disinfectants
postmilking teat disinfection. No chapping or           only. Results of that study showed that
imtation of teats was observed, and no adverse          premilking teat disinfection significantly
effects were noted. Results of the present study        reduced the rate of new IMI by environmental
provide additional evidence to support the con-         streptococci and coliforms. The chlorous acid-
cept that premilking teat disinfection is an            chlorine dioxide teat disinfectant evaluated in
effective management procedure for the                  the present study was significantly more effec-
prevention of I M I during lactation.                   tive as a premilking and postmilking teat disin-



TABLE 3. Distribution of Gram-negative bacteria causing IMI.
                                                   Treatment
Organism                                           group                        New IMI in quarters
                                                                          (n)                          S)
                                                                                                      (b
Escherichia coli                                   Control1               16                          1.9
                                                   Predipz                 9                          1.1
Klebsiella pneumoniae                              Control                14                          1.7
                                                   W P                     9                          1.1
Klebsiella oxyroca                                 Control                 2                           .2
                                                   Predip                  4                           .5
Serratia species                                   Control                 4                           .5
                                                   Redip                   3                           .4
Enferobacfer cloacae                               Control                 1                           .I
                                                   Redip                   1                           .1
Pseudomonas aeruginosa                             Control                 0                          0
                                                   Predip                  1                           .1
All Gram-negative bacteria                         Control                37                          4.4
                                                   Predip                 27                          3.2
   In = 835 quarters available for MI.
   2n = 834 quarters available for M .
                                    I

Journal of Dairy Science Vol. 76, No. 1. 1993
                                              OUR INDUSTRY TODAY                                           29 1
TABLE 4. Pathogens isolated from mammary glands of          different teat disinfectants need to be con-
cows with clinical mastitis.
                                                            ducted in herds with a high prevalence of
                                       Experimental         contagious mastitis to verify results observed
                                            PrOUD           in the present trial and to provide additional
Organism                          Control           pndip   data documenting the effectiveness of this
Escherichia coli                  13                 8      procedure against Streptococcus agahctiue.
Klebsiella pneumoniae              9                 6         Based on present research, premilking ud-
Klebsiella oxytoca                 2                 4      der hygiene relates critically to new IMI by
Serratia species                   3                 1      environmental and contagious mastitis patho-
Pseudomonas aeruginosa             0                 1
                                                            gens (3, 10, 11). Premilking teat disinfection in
Streptococcus uberis              11                 4
Streptococcus dysgalactiae         3                 4      association with good udder preparation likely
Staphylococcus species             4                 0      reduces bacterial populations on teat skin be-
Yeast                              3                 5      fore milking, which subsequently influences
Bacillus species                   1                 2      the ability of environmental and contagious
Streptomyces species               0                 2
Corynebacterium species            1                 0      pathogens to invade the mammary gland and
Mixed                              5                 4      to cause IMI during the milking process. Fu-
Clinical-negative'                17                12      ture studies on premilking teat disinfection
 oa
Ttl                               72                53'     could provide important information regarding
   'Different from controls (P < .lo).                      factors that are associated with mastitis patho-
   'Clinical mastitis-bacteriological negative.             gen transmission during the milking process
                                                            and could delineate possible differences be-
                                                            tween contagious and environmental patho-
                                                            gens.
fectant against Sfreprococcus species, particu-
larly Srrep. uberis. However, predipping and                                 CONCLUSIONS
postdipping treatment was no more effective
against Gram-negative bacteria than postdip-                   Premilking and postmilking teat disinfec-
ping only. Thus, different premilking teat dis-             tion with the chlorous acid-chlorine dioxide
infectants may not be equally effective against             teat disinfectant, 4XLA@, was significantly
the vast array of pathogens capable of causing              more effective in preventing new IMI by major
IMI. Furthermore, all teat dips should not be               pathogens than postdipping only. New IMI by
assumed to be safe and effective as a premilk-              Srrep. uberis and Sraph. aureus were signifi-
ing teat disinfectant.                                      cantly lower in quarters with teats predipped
    New IMI by Sraph. aureus were signifi-                  and postdipped than in quarters with teats post-
cantly lower in quarters with teats predipped               dipped only. Redipping and postdipping treat-
and postdipped in the chlorous acid-chlorine                ment was no more effective against Gram-
dioxide teat disinfectant than in quarters with             negative bacteria, coagulase-negative Sraphy-
teats postdipped only. Staphylococcus aureus                lococcus species, and C. bovis t a postdip-
                                                                                              hn
                                                            ping only. Differences in incidence of clinical
is a contagious mastitis pathogen that is trans-
mitted primarily during milking (2, 9). Post-               mastitis between treatment groups approached
                                                            significance.
milking teat disinfection has been effective
repeatedly in preventing new Staph. aureus
                                                                         ACKNOWLEDGMENTS
IMI (6, 7 , 8, 9). Results of the present study
suggest that a further reduction in Staph.                     This study was supported by Alcide Corp.
aureus IMI can be attained by premilking teat               (Norwalk, CT); the Tennessee Agricultural Ex-
disinfection in association with postmilking                periment Station; and The University of Ten-
teat disinfection, which agrees with a recent               nessee, College of Veterinary Medicine, Center
report by Sears et al. (11). Thus, premilking               of Excellence Research Program in Livestock
teat disinfection, which was developed as a                 Diseases and Human Health. We express ap-
potential method to control environmental                   preciation to personnel at the Knoxville Ex-
mastitis, would likely be effective in herds                periment Station dairy for technical assistance
with contagious mastitis. However, further                  and to Teresa Herring and Nikki Bell for cleri-
studies on premilking teat disinfection using               cal assistance.
                                                                   Journal of Dairy Science Vol. 76, No. I. 1993
292                                                OLIVER ET AL.

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