79 (101:1015-1017,October2009

IndianJoumal of AnimalSciences

Recentoutbreak of glandersin India

PRAVEENMALIKI, S K KIIURANA2, B K SINGH3 ANdS K DWTVEDIA

125 001 India

National ResearchCentre on Equines' SirsaRoad, Hisar' Haryana

Received: i6 October 2008; Accepted: 15 May 2009

Glanders' Outbreak
Key worils: Burkholderia malle v Complement fxation test' Equines'

Glanders,one of the oldest known diseasesof equines'is Table l Details of samplescollectedfor bacteriological
examinationand isolatesobtained
contagious and highly fatal disease of solipeds and has high
zoonoticsignificance. It is causedby the granr-negauve non- Total
State/Sample Nasal Lesion Pus EYe
motile bacillus Brrtlroldeia mqllei The disease in equrnes Swab
Swab Swab
is characterized by nodular lesions of the lungs and other
organs as well as ulcerative lesions of the skin and mucous Maharashtra 14 (3) 2
2
'y, 18 (4)
18(3)
membranes of the nasal cavity and respiratory passages'The Uttar Itadesh 16 (3)
Andhra Pradesh 5 (5) 5 (5)
clinical and bacteriological diagnosis of glanders is difficult ; 2 l0 (1)
HimachalPndesh 6 (l)
because of rnixed clinical picture and beaded appearance ot 21(l) ; 30 (l)
Uttrakhand
the organismsin tissue sections.Human is accidentalhost Punjab 3 3
and the disease usually results from occupational contact naryana ; ; 9
Though the organismis susceptibleto various antibiotics."x Delhi 13 l3
vilro, fieatment is difficult and needs longer course wltn Jammuand Kashmir 13 13
combination of antibiotics upon early diagnosis No vaccine Total 98 (13) ; sat ; r 1 9( 1 4 )
is available. The diseaseis notifiable in India under Glanders
indicatenumberof B' mallei isolates'
Figuresin theparantheses
and Farcy act 1899.
Glanders is reported from Middle Eastern countries, the
Indian subcontinent, SoutheastAsia, parts of China and serum sampleswele collected./receivedfor serology including
Mongolia, Africa and South America. The organism has got those received in the lab for follow up surveillance till March
2008.
a clear scope of bio-terrorism and was believed to have been
Isola(ion and characterizaLionof the causativeagent was
used during World War I and World War II on horses and
done as per standardprotocol. The serum sampleswere
humans. In last two decades,the occurrenceof outbreaks
amongstequinesis steadilyincreasingand thus is currently subjected to standard complement fixation test (CFf) as per
consiJered as a re-emerging disease(Wiftig et al 2OO6)'The OIE protocol. Intrapalpebrodermal (IPD) mallein test using
occurrence of the disease leads to international trade mallein PPD (IVRI) was performed on the reactor arumals
for confirmation of disease before elimination The crude
restrictions. Considering that the diseasein India was
historically restrictedto certain pocketswith sporadiccases' extracts of the isolated organisms were analyzed by standard
the preseni report accounting the re-emergenceof the dreaded Sodium clodecyl sulphate-polyacrylamide gel electrophoresis
(SDS-PAGE).Thesewere also subjectedto irnrnunoblotting
diseaseamongstequinesin India as outbreakduring 2006- (diluted to l:750)
08 is very pertinent for applying early mitigation and confiol iwestem blotting) using the horse sera
collected from a horse naturally infected from B' mqltei
strategy. IgG-HRPO
infection and were developed by anti-horse
Ninety eight nasalswabs,7 abscessswabs,5 pus samples oride
conjugate using Diaminobenzidine tetrahy&oci @AB)
and g eye swabs(Table lt were cotlectedftom equinesin
different parts of the country following detailed clinical as substrate.
Several clinical cases showing typical nasavcutaneous
examinationduring 2006-2008. In addition,a total of 13,632
form of glanders were found in Maharashtra, Punjab, Uttar
Pradesh,Uttrakhand, Andhra Pradesh,Himachal Pradesh and
Present address: lsenior Scientist (email: malik-p Haryana. Major clinical symptoms include nasal ulcers'
@indiatimes.com), 25eniorScientist,3PrincipalScientistand
aFormerDirector, Plot No 79, Flat nodular swellings all over the body, edematous swelling or
Incharge,Equine Health Unit,
Sector-56'Gurgaol,Haiyana wounds on legs. cough. dyspnoea.reducedappetiteand
NO. B-3 Kiral Apartments,
1016 MN,IK ET AL. llnd.ianJoumat of Animal Sciences79 (10)

containment and eradication of the diseasefrom the country.

Y westem blots of the isolates with positive serum of affected

horse revealed reactive protein at about 40 kD in all the
isolateswhile negative seradid not show thesebands.
On serological examination of the samples employing

t
CFT, 140 equinesfrom Maharashtra(23), Uttar Pradesh(70),
Uttrakhand (21), Punjab (3), Himachal Pradesh(6), Andhra
Pradesh(16) and Haryana (1) were declaredpositive (Table
2). The titre in the positive animalsrangedfrom l:16 to 1:256.
For eradication of glanders, CFT has been reported to be
highly sensitiveand 9495Vo acctvate.The test hasbeen the
preferredand most extensivelyuseddiagnostictool because
of its capacity to detect clinically inapparent carriers and
chronically infected hones. It is widely accaptabletest for
international nade (Anonymous 2006).
Though the animals were declared glandered based on
clinical, microbiological and serologicalfindings, thesewere
finally testedby mallein IDP test using mallein PPD before
elimination by the local animal husbandry authorities in
Maharashtra and Himachal Pradesh.No human case was
repofied with any symptom simulating glanders.
In India, casesof glandershave earlierbeenreportedfrom
different parts in post independencaem, most of them from
north India (Gulati and Gautam 1962, Singh 1964). Verma
( 1981) feported incidence and epidemiology of glanderson
rnilitary farms in India and found high morbidity in thickly
populated equines. Sporadic incidences in certain pockets
Photo1: A horseexhibitingclassicallesionsof glanders(nasal of India have been reported (Anonymous 1988, 1989-90,
and cutaneousfoms) lMisra et al. 1985). Kumar et al. (1999) reported the last
Table2. Resultsof ComplementFixationTeston serumsamples
wasting (Photo 1). Three forms i.e. cutaneous,nasal and collected/received
pulmonary forms of glanders described in the literature were
not distinct in different animals in the present outbreak as Stateru.T. Number of samples
reported earlier.
2006-0'7 2007-08 Total
Fourteen samplesyielded isolatesindistinguishablefrom
Burkholderia mallei, including 13 ftom nasal swab and 1 Andhra Pradesh 50 1'771(t6) l82l (16)
from pus sample (Table 1). On Glycerol Dextrose Agar Chandigarh 25 l6 41
(GDA), there was a confluent, slightly cream-coloured Chhattisgarh 105 "1 l12
growth that was smooth, moist and viscid after 24 h Delhi 139 949 1688
Gujrat 305 246 551
incubation.With continuedincubation,the growth thickened
Haryana 311 240(t) 5 5 1( 1 )
and became darker and tough. The organisms were small 252 (6)
Himachal Pradesh 0 2s2 (6)
gram-negativerods appearedbipolar, especially in shorter Jammu and Kashmir 11 319 330
rods. No hemolysis on blood agar and no growth on Kamataka r37 6t2 749
McConkeysAgar and at 42'C was observed.All isolateswere Madhya Pradesh t66 2t'7 383
'720(23)
non-motile and were catalaseand oxidase positive but did Maharashtra 3s7 (23) 363
not reduce nihates. Except weak fermentation of glucose, Manipur 50 12 62
'723(3) 2505 3228(3)
fructose and mannitol. the isolates looked inert to vadous Punjab
sugar fermentation reactions (sucrose, lactose, galactose, Rajasthan 535 308 843
Tamil Nadu 156 359 515
sorbitol, mannose,maltose,dbose and rhamnose).
Ultar Pradesh 548 (70) 664 r2\2 (10)
PAGE profile revealed a large number of bands ranging
Uttarakhand 1 6 4( 1 ) 273 (20) 43',7(21)
from <20 kD to >150 kD. However, the profile indicated all West Bangal 13 124 t3'7
the isolates to be similar indicating a common source of Total 4395(97) 923'7(4!) 13632(140)
infection in all the states. This was important from
epidemiologicalpoint of view and would be helpful in Figures in the paranthesesindicate number of positive samples.
October20091 RECENT OUTBREAK OF GLANDERS IN INDIA
confirmed case of glanders in a mule from Rohtak, Haryana
before the curent outbreak.
Authorities of state and central animal husbandry
departments were appropriately notified. The veterinarians
and animal owners were given guidelines on sanitary
measuresandhygienic precautions.Strategiesfor prevention
of further spread of the disease to other areas and
neighbouring states and action as per Glanders and Farcy
Act (1899) were also suggested.Follow up ofthe occurrence
revealednegativestatusbasedon testingon more than 13500
samples from different statesthereafter (Table 2). The disease
appearedto have startedsubsequentto movementof animals
by traders coming ftom northern pads of India, thus it is
considered that the nidus of infection may be present
somewhere in North India.
The Centre is actively monitoring the situation nation-
wide though intense follow ups on samplesfrom different
states of the country along with prompt and dedicated action
taken by the state authorities following notification.
Systematictesting by serology and physical surveillanceby
local authorities in various stateswas important in view of
the possibility of latent or clinically inapparent casesand
thereafteris being continued.The diseasewas successfully
contained in the country mainly due to efforts of NRCE in
coordination with state animal husbandryauthorities.
SUMMARY
Glanders,a notifiable highly contagiouszoonotic disease
primarily of equids, is causedby Burkholderia mallei. The
diseasein India was restricted to certain pockets with sporadic
cases.A major outbreakof glandersoccurredin India starting
in 2006 from Maharashtra(26 cases).The diseasehas been
reported later on from six other states namely Uttar Pradesh
(70), Uttrakhand(21), Punjab (3), Andhra Pradesh(16),
Himachal Pradesh(6) and Haryana(l) till 2007.The disease
was confirmed among equines based on clinical signs and
symptoms, microbiological and serological investigations.
A total of 14 isolatesindistinguishablefrom B. mallei werc
obtained ftom clinical sarnpleswhile serum samplesof 140
l0l;
equineswere found positive by Complement Fixation Tesr
Strategies for containment of the disease to other areas a.od
action as per Glandersand Farcy Act, 1899were suggested
Follow up revealed negative statusin various statesindicating
that the diseasewas successfullycontainedin the country'.
ACKNOWLEDGEMENTS
The authors are thankful to Director. NRCE. Hisar for
extending necessaryfacilities; Joint Director and his team,
WRDDL, Pune for cooperationand assistancein collection
of samples; Joint Director (HCF), DRDE, Gwalior for
providing containment facilities. The untiring help by Sh.
Sita Ram and Sh. Guru Dutt is highly appreciated and
acknowledged.
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