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, 1993,12 (3), 909-922
Successful therapeutic regimens for treating Brucella melitensis and Brucella abortus infections in cows
A.I. RADWAN *, S.I. BEKAIRI *, A.M. AL-BOKMY *, P.V.S. PRASAD *, O.M. MOHAMED ** and S.T. HUSSAIN **
Summary: Three therapeutic regimens were evaluated in 121 cows naturally infected with Brucella melitensis or Brucella abortus, using a combination of long-acting Oxytetracycline (LA-OTC), streptomycin (ST) and OTCintramammary infusion (IMI). Cessation of shedding of b r u c e l l a in udder secretions and absence of Brucella in selected tissues were considered criteria for successful treatment. Regimen A (tested on 35 cows) consisted of LA-OTC 25 mg/kg administered intramuscularly (i.m.) every 3 days for 42 days, ST 25 mg/kg i.m. daily for 8 days, and OTC-IMI 20 ml/teat daily for 4 days. Regimen B (tested on 53 cows) was similar to regimen A, except that ST was administered every 2 days for 16 days and OTC-IMI every 2 days for 8 days. Both regimens were equally effective in eliminating Brucella organisms from all cows involved in the tests and no relapses were recorded. However, regimen C, which was similar to regimen A, except that ST was administered every 3 days for 24 days and OTC-IMI every 3 days for 12 days, resulted in the elimination of Brucella organisms from only 30 (91%) of 33 cows. Before commencement of the therapeutic regimens, B. melitensis biovar 1 or 2 had been repeatedly isolated from udder secretions of 103 cows and B. abortus biovar 1 from mammary secretions of 18 cows. KEYWORDS: Antibiotics - Brucella abortus - Brucella melitensis - Cattle Oxytetracycline - Streptomycin - Therapy.
INTRODUCTION Several c h e m o t h e r a p e u t i c agents have b e e n employed in recent decades for the treatment of Brucella abortus infection in cows; however, n o n e of these has been entirely successful. N u m e r o u s chemical agents, general antimicrobials (phenols or dyes), trace elements, minerals and mixtures of vitamins (A and E ) have b e e n tried unsuccessfully. F u r t h e r m o r e , a t t e m p t s using antibiotics such as penicillin or sulfonamides failed to cause cessation of the shedding of B. abortus from the mammary secretions of infected cows, or caused only temporary cessation (7,11,37).
* Animal Production and Health Section, National Agriculture and Water Research Centre, Ministry of Agriculture and Water, P.O. Box 17285, Riyadh 11484, Saudi Arabia. ** National Agriculture Development Co., P.O. Box 2557, Riyadh 11461, Saudi Arabia.
melitensis. However. the regimens previously tried involved small doses and few injections of the antibiotics used.m. the present study was undertaken to evaluate the efficacy of three long-term therapeutic regimens using a combination of LA-OTC. using high doses of L A . the use of these agents alone or in combination with ST has succeeded in eliminating the symptoms of this disease and reducing the shedding of brucellae by infected cows. Furthermore. In one of these studies. without side-effects and relatively inexpensive. In this trial. alone or in combination with ST. . In a n o t h e r study.13. terramycin. The aim of the study was to obtain a therapeutic regimen which is effective. Radwan and colleagues (35) evaluated six different long-term treatment regimens on 480 sheep and goats naturally infected with B. Moreover. injections of L A .O T C (20 mg/kg intramuscularly [i. the presence of antibiotic residues in milk and the failure to cure udder infections in many cases have led to the general conclusion that such treatment is unsuitable for the control of bovine brucellosis. Only one sheep was found to be bacteriologically positive at slaughter. singly or in combination. daily for 6 weeks) combined with ST (1 g i. has resulted in the reduction of abortions in infected herds or individual cows (20. inoculation with OTC. In addition.O T C (20 mg/kg i. tetracyclines and streptomycin (ST). the cost of therapy.m. every 2 days for 2 weeks) proved to be the most practical.29). such therapeutic regimens have been applied in infected herds to prevent abortions and reduce the spread of brucellosis.] or i.m.4%) of 14 cows (24).) inoculations with O T C (12. safe. practical. In addition. daily for 3 weeks) showed a synergistic effect.p. melitensis or B. serious local reactions were reported in cows.p. every 3 days for 6 weeks. these regimens have failed to result in complete cure. One of these regimens ( L A . increasing the percentage of Brucella-tree sheep to 82%.V. on 118 sheep naturally infected with B.] every 3 days for 2 weeks) combined with ST (20 mg/kg i. by the i.O T C 25 mg/kg i. abortus were injected daily for 6 days with 1 g Chlortetracycline.000 mg LA-OTC i. However.500 or 1.m. 46 days after infection (30). every 2 days for 4 weeks combined with ST 20 mg/kg i. effective and least expensive means of achieving complete elimination of B. 69% and 100% of the sheep were found to be Brucella-îre-c at the end of the trial. With regard to brucellosis in sheep.O T C combined with ST. Radwan and colleagues (34) conducted a long-term treatment trial using a variety of doses of O T C . then 20 days later with 1.p. melitensis in the 80 sheep and goats treated (35).m. In this trial.910 The use of broad-spectrum antibiotics such as aureomycin. 52%. 6 sheep were Brucella-iiee. daily for 1 week) resulted in cessation of shedding of B. abortus in 10 (71. when L A . However. ST and OTC intramammary infusion (IMI) in eliminating B. melitensis. Furthermore. Consequently. 9 sheep experimentally infected with B. when 8 infected Najdi sheep were inoculated with 1. In view of the encouraging results in cows (24.000 mg O T C daily for 6 weeks. in order to make treatment of Briicella-infected cows with superior genes a viable alternative to slaughter.p.21). groups of infected Najdi sheep were injected with 250. daily for 1 week) 14 (67%) of 21 cows w e r e successfully treated (25).m. abortus from naturallyinfected cows. every 3 days for 2 weeks) alone was successful in curing 3 (21%) of 14 cows.O T C was combined with ST (20 mg/kg intravenously [i. In the respective groups.m. due to repeated intraperitoneal (i. With the development of Oxytetracycline (OTC) and long-acting (LA)-OTC. route. treatment with OTC (250 mg i.4 g t o g e t h e r with i m m u n e serum for 3 days. the majority of the treated sheep developed subcutaneous sterile abscesses in the flank at the site of repeated i.24. Recently.25) and the successful results in sheep and goats (35).p. inoculation of L A .25. at necropsy (34).
In the third herd.22. particularly because L A . date of recent pregnancy and n u m b e r of previous abortions due to Brucella. B and C).p r e g n a n t cows were artificially inseminated. The 121 cows were divided into three treatment groups (A. B. These animals were selected to provide a m o r e conclusive evaluation than that obtained in previous studies. melitensis. and selected tissue samples were cultured. these were placed in g r o u p B together with 35 cows with B. The reason for the division into three treatment schedules was to choose the most practical and effective regimen. melitensis was isolated from udder secretions of 103 cows (biovar 1 from 97 cows and biovar 2 from 6 cows). eight years prior to the present study. 89 cows were found not to be pregnant (having already aborted or calved 4-8 months before t r e a t m e n t ) and 32 cows were in various stages of pregnancy. abortus was small. T h e animals belonged to two breeds (108 Holstein x Friesian and 13 Jersey).28). Three months later. all n o n . Shortly before initiation of the present treatment regimens. number of previous pregnancies. melitensis (biovars 1 and 2) which had been introduced to the two farms 8-12 m o n t h s before bovine brucellosis occurred in the two cattle herds (unpublished findings). Therapeutic agents The pharmaceutical products used in this study were as follows: . respectively .O T C is a long-acting antibiotic. All cows in the study were isolated from other animals. in contrast to ST and OTC-IMI which are not. Non-treated cows were not included as controls. each containing l g or 4 g. immediately after birth. as several previous studies have indicated that Brucella infections persist in cattle and s p o n t a n e o u s recovery is uncommon (10.911 MATERIALS AND METHODS Animals A total of 121 cows were selected. without regard to age. The three herds were composed of highly valuable breeding cows originally imported from the United States of America (USA) and Europe.ST sulphate (from Giza. In two herds. a total of 3 calves (1 from a cow in each treatment group) were sacrificed.O T C injectable solution (from F i a n c e ) containing 200 mg O T C base p e r millilitre . One m o n t h after t r e a t m e n t . In addition.L A . and selected tissues were collected for bacteriological examination. E g y p t ) supplied in vials. dissolved (just before use) in 3 ml or 12 ml sterile distilled water. approximately four years prior to this study. The cows were aged 3-10 years and weighed between 450 kg and 750 kg. abortus biovar 1 was isolated from the mammary secretions of 18 cows. B. a total of 12 pregnant cows (4 from each t r e a t m e n t group) were sacrificed. which gave positive results for brucellosis after serological and bacteriological tests. T h e cows contracted brucellosis from sheep and goats infected with B. These two herds originally tested seronegative on arrival in Saudi A r a b i a . Milking was ceased in lactating cows. Herd records indicated that none of the cows had been vaccinated during calfhood. Since the number of available cows infected with B. The cows originated from three large commercial dairy herds raised in the central and eastern provinces of Saudi Arabia. This herd contained few seropositive cows on arrival in Saudi A r a b i a .
brain. 5 mg Prednisolone and 10 ml special excipient. Bacteriological examination T h e m e d i u m for Brucella culture was p r e p a r e d as described previously (35). 100 mg neomycin sulphate. Each tissue specimen was individually homogenised in a tissue grinder and aliquots were spread with sterile cotton swabs on several (4-8) freshly prepared plates of culture medium. Iowa. cultures of udder secretions were m a d e from the selected 121 positive cows to confirm shedding of Brucella organisms. Therapeutic regimens The treatment regimens of the three groups of infected cows are shown in Table I. New Haw. In addition. milk samples from all seropositive cows were subjected to bacteriological examination for identification of Brucella shedders. Furthermore.OTC-IMI (from Athens. from the 12 pregnant sacrificed cows (4 cows from each treatment group which were slaughtered at 4 months post-treatment) the following tissue specimens were aseptically collected for bacteriological examination: supramammary. The L A . udder secretion. USA. iliac. uterus. 100 mg oleandomycin p h o s p h a t e . Similar selected tissue specimens were cultured from 3 sacrificed newborn calves (1 from a cow in each treatment group). thigh and shoulder muscles. In addition. animals were initially screened for the presence of Brucella agglutinins by the rose bengal test (3). precrural. T h e rose bengal test antigen was obtained from the Central Veterinary Laboratory. Immediately before initiation of the therapeutic regimens. lung and spleen of foetuses from the sacrificed animals were cultured. A t each examination. each containing 200 mg OTC hydrochloride. Agglutination at 1:100 or greater was considered positive. Both ST and OTC-IMI were administered with L A . every month after completion of treatment until calving and again every 2 weeks after calving for 6 m o n t h s . at a dosage of 20-30 ml per site. liver. mesenteric. liver and spleen. United Kingdom. in the cervical. mediastinal.O T C and ST were injected i. and sections of udder. Aliquots from each udder secretion sample were spread with sterile cotton swabs on 2-4 freshly prepared plates of culture medium.m. ovary. popliteal and head lymph nodes. In addition. samples of stomach contents. abortus strain 1119-3) was obtained from the United States Department of Agriculture in Ames. Serological testing All animals were subjected to three successive serological examinations at monthly intervals.912 .and bacteriologically-positive cows were serologically tested again immediately before initiation of the present treatment regimens and at monthly intervals after completion of treatment (for 4 months in 12 sacrificed cows and for 16 months in the remaining 109 cows). prescapular. sera from the selected 121 serologically. Separate 30 ml samples of udder secretions from each quarter were also cultured every week during treatment. The plates were incubated at 37°C for 7 days in the presence and absence of 10% C 0 a t m o s p h e r e .O T C at the c o m m e n c e m e n t of therapy. The standard Brucella plate antigen (made of B. Following the three serological examinations. Animals testing positive to the rose bengal test were retested by the standard US plate agglutination procedure for the determination of Brucella antibody titres (3). sera from all calves born from treated cows were examined immediately after birth. The date of previous abortions in relation to the initiation of treatment was not recorded on the farms involved. Weybridge. T h e isolated 2 . Greece) in 10 ml syringes.
No of Success/treated Tissuesa)Udder secretions < > b e i.m.m.m. 25 mg/kg IMI 20 ml/teat 1 e B 8 14 49/49(100%) 35(c) ST(f) 18( ) 14 8 4 3/4 d LA-OTC( ) i. 25mg/kg IMI 20 ml/teat 3 3 27/29(91%) i.m.: IMI: a) b) c) d) e) f) g) intramuscular (injection) intramammary infusion number of sacrificed cows from which selected tissues were cultured for Brucella number of cows from which mammary secretions were repeatedly cultured for Brucella cows naturally infected with B. melitensis cows naturally infected with B.m. 25mg/kg &OTC-IMlig) IMI 20 ml/teat 25mg/kg 2 2 3 4/4 4 e C 3 33W LA-OTC( ) STW &OTC-IMl(s) i.m. abortus from naturally-infected cows Regimen FA v 3 No.TABLE I Efficacy of three treatment regimens in eliminating Brucella melitensis or B. abortus long-acting Oxytetracycline injectable solution (from France) 200 mg/ml streptomycin sulphate (from Egypt) Oxytetracycline intramammary infusion (from Greece) 913 .of Periodicity treated Therapeutic agents cows Route Dosage E ™° Y ^¿J.m. i. 1 4 8 4/4 31/31 (100%) 3 14 A 35(c) LA-OTC( ) ST( ) & OTC-IMI(g) f 25mg/kg i. 25mg/kg i.
The biotyping of the identified isolates was performed at the Central Veterinary Laboratory.O T C and ST. aborted foetuses from the three cattle herds involved were cultured for Brucella isolation. microscopically. Furthermore. there were unusually high rates of metritis. However. New Haw. Moreover. Weybridge. . The abortion rate in the three herds ranged between 5% and 1 5 % and the infertility r a t e was approximately 1 5 % . retained placenta and mastitis. B and C experienced n o r m a l calving within 4 m o n t h s following completion of the treatment. United Kingdom. Of the cows treated with regimens A. salmonellosis. Side-effects of the therapeutic agents In 8 cows. F u r t h e r m o r e . or the use of c h e m o t h e r a p e u t i c measures. there was very light swelling and signs of painful sensations at the site of r e p e a t e d L A . with milk production reduced by 10-15%. in addition to routine bacteriological examination for listeriosis. the cows had b e e n living in an infected environment for periods of between 1 and 8 years. mycoplasmosis and mycotic abortion. On the whole. particularly in the thigh muscles. biochemically and serologically (3). in the breeding season following treatment. the respective cow was considered to be cured (successful t r e a t m e n t ) . and the average cost of the therapeutic agents alone used (regimen A.914 Brucella cultures were identified morphologically. RESULTS Breeding performance A comparison was made between breeding performance in the three herds prior to identification of the infected cows and that in the treated animals in the breeding season following treatment. No abortions were observed a m o n g the treated cows. Before t r e a t m e n t . no systemic reactions were observed in any of the treated cows during the treatment period. When Brucella organisms could no longer be recovered from udder secretions nor from any of the selected tissue specimens collected at necropsy. In addition. B or C) in treating a cow weighing 600 kg was US$186 (SR700). B and C. Cost of therapeutic agents and animals The weight of the treated cows ranged between 450 kg and 750 kg. In this study. the cows were living in an uninfected environment. and milk production was within the normal range for uninfected animals of the same breed. C o n t i n u e d or r e s u m e d shedding of Brucella organisms in udder secretions or the isolation of Brucella organisms from any of the tissue specimens obtained at necropsy were considered as conclusive evidence of treatment failure. the 32 cows which were in various stages of pregnancy at the time of initiation of the therapeutic regimens A. T h e s e slight local reactions disappeared within 36 hours.O T C injections. Meat inspection of the twelve cows which were slaughtered (4 months post-treatment) revealed complete absence of any detectable abnormalities in the muscles at the site of intramuscular inoculations of L A . there were no cases of metritis or mastitis in the treated cows. the treated cows expelled placenta from the womb without requiring any special manual manipulation. 94% b e c a m e p r e g n a n t and experienced n o r m a l calving.
including the cost of air freight to Saudi Arabia. they ceased shedding Brucella. abortus from 121 naturally-infected cows is shown in Table I. melitensis and 18 infected with B. all selected tissue samples from 4 slaughtered cows and the repeated udder secretion samples were found to be Brucella-free. All selected tissue specimens from the 3 newborn calves (1 calf from a cow in each treatment group) were found to be free from Brucella organisms. abortus w e r e isolated. udder tissue and supramammary lymph nodes of only 1 of 4 sacrificed cows (at 4 months post-treatment). when these 2 Brucella-shedding cows were re-treated with regimen B. However. However. Bacteriological findings In the present study. in the 3 cows of regimen C where t r e a t m e n t failed. There was a decrease of two. shedding of Brucella organisms in mammary secretions had ceased when these secretions were cultured (1 week after initiation of the antibiotherapy) and did not recommence. melitensis or B.600 each. In all successfully treated cows in the present study (118 cows treated with regimen A. When 53 cows (of which 35 were naturally infected with B. no Listeria. at a cost of US$3. only B. B or C). abortus) were treated with regimen B. B. when 33 cows (naturally infected with B. the selected tissue specimens from 4 sacrificed cows (at 4 months posttreatment) and all repeated udder secretion samples from the remaining 31 cows (for 16 months post-treatment) were all found to be free from Brucella organisms. T h e efficacy of the t h r e e t r e a t m e n t regimens applied in eliminating B. Serological findings Among the treated animals. there was a decrease of only one dilution in the US plate agglutination test (from 400 to 200) from the beginning of t r e a t m e n t until 4 months post-treatment in 4 of 12 slaughtered cows. However. the amount of shedding declined after the initiation of treatment and remained low until 4 months post-treatment in 1 slaughtered cow. In regimen A. Salmonella. melitensis biovar 2 was isolated (1-5 colonies per plate) from mammary secretions. . while all calves born from treated cows were seronegative for brucellosis. The Brucella agglutinin titres w e r e also c o m p a r e d with results o b t a i n e d p r e treatment and 16 months post-treatment in the remaining 109 treated cows. melitensis biovar 1 was isolated from the repeated udder secretion samples from only 2 of 29 cows. only 5% of the 109 cows became seronegative 16 months posttreatment. melitensis) were treated with regimen C.915 The treated cows were imported a few years previously as pregnant heifers. melitensis or B. The average number of Brucella colonies recovered per plate from the udder secretion samples of these 2 cows prior to treatment was approximately 30 times higher than the number recovered following treatment. Mycoplasma or pathogenic fungi were isolated from the aborted foetuses examined in the three herds. t h r e e and four dilutions in 4 2 % . In addition. which ranged between 2 and 8 colonies per plate. 4 3 % and 10% of the treated cows respectively. In addition. or after calving in the other 2 cows (these animals were then re-treated with regimen B). B. T h e remaining 8 slaughtered cows did not show any decrease in titre over the same period.
none has been fully successful.916 DISCUSSION In order to increase milk production. with the development and widespread use of techniques such as artificial insemination and embryo transfer. bone marrow. abortus. A n effective. However. 2 and 3 were responsible for all infections in sheep. abortus for 8%. Such infection was directly responsible for the high rate of abortions. B. except cows in one dairy herd which had B. particularly in lymph nodes. liver. Although this has been the goal of several workers. melitensis biovars 1. but from the subsequent problems or irregular breeding. 35. metritis. infertility. melitensis infection from sheep and goats raised on the same farms. On the other hand. namely: the Netherlands. T h e infected vaccinated animals continue to present a serious public health risk. camels and dairy cattle. Slaughter of infected animals with superior genetic potential has serious economic and genetic consequences (24). melitensis accounted for 92% of human cases and B.31.33. An eradication campaign through slaughter and compensation is not within the economic scope of developing countries. the . mass vaccination of infected herds protects only uninfected animals without altering the course of infection. B.32. In addition.16. loss of milk production. in B. however. Germany and France.23. abortus. Consequently.39). while B. it is impossible to apply only one method which would suit every country. In the present study. cattle with superior genetic potential have become increasingly valuable. the cattle were raised in large n u m b e r s u n d e r intensive management systems and sometimes together with small ruminants. Brucellosis has been reported and confirmed in both livestock and humans in Saudi Arabia. Livestock producers in Saudi Arabia and in many other developing countries cannot afford the traditional "test and slaughter" approach used in developed countries. not just for abortion or stillbirth. leading to serious consequences for cattle performance and cattle handlers. suis was not found in animals or humans (1. two of the cattle herds contracted B. T h e following explanations were given for the previous t r e a t m e n t failures and difficulties: a) Brucella organisms are known to survive within phagocytic cells of the reticuloendothelial system. one worker assessed the average cost of an infected cow at approximately US$3. abortus infection.34.5. The "test and slaughter" m e t h o d combined with calfhood vaccination no doubt contributes to considerable reduction in the number of infected herds. On arrival in Saudi Arabia. practical and safe antibiotherapy would be of e n o r m o u s benefit to producers in these countries as an alternative to slaughter of infected animals of high value.19. goats.18. spleen. especially where it is difficult to change agricultural customs or social habits. In a campaign against brucellosis.6. commercial dairy production was introduced to Saudi Arabia through the importation of valuable and highly productive exotic cattle at a very high cost. These cattle were originally imported as p r e g n a n t heifers from the USA and some E u r o p e a n countries. retained placenta and mastitis with reduced milk production.4.2. Moreover.200. it is recognised that the economic consequences of infection are e n c o u n t e r e d principally during initial infection and become less prominent in further pregnancies. Transmission of this infection to cattle p r o d u c e d an infection similar to that p r o d u c e d by B. for economic reasons this method cannot be applied in all cases. and the reduced h u m a n productivity resulting from a spread of infection (38).
e) Use of the intraperitoneal route for administration of antibiotics created serious reactions in some cases (17.37). O T C proved to be effectively bactericidal when combined with 10 µg/ml of ST (15. p e r m i t t e d growth or was bacteriostatic for brucellae within tissue cells. This may be due to the'fact that both ST and OTC-IMI are .0 µg/ml. L A .O T C or ST was used alone. abortus.O T C combined with ST was used for the t r e a t m e n t of cows naturally infected with B. when administered every 3 days in regimen C.34).26). OTC-IMI was used locally in combination with systemic (i. Since successful therapy in human beings appears to depend on the permeability of cell walls to drugs (24).25. g) High cost of antibiotherapy (24.25. in order to decrease the n u m b e r of Brucella organisms in u d d e r tissues and secretions.26.25.26. The present study aimed to overcome the above problems and involved work under particularly difficult conditions with regard to the level of infection among the cows used. This c o m b i n a t i o n p r o v e d to be highly effective in eliminating Brucella organisms from all infected cows in regimens A and B.11.13.29).24). c) Insufficient doses of effective antibiotics (12. a combination of L A . and the possible harmful effect on consumers (27). 25. F u r t h e r m o r e .) inoculation of L A .25.12. b) Incorrect choice of antibiotics (7. However. In addition.917 mammary glands. Consequently. h) Failure to cure udder infections in many cases (24. ST acts synergistically with O T C to inhibit growth of B.13. ST was also used. i) Presence of antibiotic residues in milk and m e a t of t r e a t e d animals.O T C and ST. melitensis (35). O T C at 0. as one injection gave an effective concentration of 0. O T C . and are therefore p r o t e c t e d from antibodies. which were all serologically and (particularly) bacteriologically positive. For these reasons. c o m p l e m e n t and antibiotics. reproductive organs and tissues with a weak blood supply. Consequently. d) Insufficient duration of treatment (8. For this reason. T h e ability of Brucella organisms to survive intracellularly can result in long-term chronic infection.5-1. abortus within bovine cells cultured in vitro.6 ug/ml for three days (26).24.O T C and ST proved very effective in curing sheep and goats naturally infected with B.24.24.25. the O T C base was used because it is capable of penetrating intracellularly and inhibits bacterial protein synthesis at the level of the ribosomes.n e g a t i v e bacteria.24.25. relapses after antimicrobial t r e a t m e n t w e r e also explained by the intracellular location of brucellae (9.25) on the treatment of bovine brucellosis indicated poor success rates when L A .I M I and ST.m. f) Use of only one antibiotic (20. In addition. melitensis or B. in infected cows. the following points were taken into account when attempting to obtain more successful results. 36). were less effective in eliminating Brucella organisms in infected cows.34).O T C and ST.26). 34. despite the use of a combination of L A . However. because it is known to inhibit p r o t e i n synthesis of G r a m .29). However.21. early studies (20. The L A .O T C variant was selected to save time and effort and to provide long-lasting O T C concentration in blood plasma.26). before initiation of the present treatment regimens. milking was stopped in lactating cows and udders were completely evacuated before each injection. several authors failed to cure udder infections in many cases (24. Furthermore.
relapses did not occur in any of the 88 cows treated with regimen A or B (provided that the treatment protocol is strictly observed). inoculation of L A .75).O T C did not result in harmful systemic effects or any obvious lasting local reactions at the site of inoculation. inoculation of OTC was not safe. In Saudi Arabia. Even in relapsed cows (3 animals in regimen C).12. only a small n u m b e r of Brucella colonies were recovered from large tissue samples. In the present study. Also. as animals were only monitored for 16 months post-treatment. In conclusion. the cost of supplies and intervention should be considered. however.29). further research may yield a regimen . This did not include the cost of e q u i p m e n t or interventions. although O T C has been considered the most effective antibiotic for treatment of brucellosis.600. However. a small number of relapses do occur. T h e average cost of the antibiotics used in regimen A. the majority (approximately 90%) of the cows treated were monitored bacteriologically for 16 months post-treatment in order to detect possible relapses. this treatment would probably be limited to valuable breeding animals. relapses occurred in only 3 of 33 cows treated with regimen C. The cost of successful treatment of a cow represents approximately 5% of the original price of the animal. In the present trial. 34) concluded that i. Relapses have also been reported in cows (14. consequently.25. However. which is considered economically feasible in Saudi Arabia.13. without side-effects and relatively inexpensive in causing cessation of the symptoms of brucellosis and eliminating Brucella organisms from infected cows. However. and the protection of Brucella organisms in the presence of milk. The cost of antibiotics used was calculated according to sale prices in the Saudi market at the time of the experiment (US$1 = SR3. the treated cows were originally imported as pregnant heifers at an average cost of US$3. regimens A and B proved to be effective. practical. The present results of serological examinations were of some use in evaluating the effectiveness of the treatment regimens. all t r e a t e d cows were dried before initiation of treatment.p.24. there was no risk of harmful effects on h u m a n beings from antibiotic residues in the milk from these cows during and after treatment.24. In the present study.26. i. and t h e r e was no nontreated control group for comparison.I M I alone would be effective for the treatement of lactating cows due to the continuous flushing of OTC by milking. However. the residues of antibiotics in milk and meat of treated animals should be investigated to determine accurately when milk and/or meat can be safely consumed following completion of the treatment regimens. B or C for treating a cow of an average weight of 600 kg was US$186 (SR700). because all serological and bacteriological examinations were performed free of charge at Government institutions. Previous studies in cows and sheep (17. reported that serological findings were not useful in evaluating the effectiveness of treatment regimens (24.25). at the present time. when treatment protocols were strictly observed. due to the development of several serious local reactions at the site of injection. However. Consequently. it seems unlikely that O T C . The doses and duration of application of selected antibiotics in the present study indicate that the long-term therapy and the drug combinations used yielded better results than several other regimens (8. including the cost of air freight.m.34). Other investigators. T h e results obtained were in agreement with findings on successful treatment of brucellosis in sheep and goats when monitored for 8 months post-treatment (35).918 not long-acting antibiotics. In humans. in other situations. the results were not conclusive. In the present study.
for his encouragement and co-operation.T. it is concluded that the failure of a small number of cows to respond to regimen C cannot be accounted for by t h e d e v e l o p m e n t of resistance by Brucella. S.M. abortus dans celles de 18 vaches. Prasad.S. A. Bekairi. Quant au protocole C. pratiquée également tous les trois jours pendant 12 jours. Hussain.) quotidiennement pendant huit jours et en une OTC-IMI quotidienne de 20 ml par trayon pendant quatre jours. combinant à la fois l'Oxytétracycline à action prolongée (long-acting Oxytetracycline : LA-OTC). il n'a permis l'élimination des brucelles que sur 30 (91 %) des 33 vaches traitées. D i r e c t o r General of the Saudi National Agriculture and Water Research Centre and National Director of Project 016/UTFN/SAU/FAO. Deux critères ont été jugés déterminants pour conclure au succès du traitement : l'arrêt de l'excrétion de brucelles dans les sécrétions mammaires et l'absence de brucelles dans des tissus spécifiques. .M. Le protocole B (appliqué à 53 vaches) était similaire au précédent. similaire au protocole A. Thanks are also extended to the owners and veterinary staff of the three dairy farms involved. hormis pour la ST qui était administrée tous les trois jours pendant 24 jours et l'OTC-IMI. Avant l'application de ces trois protocoles thérapeutiques. Mukayel and A . ACKNOWLEDGEMENTS The a u t h o r s wish to express their sincere t h a n k s to M. Al-Bokmy. The authors also acknowledge the technical assistance of A .I.m. de 25 mg/kg de ST (i. melitensis ont été régulièrement isolés dans les sécrétions mammaires de 103 vaches et le biotype 1 de B.S. for their s u p p o r t and c o . pratiquée un jour sur deux pendant huit jours. Radwan.) de 25 mg/kg de LA-OTC tous les trois jours pendant 42 jours. * * * DES PROTOCOLES THÉRAPEUTIQUES EFFICACES POUR LE TRAITEMENT DE L'INFECTION PAR BRUCELLA MELITENSIS ET BRUCELLA ABORTUS CHEZ LES VACHES.A. Le protocole A (appliqué à 35 vaches) consistait en une administration intramusculaire (i. Furthermore. les bio types 1 ou 2 de B.919 which could b e used for the t r e a t m e n t of all infected animals in a h e r d and for prophylaxis of those animals recently exposed to Brucella infection. sauf pour la ST qui était administrée un jour sur deux pendant 16 jours et pour l'OTC-IMI. Al-Taher. P. .I. Les deux modes de traitement se sont révélés aussi efficaces l'un que l'autre pour l'élimination totale des brucelles des vaches objet de l'expérimentation et aucune récidive n'a été constatée. T h e t r e a t m e n t of seropositive animals should be performed together with vaccination of all seronegative animals in the same herd to achieve effective control and minimise public health risks. Bin Salamah.V.o p e r a t i o n in t h e i m p l e m e n t a t i o n of the p r e s e n t treatment regimens. O. Mohamed et S. la streptomycine (ST) et l'infusion intramammaire d'OTC (OTC-intramammary infusion : OTC-IMI). Résumé : Trois protocoles thérapeutiques ont été évalués chez 121 vaches atteintes d'une infection naturelle par Brucella melitensis ou Brucella abortus.m.
que fue experimentado en 35 vacas. .Brucella abortus . . . (1988).Brucella melitensis .G. A.m. .Brucella abortus .Brucella melitensis . 7. Institut National de la Recherche Agronomique.E. & V E R G E R J .D.) de 25 mg/kg de LA-OTC cada tres días durante 42 días. SINOUSSI Y .Terapéutica.351-357.920 MOTS-CLÉS : Antibiotiques .M.T. CHEN M .Streptomycine . un 91%. abortus en las secreciones mamarias de 18 vacas. a partir de una combinación de oxitetraciclina de acción prolongada (long-acting Oxytetracycline: LA-OTC). A N G U S R.. Prasad. que se experimentó en 53 vacas. 2. Radwan. Al-Bokmy. sólo permitió eliminación de brucelas en 30 de las 33 vacas tratadas. 190 pp. ALTON G..Oxytétracycline .S.. * PROTOCOLOS TERAPÉUTICOS EFICACES PARA EL TRATAMIENTO DE LA INFECCIÓN POR BRUCELLA MELITENSIS Y BRUCELLA ABORTUS EN VACAS. Uno y otro tratamiento se mostraron equivalentemente eficaces para la eliminación total de las brucelas en las vacas tratadas y no se constató ninguna recidiva.. Antes de aplicar estos tres protocolos terapéuticos. S. Por último. & HARLAND P. que se administró cada dos días durante 16 días.S. M .Brucellosis in children. 6. El protocolo A. Hussain. es decir. .I. C . Saudi biol.m.. Ann.I.A study on brucellosis among animals of some farms and rural areas of Saudi Arabia.) diarios durante ocho días y en una OTC-IMI diaria de 20 ml por pezón durante 4 días. era similar al anterior excepto en cuanto a la ST. Mohamed y S. Paris.R.H. PALABRAS CLAVE: Antibióticos .G.M E Z A I N I S. . (1984). HAWARI A.Y.Bovinos . AL-MARSHEDY A. Para considerar el tratamiento exitoso se tuvieron en cuenta dos criterios: el cese de la excreción de brucelas en las secreciones mamarias y la ausencia de brucelas en tejidos específicos. (1986). P. de 25 mg/kg de ST (i. Bekairi. CHANG S. O. melitensis fueron regularmente aislados en las secreciones mamarias de 103 vacas y el biotipo 1 de B.Bovins . El protocolo B. REFERENCES 1. estreptomicina (ST) e infusión intramamaria de OTC (OTC-intramammary infusion: OTC-IMI). Proc.Oxitetraciclina . . A L .25-27. los biotipos 1 o 2 de B. Soc. WANG C F . el protocolo C.Estreptomicina .M.V. y ala OTC-IMI.Thérapie. & CHOU N.A. Resumen: Se evaluaron tres protocolos terapéuticos en 121 vacas infectadas por vía natural por Brucella melitensis o Brucella abortus. consistía en la administración intramuscular (i. que se administró cada dos días durante ocho días. 3. M . JONES L .Techniques for the brucellosis laboratory. similar al protocolo A salvo en que la ST se administraba cada tres días durante 24 días y la OTC-IMI también cada tres días durante 12 días. Saudi Med.
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