Investigation of

Outbreak
Acute Sudden Death
Syndrome in Mule herds
of Udayapur 2063

Dr.Kedar Karki
Dr. Purnima Manandhar
Dr.Pragya Koirala
 Objective of Study
 Identification of the possible cause of the
Syndrome.

 To identify the causative agent involved.

 To correlate similar outbreak

 To Suggest the possible treatment and

prevention Methods.
 Material and Methods
 Field investigation.

 Laboratory investigation of the relevant

sample.

 Review of clinical and laboratory finding.

History/Epidemiolog
y
The DLSO Udayapur reported
the periodic acute sudden
death of total 31 adult mules in
a herd of 900, during May –
October,2006
History: Symptoms
Reported before death
Increased respiratory rate,
Trembling, Tremor,
Excessive , Sweating,
Sudden death,
 Review of World wide
Epidemiology of Syndrome
 In 1995, 38 horses died to moldy corn poisoning in
Kentucky and Virginia (U.S.A).

 From 1995-2004, 75 horses affected by

neurological disorder in Norway due to feeding of
moldy silage

 Purdue Indiana,(U.S.A.)1/6 horse died due to

moldy corn poisoning from1997-2004
 World wide Epidemiology of
syndrome

Southern Brazil (spring of 1996)

 3 males and 1 female; died (3 to 4 years
old thoroughbred horses)
 native grass supplemented with 2 kg of
corn/horse/day.
World wide Epidemiology of
syndrome
Brazil (1994)
 41/58 died
 caused by fumonisin concentration lower
than 10 ppm.

Oaxaca, Mexico (1998)

 donkeys died showing a neurological
syndrome,
 World wide Epidemiology of
syndrome
 Iran (2003),
Equine Leukoencephalomalacia

 Spain(1988)
leukoencephalomalacia (eaten corn with a pinkish fungal
growth)

 The fumonisins were discovered in 1988 following the

1970 outbreak of Equine Leukoencephalomalacia in
South Africa.
World wide Epidemiology
of syndrome
 North Carolina (1989)
white-tailed deer
Leukoencephalomalacia
moldy field corn (Fusarium moniliforme)
 Preliminary outbreak
Investigation
On field investigation/ physical
examination
 mule/horses - normal health,
 fed 20-25% cracked and moldy corn.
Aspergillus affected Maize
MOLDY CORN
Preliminary Field
Investigation

Post-mortem examination
 Severe congestion of liver, lung, spleen,
heart,
 Serosangqinus fluid in thoracic cavity.
 Hemorrhage in stomach mucosa.

Preliminary cause of sudden death was

suspected for moldy grain poisoning.
 Samples for Laboratory
from dead/healthy mule
 Bacteriological examination
Nasal swab, blood (healthy mules)

 Histopathology examination
Liver, lung, spleen, heart tissue

 Parasitological examination
Blood, Fecal samples (healthy mules)
 Samples for Laboratory
from dead/healthy mule
 Toxicological analysis
Liver, lung, spleen, heart, intestine (dead)

 Serological examination
Serum (healthy mules)

 Mycological culture
Feed grains: Maize, Gram
 Result and Discussion
 Bacteriological culture of tissue specimen,
swab blood - revealed no growth of any bacteria.

 Blood parasite - Negative.

 Chemical toxin in tissue – Negative

 Intestinal parasite - Mixed Stongyles spp.

 Laboratory Investigation:
Results
 Penicillium, Aspergillus, Candida spp
on mycological media (CVL).

 60-110 CFU/gm Penicillium colonies

recorded in feed samples.
(Department of Food Tech and Quality
Control Kathmandu)
 Unacceptably High Fungal Levels
Identified in Horse Feeds
Common fungus
(Aspergillus, Fusarium, and Penicillium) –
Equine feed
(proposed limit of 104 cfu per gram).
 The Mycobiota and Toxicity of
Equine Feeds
 Feed mould counts - <1× 102 to 1×
105 cfu/g.
 Frequent isolated genus - Aspergillus
(40.54%), Penicillium (18.38%) and
Fusarium (16.22%).
Mycoflora of the toxic feeds associated with
moldy corn poisoning in equine outbreaks
in Brazil
 The Mycoflora of 39 feed samples associated with
29 EquineLeukoencephalomalacia (ELEM)
outbreaks was studied from in Brazil. most
frequent mold which occurred Fusarium spp. in
97.4% of samples followed by Penicillium spp. in
61.5% and Aspergillus spp. in 35.9%, (1988 to
1990,)
 Molds, Mycotoxins and Their Effect
on Horses
Pasture grasses, hay, grain, straw and stubble can
all support the growth of various fungi.
Aspergillus, Claviceps, Stachybotrys, Fusarium
and Penicillium. Ontario Canada (September
2003 )
 Horse Feed Recall Affects East
Coast U.S.A.

 The products might contain an ingredient

with an unacceptably high level of
aflatoxin. Aflatoxins are a form of
mycotoxins, and are secondary
metabolites produced by Aspergillus spp.
fungi. by: Erin Ryder, TheHorse.com
News Editor
May 05 2008, Article # 11810
Histopathological changes
Liver
• Fatty degeneration of hepatocytes
• Mononuclear cells infiltration in the form of
few nodules.

Lungs
• Perivascular cuffing
• Infiltration of mononuclear cells in the form
of nodules.
Histopathological changes
Kidney
• Deep medullary region reveals infiltration
of mononuclear cells.

Spleen
• The number of white cells seems to be
increased in the area of white pulp.
Suggestive of chronic/viral disease.
(C.V.L.Patholigy Unit)
Animal Disease Diagnostic Laboratory
Purdue University
 Histological description of hepatocytes necrosis
and vacuolization with bile duct proliferation is
more characteristic of poisoning by the
mycotoxins, Aflatoxins, also produced primarily
by Aspergillus sp. molds. In many species other
than horses, fumonisin does cause liver damage,
however in horses, the lesions are primarily in the
brain and can often be seen at post-mortem and
histologically..
Animal Disease Diagnostic Laboratory
Purdue University

 It is unfortunate that there still is not a post-

mortem test for fumonisins in tissues and that we
still must rely on analysis of feed for confirmation
of the presence of fumonisins.
Steve Hooser

Stephen B. Hooser, DVM, PhD, DABVT

Head, Toxicology Sect. & Asst. Director,
Animal Disease Diagnostic Laboratory
Purdue University
 Cardiovascular change with
Moldy corn poisoning
 Interfere the metabolism of fat -sphingolipids.

 Increased serum, sphingolipids, decreased cardiac

function preceding neurological signs.

 Uncontrolled dilatation of blood vessels of

brain,swelling,increased itracranial pressure, brain
damage
 Laboratory Investigation:
Results
Serum sample-
 Revealed positive for Japanese's
Encephalitis (Elisa)
 Treatment and Control Measure
advised
 Advised for proper drying of grain before
feeding.

 Commercial toxin binders, (Varishta,

Toxicurb) @ 1kg/tone of grain for 15days
along with Liver tonic, (Hepatocare,bioliv),
Immunomodulaters, (immunocare),
Promin,Vitamin B complex).
 Final Diagnosis
 Moldy Corn Poisoning in Mule

 Situation Today- Mortality under control

 1200 mules plying on Udayapur Khotang

Rout
 Conclusion
 Findings of this investigation indicates that moldy feed
grains and ingredients are infested with toxic fungus is
emerging as a new health hazard for livestock and
poultry. Seasonal prevalence of mycobiota of feed
forage need to be assessed.

 Simultaneous use of toxinbinder,adaptogen like

livertonic,mineralmixture and Immunomodulaters drugs
promises to help in minimizing health risk in livestock
and poultry production should be looked into.
 Acknowledgement
 Dr.Rebatiman Shrestha,
 Dr.Sadananda Deo,
 Dr.Ganesh Raj Panta,
 Dr.Salina Manandhar,
 Dr.Vinaya Kumar Karna,
 Dr.Keshab P.Shah,
 Dr.Shanker Yadav
 Mr.Sonfi Shah,
 Mr. .B.B.Kunwar,
 Mr.Tek B.Air.
 Mr.Gyan B.Bogti.
 Mr.Samir Gurung Polchem Hygiene lab Pune
 Mr.Nishant Bhattarai Varsha Multi tech.Banglore

For their contribution in Field and laboratory investigation and

control of this disease syndrome