Use of dry period intervention is one of the of import constituents of an effectual mastitis control programm. The present survey was planned to measure the comparative efficaciousness of Terramycin and enrofloxacin as systemic dry period therapy in the control of bubaline mastitis. For this intent, twenty seven pregnant dry American bisons were selected and divided into three equal groups. Animals in group G1 were treated with Enrofloxacin ( Inj. Encure10TM ; Nawan Laboratories, Pakistan ) @ 2.5 mg/kg organic structure weight intramuscularly at 14 yearss and 7 yearss prior to expected day of the month of birth. Group G2 were treated with Oxytetracycline-HCl ( Inj. Oxy-KakTM LA ; Kaksian, Pakistan ) @ 11 mg/kg organic structure weight intramuscularly at 14 yearss and 7 yearss prior to expected day of the month of birth and group G3 served as non medicated control. Samples of mammary secernments were collected aseptically 14 yearss prior to expected calving and milk samples from each one-fourth aseptically collected at twenty-four hours 7 and 14 station calving. The efficaciousness of interventions was evaluated through prevalence of clinical and subclinical mastitis before, at and after birth and bacteriological remedy rate. Prevalence of clinical mastitis and one-fourth wise prevalence of subclinical mastitis station calving after systemic dry period therapy with enrofloxacin and Terramycin were lower than control group. Postpartum remedy rate of septic quarters at twenty-four hours 14 were 91.67 % with enrofloxacin intervention groups, 70 % with oxytetracycline intervention groups and 21 % with control groups. The remedy rate among the groups was significantly different ( p & lt ; 0.05 ) . The highest remedy rate belonged to enrofloxacin group which was significantly higher than that of control ( p & lt ; 0.05 ) , followed by Terramycin group which was besides significantly higher than that of control ( p & lt ; 0.05 ) ; nevertheless there were no important difference between enrofloxacin groups and oxytetracycline groups ( P & gt ; 0.05 ) . Results suggested that dry period antibiotics therapy offers the best chances to take bing, relentless intramammary infections and forestalling new intramammary infections.
Cardinal Wordss: American bison, Mastitis, dry period therapy,
Mastitis is one of the most economically of import diseases of milch animate beings and besides causes the alterations in glandular tissues impacting quality and measure of milk ( Ullah et al. , 2005 ) .
Mastitis can be prevented when an effectual control plan is adopted. Dry period therapy is an indispensable portion of this control plan. The end of mastitis control during the dry period is to hold as few septic quarters as possible at the following calving ( Eberhart, 1986 ) . At present, the most effectual agencies of accomplishing these aims is to administrate dry period antibiotic therapy during dry period. ( Cousins et al. , 1980 ; Radostitis et al. , 2007 ) . The usage of antibiotic dry cow therapy and the intervention of intramammary infection during dry period has been a footing for mastitis direction and control ( Bradley, 2002 ) . Dry cow therapy eliminates bing intra-mammary infections about 70 to 98 % during the dry period and forestalling new intra-mammary infections by 50 to 75 % ; a cardinal portion of a successful mastitis control programme ( Eberhart, 1986 ; Janosi and Huszenicaza, 2001 ; Petzer et al. , 2009 ) .
Remedy rate is higher in dry period than during lactation, higher concentration of drugs can be used, cut down new intramammary infections during dry period, damaged tissue may renew anterior to break uping, incidence of clinical mastitis at caving is besides reduced ( Nickerson and Owens, 1994 ) . During the dry period, riddance of infection with an antibiotic is easier than during lactation as the drug is non milked out and a higher and more unvarying concentration of antibiotics is maintained in the bag. In add-on there are no economic losingss due to flinging of antibiotic incorporating milk ( Sandholm et al. , 1995 ) . Dry cow antibiotic intervention is more efficacious than lactation therapy and has less hazard of milk residue ( Berry and Hillerton, 2002 ; Bradley and Huxley, 2003 ) .
The intramammary path is the path of pick for bringing of dry period therapy and has the advantages of being good reached. Its disadvantages are the hazards of both physiological and anatomical harm to the streak canal and vaccination of beings at the clip of extract ( Bradley and Huxley, 2003 ) . Systemic dry period therapy may hold advantages, better distribution of drug in the bag tissue which may take to better remedy of intra-mammary infections ( Ziv, 1980 ) and turning away of new infections which is possible hazard at disposal of intramammary extract ( Boddie and Nickerson, 1986 ) . Systemic disposal could simplify dry cow therapy modus operandi. It would besides extinguish the hazard of presenting infections through non-sterile intramammary injection ( Soback, 1988 ) . Systemic disposal of antibiotics at drying off or some hebdomads before birth seems to be effectual intervention for intra-mammary therapy, which may advisable for pattern ( Nickerson et al. , 1999 ; Bolourchi et al. , 1995 ; Zecconi et al. , 1999 ) . The most efficacious clip for intervention seems to be 7 to 14 yearss prior to the expected calving. Treatments earlier than 14 yearss prior to break uping may let clip after the intervention for new intramammary infections to happen prior to break uping ( Oliver et al. , 2003 )
Materials and Methods
A sum of 20 seven dry pregnant American bisons were selected from the different farm animal farms and indiscriminately allocated to three equal groupaa‚¬a„?s viz. G1, G2 and G3. Samples of mammary secernments were collected aseptically 14 yearss prior to expected calving. Collection of mammary secernments from each one-fourth was done aseptically harmonizing to National Mastitis Council guidelines ( Hogan et al. , 1999 ) . Each teat terminal was scrubbed smartly with cotton gauze soaked with 70 per centum ethyl intoxicant. Immediately following mammary secernments aggregation, antibiotic interventions were given to the animate beings. Animals in group G1 were treated with Enrofloxacin ( Inj. Encure10TM ; Nawan Laboratories, Pakistan ) @ 2.5 mg/kg organic structure weight intramuscularly at 14 yearss and 7 yearss prior to expected day of the month of birth. Group G2 were treated with Oxytetracycline-HCl ( Inj. Oxy-KakTM LA ; Kaksian, Pakistan ) @ 11 mg/kg organic structure weight intramuscularly at 14 yearss and 7 yearss prior to expected day of the month of birth and group G3 served as non medicated control.
Ten milliliter of milk samples from each one-fourth aseptically collected at twenty-four hours 7 and 14 station calving. The gathered samples will be shifted to microbiological research lab, college of Veterinary and Animal Sciences, Jhang for isolation and biochracterisation of prevailing mastitis pathogens. Procedure described by National Mastitis council Inc. , USA ( 1990 ) was followed for the aggregation of milk sampling stations. Sterile phial of 15 milliliters capacity were used. Each teat terminal was scrubbed smartly with cotton gauze soaked with 70 per centum ethyl intoxicant. A separate swab was used for each nipple. While keeping the phials every bit horizontal as possible, the cap was removed without touching interior surface and held with the interior surface facing downwards.
Precalving secernment samples and postcalving milk samples were processed for bacteriology within 24 hr of aggregation following storage at 4AA°C. Microbiological process described by National Mastitis Council Inc. , USA ( 1990 ) was followed for culturing the samples and designation of mastitis pathogens. The mastitic milk samples were gently shaken 8 times to acquire a unvarying scattering of the pathogens. Using a platinum-rhodium cringle, 0.01 milliliter of milk sample was streaked onto blood agar home bases. Four one-fourth milk samples were cultured on a 100 millimeter home base by plating single one-fourth sample on one quarter-circle of home base and were incubated at 370 C for 48 hours. A one-fourth was considered to be infected if 5 or more similar settlements were present on home base. ( Roberson et al. , 1988 ) The one-fourth should be considered septic if milk secernments samples contain one of mastitis pathogens before intervention. The absence of the same bacterium at trying 7 and 14 yearss following birth were interpreted as a bacteriological remedy otherwise in the presence of the bacteriums the mammary one-fourth remained uncured. An clean one-fourth at drying off that was infected at calving was considered to bespeak new intramammary infections. A one-fourth that was infected at drying off but septic with another being at break uping besides indicated new intramammary infections.
The cultural and morphological features of primary growing were studied by scrutiny of settlement features and readying of vilifications from different settlements. These vilifications were stained with Gramaa‚¬a„?s staining method and examined under the microscope. The primary growings were purified by frequent bomber culturing on selective and differential media. The selective and differential media used were MacConkeyaa‚¬a„?s agar ( for streptococcic species ) and blood agar ( haemolytic species ) .
Each of the isolate was identified on the footing of cultural and morphological features, motility, haemolytic and biochemical belongingss as described ( Cruickshank et al. 1975 ) . Catalase positive. Gram positive coccal isolates presumably identified as staphylococci or Micrococcus. The genus of bacteriums determined on the footing of settlement morphology, Gram discoloration, haemolysis form, catalase trial, and esculin reaction. Gram-positive, catalase-positive isolates were farther tested utilizing a tube-coagulase trial. Coagulase-positive isolates were defined as Staphylococcus aureus and coagulase-negative isolates as CNS. Gram-positive, catalase-negative isolates were CAMP-tested: esculin-positive and CAMP-positive or negative isolates were defined as Strep. uberis ; esculin-negative and CAMP-negative isolates were defined as Streptococcus dysgalactiae ; and esculin negative and CAMP-positive isolates were defined as Streptococcus agalactiae. Gram-negative rods were subcultured on MacConkey. Corynebacterium spp. was defined as little Gram-positive rods organized in typical “ pallisades ” . Organism other than staphylococcuss identified by everyday biochemical trials ( National Mastitis Council Inc. , 1990 ) .
Percent prevalence of mastitis was calculated in all groups. Cure rate of septic one-fourth among groups were calculated utilizing chi square trials comparing treated groups and the control. Each intervention group was besides compared with other group and with the control group utilizing two relative Z trial. P & lt ; 0.05 considered as important and P & gt ; 0.05 considered as non important ( Steel et al. , 1997 ) .
Prevalence of clinical mastitis at birth
Out of entire 27 animate beings 3 ( 11 % ) were clinically mastitic at birth. Among these no animate being were positive in enrofloxacin group ( 0 % ) , one in oxytetracyclin group ( 11.11 % ) and two in control group 2 ( 22.22 % ) were positive. Ocular scrutiny of bag and nipples followed by tactual exploration revealed that udder one-fourth were asymmetric due to swelling clinical symptoms swelling of udder presence of seeable alterations ( e.g. , coagulums or stain ) of mastitic milk, hurting, heat, and soundness were observed. Out of 144 quarters 7 ( 9.72 % ) were infected. No one-fourth was infected at birth in enrofloxacin group ( 0 % ) , two quarters were infected in oxytetracyclin group ( 5.56 % ) , and in control group 5 ( 13.89 % ) were positive for mastitis. Percent Prevalence of clinical mastitis at birth is higher in control group as compared to dry period antibiotics intervention groups.
4.2 Prevalence of subclinical mastitis after birth
Out of 27 animate beings 7 ( 25.93 % ) were subclinical mastitic after birth at twenty-four hours 7 station calving. Among these with enrofloxacin intervention were 1 ( 11.11 % ) , with oxytetracycline 2 ( 22.22 % ) and in control group 4 ( 44.44 % ) as depicted in Table 4.3. One-fourth wise prevalence of subclinical mastitis was 14 ( 38.89 % ) after birth at twenty-four hours 7 station calving. Among these with enrofloxacin intervention group were 1 ( 2.78 % ) , with Terramycin were 3 ( 8.33 % ) and in control group 10 ( 27.78 % ) quarters were infected. Consequences showed that prevalence of subclinical mastitis is higher in control group as compared to intervention groups. Similarly one-fourth wise prevalence of subclinical mastitis at twenty-four hours 14 station calving was 11 ( 30.56 % ) . Among these with enrofloxacin intervention group were 1 ( 2.78 % ) , with Terramycin were 3 ( 8.33 % ) and in control group 7 ( 19.44 % ) quarters were infected. Animals that had no discernible mastitis symptoms in milk or mammary lobes while one or more querters detected as infected with mastitis pathogens were classified as holding subclinical mastitis
4.3 Prevalence of clinical mastitis after birth
Out of 27 animate beings merely 2 ( 7.41 % ) were holding clinical mastitis after birth at twenty-four hours 7. Among these no animate being was infected with clinical mastitis in enroflaxacin and oxytetracycline intervention group. No one-fourth was infected clinically in these groups. In control group 2 ( 8.33 % ) animate beings clinically positive. Neither any carnal nor any one-fourth was found to be infected at 14 yearss post break uping.
4.4 Postpartum remedy rate of septic quarters
Postpartum remedy rate of septic quarters at twenty-four hours 14 were 91.67 % ( 11/12 ) with enrofloxacin intervention groups, 70 % ( 7/10 ) with oxytetracycline intervention groups and 21 % ( 3/14 ) with control groups. The remedy rate among the group were significantly different ( p=0.001, P & lt ; 0.05 ) . The highest remedy rate belongs to enrofloxacin group which was significantly higher than that of control ( p=0.0002 P & lt ; 0.05 ) , followed by Terramycin group which was besides significantly higher than that of control ( p=0.008 P & lt ; 0.05 ) ; nevertheless there were no important difference between enrofloxacin groups and oxytetracycline groups ( p=0.90 P & gt ; 0.05 ) as depicted in Table 4.8
The present survey was the first, to the best of our cognition designed to compare the efficaciousness of Terramycin and enrofloxacin as systemic dry period therapy in the control of bubaline mastitis. This survey was performed on 20 seven dry pregnant American bisons. This was based on aims that dry period antibiotics therapy before birth eliminates the bing intramammry infections and forestalling new intramammary infections. Prevalence of clinical mastitis and one-fourth wise prevalence of subclinical mastitis station calving after systemic dry period therapy with enrofloxacin were 0 % and 2.78 % which was lower than control group. Prevalence of clinical mastitis and one-fourth wise prevalence of subclinical mastitis station calving after systemic dry period therapy with Terramycin were 11.11 % and 8.33 % which is lower than control group. In control group, prevalence of clinical mastitis and one-fourth wise prevalence of subclinical mastitis station calving were 22.22 % and 27.78 % which is higher than systemic dry period intervention groups. Cure rates of septic quartes after parental enrofloxacin were 91.67 % , with Terramycin were 70 % and in control group self-generated were 21 % . These findings are consistent with Soback et Al. ( 1990 ) who observed similar findings that systemic dry cow therapy utilizing norfloxacin nicotinate, which possesses big distribution volume, long half life, and is extremely active against the pathogen involved, was more effectual than the other interventions. Both enrofloxacin and oxyteracycline are good distributed into the organic structure fluids and should be able to make the site of infection. But enrofloxacin has better consequences than Terramycin. In other survey, Hovareshti et Al. ( 2007 ) acquire similar consequences with injection of tylosin intramuscularly 83 % , with intramuscularly enrofloxacin injection 70 % while comparing with dry cow intramammary readyings 93 % . However dry cow intramammary readyings were the most effectual methods to bring around the intramammary infections. However in big dairy herds, it is about impractical, tendios, unsafe and non expectable by the husbandmans. Additionally, this may present environmental bacteriums and fungi into the quarters by insanitary use of bag. The hazard of antibody residue was lower in systemic intervention as compared to intramammary extracts. Similarly better efficaciousness obtained with systemic dry cow therapy by ( Smith and Hogan 1995 ; Marco, 1996 ) . There was no important difference on remedy rate % of intramammary infections between intramammary and systemic antibiotics combinations, systemic antibiotic merely or between systemic antibiotic and intramammary antibiotic merely.
Systemic antibiotics dry period therapy can be cost effectual particularly in herds where subclinical mastitis is a chronic job. Our consequences, nevertheless, agree with old studies utilizing a macrolide antibiotic, tylosin, 2 hebdomads before the expected twenty-four hours of calving by ( Zecconi et al. 1999 ) . Systemic disposal of antibiotics could simplify dry period therapy modus operandi. It would besides extinguish the hazard of presenting infections through non-sterile intramammary injection. Systemic dry period therapy had so far reported inconsistent consequences with ( Soback et al. 1990a ; Erskine et Al. 1994 ; Smith and Hogan, 1998 ; Nickerson et Al. 1999 ; Zecconi et Al. 1999 ) . In contrary to this survey Shpigel et Al. ( 2006 ) observed really low remedy rate after systemic cefquinome intervention was comparable to the self-generated remedy rate observed in untreated controls in old surveies. The unfavourable consequences of the cefquinome systemic dry period therapy might reflect unequal pharmacokinetic belongingss of the drug sing hapless udder incursion in subclinical mastitis and short antimicrobic consequence. In another survey Erskine et Al. ( 1994 ) compared efficaciousness of intramuscular Terramycin as a dry period intervention for Staphylococcus aureus mastitis. Cure rate of septic one-fourth with oxytetracycline intervention group were ( 34 quarters, 10 remedies ) 29.4 % ) 30 twenty-four hours station calving and 21.2 % ( 7 of 33 ) of the quarters were considered to be cured, 60 yearss station calving and got inferior consequence from the present survey. One ground might be, they used dry period antibiotics therapy at drying of, dry period length may be long and carnal regain infections during dry period. Similarly Nickerson et Al. ( 1994 ) compared parenteral disposal of dry cow therapy by hypodermic injection of tilmicosin at 5 mg/kg of organic structure weight at dry-off and once more 4 yearss subsequently was uneffective against intramammary infections caused by Staphlococcus aureus ( 9.1 % remedy rate ) . Parker et Al. ( 2008 ) used of an internal nipple canal sealer and nipple dipping as dry period therapy in heifers to cut down the station break uping intramammary infections prevalence and pathogen associated with clinical mastitis by diminishing the incidence of new infections over high hazard peripartum period. It may be a utile method for forestalling new intramammary infections but it can non bring around bing intramammary infections.
In the present survey prevalence of staphylococci aureus, Streptococcus agalactiae, E. coli, coagulase negative staphylococcus and corynebacterium were at twenty-four hours 7 and 14 postpartum were 0 % 0 % , 0 % , 2.78 % and 0 % severally in enrofloxacin groups. Cure rate of staphylococci aureus, Streptococcus agalactiae, E. coli, and corynebacterium were 100 % while in coagulase negative staphylococcus was 66.66 % . In oxytetracycline groups prevalence of staphylococci aureus, Streptococcus agalactiae, E. coli, coagulase negative staphylococcus and corynebacterium station calving were 5.56 % , 2.78 % , 0 % , 2.78 % , and 0 % severally. Remedy rate staphylococci aureus was 66.66 % , Streptococcus agalactiae 0 % , E. coli 100 % , and coagulase negative staphylococci 50 % . In the control group prevalence of staphylococci aureus, Streptococcus agalactiae, E. coli, coagulase negative staphylococcus and corynebacterium station calving were 19.44 % , 8.33 % , 5.56 % , 5.56 % and 2.78 % severally. Spontaneous remedy rate of staphylococci aureus was 12 % , Streptococcus agalactiae 0 % , E. coli 0 % , coagulase negative staphylococci 33.33 % , and corynebacterium 0 % . Almost similar remedy rate obtained Petzer et Al ( 2009 ) utilizing intramammary merchandise containing ( Cephalexin 250 milligram and neomycin sulfate 250 milligram ) . The remedy rate of varied from 94.4 % for Staphylococcus aureus, 100 % for Streptococcus agalactiae and Streptococcus dysgalactiae, 78.1 % for coagulase negative staphylococcus and 100 % for the other minor pathogens. The one-fourth new infection rate during the dry period was 17.44 % in control groups. These consequences are about similar with the present survey.
In the present survey, four new infections ( 11.11 % ) were besides occurred in the control group but no new infections appeared in the treated group. These findings consistent with Natzke ( 1971 ) that the development of new one-fourth infections during the dry period without dry period therapy, are 10 to 15 % . These infections can cut down milk production by 36 % during the first month of the subsequent lactation. Approximately 33 to 50 % of the subclinical infections rates, postpartum consequence in clinical symptoms of mastitis during the lactation ; 50 % of these new infections occur in cattles that were antecedently infected in at least one one-fourth ( Healed et al. , 1977 ) . Osteras et Al. ( 1991 ) recorded new intramammary infections rates during the dry period of between 13.1 % and 24.0 % . However, most other surveies which were in understanding with the findings of present survey described lower rates of between 4 % and 14 % ( Natzke, et al. , 1975 ; Oliver, 1988 ; Schukken, et al. , 1993 ; Smith et al. , 1985 ; Williamson et al. , 1995 ) . The expected rate of new intramammary infections during the dry period in bacteriums negative quarters that were untreated was reported by Eberhart ( 1986 ) to change between 8 % and 12 % while Berry and Hillerton ( 2002 ) reported new infection rates of 34.4 % in untreated cattles compared with 10.3 % in treated cattles. The decrease of new intramammary infections during the dry period with dry period therapy has been estimated at between 50 % to 80 % ( Eberhart, 1986 ) .
The optimal production and maximal day-to-day output of milk can merely be achieved if mastitis is prevented at herd degree by following guidelines of mastitis control plan. The dry period offers the best chance during the lactation rhythm to take bing, relentless intramammary infections, though unluckily this chance is tempered by an increased hazard of new intramammary infections. Dry period antibiotics therapy reduces this hazard. Dry period antibiotics therapy has the double intent of intervention and bar. Systematic dry period antibiotics therapy is recommended for herds with a high infection rate. It is better, easy to follow and friendly to husbandmans. Dry period antibiotic therapy is besides suggested in herds with low bodily cell counts and low prevalence of contagious mastitis pathogens, to minimise the new dry period infections by environmental pathogens which can ensue in a high incidence of clinical mastitis in the early lactation. Dry period antibiotic therapy administered within 1 to 3 wk before expected calving might hold best consequences in our conditions. In order to accomplish the best remedy rates it is of import to choose an appropriate antibiotic that have high soaking up in bag.
Table 4.8: Day 14 station break uping one-fourth based remedy rate of infections treated with enrofloxacin and oxytateracycline
Entire Number of Quarter
Number of Quarter Infected Before Treatment
Number of Quarter Cured
Cured Rate % age