Rev. sci. tech. Off. int. Epiz.

, 1993,12 (3), 909-922

Successful therapeutic regimens for treating Brucella melitensis and Brucella abortus infections in cows

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.

treatment with OTC (250 mg i.O T C combined with ST. abortus from naturallyinfected cows.m. the present study was undertaken to evaluate the efficacy of three long-term therapeutic regimens using a combination of LA-OTC. 52%.m. the majority of the treated sheep developed subcutaneous sterile abscesses in the flank at the site of repeated i. inoculation of L A . Furthermore. The aim of the study was to obtain a therapeutic regimen which is effective.4%) of 14 cows (24). every 2 days for 2 weeks) proved to be the most practical. every 2 days for 4 weeks combined with ST 20 mg/kg i. daily for 6 weeks) combined with ST (1 g i. every 3 days for 6 weeks.m. terramycin. the regimens previously tried involved small doses and few injections of the antibiotics used. . In the respective groups. safe.] or i.m. ST and OTC intramammary infusion (IMI) in eliminating B.m. groups of infected Najdi sheep were injected with 250. In a n o t h e r study. the cost of therapy. With regard to brucellosis in sheep. daily for 1 week) 14 (67%) of 21 cows w e r e successfully treated (25). 6 sheep were Brucella-iiee.m.000 mg LA-OTC i. Moreover. In addition.p. However. such therapeutic regimens have been applied in infected herds to prevent abortions and reduce the spread of brucellosis.p. Furthermore. With the development of Oxytetracycline (OTC) and long-acting (LA)-OTC. melitensis or B. when 8 infected Najdi sheep were inoculated with 1.p. daily for 1 week) resulted in cessation of shedding of B. In this trial. serious local reactions were reported in cows. then 20 days later with 1.000 mg O T C daily for 6 weeks. 9 sheep experimentally infected with B. Recently.21). In one of these studies. by the i. using high doses of L A . melitensis. without side-effects and relatively inexpensive.) inoculations with O T C (12. Only one sheep was found to be bacteriologically positive at slaughter. route. due to repeated intraperitoneal (i.500 or 1. In view of the encouraging results in cows (24. has resulted in the reduction of abortions in infected herds or individual cows (20. In addition. 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. Radwan and colleagues (35) evaluated six different long-term treatment regimens on 480 sheep and goats naturally infected with B.4 g t o g e t h e r with i m m u n e serum for 3 days. tetracyclines and streptomycin (ST). increasing the percentage of Brucella-tree sheep to 82%. when L A .p. One of these regimens ( L A . 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.910 The use of broad-spectrum antibiotics such as aureomycin. inoculation with OTC. 69% and 100% of the sheep were found to be Brucella-îre-c at the end of the trial. on 118 sheep naturally infected with B. practical. alone or in combination with ST. at necropsy (34). In this trial. in order to make treatment of Briicella-infected cows with superior genes a viable alternative to slaughter.] every 3 days for 2 weeks) combined with ST (20 mg/kg i. melitensis. Radwan and colleagues (34) conducted a long-term treatment trial using a variety of doses of O T C .V. abortus were injected daily for 6 days with 1 g Chlortetracycline. Consequently. these regimens have failed to result in complete cure.25) and the successful results in sheep and goats (35). daily for 3 weeks) showed a synergistic effect. abortus in 10 (71.O T C (20 mg/kg i.O T C (20 mg/kg intramuscularly [i.m.25. However.O T C 25 mg/kg i.13. singly or in combination.p.24. melitensis in the 80 sheep and goats treated (35). every 3 days for 2 weeks) alone was successful in curing 3 (21%) of 14 cows.29). effective and least expensive means of achieving complete elimination of B. 46 days after infection (30). injections of L A .O T C was combined with ST (20 mg/kg intravenously [i. However.

all n o n .22. approximately four years prior to this study. Since the number of available cows infected with B. T h e animals belonged to two breeds (108 Holstein x Friesian and 13 Jersey). One m o n t h after t r e a t m e n t . T h e cows contracted brucellosis from sheep and goats infected with B. each containing l g or 4 g. these were placed in g r o u p B together with 35 cows with B. date of recent pregnancy and n u m b e r of previous abortions due to Brucella. In the third herd. immediately after birth. Therapeutic agents The pharmaceutical products used in this study were as follows: . a total of 3 calves (1 from a cow in each treatment group) were sacrificed. 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).p r e g n a n t cows were artificially inseminated. number of previous pregnancies. Shortly before initiation of the present treatment regimens.L A . B and C). These two herds originally tested seronegative on arrival in Saudi A r a b i a . The three herds were composed of highly valuable breeding cows originally imported from the United States of America (USA) and Europe. In two herds. This herd contained few seropositive cows on arrival in Saudi A r a b i a . without regard to age. Non-treated cows were not included as controls. in contrast to ST and OTC-IMI which are not. and selected tissue samples were cultured.911 MATERIALS AND METHODS Animals A total of 121 cows were selected.O T C injectable solution (from F i a n c e ) containing 200 mg O T C base p e r millilitre . respectively .28). which gave positive results for brucellosis after serological and bacteriological tests. Three months later.O T C is a long-acting antibiotic. melitensis. abortus was small. Herd records indicated that none of the cows had been vaccinated during calfhood. 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). abortus biovar 1 was isolated from the mammary secretions of 18 cows. B. eight years prior to the present study. All cows in the study were isolated from other animals. E g y p t ) supplied in vials. a total of 12 pregnant cows (4 from each t r e a t m e n t group) were sacrificed. These animals were selected to provide a m o r e conclusive evaluation than that obtained in previous studies. B. particularly because L A .ST sulphate (from Giza. 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. dissolved (just before use) in 3 ml or 12 ml sterile distilled water. The cows originated from three large commercial dairy herds raised in the central and eastern provinces of Saudi Arabia. In addition. The 121 cows were divided into three treatment groups (A. 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. The cows were aged 3-10 years and weighed between 450 kg and 750 kg. Milking was ceased in lactating cows. and selected tissues were collected for bacteriological examination.

O T C and ST were injected i. Agglutination at 1:100 or greater was considered positive. Both ST and OTC-IMI were administered with L A . cultures of udder secretions were m a d e from the selected 121 positive cows to confirm shedding of Brucella organisms. in the cervical. abortus strain 1119-3) was obtained from the United States Department of Agriculture in Ames. The standard Brucella plate antigen (made of B. The L A . at a dosage of 20-30 ml per site. Therapeutic regimens The treatment regimens of the three groups of infected cows are shown in Table I. USA. samples of stomach contents. 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. 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). and sections of udder. sera from the selected 121 serologically. 100 mg oleandomycin p h o s p h a t e . liver and spleen. liver. lung and spleen of foetuses from the sacrificed animals were cultured. New Haw. 5 mg Prednisolone and 10 ml special excipient. A t each examination. Iowa. prescapular.m. ovary.912 . mesenteric. every month after completion of treatment until calving and again every 2 weeks after calving for 6 m o n t h s . 100 mg neomycin sulphate. 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 . T h e rose bengal test antigen was obtained from the Central Veterinary Laboratory. brain. 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. mediastinal. Weybridge. udder secretion. Following the three serological examinations. Serological testing All animals were subjected to three successive serological examinations at monthly intervals. The date of previous abortions in relation to the initiation of treatment was not recorded on the farms involved. Furthermore. thigh and shoulder muscles. United Kingdom. In addition.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). In addition. each containing 200 mg OTC hydrochloride. Similar selected tissue specimens were cultured from 3 sacrificed newborn calves (1 from a cow in each treatment group). iliac. T h e isolated 2 . Aliquots from each udder secretion sample were spread with sterile cotton swabs on 2-4 freshly prepared plates of culture medium.OTC-IMI (from Athens. popliteal and head lymph nodes. milk samples from all seropositive cows were subjected to bacteriological examination for identification of Brucella shedders. Greece) in 10 ml syringes. sera from all calves born from treated cows were examined immediately after birth. In addition.O T C at the c o m m e n c e m e n t of therapy. animals were initially screened for the presence of Brucella agglutinins by the rose bengal test (3). 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). Separate 30 ml samples of udder secretions from each quarter were also cultured every week during treatment. precrural. Immediately before initiation of the therapeutic regimens. uterus.

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.m.m.m. 25mg/kg i. No of Success/treated Tissuesa)Udder secretions < > b e i.m.m.m.TABLE I Efficacy of three treatment regimens in eliminating Brucella melitensis or B.m. abortus long-acting Oxytetracycline injectable solution (from France) 200 mg/ml streptomycin sulphate (from Egypt) Oxytetracycline intramammary infusion (from Greece) 913 .: 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. abortus from naturally-infected cows Regimen FA v 3 No. melitensis cows naturally infected with B.of Periodicity treated Therapeutic agents cows Route Dosage E ™° Y ^¿J. 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. i. 25mg/kg IMI 20 ml/teat 3 3 27/29(91%) i. 1 4 8 4/4 31/31 (100%) 3 14 A 35(c) LA-OTC( ) ST( ) & OTC-IMI(g) f 25mg/kg i.

there were unusually high rates of metritis. 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 . or the use of c h e m o t h e r a p e u t i c measures.O T C injections. When Brucella organisms could no longer be recovered from udder secretions nor from any of the selected tissue specimens collected at necropsy. B and C. However. . the 32 cows which were in various stages of pregnancy at the time of initiation of the therapeutic regimens A. In addition. the respective cow was considered to be cured (successful t r e a t m e n t ) . in the breeding season following treatment. Moreover. Before t r e a t m e n t . mycoplasmosis and mycotic abortion. New Haw. In this study. F u r t h e r m o r e . there was very light swelling and signs of painful sensations at the site of r e p e a t e d L A . in addition to routine bacteriological examination for listeriosis. and the average cost of the therapeutic agents alone used (regimen A. On the whole. the cows were living in an uninfected environment. United Kingdom.914 Brucella cultures were identified morphologically. Weybridge. particularly in the thigh muscles. aborted foetuses from the three cattle herds involved were cultured for Brucella isolation. the treated cows expelled placenta from the womb without requiring any special manual manipulation. microscopically. no systemic reactions were observed in any of the treated cows during the treatment period. B and C experienced n o r m a l calving within 4 m o n t h s following completion of the treatment. Cost of therapeutic agents and animals The weight of the treated cows ranged between 450 kg and 750 kg. with milk production reduced by 10-15%. the cows had b e e n living in an infected environment for periods of between 1 and 8 years. Of the cows treated with regimens A. biochemically and serologically (3). T h e s e slight local reactions disappeared within 36 hours. 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. B or C) in treating a cow weighing 600 kg was US$186 (SR700). 94% b e c a m e p r e g n a n t and experienced n o r m a l calving. there were no cases of metritis or mastitis in the treated cows.O T C and ST. Furthermore. Side-effects of the therapeutic agents In 8 cows. 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 biotyping of the identified isolates was performed at the Central Veterinary Laboratory. and milk production was within the normal range for uninfected animals of the same breed. salmonellosis. retained placenta and mastitis. The abortion rate in the three herds ranged between 5% and 1 5 % and the infertility r a t e was approximately 1 5 % . No abortions were observed a m o n g the treated cows.

when these 2 Brucella-shedding cows were re-treated with regimen B. t h r e e and four dilutions in 4 2 % . or after calving in the other 2 cows (these animals were then re-treated with regimen B). When 53 cows (of which 35 were naturally infected with B. 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. 4 3 % and 10% of the treated cows respectively. 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.915 The treated cows were imported a few years previously as pregnant heifers. melitensis) were treated with regimen C. melitensis biovar 1 was isolated from the repeated udder secretion samples from only 2 of 29 cows. in the 3 cows of regimen C where t r e a t m e n t failed. they ceased shedding Brucella. which ranged between 2 and 8 colonies per plate. including the cost of air freight to Saudi Arabia. B or C). . melitensis biovar 2 was isolated (1-5 colonies per plate) from mammary secretions. all selected tissue samples from 4 slaughtered cows and the repeated udder secretion samples were found to be Brucella-free. However. 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. only 5% of the 109 cows became seronegative 16 months posttreatment. at a cost of US$3. There was a decrease of two. Mycoplasma or pathogenic fungi were isolated from the aborted foetuses examined in the three herds. Bacteriological findings In the present study. In regimen A. udder tissue and supramammary lymph nodes of only 1 of 4 sacrificed cows (at 4 months post-treatment). no Listeria. 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. However. T h e remaining 8 slaughtered cows did not show any decrease in titre over the same period. 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. melitensis or B. while all calves born from treated cows were seronegative for brucellosis. 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. B. However.600 each. In addition. In addition. abortus w e r e isolated. only B. In all successfully treated cows in the present study (118 cows treated with regimen A. 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. the amount of shedding declined after the initiation of treatment and remained low until 4 months post-treatment in 1 slaughtered cow. Salmonella. abortus from 121 naturally-infected cows is shown in Table I. when 33 cows (naturally infected with B. melitensis and 18 infected with B. abortus) were treated with regimen B. Serological findings Among the treated animals. B. melitensis or B.

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. melitensis accounted for 92% of human cases and B. Although this has been the goal of several workers. two of the cattle herds contracted B.32. in B. the . commercial dairy production was introduced to Saudi Arabia through the importation of valuable and highly productive exotic cattle at a very high cost. while B.19. Brucellosis has been reported and confirmed in both livestock and humans in Saudi Arabia. Germany and France. An eradication campaign through slaughter and compensation is not within the economic scope of developing countries. liver. loss of milk production.6. 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. but from the subsequent problems or irregular breeding. especially where it is difficult to change agricultural customs or social habits.18.2. one worker assessed the average cost of an infected cow at approximately US$3. On the other hand. However. 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. particularly in lymph nodes. except cows in one dairy herd which had B.4.916 DISCUSSION In order to increase milk production. melitensis infection from sheep and goats raised on the same farms.33. 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. infertility. abortus infection. 35. abortus. for economic reasons this method cannot be applied in all cases.23.5. bone marrow. mass vaccination of infected herds protects only uninfected animals without altering the course of infection. melitensis biovars 1. it is impossible to apply only one method which would suit every country. Slaughter of infected animals with superior genetic potential has serious economic and genetic consequences (24).200. Such infection was directly responsible for the high rate of abortions. 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. spleen. none has been fully successful. retained placenta and mastitis with reduced milk production. abortus for 8%. B. 2 and 3 were responsible for all infections in sheep. goats. A n effective. namely: the Netherlands. however. On arrival in Saudi Arabia.16. Consequently. leading to serious consequences for cattle performance and cattle handlers. suis was not found in animals or humans (1. Livestock producers in Saudi Arabia and in many other developing countries cannot afford the traditional "test and slaughter" approach used in developed countries. abortus. 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. In addition. not just for abortion or stillbirth. Moreover. 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. metritis. cattle with superior genetic potential have become increasingly valuable. with the development and widespread use of techniques such as artificial insemination and embryo transfer. In the present study. B. T h e infected vaccinated animals continue to present a serious public health risk. In a campaign against brucellosis.31. camels and dairy cattle.34.39). and the reduced h u m a n productivity resulting from a spread of infection (38).

For this reason. despite the use of a combination of L A .24. in order to decrease the n u m b e r of Brucella organisms in u d d e r tissues and secretions.24.26.11. 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. which were all serologically and (particularly) bacteriologically positive. as one injection gave an effective concentration of 0. b) Incorrect choice of antibiotics (7. However. when administered every 3 days in regimen C. Furthermore.37). g) High cost of antibiotherapy (24. However.) inoculation of L A . d) Insufficient duration of treatment (8. several authors failed to cure udder infections in many cases (24.n e g a t i v e bacteria. and are therefore p r o t e c t e d from antibodies.O T C and ST. ST acts synergistically with O T C to inhibit growth of B.25. 25. early studies (20. abortus within bovine cells cultured in vitro.6 ug/ml for three days (26). milking was stopped in lactating cows and udders were completely evacuated before each injection. and the possible harmful effect on consumers (27).26. ST was also used. 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. OTC-IMI was used locally in combination with systemic (i.O T C and ST.24).13.O T C variant was selected to save time and effort and to provide long-lasting O T C concentration in blood plasma. 34. i) Presence of antibiotic residues in milk and m e a t of t r e a t e d animals. because it is known to inhibit p r o t e i n synthesis of G r a m . In addition. reproductive organs and tissues with a weak blood supply. Since successful therapy in human beings appears to depend on the permeability of cell walls to drugs (24). F u r t h e r m o r e .25) on the treatment of bovine brucellosis indicated poor success rates when L A . However. O T C proved to be effectively bactericidal when combined with 10 µg/ml of ST (15.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. relapses after antimicrobial t r e a t m e n t w e r e also explained by the intracellular location of brucellae (9.34). This may be due to the'fact that both ST and OTC-IMI are . c) Insufficient doses of effective antibiotics (12. e) Use of the intraperitoneal route for administration of antibiotics created serious reactions in some cases (17.29). L A .O T C or ST was used alone.5-1.26). T h e ability of Brucella organisms to survive intracellularly can result in long-term chronic infection.I M I and ST. The L A . 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. c o m p l e m e n t and antibiotics. melitensis (35). 36).24.25. the following points were taken into account when attempting to obtain more successful results. a combination of L A .24.25.34). in infected cows. For these reasons. O T C at 0. abortus.O T C and ST proved very effective in curing sheep and goats naturally infected with B. before initiation of the present treatment regimens. f) Use of only one antibiotic (20.26).m.13.26). In addition.25.12. p e r m i t t e d growth or was bacteriostatic for brucellae within tissue cells.917 mammary glands.25.21. were less effective in eliminating Brucella organisms in infected cows. Consequently.0 µg/ml.29).25. melitensis or B. Consequently. h) Failure to cure udder infections in many cases (24. O T C .

and t h e r e was no nontreated control group for comparison. 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. because all serological and bacteriological examinations were performed free of charge at Government institutions. including the cost of air freight.26. Other investigators. the majority (approximately 90%) of the cows treated were monitored bacteriologically for 16 months post-treatment in order to detect possible relapses.25. reported that serological findings were not useful in evaluating the effectiveness of treatment regimens (24. However. relapses occurred in only 3 of 33 cows treated with regimen C. although O T C has been considered the most effective antibiotic for treatment of brucellosis. In conclusion. i.O T C did not result in harmful systemic effects or any obvious lasting local reactions at the site of inoculation. without side-effects and relatively inexpensive in causing cessation of the symptoms of brucellosis and eliminating Brucella organisms from infected cows. only a small n u m b e r of Brucella colonies were recovered from large tissue samples.29). relapses did not occur in any of the 88 cows treated with regimen A or B (provided that the treatment protocol is strictly observed). a small number of relapses do occur. 34) concluded that i. Relapses have also been reported in cows (14. In Saudi Arabia. This did not include the cost of e q u i p m e n t or interventions.34).918 not long-acting antibiotics. Previous studies in cows and sheep (17.25).600. 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. Even in relapsed cows (3 animals in regimen C). However. B or C for treating a cow of an average weight of 600 kg was US$186 (SR700). however. However. 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 results were not conclusive.13. inoculation of L A . In humans. this treatment would probably be limited to valuable breeding animals.p. regimens A and B proved to be effective. the cost of supplies and intervention should be considered.I M I alone would be effective for the treatement of lactating cows due to the continuous flushing of OTC by milking. all t r e a t e d cows were dried before initiation of treatment. Also. 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. T h e average cost of the antibiotics used in regimen A. inoculation of OTC was not safe. at the present time. it seems unlikely that O T C . as animals were only monitored for 16 months post-treatment. 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). However. consequently. practical. The present results of serological examinations were of some use in evaluating the effectiveness of the treatment regimens. the treated cows were originally imported as pregnant heifers at an average cost of US$3. when treatment protocols were strictly observed. Consequently. further research may yield a regimen .12. However. which is considered economically feasible in Saudi Arabia.m. In the present trial. and the protection of Brucella organisms in the presence of milk. In the present study. In the present study.75). In the present study.24. The cost of successful treatment of a cow represents approximately 5% of the original price of the animal.24. in other situations. due to the development of several serious local reactions at the site of injection.

for his encouragement and co-operation. de 25 mg/kg de ST (i. similaire au protocole A.) de 25 mg/kg de LA-OTC tous les trois jours pendant 42 jours. abortus dans celles de 18 vaches. Le protocole B (appliqué à 53 vaches) était similaire au précédent.S. 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.I. les bio types 1 ou 2 de B. The authors also acknowledge the technical assistance of A . 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. Avant l'application de ces trois protocoles thérapeutiques.A.M.) quotidiennement pendant huit jours et en une OTC-IMI quotidienne de 20 ml par trayon pendant quatre jours. pratiquée également tous les trois jours pendant 12 jours. Mukayel and A . la streptomycine (ST) et l'infusion intramammaire d'OTC (OTC-intramammary infusion : OTC-IMI). Furthermore. 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. Thanks are also extended to the owners and veterinary staff of the three dairy farms involved. il n'a permis l'élimination des brucelles que sur 30 (91 %) des 33 vaches traitées. Le protocole A (appliqué à 35 vaches) consistait en une administration intramusculaire (i. melitensis ont été régulièrement isolés dans les sécrétions mammaires de 103 vaches et le biotype 1 de B.V. hormis pour la ST qui était administrée tous les trois jours pendant 24 jours et l'OTC-IMI. O. Al-Bokmy. Mohamed et S. Radwan.T. Quant au protocole C. Prasad.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. A. combinant à la fois l'Oxytétracycline à action prolongée (long-acting Oxytetracycline : LA-OTC). 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. Bekairi.S. .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. for their s u p p o r t and c o .I. S. Hussain. ACKNOWLEDGEMENTS The a u t h o r s wish to express their sincere t h a n k s to M. Al-Taher.M. * * * DES PROTOCOLES THÉRAPEUTIQUES EFFICACES POUR LE TRAITEMENT DE L'INFECTION PAR BRUCELLA MELITENSIS ET BRUCELLA ABORTUS CHEZ LES VACHES. Résumé : Trois protocoles thérapeutiques ont été évalués chez 121 vaches atteintes d'une infection naturelle par Brucella melitensis ou Brucella abortus. pratiquée un jour sur deux pendant huit jours. Bin Salamah. P.m. 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. sauf pour la ST qui était administrée un jour sur deux pendant 16 jours et pour l'OTC-IMI.m. .

sólo permitió eliminación de brucelas en 30 de las 33 vacas tratadas. Ann.Oxytétracycline . que se administró cada dos días durante ocho días.. 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.920 MOTS-CLÉS : Antibiotiques .Thérapie.S. & HARLAND P. A L .Y. . 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.Brucella abortus . JONES L .25-27. que fue experimentado en 35 vacas.. ALTON G. abortus en las secreciones mamarias de 18 vacas. .A. .Oxitetraciclina .Estreptomicina . Saudi biol.351-357. consistía en la administración intramuscular (i. (1988). Proc. .E. & V E R G E R J . CHEN M . es decir.Brucella melitensis .S.T. PALABRAS CLAVE: Antibióticos . Prasad. 7. (1986). O. . un 91%. Mohamed y S.) diarios durante ocho días y en una OTC-IMI diaria de 20 ml por pezón durante 4 días.. que se experimentó en 53 vacas.G. HAWARI A.I. El protocolo B. Por último.Brucella abortus . Al-Bokmy.) de 25 mg/kg de LA-OTC cada tres días durante 42 días. que se administró cada dos días durante 16 días. * PROTOCOLOS TERAPÉUTICOS EFICACES PARA EL TRATAMIENTO DE LA INFECCIÓN POR BRUCELLA MELITENSIS Y BRUCELLA ABORTUS EN VACAS. era similar al anterior excepto en cuanto a la ST. . WANG C F . 2. (1984).V. REFERENCES 1. Hussain. Soc.M.H. Radwan. y ala OTC-IMI. AL-MARSHEDY A. melitensis fueron regularmente aislados en las secreciones mamarias de 103 vacas y el biotipo 1 de B.A study on brucellosis among animals of some farms and rural areas of Saudi Arabia.G. & CHOU N.Brucella melitensis . 3. M . Resumen: Se evaluaron tres protocolos terapéuticos en 121 vacas infectadas por vía natural por Brucella melitensis o Brucella abortus. a partir de una combinación de oxitetraciclina de acción prolongada (long-acting Oxytetracycline: LA-OTC).m.Bovins . Antes de aplicar estos tres protocolos terapéuticos. el protocolo C. 6. A N G U S R. Paris. . El protocolo A.M E Z A I N I S. de 25 mg/kg de ST (i. S. SINOUSSI Y . 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.Bovinos .. Saudi Med. estreptomicina (ST) e infusión intramamaria de OTC (OTC-intramammary infusion: OTC-IMI).. CHANG S. P. 190 pp.Streptomycine . C . M . Bekairi.M.Brucellosis in children. los biotipos 1 o 2 de B.I.R. Institut National de la Recherche Agronomique.D. A.Terapéutica.Techniques for the brucellosis laboratory.m.

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