African Journal of Cellular Pathology 8: 15-20 (2017)
The Official Journal of the Society for Cellular Pathology Scientists of Nigeria
ISSN 2449 - 0776
www.ajcpath.com
SEROPREVALENCE AND HAEMATOLOGICAL STUDIES OF SOME
WILD AND SEMI-DOMESTICATED BIRDS NATURALLY INFECTED
WITH AVIAN POX VIRUS IN ZARIA, NGERIA
Andamin AD1, AkadeFT2, Samuel FU3, Enam SJ4, Aliyu HB5, Abdu PA6, Oladele SB4
1. Department of Agriculture and Natural Resources, Jalingo Local Government, Taraba State,
Nigeria
2. Department of Animal Health, College of Agriculture Jalingo, Taraba State, Nigeria
3. Department of Biotechnology, National Animal Production Research Institute/ABU, Zaria,
Nigeria
4. Department of Pathology, Faculty of Veterinary Medicine, Ahmadu Bello University
(A.B.U), Zaria, Nigeria
5. Avian Unit, Veterinary Teaching Hospital, A.B.U., Zaria, Nigeria
6. Department of Medicine, Faculty of Veterinary Medicine, A.B.U., Zaria, Nigeria
Corresponding Author: Andamin AD
Email: [email protected]
Abstract
Aim: To determine seroprevalence and haematological parameters of some wild and
semidomesticated birds naturally infected with avian pox virus (APV).
Methods: A total of 160 birds belonging to 12 species were used for the study. Serum samples
obtained from these birds were analyzed for antibodies to Avian Pox Virus (APV) using agar
gel precipitation test. Natt-Herricks methods and thin blood smear technique were used for
the haematological analysis.
Results: APV serum antibody positivity was 90%, 100%, 80%, 100%, 70%, 80%, 90%, 70%,
100%, 90%, 80% and 80% for Speckled pigeons, domestic pigeons and Mourning collar
dove, Laughing dove, Village weaver, Cut throat fire finch, Cattle egret, Helmeted guinea
fowl, Rose-ringed parakeets, African silver billed, Senegal parrots and Red-billed quelea,
respectively. The highest PCV of 51.0±4.0%, Hb concentration of 16.7 ± 0.8 g/dl and T.P (5.3
± 0.2 g/dl) were obtained from Rose-ringed parakeet, African silver billed and Laughing dove
respectively. The mean range values for PCV, Hb, MCV, MCH, MCHC and TP of all the
birds in this study were between 28.0 ± 2.3 to 51.0 ± 4.0 %, 3.8 ± 0.4 to 16.4 ± 0.8 g/dl and
2.07 ± 2.02 to 5.3 ± 0.2 g/dl respectively. Also, the mean range values for mean corpuscular
volume (MCV),mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin
concentration (MCHC) of all the birds were between 78.9 ± 4.2 to 138.0 ± 22.3 fl, 28.2 ± 2.5
to 49.1 ± 7.9 pg, 78.9 ± 4.2 to 138.0 ± 22.3 fl, 28.2 ± 2.5 to 49.1 ± 7.9 pg and 31. 6 ± 1.19 to
37.7 ± 1.7 g/dl and 31. 6 ± 1.19 to 37.7 ± 1.7 g/dl, respectively.
Conclusion: Avian pox virus is present in some wild and semi-domesticated birds in Zaria
and could spread to commercial poultry.
Key words: Seroprevalence, Haematology, Wild birds.
consumption. (Newathe and Lamorde, 1982;
Oladele et al., 2001). At the moment, no
information on the estimated number of wild
birdsin Nigeria.Guinea fowls and pigeons are semi
domesticated birds that are kept in towns and
villages; and are also abundant in the wild in
African continent, (Oladele et al., 2005).Domestic
and wild birds have been documented to be
susceptible to some viral diseases. Conventional
disease control programmes are easier to supply to
INTRODUCTION
Birds serve numerous functions in the economy of
many nations in terms of meat and egg supply apart
from their value as a source of income among
peasant (Nuru, 1992).In Nigerian, poultry consist
of about 10 million local chicken, 45 million
guinea fowls, and 1 million each of turkeys, pigeon
and ducks. It is estimated that poultry supplied
about 10% of the total meat for human
15
ANDAMIN ET AL
AVIAN POX VIRUS
birds on organized farms, but are very difficult
where movements are unrestricted. It is these
unrestricted domestic and wild birds that maintain
the virulent viruses and bacteria in circulation,
challenging continuously birds on the commercial
poultry farm, (Newathe and Lamorde, 1982;
Olayemi and Fagbohun, 2006).Avian pox, a viral
disease of birds is caused by one of the larger
viruses of the pox virus group. This relatively
slow-developing disease is characterized in birds
by discrete, proliferative lesion on the skin of the
toes, legs or head and or mucous membranes of the
mouth and upper respiratory tract. Systemic
infections may also occur in birds, (Tripathy and
Reed, 2003). Avian pox is comparable to the pox
infections of wild mammals (Robinson and Kerr,
2001), domestic mammals (for example, sheep
sore-mouth, swine poxes and those of man (small
pox). The subgroup of avipox viruses contain a
number of species and strains that vary in their
pathogenicity and host specificity, (van Riper and
Forrester, 2007).The widespread avian disease has
been found in a large number of bird families, with
Phasianidae,
Emberizidae
seeming
more
susceptible than others. In most birds, avian pox
infections are mild and rarely result in death.
However, when lesions are on the eyelids or
mucous membranes of the oral and or respiratory
cavities, mortality can be high, (Tripathy et al.,
2000). As with many other diseases that are density
dependent, avian pox transmission is enhanced
with increasing vector and or host densities,
(Vargas, 1987; van Riper et al., 2002). In the wild,
the warmer and mesic regions of the world support
more potential vectors, thus in these areas the
prevalence of avian pox is higher, parricularly in
flocking wild birds (Annuar et al., 1983; van Riper
and Forrester, 2007). Rural poultry production in
Nigeria is faced with problems of improper
management practices, such as lack of vaccination
and routine treatment or deworming. This usually
results in a serious problem that lead to decrease in
egg production, low quality of meat and poor
productivity performance by the bird, which could
make the birds more susceptible to avian pox
infections, (Olayemi et al., 2002).To the best of our
knowledge, there is no information on
seroprevalence and haematological studies of some
wild and semi domesticated birds naturally
infected with avian pox virusin Zaria, Northern,
Nigeria. Therefore it is pertinent to determine the
seroprevalence and haematological parameters of
these important avian species in relation to avian
pox, which is equally an important disease
MATERIALS AND METHODS
Study Area
The experiment was conducted in Zaria, Northern,
Nigerian, which is located in the Northern Guinea
Savannah zone (11o 10 N, 07O 38’E).Trees and
grasses characterize the vegetation of this zone
with average rainfall, ranging from 1000mm to
1250mm, and temperature of 17oC to 33oC (Sa’id
et al., 1994; Oladele et al.,2003). Different species
of domestic and wild birds have been identified in
this geographical zone (Oladele et al., 2012).
Experimental Birds
A total of 160 birds consisting of 40 helmeted
Guinea fowls, 20 domestic pigeons, 10 Speckled
pigeons, 10 Mourning Collar doves, 10 Laughing
doves, 10 Village weavers, 10 Cut throat fire
finches, 10 Cattle Egrets, 10 Rose-ringed
parakeets, 10 African silver billed, 10 Red-Billed
Quelea, 10 Senegal parrots were used for the
purpose of the study. Both the wild and semidomesticated birds were sampled from Zaria
metropolis, Kaduna State, Nigeria.
Collection of Blood and analysis
Two to five milliliters of blood were collected from
each bird using sterileplastic disposable 5ml
syringe and 25 x 7 mm gauge needle via brachial
vein. Half of the blood was put in an EDTA bottle
and the remaining half of the blood from each bird
was allowed to coagulate, then centrifuged at 313
xg for 15minutes to obtain serum and stored at 20oC until required.
PCV, haemoglobin,
differential cell count and total protein were
performed according to standard procedures
Serological Procedure
The agar-gel precipitation test was carried out on
the serum samples for the detection of Avian Pox
antibodies using the methods described by
Dhinakar Raj et al. (1995). Agar-gel precipitation
test was carried out using immuno-diffusion plate
at 7.2±0.1 pH and the template was incubated at
37oC in humid chamber to prevent drying of the
agar and read at 24, 48, and 72 hour interval under
diffused light. The result was compared between
the positive and the negative test serum. Positive
serumsamples show a line of precipitin between
the serum antigens well while the negative serum
samples show no line of precipitation.
Data Analysis
Descriptive statistics was performed for all
analysis. kruskal-wallis one way ANOVA was
performed to identify differences in analyses
between species. A value of P< 0.05 was
considered significant.
16
ANDAMIN ET AL
AVIAN POX VIRUS
RESULTS
Ninety per cent of the serum samples obtained
from Speckled pigeons were positive for
antibodies to avian pox virus, 20 (100 %) serum
samples obtained from domestic pigeons was
positive, while 8 (80 %) serum samples out of 10
obtained from Mourning Collar doves were also
positive, 10 (100 %) serum samples out of the10
obtained from Laughing doves were positive for
avian pox virus antibodies, 7 (70 %) were positive
from the 10 sera obtained from Village weavers,
while 8 (80 %) were positive from the 10 sera
Obtained from Cut throat fire finches. Nine (90 %)
serum samples out of 10 obtained from Cattle
Egrets were positive, while out of40 serum samples
obtained from helmeted guinea fowls, 37 (70 %)
tested positive. On the other hand, 10 (100 %) sera
obtained from Rose-ringed Parakeet all tested
positive, while out of 10 sera obtained from
African silver billed 9 (90 %) were positive. All the
10 (100 %) sera obtained from Senegal parrots
were positive, while only 8 (80 %) were positive
out of the 10 sera obtained from Red-billed quelea
(Fig. 1).
120
Seroprevalence (%)
100
80
60
40
% POSITIVE
% NEGATIVE
20
RBQ
SPA
ASB
RRP
HGF
CE
CTFF
VW
LD
MCD
DP
SP
0
SPECIES OF BIRDS…
Fig. 1:Seroprevalence of avian pox virus antibodies of some wild and semi-domesticated birds
Key:
SP-Speckled Pigeon, CTFF-Cut Throat Fire Finch,SPA-Senegal Parrot, DP-Domestic Pigeon, CE-Cattle Egret,
RBQ-Red-Billed Quelea, MCD-Mourning Collar Dove, HGF-Helmented Guinea Fowl, LD-Laughing Dove RRPRose-ringed Parakeet, VW-Village Weaver, ASB-Africa Silver Billed
The mean values for PCV, Hb, RBC and TWBC of
birds that were positive to avian pox virus
antibodies and those without avian pox antibodies
are shown in Table 1. The mean value for PCV of
40.0±1.3% obtained from Helmeted Guinea
fowlthat were positive to avian pox antibodies is
lower than the mean value of 42.0±3.6% recorded
from the same species of bird that were negative to
avian pox antibodies. On the other hand, the mean
values of 3.30±0.3x1012/L of RBC obtained from
Senegal Parrot that were positive to avian pox virus
antibody is higher than the mean value of
2.80±1.4x1012/L of RBC recorded from the same
species of birds that were negative to avian pox
virus antibodies (Table 1). Table 2, shows the mean
differential absolute leucocyte count and total
plasma protein of birds that were positive to avian
pox virus antibodies and birds that were negative
to avian pox antibodies.The mean heterophil
values of 0.2±0.1x109/L obtained from Cut throat
Fire Finch that were positive to avian pox virus
antibodies is lower than the mean heterophil value
of 0.9±0.04x109/L recorded from the species of
birds that were negative to avian pox antibodies.
Similarly, the mean value for basophil
(0.02±0.0x109/L) obtained from Mourning Collar
Dove that were positive to avian pox virus antibody
was lower than the value of 0.05±0.03x109/L
recorded from the same species that were negative
to avian pox antibodies (Table 2).
17
ANDAMIN ET AL
AVIAN POX VIRUS
Table 1: Comparative values of blood cellindices between positive and negative cases of avian pox
antibodies of some wild and semi domesticated birds
Parameters
Weight ( g)
PCV(%)
HB (g/dl)
12
RBC(X10 /L)
MCV (fl)
SP
( n=9/1)
DPG
( n=20)
MC
(n=8/2)
LD
( n=10)
VW
( n=7/3)
278±8.9
161.0±4.8
88.3±1.9
73.8±3.9
250.4±0.0
-
81.7±7.3
-
39.3±1.2
31.9±1.7
35.4±2.7
38±0.0
-
34.7±2.2
12.8±0.4
11.8±0.6
12.7±0.9
23.9±13.8 8.9±0.3
252.2±11
617.6±57 77.9±1.1
9.1± 0.3
115.3±5.7 10.5±0.4
8.87±0.6
9.4±0.0
233.1±0.0
673±246
-
8.6± 0.0
100.±2.3
12.2±1.1
36.3±2.7
37±3.1
37.7±2.8
27.3±2.5
40.±1.3
51.0±4.0
48.8±2.4
40.6±4.8
31±3.4
-
36.3±0.9
38±2.00
34±0.0
42.±3.6
-
41±0.0
28.±16.
44±2.5
12.6±1.
12.8±0.9
10.2±0.6
13.3±0.4
3.8±0.4
16.7±0.8
13.8±1.6
9.8±1.19
12.9±0.4
12.9±0.9
10.7±0.0
13.9±1.2
-
14.±0.0
10.9±4.8
14.5±0.85
ASB
(n=9/1)
SPA
( n=8/2)
RBQ
(n=8/2)
13.7±2.5
3.1±0.2
3.4±0.3
4.5±0.4
2.9±0.2
3.9±0.3
4.9±0.5
2.7±0.1
4.4±0.3
3.8±0.4
5.4±0.7
3.3±0.3
3.7±0.3
3.6±0.0
-
4.9±1.4
-
3.8±0.2
4.4±0.0
1.9±0.0
3.9±2.4
-
3.8±0.0
2.8±1.4
3.8±0.3
130.0±8.7
95.7±5.5
80.1±4.4
128±13.3
99.2±13.8 82.3±9.8
97.8±6.6
102.4±8
139±22.3
98.6±11
119.8±8
83.±5.3
95.5±6.6
87.3±3.2
183.8±0
212.7±8
-
108.1±0
97.9±11.6 117.2±1.3
34.2±4.4
27.9±3.2
37±1.9
35.1±2.4
49.1±7.9
33.3±3.5
40.5±2.9
26±2.2
95.5±6.6
29.6±1.6
57.8±0.0
70.7±29.3 -
36.9±0.0
40.8±3.0
38.±0.4
34.3±0.6
34.±0.5
38.4±1.8
33.2±0.0
34.2±1
33.9±0.5
31.2±1.4
34.±2.6
33.9±0.6
31.4±0.0
33.2±0.0
34.1±0.0
42.6±8.2
33±0.05
2.7±0.3
3.2±0.2
3.8±0.4
4.9±0.3
2. 5±0.3
3.3±0.2
3.7±0.3
35.5±0.6
1.6±0.2
1.1±0.4
1.8±0.2
1.1±0.0
2.1±0.03
1.5±0.2
-
44.8±2.7
35.1±1.6
-
33.6±1
37.1±0.6
33.9±0.0
74.5±16
29±1.9
28.4±2.7
36.2±0.4
-
RRP
(n=10)
-
42.2±0.0
MCHC(g/l)
HGF
( n=37/3)
12.6±0.0
124.2±0.0
MCH(pg)
CE
( n=9/1)
12.5±0.9
CT
( n=8/2)
39.9±2.1
34.6±0.9
39.1±4.7
TWBC(X109/L) 3.1±0.2
3.6±0.3
0.5±0.0
2.9±0.2
3.25±0.2
2.1±0.5
35.2±0.7
3.2±0.3
KEY:
n=Total number of birds sampled positive/negativeASB= African Silver Billed, - = Species of birds with only positive case, SP=Speckled Pigeon, SPA=Senegal
Parrot, PCV=Packed Cell Volume, DPG= Domestic Pigeon, RBQ= Red Billed Quele, HB= Haemoglobin Concentration, MC=Mourning Collar Dove,RRP= RoseRinged Parakeet,RBC=Red Blood Cell Count, LD= Laughing Dove,MCV= Mean Corpuscular Volume, VW= Village Weaver,MCH=Mean Corpuscular
Haemoglobin, CT= Cut throat Fire Finch,MCHC=Mean Corpuscular Haemoglobin Concentration, CE= Cattle Egret,TWBC=White Blood Cell, HGF= Helmeted
Guinea Fowl
Table 2: Comparative values of differential leucocyte count and total plasma protein between positive
and negative cases of avian pox antibodies of somewild and semi domesticated birds
Parameters
H(X109/L)
L(X109/L)
SP
(n=9/1)
DPG
(n=20)
0.2±0.0
0.4±0.1
0.3±0.1
0.01±00
-
0.2±0.1
2.5±0.2
2.8±0.3
2.6±0.2
0.5±0.0
B(X109/L)
0.02±0.0
0.04±00
0.0±0.0
9
E(X10 /L)
9
M(X10 /L)
-
0.02±0.0
0.05±0.03
0.1±0.0
0.03±0.0
0.02±0.0
-
0.06±0.01
0.1±0.0
4.3±0.3
6±0.0
0.1±.0.0
-
0.01±0.0
0.0±0.05
4.4±0.2
-
LD
(n=10)
0.1±0.0
2.9±0.2
1.7±0.3
0.03±00
0.02±0.0
TP(g/dl)
MC
(n=8/2)
4.1±0.7
4.0±0.5
0.03±0.0
0.06±0.0
0.05±0.0
5.3±0.2
-
VW
(n=7/3)
CT
(n=8/2)
CE
(n=9/1)
HGF
(n=37/3)
0.09±0.0
0.2±0.1
0.8±0.2
0.8±0.0
0.2±0.1
0.9±0.04
0.15±0.0
2.3±0.4
2.9±0.2
2.7±0.2
0.6±0.4
1.4±0.2
0.7±0.0
0.01±0.0
0.01±0.0
0.06±0.0
0.01±0.0
0.01±0.0
0.01±0.0
0.2±0.1
0.02±0.0
0.01±0.0
0.2±0.0
0.02±0.0
0.01±0.0
0.13±0.0
0.03±0.0
0.02±0.0
0.02±0.01
0.04±0.0
2.21±0.11
2.4±0.3
3.3±0.2
1.7±0.8
2.4±0.7
4.4±0.0
RRP
(n=10)
ASB
(n=9/1)
SPA
(n=8/2)
RBQ
(n=8/2)
0.4±0.1
0.5±0.1
0.9±0.2
1.0±0.04
0.7±0.5
-
0.12±0.0
0.2±0.0
0.05±0.0
3.6±0.2
2.4±0.3
1.2±0.2
2.1±0.1
2.5±0.3
1.1±0.6
-
0.92±0.0
1.8±0.0
1.24±0.3
0.1±0.0
0.1±0.2
0.02±0.0
0.02±0.0
0.03±0.0
-
0.01±0.0
0.0±0.0
0.03±0.02
0.2±0.0
0.1±0.5
0.02±0.01
0.1±0.04
0.04±0.01
0.1±0.3
-
0.02±0.0
0.0±0.0
0.05±0.03
0.1±0.0
0.2±0.0
0.1±0.04
0.02±0.0
0.0±0.02
0.03±.0.01
0.04±0.0
0.1±0.1
-
0.04±0.0
0.04±0.01
0.01±.0.0
4.6±0.2
4.4±0.4
2.6±0.2
2.9±0.2
2.5±0.2
3±0.0
2.5±0.5
3.4±0.4
4.1±0.6
-
KEY:n=Total number of birds sampled positive/negative SP=Speckled Pigeon HGF= Helmeted Guinea Fowl
- = Species of birds with only positive case, DPG= Domestic Pigeon RRP= Rose-Ringed Parakeet, H= Heterophil,MC=Mourning Collar
Dove,ASB= African Silver Billed, L=Lymphocyte, LD= Laughing Dove,SPA=Senegal Parrot, B=Basophil, VW= Village Weaver,RBQ= Red
Billed Quelea, M=monocyte, CT= Cut throat Fire Finch, TP= Total Protein, CE= Cattle Egret, E= Eosinophil
18
ANDAMIN ET AL
AVIAN POX VIRUS
DISCUSSION
The results of this study demonstrated serological
evidence of avian pox virus (APV) antibodies in all
the wild and semi domesticated birds sampled in
this study. This finding agrees with the report of
Clements (2000) that if the birds are adequately
exposed to APV they are susceptible to one or
more of the avian pox virus strains. This shows that
these species of bird in this study were already
exposed to APV prior to sampling and hence, could
be a source of infection to commercial
poultry.Most of the sampled birds, such as
pigeon’s, doves, weaver birds and guinea fowls
that were positive for APV antibodies were those
usually found around human habitations and
commercial poultry houses. It is therefore,
important to note that they can serve as means of
transmission of avian pox infection as they interact
with the domestic fowls. This could be possible
through contamination of feed and water of
domestic birds with their faeces or scabs formation
or direct contact with the susceptible host and
mechanical means via biting insects.Since no
clinical signs or lesions of avian pox were found in
all the birds where APV antibodies were detected,
these wild birds can serve as reservoirs of avian
pox infection, and consequently, shed the virus in
and around poultry houses, thereby becoming
threat to poultry production in Zaria, Nigeria. The
differences
observed in some of the parameters (Lymphocyte,
heterophil, total protein, monocyte) in this
presented study may be due immunological
response to bring the infection the level of sterile
immunity.The normal PCV values for many bird
speciesranges between 29% and 58.5% (Thrall,
2004). Most of the birds sampled in this study had
values within the normal range values irrespective
of whether they were positive or negative for avian
pox virus antibodies. This finding agrees with the
report of Forrester and Spalding (2003).The lowest
TP values of 2.07±0.2g/dl obtained in this study
from Village Weaver (Ploceuscycullatu) is less
than the lower value of 3.00g/dl established for
both apparently healthy wild and domestic birds,
while the highest value of 5.3±.2g/dl obtained in
this
study
from
Laughing
dove
(Streptopeliasenegalensis) were within the normal
range of 6.00g/dl established for wild and domestic
birds (Campbell and Coles, 1986).
Conclusion
This study demonstrated theserologic evidence of
avian pox virus antibodies in some wild and semi
domesticated birds in Zaria, Nigeria.The
predominant leukocytes of the sampled birds in
this study were the lymphocytes and heterophils,
accounting for up to 95% or more of the WBC on
the average.
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