BcniSnef,Vel.Rlecf.Res.Vol.VIII,.Januarv,No.l,1998 -
CLINICO-EPIZOOTIOLOGICAL STUDY ON
BRUCELLOSIS IN DONKEYS
By
Hamoda, F.K. and Montaser, A.M.*
The present investigation Was carried out on
206 male and 52 _`anale donke y s at haluobia (Jovernorate. These animals Were examined _linically . serolo,ically using different iimnunoassays as well as cieriologica1Iy for detection of brucella organisms.
Clinical examination revealed no characteristic clinical signs of xucellosis in donke y s . Results of serotests indicated that percentages of roreactors in complement fixation (('PT) , serum tube agglutination Ski) _ buttered acidified plate agglutination f13:\P:\.'fi.Rose-Bengal RBPT) . Rikanol (RIVI) tests and competitive enz y me Iinked mmunosorbent assa y (cEl_IS.A) were 16.5. 7.29. 19.42 . ID.
19. 10.19
=nd 7.9-°i, in male donke y s and 17.30. 11.54. 21.1. , I Io . I I.>•4 and in female donkeys respectively. Application of in
\IRT) on milk of 52 site-donkeys. revealed 7.69%positi\e reactors. rim_ test
Trials fix brucella isolation from milk samples. s\Nabs from fistulous withers lesion and specimens of testicles revealed two isolates of
Brtieellal nteltenesis biovar 3 loin milk.
Brucellosis is still one of the most iinporlanl diseases affecting
7nimals and man in rni st count r ies of the world ( Ratloslits ci al.. 1994 nd Benkiratie. 1997)
I
'
Dept. of .Animal Med.. l-ac.of Vet.Med., Zagazig t_
1 niv. I enha.
Dept. of f3rucellosis.:\nilnal Health Research Institute. 0okki. Ciiza.
105 iicni-gn f
\e4: ied.kles.VoI.VIII .l.luar ,No.7,i998
Although brucellosis among cattle. sheen . goats acid camels been studied in H ypt by several workers (Fayed ct al.. 1982 : Sale  al.. 1987 : Refai , 1989
Farid et al., 1995 and El-Sa\eaIlly et al.. 19' Yet, this disease received little attention in equine especiall y donkeys
4
Equine brucellosis is important not only its a clinical entlq' also as a potential source of infection for man and other animal spe (i)enny. I973). Incidence of brucellosis in donke y s using semi lo i:u noslic tools were reported as i 61% in India Cl)ralgonas

cl al.. IS .Ind 7.5
o b in (irececc ( l ada\ cl a!.. 1991). In I :,_v pt. I'almmn and Sa
(f 97-t) .
Salem cI al.. ( 1975) and Fl-Ruby (197`)) recorded the iucidc
UI I)IUCCIlosis III d oIIke
\s 4(10(1 .
16.5'% and 27.05
% % I'Cspeeii\
Rcccull\ .\bcicl-halIer ei al., ( 1995) reported lime incidence of doni 111'ticcllosis as 2tl 61°n . l2.97° .
13.354481Ul f.+.35°b lloslll\e reac using lute at,uliltln;uiom
(TAT) Rose-Ren,!cll pl,llc test (RBI buttered acidified plate test (13AP.A) and
(bombs test. In horses Alt[ .;id Mtinii (1995) demonst r ated that incidence of horse brireellosisI
5.78% ader et <aI.• (1995) cited that the serological incidecici c:.:cellosis in horses was 5.88% by application ol • the three scrolol tests ('FAT.
RI3P 1 , 13APA) , while Gslnat (1996) recorded that 8.0 4.51 and 6.5% of horses sera were positive in Rose Bengal Plate Serum agglutinui:on test and
Complement Fixation lest and isol Bence/!u uhorm , biotylie (I) from affected fetlock joint of two horse
I\lc( a11ghe y and Kerr ( 1967) found that when horses runt inLcled cattle, a relatively high proportion become inlceledi developed a positive reaction by serum agglutination test wit showing clinical signs. Pregnant mares would about on grazing cattle inFected hrucclla ( (.'t'ossnrty and 13onson,
1968). I-low Robertson et A. ( 197;) mentioned that brucellosis rarely cause aim of' mares Ilinloo et al., (1978) reported that linuellu cd>I is associated with chronic bwsal enlargement of the neck. withers
(06
Beni-Suet Vct.i%1ed.Res
.Vol.VIII,.latnuarc.No.I.!998 -- — navicular bursae caruin_u intermittent lameness, \Ia.\lillan (1985) mentioned that horses infected w+ith brucellosis suffered generalized infection with signs including stillness. fluctuatine temperature and lethargy. Moreover Knottenbelt and Pascoe ( 1991) and Kobluk et al., (1995) reported that brucellosis are probably less significant and is declining as a cause of fistulous withers and a wide variety of bacterial species invol v ed in etiology of this problem among equidac. Recently Ahdel-Kader ct al., (1995) observed that horses . mules and donkeys did not show any sign of brucellosis, meanwhile they detected antibodies against brucella abortus using, diftcrent serological tests . While lismat ( I996) reported that brucella infected horses showed febrile reaction in some cases with normal appetite and good body condition. panful swelling on supraspinous bursa or atlantal bursa, ulcreativc wounds, lameness and abortion in 3 mares. Moreover Blowey (19)6) declared that brucellosis ma y cause chronic joints or tendons infcc(ion and (istulous withers.
In view of the lack of literature about donke y s hrucellosis and also due to the continuing incidence of brucellosis in rwiiinants and human-beings . the aim c f this study was to throw some light on rucellosis in donkeys including clinical abnormalities . serological and bacteriological examination.

I- :>,nintals :
A total of 258 donke y s of which 206 males and 52 females from Moshtohor and seven adjacent villages at Kahrobia governorate were used in this investigation . Out of them seven donkeys ( 2 Females and five males) were showed fistulous withers and ten males exhibited slight enlargement of testes. All of these animals have history of contact with ruminants (cattle . buffaloes, sheep , goats and camels) and grazed alongside.
107
Reni-Suct,V et.Med,Res.VuLVIII,.Janua ry,No.1,1998
2- Samples and procedures a- Serum samples were collected from all donke y s under inv( and stored at -20 until being, used for serotesting. h- Milk samples N+erc collected under hygienic measures in MacCrteny bottles from lactating she-donkeys for perkriuin g in test (MART) and bruceIla isolation (.Alton ct al..
1988). c'I esticles . were surgically taken front three male donkeys (castrat for brucella isolation.
(.Icon et al.. 1988) . dSwabs , seven swabs were collected from lesions of fistulous under hygienic measures in closed sterile test tube and sent imm (u lahoralorn liw bacieriological examination (Alton et al.. 1988) .
3- Imnumo-assac fur dia2nuxis of hruccllosis
('omplemeut fixation test (('I' I'), Standard tube agglutination (SAT) . Rose-Bengal
Plate Test ( R13l' I) . Buttered acidified ph agglutination test (BAI':\) and Rivanol test were applied according .Alton et al., (1988) . While competiti\e enzyme linked inutiunosorbt assa y
(cF:I.ISA) was used after Davis et al., ( 1980) .
Results arc shown in tables (I,2 & 3) ..
Control of brucellosis depends largely on elimination of ani reservoirs (Radostils ct al.. 1994). In ected horses may act as carriers infection and upset the control of brucellosis in free herds (I lungerfi 1990) . ('linical examination of investigated donkeys revealed that ne all of caanuned donkeys were apparent normal clinically except some
108
Kcni Sucl',Vet.(<lcd.ltes.Vol.\ III,.lanu:Irv.No,I.1998 seropositive animals showed Cstulous withers or sligfu enlargement of one or both testicles , moreover although some of prcguaui Ienriles were reacted positively . they did not show

abortion. I hese results (Ii let from those of Fl-[3ohv ( 1979) who observed capped knee . capped elbow. and abscess on the bod y and are similar to the Findings of Fahni'\ and Salerii
1974) Moreover these results +ere neativ justi lied by prior work of Rankin (
1973) who sttaed that equine brucellosis tends to be cluonic and subelinical and Denny (1973) who reported latent or dormant infection are the most probable form of brucellosis in horses and
Salem et al.,
(1975) who observed that seropositive females donke y s did not show abortion.
Recently, Abdel-Kader et al.,
(1995) declared that exposure of donkeys. mules and horses to hrucella infection ith consequrnl development of anti-brucella antibodies appeared to lc a Common occurrence ot'donkeys brucellosis in I.gypl wilhuul evidence ol':IppICIII Clinical signs.
Scrolouical iliaitio>is ul hruccllosis is time main tool liar (IeIccIi('n of infected animals
. but it cmi not idcnlil y all inlecled ,uiini;lls at all Stages ofthe disease includiim acute and chronic pliuses:i. aiII :u talent infection:. Reviewing table
I
I) . the prevalence of brucellosis using ('H
S;\ I . 13:\PA 1 . IMP I . R11 and clil.ISA were
16.5. 7.2S , 19.42,
10.19 and
7.94% in rnafe donkeys serum respectively. i lie obtained results concerning (TT showed high sepcificity and sensitivity . this goes in perfect ltarrnonv with conclusion of Sayotr (1995) who mentioned that (FT is the best definitive test which is cu rently available because it gives maximum balance of sensitivit y and specificity . Regarding results of SA I it presented lower sensitivity (7.2894) seroreactors percentage than that recorded b y I:I-l3oh\
(1979):
Salem et al..
(l9 75) and AbdelKader el al.. (1995) . this might attributed to the difference ill the locality and the techniques employed.
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Beni-Suef,Vet.NIed.Res.Vol.N AIl,Januarv,'So.1.1998 ---
Concetnittg results of I3AP.\ f . one can
It t c highest setIsiIkit%
(19.42"x) over other serotests . These findings were higher than to that recorded to Abdel-Kader
(1995) who detected
13.35% using the saint test. The high sensitivity of'f3AI'.AT is mflltloly attributed to the low final antigen concentration (3%) after the addition of serum (Alton et aL 1988).
Regarding to results of RBP f . it gave 10.19°,-0. this percentage was less than that recorded by ci r because (F1 is more sensitive to lg(i I than the RI 1 r 1, "l hest results could also be explained by Morgan et al..
(1969) and Davies
(1971) who noticed the ability of R13P l' to detect infected animals missed by SAT . The data recorded on using of RIVT (10.19°,•x)
%vas similar to that of RI3P'I .this might be attributed to equal sensitivity of both tests in donkeys.
Concerning results cf?1.ISA (7.28%) , the lower percentage recorded in this study mi llt be attributed to the filet that this test was especial I\ stand.udi/ed to work with bovine sera. Nloreo .
rr positive cli IS.A results are only associated with active inflection with brucellosis so that serum antibodies can compete well with the monoclonal wgluuatc ol'Ihe

test.
The coiielatiott between different inmuu o-assa y s in this stud indicated that RAP:\ I and C'F'I' appeared to be more sensitive butt ('I was more specific in detection of ' seroreactors to brucella infection meanwhile 13ARA I is not specific (Iuc to cross reaction with otht organisms sharing antigenic .
structure . Although ;locus and f3arto (198 .
1) reported that BAPAT was used as a screening test for detection brucellosis . yet must be confirmed b y other serological tests . therefore battery of' itnmunoassays should be applied (Savour . 1995 an Ihrahitun.
1996).
Beni-Suet,Vet.1
1 ed.l2es.VoLVItl,.lanuarv,ho. 1,1998 --.. --
Concerning results ol'serotests on sera of females (table. 2) , these alts were slightly higher than those recorded in males. The higher o-reactors in tetnales ma y be due to higher exposure of females to .lection (El-13ohv.
1979) . these results are in accordance with those observed by Salem et al., ( 1975) and I{If3oln c1979) w+ho declared higher suscepility of females over males donkey of horse to b rucellosis.
On the other hand these findings contradicted to those of (iibbons and \tanning. ( 1969) who mentioned that there was no relation between the sex and incidence of brucellosis. Application of
11111k riot test on the 52 she-donkeys . gave (8.5%) positive reactors .
Ibis finding was in a similarity to that repotted by I L .I-f3ohy (1979) and
(lie test can be considered a useful lest in diagnosis of she-donke y s hrucelhosis . so repeated testing of donkeys milk beside serotesting should be put in consi(Irration in diagnosis of donke y s brucellosis.
I're\'alrnre oI anti-htnrella antibodies in dt1nke\s in close contact with ruminants suggest that donke y s might be exposed to brucellosis \\ithout appearance of clinical e~ .
idence. This is Itirhh au,menlrcd by explanation of ('open cl ;tl.. ( 1992) who stated scioposilivit\ indicates evidence of exposure to an organism but not necessarily current infection.
Bacteriological examination of ml
IL. revealed detection of two isolates of brucella mcltensis biovar 3. while swabs loom fistulous withers and specimens of testicles showed negative results . Difference between the results obtained in this study and those reported by
Fahrm
Salem (1974) who isolated 13r.abortus 1 110111 dunke\ manifested fistulous withers ma y be due to the wide pre\alence of 13r.mrltensis biovar .; In cattle nowadays and the chance of contact between them and the investigated donkeys.
— Beni-Suef,\'ct.%led.Res.Vol.VIll,Janu:u'v,No.1,1998
From the epidemiological point of view. it is important to appreciate that application of hygienic measures however good. are not effective against the disease where synlptonlless carriers which reacted positively or negatively serologicall y can disseminate the organism in large nunlbers and become it source of infection and Conlanlinati011l to other animals . human-beings as well as the surrounding env itonnlent (7.aghloul and Kanlel. 1985) , therefore periodic serological nlonitorittg of equidac especiall y those in close contact with

brucellosis Fire herds should be adopted (Blaha, 1989 and Radostits ci al our grad possihilil) al.. 1995) . since there ol , hruccllac shedding b\ eyuidac (Abdel-Kader cl
In conclusion it is obvious from clinical . serological and bncmc uol4},fical crilcriat
Illal donkc\'s \\rrc e\pu,rd Io brucella inti•ction and sub
,c-(JtW tl\ reacted serologically without obvious cll:lrtctcristic clinical evidence, also donkeys might be secrete the brucellae organisms in Secretions as milk and act as source of infection to other incontact animals and humans. Periodical serum and milk testing of donkeys with highly sensitive and specific imnluno-assays as ('Fr or 13APA'I' should he recommended . but rather a battery of assn s are necessar y for final drlinitivc climinauon of reservoirs or carriers.
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