Indian Vet. J.
, March 2017, 94 (03) : 79 - 80
Ultrastructural Changes in Low Grade Ochratoxicosis in Broiler Chicken*
N. Sailaja, C. Balachandran1, B. Murali Manohar, R. Sridhar, K. Kumanan and M. Lakshman
Department of Veterinary Pathology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai- 600 007.
(Received : 08-06-2016 235/16
Abstract
An ultrastructural study was conducted on the
kidneys of broiler chickens fed with low levels of
Ochratoxin (OA) [30, 60 and 90 ppb] from 0 to 5
weeks of age. The proximal convoluted tubular
epithelium showed cellular swelling, loss of
brush borders, loose intercellular junctions,
karyolysis and mild swelling of mitochondria
at 30 ppb OA, irregular/ granular nucleoplasm
with electron dense marginated chromatin and
disrupted nucleoli at 60 ppb OA level and the
swollen nuclei, fused inner and outer nuclear
membrane and disrupted chromatin material at
90 ppb OA level. In all these cases OA caused the
swelling, condensation, disrupted cristae, loss of
matrix and small vacuolations in mitochondria
and cytoplasm. The study showed that even 30
ppb levels of OA could induce ultrastructural
changes in the kidney of broiler chicken.
Key words: Ochratoxicosis,Broiler chicken,
Kidney, Ultrastructural changes
Among the various types, ochratoxin A
(OA) is most toxigenic and found as a natural
contaminant of food grains and is responsible
for growth suppressive effects in avian and
mammalian species. The study was focused
on ultrastructural changes in kidney due to
exposure of low concentration of OA in the diet.
Materials and Methods
The OA was produced by using Aspergillus ochra-
ceus NRRL 3174 culture in wheat substrate. The
representative feed samples and wheat culture
powder were quantied for ochratoxin by thin
layer chromatography at Pharmacovigilance
*Part of Ph.D. thesis of rst author approved by Tamil Nadu
Veterinary and Animal Sciences University, Chennai.
1
Corresponding author : Email : cbalachandran2000@yahoo.com
The Indian Veterinary Journal (March, 2017)
Accepted : 29-07-2016)
Laboratory for Animal Feed and Food Safety,
Chennai. The feed pretested for aatoxin, ochra-
toxin-A, T-2 toxin, citrinin, cyclopiazonic acid,
penicillic acid and zearalenone and found free
of these toxins was procured from then Insti-
tute of Poultry Production and Management,
Chennai. Forty eight newly hatched Vencobb
broiler chicks supplied by M/s.Venkateshwara
Hatcheries Pvt., Ltd. Chennai were randomly
allotted into four groups of 12 each (0, 30, 60
and 90 ppb OA in the feed comprising of group
1 to 4 respectively), housed in battery brooders
with ad libitum supply of feed and water and
vaccinated on 7th day with commercially avail-
able vaccine of NDV-F manufactured by M/s
Ventri Biologicals, Pune. Six birds from each
group were sacriced by the end of 5th week of
age. Kidney tissues were xed in 2.5 per cent
glutaraldehyde in 0.1 M phosphate buffer (pH
7.2), processed as per the standard procedures
(Bozzola and Russell, 1998) at RUSKA Labs,
College of Veterinary Science, Rajendranagar,
Hyderabad.
Fig.1 Broiler chick-5w-OA-30 ppb: Transmission electron mi-
crograph-Kidney-PCT-Epithelial cell-Loose intercellular junc-
tions-Cytoplasmic vacuolations-Swollen mitochondria-Loss of
cristae and vacuolations. Uranyl acetate-Lead citrate stain x
9450
79
 Ultrastructural Changes in Low ...
Fig.2 Broiler chick-5w-OA-30 ppb: Transmission electron mi-
crograph-Kidney-PCT-Epithelial cell-Loose intercellular junc-
tions-Cytoplasmic vacuolations-Swollen mitochondria-Loss of
cristae and vacuolations. Uranyl acetate-Lead citrate stain x
18900
Results and Discussion
The proximal convoluted tubules revealed
swelling of epithelial cells, loose intercellular
junctions, loss of brush borders, varied sizes
of nuclei and karyolysis, mild swelling and
vacuolations in the mitochondria with loss of
cristae in a few mitochondria in group 2 (Fig.1).
In addition, dense nuclear membranes, clear
inner and outer nuclear membranes, irregular/
granular nucleoplasm with electron dense
marginated chromatin and dense dark disrupted
nucleoli, condensed mitochondria with disrupted
cristae, disrupted endoplasmic reticulum and
cytoplasmic vacuolations were noticed in group
3 (Fig.2). Apart from above changes, fusion of
inner and outer nuclear membrane, loss of
mitochondrial matrix and cristae were noticed
in group 4 (Fig.3).
The ultrastructural changes observed in
the present study at lower levels of OA indicated
degenerative and necrotic processes which are
dose dependent. These results suggest that
the mitochondria in the proximal convoluted
tubules of kidney were most sensitive to OA
toxicity. The above ndings are in accordance
with the reports of feeding OA to broiler chicken
but at higher levels of 2.0 and 4.0 ppm for 20
days (Dwivedi et al.,1984), 1.0, 9.0 and 20.0 ppm
for 21 days (Solcan et al., 2008) and 1.0 ppm
for 35 days (Jayaramu et al., 2012). Likewise,
abnormal forms of mitochondria in the proximal
convoluted tubules were reported by feeding 50
μg/kg body weight OA to broiler chicken from
6 to 21 days (Solcan et al., 2013). The loss of
mitochondria membrane and integrity of other
80 The Indian Veterinary Journal (March, 2017)
Fig.3 Broiler chick-5w-OA-90ppb-Transmission electron
micrograph-Kidney-PCT-Epithelial cell-Cytoplasmic vacuola-
tions-Fusion of inner and outer nuclear membranes-Disrupted
chromatin material; Mitochondria-Disappearance, disintegra-
tion, vacuolations, loss of matrix and cristae. Uranyl acetate-
Lead citrate stain x 7650
organelles can be due to a competitive inhibition
of protein synthesis by OA and lipid peroxida-
tion. These ultrastructural changes reect
altered mitochondrial function and protein
synthesis (Solcan et al., loc.cit.).
Summary
Feeding of 30, 60 and 90 ppb ochratoxin A
to broiler chickens from o to 35 days caused
swelling, loss of brush borders, variable sized
mitochondria showing vacuolations and loss of
cristae and chromatin margination and disrup-
tion of nucleoli and chromatin disintegration
in the nuclei in the lining proximal convoluted
tubular epithelium.
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