Tropical Animal Health and Production, 32 (2000) 165^171

# 2000 Kluwer Academic Publishers. Printed in the Netherlands

An Outbreak of Gangrenous Syndrome among Bu¡aloes and

Cattle in West Bengal: Clinicopathological Studies
A. Sikdar*, G.C. Chakraborty, D. Bhattacharya, S. Bakshi, D.K. Basak,
A. Chatterjee and S.K. Halder
Eastern Regional Station, Indian Veterinary Research Institute, 37 Belgachia Road,
Calcutta 700 037, India
*Correspondence

Sikdar, A., Chakraborty, G.C., Bhattacharya, D., Bakshi, S., Basak, D.K., Chatterjee, A. and Halder, S.K.,
2000. An outbreak of gangrenous syndrome among bu¡aloes and cattle in West Bengal: clinicopathological
studies. Tropical Animal Health and Production, 32(3), 165^171

ABSTRACT

Gangrenous syndrome/Degnala disease was recorded in a large number of bu¡aloes and cattle in
Murshidabad district of West Bengal, India. Fusarium spp. had been isolated from the mouldy paddy
straw which were fed to the animals. There was a reduction in the incidence of the disease following
withdrawal of the mouldy paddy straw. Histopathological examination showed necrosis and loss of
architectural details in the skin.

Keywords: bu¡alo, cattle, Degnala disease, Fusarium, gangrenous syndrome, mould, pathology

Abbreviations: BOD, biological oxygen demand; H&E, haematoxylin and eosin; PAS, periodic acid^
Schi¡; SDA, Sabouraud's dextrose agar; SDACC, SDA-cycloheximide and chloramphenicol

INTRODUCTION

Since the ¢rst report by Shirlaw in 1939 from Sheikhpura (now in Pakistan), gang-
renous syndrome/Degnala disease in bu¡aloes and cattle has been reported from
various states in India, including Punjab, Haryana, Kerala and Uttar Pradesh (Sastry,
1983). Even though the disease has been known to exist for about 50 years, its aetiology
is still undetermined. Some workers claimed it to be associated with selenium toxicity
(Basak et al., 1994), while others have postulated that the syndrome is due to
mycotoxicosis or ergot poisoning (Kwatra, 1980; Mallick et al., 1990).
This communication records an outbreak of gangrenous syndrome among bu¡aloes
and cattle in West Bengal, India. Various possible causes are discussed and an attempt
is made towards identi¢cation of the aetiological agent/factor.

165
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MATERIALS AND METHODS

The investigation was carried out in the district of Murshihabad in India, bordering
Bangladesh, during the last week of December 1997.
Out of a total population of 844 188 cattle and 84 001 bu¡aloes, 21 846 animals were
a¡ected, comprising 420 cattle and 21 426 bu¡aloes of either sex. Animals of more
than 2 years to 8 years of age su¡ered from the disease, exhibiting gangrenous lesions
of the extremities, e.g. legs, tail, ears and, in some cases, the tongue. Some of the
a¡ected animals also showed cracks on their hooves. In severe cases there was
sloughing of the necrosed skin.
The following materials were taken:

(1) Fifty sloughed-o¡ pieces of skin (41 from bu¡aloes and 9 from cattle) and 10
biopsies (5 each from cattle and bu¡alo) were collected from lesions on the legs,
tail and ears, both into 10% formalin and into sterile test tubes. The materials
collected into 10% formalin were processed for histopathological examination by
H&E stain and periodic acid^Schi¡ stain (PAS). The samples collected in sterile
test tubes were brought on ice to the laboratory and subjected both to
bacteriological examination by culture at 378C on nutrient agar, MacConkey
and blood agar media in a bacteriological incubator and to mycological
examination by culture on Sabouraud's dextrose agar medium (SDA) and
SDA^cycloheximide and chloramphenicol agar (SDACC) at 278C and 378C in
a biological oxygen demand incubator (BOD incubator).

(2) Skin scrapings were collected from a¡ected tails for examination for mange
infestation and mycotic infection. Twenty-two samples were collected from
bu¡alo and 5 samples from cattle.

(3) Samples of the rice straw which was being fed were collected from households
where the animals were a¡ected. Samples of straw were also brought from South
24-Parganas district, West Bengal, India, from households where the animals had
not developed signs of the disease. Scrapings from mould-infested areas of the
straw were subjected to direct cultural examination on SDA and SDACC at 278C
and 378C.

(4) Rectal faecal samples from 41 a¡ected bu¡aloes and 9 infected cattle were
collected into 10% formalin for examination for coccidian oocysts and the eggs
of gastrointestinal nematodes.

(5) Blood smears were prepared from samples taken from an ear vein from the same
animals from which the pieces of skin and rectal faecal samples were collected.
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RESULTS

On enquiry from the animal owners, it was learnt that the disease syndrome had been
observed since the last week of November 1997, following a few days of intermittent
winter rain.
All the a¡ected animals had been stall fed since October 1997, being mainly o¡ered
paddy straw. The numbers of animals were 2^6 per household, as is general under rural
conditions in West Bengal.
The gangrenous syndrome was characterized by necrosis and gangrene of the tail,
the lower part of the limbs (Figure 1), the ears and, in some cases, the tongue. In severe
cases, sloughing of the tail had occurred.

Figure 1. Gangrene of lower part of limbs

Bacteriological and fungal cultural examination of the samples did not yield any
pathogenic organisms of importance. Blood smears were negative for any haemopro-
tista. Examination of skin scrapings for fungus and parasites after treatment with 10%
KOH failed to produce any positive results. Examination of faecal samples from
a¡ected animals were negative for coccidian oocysts and helminth ova.
Scrapings from the mouldy straw examined under the microscope revealed typical
1^2-celled microconidia (9.89+0.89 mm) and multicelled macroconidia (21.94+2.66
mm), which resembled those of Fusarium spp. (Figure 2).
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Figure 2. Microconidia and macroconidia of Fusarium (original magni¢cation 6400). m,

microconidia; M, macroconidia

Samples of straw collected from households where cattle and bu¡aloes were a¡ected
yielded typical Fusarium-like colonies from 48 h onwards on culture on SDA at
278C+28C, whereas typical colonies only developed from 96 hours onwards on
SDACC. Microscopic examination of these cultures revealed similar macroconidia
and microconidia to those in the scrapings from the mouldy hay.
The sections from the tail showed necrosis in which the architectural detail was
completely lost. The super¢cial layer was disrupted and exfoliated (Figure 3). Sweat
glands and sebaceous glands could not be identi¢ed. However, in a few areas, dead hair
follicles could be recognized (Figure 4). No PAS positive bodies could be detected in
these sections.

DISCUSSION

The morbidity of this disease was 25.5% in bu¡aloes compared with only 0.04% in
cattle. The overall mortality in cattle and bu¡alo was 0.01%. It was not possible to
calculate separate mortality rates for cattle and bu¡alo owing to non-availability of
species-speci¢c mortality reports. Although the mortality was not great, enquiry
revealed that a¡ected male bu¡aloes could not be used for ploughing for at least 30
days. The local rate for ploughing is Rs 100 per day, so the episode caused heavy
economic losses to the farmers in the area.
 169

Figure 3. Exfoliation and disruption of super¢cial layer of tail (original magni¢cation 6200,
H&E)

Figure 4. Photograph showing presence of dead hair follicles (original magni¢cation 6400,
H&E). H, hair follicle
170

The causative agent of Degnala disease is still debatable and various workers from
several parts of the country have tried to discover it. In India two schools of thought
exist (Sastry, 1983). Some workers have considered that the cause of the gangrenous
syndrome is a mycotoxin produced by Fusarium spp. (Charan et al., 1973; Kwatra,
1980; Mallick et al., 1990). All of these three groups were able to isolate Fusarium from
the rice straw. Kwatra (1980) successfully produced the typical gangrenous syndrome
after feeding a broth culture of Fusarium equisetti and potassium oxalate.
In some other countries, a more or less similar condition known as `fescue foot' has
been reported. The disease was described in animals fed on hay and an active principle
could be extracted from the hay. The possible role of a mycotoxin produced by
Claviceps, Fusarium, Aspergillus terreus and Epichloe typhina has been suggested
(Blood et al., 1983). Conversely, it has been claimed by various workers (Dhillon,
1973; Prasad et al., 1982; Basak et al., 1994) that chronic selenosis is responsible for
Degnala disease. However, these two situations, selenosis and mycotoxicosis of
Fusarium origin, are very much dependent upon external conditions.
Selenium toxicity mostly occurs in arid zones and particularly in areas where the
mean annual rainfall is less than 500 mm. Otherwise excessive rainfall may promote
leaching out of the available selenium from the soil (The Merck Veterinary Manual,
1973; Jones et al., 1981; Blood et al., 1983). On the other hand, Fusarium contamina-
tion appears to be associated with cool and wet weather. Ideal conditions are said to be
intermittent rain in the winter season (Bains, 1979). During the present investigation it
was observed that there had been a few days of rain during the winter season in
Murshidabad district.
Apart from selenium toxicosis and Fusarium-associated mycotoxicosis, other condi-
tions such as foot rot, ergot poisoning, and sequelae of enteric salmonellosis may
produce similar lesions. However, the possibility of foot rot may be ruled out because
the lesions of that disease are restricted to the foot and the causative organism can be
found in the a¡ected feet. Ergot poisoning occurs when seeds are infected with
Claviceps purpura during warm and wet seasons (Blood et al., 1983). In the present
investigation, ingestion of such seeds was out of the question because the animals were
fed only on rice straw and, in addition, the weather during the reported outbreak was
wet and cold. Finally, gangrenous dermatitis as a sequel to enteric salmonellosis only
develops after the onset of enteric signs (Blood et al., 1983). The bu¡aloes and cattle
involved in the present investigation never developed enteric signs before the develop-
ment of the gangrenous syndrome.
Thus, the history, laboratory investigation, climatic conditions and clinicopatholo-
gical studies in the present outbreak of gangrenous syndrome or Degnala disease
among cattle and bu¡aloes support the possibility of Fusarium-associated mycotox-
icosis. However, further laboratory investigations, such as estimation of mycotoxin
concentrations and experimental feeding of Fusarium-contaminated straw to suscep-
tible animals, are needed to provide substantial con¢rmation of these observations.
 171

REFERENCES

Bains, B.S., 1979. A Manual of Poultry Diseases, (Editions Reche, Basle), 179^182
Basak, D.N., Banerjee, L.N., Mitra, M., Roy, P. and Chakraborty, A., 1994. Incidence of spontaneous
degnala-like disease in bu¡aloes in and around Naksipara P.S. of Nadia district, West Bengal. Indian
Veterinary Journal, 71, 1225^1228
Blood, D.C., Radostits, O.M. and Henderson, J.A., 1983. Veterinary Medicine: A Text Book of the Diseases
of Cattle, Sheep, Pigs, Goats and Horses, 6th edn, (The English Language Book Society and Ballie©re-
Tindal, London), 581, 1157, 1169
Charan, K., Singh, S.P. and Singh, K.P., 1973. Chronic ergot poisoning-like syndrome (Degnala disease) in
bu¡aloes and cattle in Haryana and Punjab States. Indian Veterinary Journal, 50, 484^486
Dhillon, K.S., 1973. Preliminary observations on the treatment of Deg (Dek)nala disease in bu¡aloes and
cattle in Haryana and Punjab States. Indian Veterinary Journal, 50, 482^484
Jones, L.M., Booth, N.H. and McDonald, L.E., 1981. Veterinary Pharmacology and Therapeutics, 4th edn,
(Oxford and IBH Publishing Co., Calcutta), 1171
Kwatra, M.S., 1980. Mycotoxicosis as a possible cause of gangrenous syndrome in bu¡aloes: a preliminary
report. Indian Journal of Animal Health, 19, 67^69
Mallick, K.P., Vanamayya, P.R. and Prasad, M.C., 1990. Gangrenous syndrome in bu¡aloes. Indian
Veterinary Journal, 67, 365^366
Prasad, T., Arora, S.P. and Chopra, R.C., 1982. Selenium toxicity as induced by feeding rice husk to bu¡alo
calves: a clinical case report. Indian Veterinary Journal, 59, 235^237
Sastry, G.A., 1983. Veterinary Pathology, 16th edn, (CBS Publishers and Distributors), 682^683
The Merck Veterinary Manual, 1973. (Merck & Co., Inc., Rahway, NJ), 947

(Accepted: 29 October 1998)

Epidëmie du syndrome gangrëneux chez des vaches et des bu¥es dans l'ouest du Bengal: ëtudes clinico-
pathologiques

Rësumë ^ Un syndrome gangrëneux (maladie de Degnala) fut observë chez un nombre important de vaches
et de bu¥es du district de Murshidabad dans l'ouest du Bengal en Inde. L'espe©ce Fusarium spp. fut isolëe de
la paille contaminëe. L'incidence de la maladie fut rëduite lorsque cette paille fut retirëe de l'alimentation
des animaux. Un examen histopathologique montra des nëcroses et une perte structurale pour la peau des
animaux.

Brote de s|¨ ndrome gangrenoso en büfalos y vacas de Bengala Occidental: estudios clinicopatolögicos

Resumen ^ Se diagnosticö s|¨ ndrome gangrenoso (enfermedad de Degnala) en una gran cantidad de büfalos
y vacas en el distrito de Murshidabad en Bengala Occidental, India. Se aislö Fusarium sp. de la paja de
arroz enmohecida con que se alimentaba a los animales. Despuës de eliminar la paja, se observö una
reducciön en la incidencia de la enfermedad. El examen histopatolögico moströ necrosis y përdida de la
arquitectura de la piel.