The Blue Cross Book. 2003, 21: 19-20.

Efficiency of fluroquinolones against repeat breeding cows.

G.N. Purohit, R.K. Chaturvedi, K.A. Gupta, K. Vyas, A.K. Gupta, N. Garg, P.K. Pareek and S.S. Sharma Department of Veterinary Obstetrics and Gynaecology College of Veterinary and Animal Science, Bikaner- 334 001 (Raj.)

ABSTRACT
Uterine flush samples of 16 (88.89%) out of 18 repeat breeding cows evincing no clinical pathology yielded more than one type of microorganisms whereas only 2 of the 6 normal breeding cows yielded (33.33%) isolates. E.Coli (29.41%) was the predominant bacteria isolated, followed by Corynebacterium and Staphylococcus sps. (17.65%, each), Pseudomonas sps. (14.70%) and Streptococci sps. in repeat breeding cows. In vitro sensitivity revealed that most of the isolates were sensitive to fluroquinolones (88% ciprofloxacin; 83.33% to pefloxacin) whereas 72.22%; 22.23%, 16.66% and 11.11% isolates were sensitive to oxytetracycline, ampicillin and penicillin. For confirmation of the laboratory results In vivo, 16 of these confirmed cases and 23 clinical cases were administered the two fluroquinolones (ciprofloxacin and pefloxacin) and the routinely used oxytetracycline under 5 day (schedule A) and single time treatment 12 hours post A.I. (schedule B). conception rates at 60 days post revealed that the best in cirpofloxacin treated animals (64.70%) in 2 groups followed by pefloxacin (60%) under 2 groups and oxytetracycline (28.57%). The conception rates were far better in treated than in untreated animals.

Introduction Sub-clinical uterine infection is thought to be one of the important cause of repeat breeding in dairy cattle. the infection often remains undiagnosed clinically but the altered uterine milieu often hampers the implantation/grown of embryo resulting in repeated estrous cycles. Singh et al.(1996) conducted trials on recent antimicrobials but found that most of the isolates were resistant to antimicrobials like cloxacillin. Fluroquinolones (ciprofloxacin, pefloxacin etc.) are an exceptionally important and rapidly developing group of antimicrobial drugs introduced into medicine for a wide variety of antimicrobial sensitivity. The present study was conducted to examine the in-vitro and in vitro efficiency of fluroquinolones against uterine microbes and their comparison with other antimicrobials. MATERIALS AND METHODS 18 Rathi and its crossbred cows belonging to LRS, Bikaner and not having any palpable or clinical pathology were included in the study. During estrus the uterus of these animals was flushed using foley catheter and Ringers lactate as flushing medium. The foley catheter was fixed at the uterine body and Ringer lactate was infused into the uterus and collected back by 2 way catheter. the flush was subjected to organism culture, identification and antibiotic sensitivity test as per method of Kirby et al.(1966). The zone of inhibition was measured and the results interpreted.

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On the basis of In-vitro sensitivity, 16 of these animals were administered the most suitable antibiotic and additionally 23 clinical cases were also assigned one of the 3 antibiotics namely ciprofloxacin, pefloxacin and oxytetracycline administered into uterine. Two schedules of treatment were used. In schedule A the animals were administered the antibiotic by intrauterine route daily for 5 days starting from the day of estrus and then inseminated during the subsequent estrus. In schedule B, the animals were inseminated and then treated with suitable antibiotic 12 hrs post Al. In all 7 animals (group-I) were given oxytetracycline treatment under schedule A using 2000 mg of the drug dissolved in 20 ml of d.w. 7 animals (Group-II) were treated using Pefloxacin (800 mg dissolved in 20 ml of d.w.) under schedule B and 8 animals (Group-III) were given pefloxacin using schedule A. 8 Animals were given ciprofloxacin treatment under schedule B (Group-IV) and 9 animals were given ciprofloxacin under schedule A (Group-V). The quantity of antibiotic used was 1000 mg dissolved in 20 ml of d.w. Six normally cycling animals formed the control group in which although some microbes were isolated but no treatment was given. All the animals treated or untreated were inseminated using semen from fertile bull and followed up for pregnancy diagnosis 60 days post-insemination. RESULTS AND DISCUSSION Bacterial Culture and Organism identification : Organisms were isolated from 16 out of 18 samples from the experimental animals while 2 were bacteriologically sterile. In control group organisms were isolated from 2 (33.33%) out of 6 samples while 4 were sterile. Out of the 34 isolates thus obtained from experimental animals the per cent frequencies of various bacterial species isolated in decreasing order were E.Coli (29.41%), Corynebacteria and Staphylococcus sps (17.65% each), Psuedomonas sps (14.70%) and Streptococci sps (11.76%) while in control group the per cent frequencies of bacterial species isolated were E.Coli (5.89%) and Staphylococcus (2.94%). Results are similar to those of Deshmukh and Markandeya (1995) and Ramakrishna (1996). It was found in general that the uterus of repeat breeding cows whether they evidence clinical signs or not harbours some aerobic microbes which may in some form alter the uterine environment leading to impaired fertility. Most of the repeat breeding cows had mixed infections with more than one type of microbes rather than a single microorganisms. This finding is similar to that of Das et al.(1996).

IN-VITRO TESTS
The results of in-vitro tests on the isolates are presented in Table 1 which reveals that 88.89%, 83.33%, 72.22%, 22.23%, 16.66% and 11.11% of the isolates were sensitive to perfloxacin, gentamycin, oxytetracycline, ampicillin and penicillin. all the isolates were sensitive to more than one antibiotic while two samples were negative to the sensitivity test. The analysis of in-vitro antibiotic sensitivity test revealed that ciprofloxacin was the most effective antibiotic while penicillin was the least effective. Money (1996) and Gupta et al.(1997) had reported similar results. Gentamicin was the second most effective antibiotic as reported earlier by Ramakrishna (1996), Singh et al. (1996) and others, while oxytetracycline was found to be least effective as reported earlier by Ramaswamy et al.(1991).

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IN-VIVO STUDIES The detailed results of antimicrobial therapy in vivo are presented in Table 2. Out of the 39 animals treated, 22 animals conceived giving an overall conception rate of 56.41% compared to 33.33% in untreated controls, in which only 2 animals out of the 6 conceived. This conforms that the treatment has a distinct advantage over non-treatment and this finding is in agreement with Sharma et al.(1988) and Satish Kumar (1990). The conception rates were best in the ciprofloxacin groups (Groups IV and Group V) 64.70% followed by pefloxacin 60% (Group II and III) and oxytetracycline 28.57%. The poor response to oxytetracycline could be because the drug is the most commonly used agent through various routes which might have led to the development of resistant strains. Ramaswamy et al.(1991) have also reported that penicillin and tetracyclines are least effective drugs in repeat breeding cows. Table 1 : Antibiotic sensitivity pattern from repeat breeder cows on the basis of zone of inhibition
SENSITIVITY PATTERN Sample No. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. Per cent Antibiotic Concentration (mcg/disc) +++ : Above ++ : Between Oxytetracycline + 0 + ++ 0 ++ 0 + + + 0 0 ++ ++ 0 + + 0 22.23 30 mcg Penicillin + 0 ++ + + ++ 0 + 0 + 0 0 + + 0 0 + 0 11.11 10 IU Ampicillin + 0 ++ + + 0 0 ++ 0 ++ 0 0 + + + + 0 + 16.66 10 mcg Gentamycin +++ 0 +++ +++ ++ ++ 0 ++ +++ +++ +++ + +++ +++ + ++ +++ + 72.22 10 mcg Pefloxacin +++ 0 +++ +++ ++ +++ 0 ++ +++ +++ +++ + +++ +++ +++ ++ +++ +++ 83.33 5 mcg Ciprofloxacin +++ 0 +++ +++ +++ ++ 0 +++ ++ +++ +++ +++ ++ +++ +++ +++ +++ +++ 88.89 10 mcg

28 mm + : Between 14-20 mm 21-27 mm 0 : Between 0-14

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Table 2 : Clinical trial results of various intrauterine antibiotic therapies in repeat breeding Rathi cows and their crossbreds harbouring subclinical infection.
Group No. of Animal Treated 7 7 8 Schedule followed A/B A B A Antibiotic used Oxytetracycline Pefloxacin Pefloxacin Overall IV V 8 9 B A Ciprofloxacin Ciprofloxacin Overall VI 6(control) NONE --No. of animals conceived in 1-2 insemination 2 3 6 9 4 7 11 2 % conception 28.57 42.85 75.00 60.00 50.00 77.77 64.77 33.33 1000 mg 1000 mg Quantity of anti-biotic used 2000 mg 800 mg 800 mg

I II III

A - treated for 5 days starting from day of estrus B - single treatment given 12 hours post AI REFERENCES Das, K.L.; Misra, P.R.; Kar, B.C. and Roy, P.K. (1996) :Aerobic bacterial isolates and their antibiogram in repeat breeding cows. Indian Vet. Jrnl. 73(8) : 900-902. Deshmukh, V.V. and Markandeya, N.M. (1995) : A study of antibiotic sensitivity pattern of bacterial isolates from repeat breeding cows. International Jrnl. Ani. Sci. 10 : 337-338. Gupta, J.; Dabas, Y.P.S. and Maurya, S.N. (1997) :Therapeutic efficacy of some antimicrobial drugs in repeat breeding cows. Indian. Jrnl. Ani. Reprodn. 18(2) : 163-164. Kirby, W.M.M.; Bauer, A.w.; Sherris, J. C. and Turak, M. (1996) : Antibiotic sensitivity tests. Am. Jrnl. clinical Path. 45: 493. Maurya, S.N.; Dabas, Y.P.S. and Gupta, R.S. (1992) : A note on bacteriological studies of cervical secretion of infertile cows and buffaloes. Indian Jrnl. Ani. Reprodn. 13(1) : 49-50. Money, I.A.L. (1996) : Efficacy of pefloxacin treatment for endometritis in a cow. Indian Vet. Jrnl. 73(10) : 1072-1073.

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Prescott, J.F. (1994) : Antimicrobial therapy in Veterinary Medicine. J. Desmond Baggot. First Indian edition. International Book Distribution Co. Lucknow. Ramakrishna, K.V. (1996) : Microbial and biochemical profile in repeat breeder cows. Indian Jr. Anim. Reprodn. 17(1) : 30-32. Ramaswamy, V.; Andrew, M.; Venugopalan, A.T.; Roy, P and Chandramohan, C.P. (1991) : Aerobic microbes of cervico-vaginal mucus from repeat breeder bovines and their antibiogram. Singapore Vet. J. 14/15 : 60-65 (Cited I Vet. Bull. (1993), 63(4) : 392 : 2623. Satish, Kumar (1990) : Gentamycin therapy for repeat breeding bovines. Indian J. Vet. Med. 10(1) : 47-48. Sharma, R.N.; singh, B.K. and Singh, M.P. (1988) : Bacteriological studies on the cervical mucus of repeat breeding crossbred cattle, their treatment and conception rate. Indian J. Anim. Reprodn. 9(2) : 105-109. Singh, N.P.; Chaturvedi, V.K. and Singh, D.P. (1996) : Bacteriological studies on repeat breeder bovines. Indian Vet. J. 73(4) : 462-463.