310
Bas.J.Vet.Res.Vol.19, No.2, 2020.
CLINICAL AND DIAGNOSTIC STUDIES OF BOVINE VIRAL
DIARRHEA IN BUFFALO CALVES AT BASRAH
GOVERNORATE ,IRAQ
Hussein A. Abdul Wahid, Kamal M. AL-Saad
Department Of Internal And Preventive Medicine, College of Veterinary Medicine,
University Of Basrah,Basrah,Iraq.
(Received 7 November 2020 ,Accepted 18 November 2020)
Keywords: BVD, Buffalo calves, Basrah.
Corresponding Author: kamalsad58@yahoo.com
ABSTRACT
Bovine viral diarrhea BVD has been detected and diagnosed in local buffalo
calve breeds of Basrah, Iraq. The study was conducted to examine (980) suspected
buffalo calves under one year old and of both sexes. One hundred sixty-eight
(168)calves give positive results with PCR test. Twenty-five (25) clinical healthy
local buffalo calves are considered as controls. Diseased calves show different clinical
manifestations belong to the disease with a significant increase indicated in the body
temperature, respiratory and heart rate, as well as the capillary refill time of diseased
buffalo calve compared with controls. Results of hematological changes indicated a
significant increase in packed cell volume in diseased buffalo calve than in controls,
Moreover, A significant leukocytopenia due to a significant lymphocytopenia was
also indicated in diseased animals compared with the control group. Results of the
clotting factor indices of diseased calves and controls show a significant decrease in
total platelet counts, However, the platelet volume and the platelet distribution width,
the clotting time, the prothrombin time, the activated partial thromboplastin time was
significantly increased in infected animals than in controls. A significant high value
was indicated in Aspartate and Alanine aminotransferase, Alkaline phosphatize as
well as the blood urea nitrogen, In BVD buffalo animals than in the control animals,
On the contrary, a significant decrease was encountered in total protein in diseased
calves than in the control group. Results of the acute phase response of the current
study revealed a significant increase in haptoglobin in BVD buffalo calves than the
311
Bas.J.Vet.Res.Vol.19, No.2, 2020.
control group, Whereas, a significant decrease in Fibrinogen time has been indicated
in BVD calves. The macroscopic examinations of the BVD carcasses revealed severe
congestion of the intestinal vessels accompanied by Ecchymotic hemorrhagic enteritis
with multiple enlargements of mesenteric lymph nodes along with most parts of the
small and large intestine with pasty fecal materials. Furthermore, atrophy of the
intestinal villi with sloughing of the epithelial lining of villi of the small intestine, as
well to hyperplasia of goblet cells, infiltration of inflammatory cells in the intestinal
mucosa, to congestion of blood vessels was also indicated via histopathological
examinations. It has been concluded that BVD has very harmful effects on domestic
ruminants, which mostly terminated by death, Therefore, applying the control
measures is the final and suitable choice to control and eliminate the disease.
INTRODUCTION
Bovine viral diarrhea is a common disease and an important infection of
livestock causes many economic losses characterized by depression, fever, diarrhea,
and immune suppression (1).
The causative agent, is a BVD virus belong to the genus Pestivirus of the
Flaviviridae. Which is considered as a single-stranded RNA genome (2). And have
another subspecies named Swine fever and Border disease virus as well as BVDV-1
and BVDV-2, according to genome sequences (3, 4). Cytopathic (Cp) and Non-
Cytopathic(Non-Cp) are other classifications according to growth characteristics in
cell culture of the two BVDV strain, Since the cytopathic type cause visual cytopathic
impact, However, the non-cytopathic could grow in cells without any harm (5, 6).
The disease leads to different forms including subclinical benign diarrhea, the
peracute type which mostly highly fatal, Thrombocytopenic, and hemorrhagic form
with hemorrhagic diathesis, Reproductive failure, persistently infected (PI) animals of
a fatal mucosal disease, Abortions form with malformations (7, 8). Moreover,
Animals that are born persistently infected could be considered viral reservoirs and
could shed copious amounts of the virus to the environment via aerosols, mucus
secretions, and fecal matters (6).
The disease causes high economic loss because of the abortion, congenital
defects of newborns, retardation of growth, abnormal reproductive diseases, high
mortalities because of mucosal form, and premature isolation, and maybe the culling
of PI animals (9, 10, 11).
312
Bas.J.Vet.Res.Vol.19, No.2, 2020.
In general, the disease can occur at all animal ages, However,The
consequences of the infection in immune-competent cattle ranges from subclinical or
mild to a highly fatal disease, Further, Most domestic animals like sheep, goats, pigs,
camels and wild animals, such as deer's and wild boars can be infected with BVD
virus and may become an important source of the infection (12, 13).
Bovine viral diarrhea is very scared in local calve buffalo breeds of Basrah
Iraq, And little information has been provided, therefore, the present work aims to
investigate the clinical and diagnostic criteria including hematological, biochemical,
histopathological and evaluation of acute phase response in diseased local buffalo
calve breeds of Basrah, Iraq.
MATERIALS AND METHODS
Examined animals and the study areas: This work was design to examine (980)
buffalo calve less than one year old and from both sexes from all Basrah province
areas, represent fifteen(15) herds with a different management system. The study was
started from, January to December. 2020. One hundred and sixty-eight (168) local
buffalo calve breeds shows signs of enteritis with bloody diarrhea. Twenty five (25)
clinically healthy local buffalo calve breeds of same ages was considered as controls.
Complete clinical examinations has been applied to all animals and fecal samples was
screened to excluded any infection with gastro-intestinal parasites(employing standard
techniques such as direct, floatation and sedimentation, Moreover, Giemsa stained
blood smears was also used to rule out the blood parasitic infection (14).
Blood Sampling and hematologic analysis: Ten milliliter of Blood samples
collected from each local buffalo calf via jugular vein. Ethylendiaminetetraacetic acid
(EDTA-mixed blood(2.5ml) was used to determine Total red blood cell count (RBC),
Hemoglobin concentration (HB), Packed cell volume (PCV), Total platelets count,
Mean platelet volume, The platelet distribution width, and total leukocyte counts
(TLC) on an automatic full digital cell counter from (Beckman, USA). Moreover,
Differential leukocyte counts were estimated on blood smears stained with Giemsa,
according to (15).
Other, 2.5 ml of blood mixed with Trisodium citrate (with a ratio of 9:1) was
used for evaluation of the prothrombin time, activated partial thromboplastin time,
and Fibrinogen time using kits from (Biolabo, France). Furthermore, Clotting time
313
Bas.J.Vet.Res.Vol.19, No.2, 2020.
was evaluated according to Bush (16). The remaining blood sample was used for
extraction of serum which was stored at -20°C until use. Serum samples extracted
from the Blood were tested spectrophotometrically according to the manufactures
instructions of (Roche Diagnostics, Indianapolis, GMBH, Germany) to evaluate,
Aspartate and Alanine aminotransferase, Alkaline phosphatize as well as the blood
urea nitrogen and Total protein. Moreover, the Estimation of Haptoglobin (Sandwich
Elisa method), As the color changes of the stop solution was measures at 450 when
using a spectrophotometer device).
RNA extraction from serum sample: The RNA of the BVDV extracted from 980
suspected buffalo calves serum samples using the QIAamp® Viral RNA kit (RNA
extraction from serum without purification) (ADIAGENE, BioX Dignostic. From
France). This procedure was adapted from the usual literature that came with the
extraction kit.560 μl of AVL buffer +Carrier RNA was added to each serum sample,
homogenized by vortex for fifteen seconds then incubated at room temperature for ten
minutes. after that, 100 μl of the supernatant was transferred to a 1.5 ml microtube.
For the binding preparations, an amount of 560 μl of ethanol 100% was added, then
homogenized by vortex for fifteen Sec..630 μl of the obtained solution was applied to
the corresponding column (Identify columns) and centrifuged for 1 minute at 10000
rpm.
The collection tube was changed and the rest of the mix was put in the
column and centrifuged for one minute at 10000 rpm. The collection tube was
changed, and 500 μl of buffer AW1 was added, followed by Centrifuging for one
minute at 10000 rpm.The collection tube was changed, 500 μl of buffer AW2 was
added and centrifuged for 1 minute at 10000 rpm.The collection tube was changed,
followed by centrifuging for 3 minutes at 10000 rpm.The column was transferred to a
microtube. Then, 60 μl of buffer AVE was added. This was followed by incubation
for 1 minute at room temperature and then centrifuging for 1 minute at 10 000
rpm.The tubes were closed and identified. They were then stored on ice immediately
at <-15°C until tested. Viral RNA is stable for up to one year when stored at –30 to –
15°C or –70°C.
cDNA synthesis and RT-PCR: For cDNA synthesis, 5 μL of extracted RNA, 1 μL of
a specific reverse primer and 14 μL of DEPC treated water were added to the
lyophilized master mix contained in Bioneer AccuPowerTM RT PreMix kit (Korea).
314
Bas.J.Vet.Res.Vol.19, No.2, 2020.
The mixture was incubated at 42°C for 60 min. cDNAsynthesis was terminated by
incubation at 95°C for 5 min. Amplification of 5ʹ UTR (288 bp) was carried out on
serum samples .RNA using the primers 324 (5´-ATG CCC WTA GTAGGA CTA
GCA-3′) and 326 (5′-TCA ACT CCA TGTGCC ATG TAC-3′) to detect pesti-virus
infection as described by Vilcek et al. (1994). In individual samples, amplification of
cDN As by PCR was performed using the primer pairs 0I 100 (5′-CAT GCC CWY
AGT AGG ACT AGC-3′)/1400R (5′-ACC AGT TGC ACC AAC CAT G-3′) as
described by Becher et al. (1999) and BD1 (5′-TCT CTG CTG TAC ATG GCA CAT
G-3′)/BD2 (5′-TTG TTR TGG TACARR CCG TC-3′) nested PCR) as described by
Vilcek et al. (1997) and BD1/BD3 (5′-CCA TCT ATR CAC ACA TAA ATG TGG
T-3′) and BD1/BD4 (5′-CCA TCC ACG CAT ACG TAG ATG TG-3′) to detect the
strains of BVDV as described by Vilcek et al. (2001).
In the nested PCR, at the first PCR, the outer primers 0I 100 and 1400R were
amplified and at the second PCR, 3 μL of the first-round PCR product was amplified
with primers BD1/BD2 using the same number of cycles and the same thermal
profile to obtain a 738 bp DNA fragment. In addition, to amplify a 428 bp amplicon to
detect the BVDV strains NADL and UK the primersBD1/BD3 and BD1/BD4 were
employed, respectively. All oligonucleotide primers were got from a commercial
origin (Bioneer, INC., Korea).
PCR was carried out in a total volume of 30 μLcontaining 3 μL of 10 x PCR
buffer, 0.5 μL of dNTPs(0.16 mM), 1 μL of cDNA, 1 μL of each primer (10pmol), 1.2
μL of MgCl2 (2 mM), 0.3 μL of Taq DNA polymerase (1.5 U) and 22 μL of
DNase/RNase free distilled water. Reactions were completed done in an automated
thermal cycler (Bio-Rad gradient Thermal Cycle) Cycle parameters for PCR were as
follows: initially 95°C for 5 min followed by thirty-five cycles in 3 continuous phases
including 94°C for 30 s, 55°C for 100 s, and 72°C for 2 min, and finally terminated by
a single cycle of a final extension at 72°C for 10 min. The RTPCR-amplified products
were examined by electrophoresis in a 1.5% agarose gel, stained with a 1% solution
of ethidium bromide, and visualized using a UV transilluminator.
Gross post-mortem and Histopathology examinations: Animals that died were
subjected to post-mortem examinations and laboratory histopathological evaluations.
The tissue samples were collected from the intestine. The collected samples were
fixed at 10% neutral buffered formalin solution for 72 hrs, then cut to appropriate
315
Bas.J.Vet.Res.Vol.19, No.2, 2020.
sizes and washed, dehydrated, cleared in xylol. Finally, it was embedded in paraffin
wax, and sectioned at the 4-5μ thickness, stained with hematoxyline and eosin, and
examined under a light microscope (17).
Statistics: The analysis of statistics between diseased and the clinically normal local
buffalo calve breeds was done using (Spss) program, The t-test (18). Data will be
presented as a mean ± standard error of the mean. P<0.05 between diseased and
controls.
RESULTS
The present work results, indicates an overall prevalence of (17.14%) in local
buffalo calve breeds at Basrah province, Iraq, As, out of (980) suspected cases (168)
found positive by PCR test.
Among the most important clinical manifestations that the diseased animals
showed was, Anorexia and unable to suck (95.23%),Clear watery diarrhea mixed
either with mucous or with blood (93.45%), Dehydration with different degrees
(93.45%), Erosive lesions present on the mouth, gums tongue and muzzle (83.92%),
Excessive salivation(81.54%), Petechial hemorrhages detected on ocular mucus
membranes (79.16%), Lacrimation (52.38%). Weakness and emaciation of diseased
animals(38.69%)Table 1.
Table .1: The clinical manifestations of diseased local buffalo calve breeds
Clinical manifestations NO. of
animals
%
Anorexia and unable to suck 160 95.23
Clear watery diarrhea mixed either with mucous or with blood 157 93.45
Dehydration with different degrees 157 93.45
Erosive lesions present on the mouth, gums tongue and muzzle 141 83.92
Excessive salivation 137 81.54
Petechial hemorrhages detected on ocular mucus membranes 133 79.16
Lacrimation 88 52.38
Weakness and emaciation of diseased animals 65 38.69
316
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Moreover, a significant (P<0.05) increase was indicated in the body
temperature, the respiratory and the heart rate, as well as the capillary refill time of
diseased local buffalo calve breeds compared with controls Table 2.
Table. 2 : Body temperature, respiratory, heart rate and capillary refill time of
diseased local buffalo calve breeds and controls
Clinical parameters Control buffalo calves
n=25
Diseased buffalo calves
n=168
Body temperature / ºC 38.4±0.28
40.6±1.32*
Respiratory rate / min 22.0±3.45 68.3±10.78*
Heart rate / min 88.4.3±6.85 133.4±12.67*
Capillary refill time/ min 1.42 ± 0.63 5.23 ± 0.72*
In the current study diagnosis of BVDV was confirmed by using PCR technique
(Fig:1), As, all selected blood samples give positive results
Figure(1): The Gel electrophoresis image showing: lane M) Exact Mark 100-1500bp
DNA ladder; Lane P) cDNA extracted from the diseased local buffalo calve breeds
used as positive control for the BVDV; Lane 1, 3, 4, 5) the Conventional PCR test
detected only BVDV in (approximately band size) ≈288bp; Lane N) cDNA extracted
from the BVDV-free animal used as a negative control.
Results of hematological changes indicated a significant increase (P<0.05) in
(PCV) in diseased buffalo calve breeds than in controls, Moreover, A significant
M P 1 2 3 4 5 N
1500 bp
500 bp
100 bp
≈288 bp
317
Bas.J.Vet.Res.Vol.19, No.2, 2020.
decrease(P<0.05) in leukocyte count due to a significant lymphocytopenia was also
indicated in diseased animals compared with control group. Table. 3.
Table.3. Alterations in hematological parameters in diseased buffalo calve breeds and
controls
Parameters Control buffalo
calves
n=25
Diseased buffalo calves
n=168
RBC x106 7.41±0.46 7.33±1.51
Hb g/dl 12.88±1.74 12.78±1.89
PCV % 34.55±4.73 44.21±6.73*
TLC x103 11.63±1.53 9.65± 0.81*
Lymphocytes /Absolutes 5990.82±300.43 4100.05± 100.62*
Nutrophiles /Absolutes 4870.33± 300.12 4800.86±2.32
Monocytes /Absolutes 320.87±10.21 318.87±30.12
Eosinophiles /Absolutes 300.34±10.23 300.62±10.9
Results concerning the indices of clotting factors of diseased buffalo local
calve breeds and control show a significant (P<0.05) decrease in total number of
platelet counts. However, the values of platelet volume and platelet distribution
width, the clotting time, the prothrombin time, as well as the values of activated
partial thromboplastin time was significantly increased (P<0.05) in infected buffalo
calves than in controls. Table 4.
Table.4: Clotting factor indices of diseased buffalo local calve breeds and controls
Parameters Control buffalo
calves
n=25
Diseased buffalo
calves
n=168
Total platelet counts × 103 443.12± 28.12 255.43± 76.32*
Platelet volume /fl 11.14± 1.21 16.15± 1.84*
Platelet distribution width % 17.66± 1.28 25.72± 3.12*
Clotting time / mint 3.21± 0.71 4.91± 1.65*
Prothrombin time /sec 14.55± 1.15 18.54± 2.32*
Activated partial thromboplastin time /sec 52.75± 3.17 68.16± 4.11*
318
Bas.J.Vet.Res.Vol.19, No.2, 2020.
A significant difference has been encountered in biochemical analysis of
diseased buffalo local calves breeds than the control animals. As significance
(P<0.05) increase was indicated in AST, ALT, ALP and BUN, In diseased buffalo
calve breeds than in controls, However, a significant (P<0.05) decrease were
encountered in total protein in infected animals with BVD than in the control
group.Table.5.
Table.5: Changes of biochemical analysis in diseased buffalo local calve breeds and
controls
Parameters Control buffalo
calves
n=25
Diseased buffalo
calves
n=168
AST U/L 58.15 ± 11.76 87.51 ± 13.04*
ALT U/L 38.53 ± 4.15 68.22 ± 8.78 *
ALP U/L 61.35 ± 7.93 79.28 ± 12.71*
BUN mg/dL 15.42 ±3.42 32.72 ± 9.15 *
Total protein g/dL 7.37 ± 0.26 6.11 ± 1.23*
Results of the acute phase response of the current study reveled a significant
increase(P<0.05) in Haptoglobin values in diseased buffalo local calve breeds than
controls, however, a significant (P<0.05) decrease in Fibrinogen time has been
indicated in BVD calves compared with controls .Table. 6.
Table. 6: Acute phase response of diseased buffalo local calve breeds and controls
Parameters Control buffalo calves
n=25
Diseased buffalo calves
n=168
Haptoglobin g/dl 0.0018±0.011 0.042±0.008*
Fibrinogen time/sec 22.24± 1.37 12.66 ± 4.41*
Ten of severely diseased local calve breeds was dying and macroscopic
examination of the carcasses revealed a severe congestion of the intestinal vessels
accompanied with Ecchymotic hemorrhagic enteritis with a multiple enlargement of
mesenteric lymph nodes along most parts of the small and large intestine with pasty
fecal materials (Fig 2, 3and 4).
319
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Figure 2: Severe congestion of intestinal blood vessels (black arrows )
Figure 3: Ecchymotic hemorrhagic enteritis with pasty fecal materials (black arrows )
320
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Figure 4: Multiple enlargement of mesenteric lymph nodes( arrows )
Furthermore, the results of histopathological analysis of those dead animals
show atrophy of the intestinal villi with sloughing of the epithelial lining of villi of the
small intestine, as well to hyperplasia of goblet cells. Moreover, infiltration of
inflammatory cells in the intestinal mucosa, and congestion of blood vessels was also
indicated .Fig. 5,6 and 7.
Figure 5.: Histopathological section of small intestine showed sloughed epithelial
lining of villi (black arrow), as well to hyperplasia of goblet cells (red arrow). H&E
stain. 10X.
321
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Figure 6: Histopathological section of small intestine showed severe sloughing of
epithelial lining of villi (black arrow), as well to atrophy of intestinal villi (red arrow).
H&E stain. 10X.
Fig.7: Histopathological section of small intestine showed infiltration of inflammatory
cells in the intestinal mucosa (black arrow), as well to hyperplasia of goblet cells (red
arrow), and congestion of blood vessels (blue arrow). H&E stain. 40X.
322
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Discussion
The infection with bovine viral diarrhoea is considered one of the most
important diseases of cattle distributed globally (1,10). However, the treacherous
feature of the causative infectious virus will contribute to substantial economic losses
in both the beef as well as the dairy animals worlds wide (19). The common sequelae
of BVDV infection in cattle are digestive disorders, respiratory, as well as
reproductive complications. Therefore, the most important losses due to the disease's
reproductive disorders might be the most economically important consequences (20).
The results of the present study indicate that an overall percent prevalence rate of
(17.14%) in local buffalo calves breeds at Basrah province, Iraq was indicated for the
first time, using PCR techniques. Results of (6%) prevalence rate were obtained by
(21) at Al-Anbar province (the western region of Iraq). Moreover, a prevalence rate of
(13.96%) was indicated by Hasan and Alsaad (22) in Nineveh in cattle. On the other
hands, it was found a different prevalence rate among different countries around the
worlds, In Iran, a prevalence rate of 18.23% (23), in Turkey 11.45% (24), In Egypt
17.2% (25), In Tunisia,2.65% (26), Moreover, in Poland 3.9% (27), and in Canada
10.44% (28).
It was documented that, Prevalence, refers to the amount of the disease in a
known population at the same time without differences between old and new cases
(29). Different biological measurements can be used for determining the occurrence
of disease infection, including, the clinical, clinical-pathological, the virus virulence,
serological methods, and production measures. However, the importance of each
measure could depend on the purpose of the investigation (1). Therefore, The
prevalence of this disease differed from country to another and from place to another
even within the same country because of the different diverse management systems,
type and number of samples, the sensitivity of the diagnostic technique which used,
availability and the efficacy of the control programs, the variations in climate, control
activity of animal movement, The variation in the animal numbers size, and presence
and persistence of the disease (30,31).
For more understanding of the disease feature which might be considered as a
continual challenge for the owners as well as the veterinarians, Researchers described
different types of Bovine viral diarrhoea, The Acute BVD form, The Severe Acute
form, The Thrombocytopenic hemorrhagic form, The Acute BVD–Bovine
Respiratory Disease Form, and The Acute–immunosuppression form of BVD.
323
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Although in most cases a significant percentage of the disease could result in
subclinical type infections. Nevertheless, whether the disease is one of the acute types
or is a subclinical form in nature, there is a specific period of the virus for shedding
(5,32).
The infection of local buffalo calves breeds with BVD reflected different
clinical signs, Although they are varied and might depend on the genetic mutation
tendency in BVDV strains and the virulence(33). However, The clinical response to
infection could be more complex and depends on different criteria including agent
factors, virulent factors, host factors, etc. which might affect the result of the clinical
disease include immune-tolerance, immune-competence, the general immune status
whether its passive or active immunity (34). Local buffalo calve breeds of the current
study show different clinical manifestations that were mentioned by others (1,12, 35,
36, 37).
It has been documented that anorexia and unable to suck occur because of the
erosive lesions that existed on the mouth, gums, and tongue due to localization of the
virus in the oral mucus membranes causing severe inflammation of these tissues
(stomatitis), which makes the animal refrain from eating or feeding because of the
severe pain resulting from the severity of inflammation with the descent of large
amounts of saliva (5,20). Moreover, different types of diarrhoea were affected
diseased buffalo calve breeds was resulting from, damage to epithelial surfaces of the
gastrointestinal system, However, more virulent strain could lead to more severe
inflammation which results in severe damage and could reach to the deep intestinal
layers causing severe damage of the intestinal blood vessels resulting in hemorrhagic
enteritis along the intestine (38, 39,40).
On the other hand (41,42) added that Acute type of the disease in newborn
animals might reflect severe enteritis and or / pneumonia. Those clinical
manifestations are related to calves suffering from a failure of passive transfer, as
passively acquired maternal antibodies are thought to be a protector. Since the clinical
disease in the face of adequate passive transfer may be related to antigenic diversity
among infecting viral strains and viral strains against which passive immunity was
developed. Lastly, inadequate passive immunity combined with immunosuppressive
effects of BVD infection may result in secondary diseases affecting various organ
systems. Also (25,27), describe a thrombocytopenic hemorrhagic type as a severe
acute form of the disease that appears to be related to a non-cytopathic form of the
324
Bas.J.Vet.Res.Vol.19, No.2, 2020.
disease, As diseased calves were suffering significant thrombocytopenia, petechial
and ecchymotic haemorrhages of mucosal surfaces, epistaxis from the nose, clear
bloody diarrhoea, fever, leukopenia, and death, same signs were indicated in buffalo
diseased calves of the current study.
Local buffalo calves of the present study show signs of excessive salivation.
This could be a usual reflection of inflammation of the oral cavity, as BVD oral
lesions characterized by the erosive type of lesions causing pain, loss of appetite,
unable to suck teats, and might terminate with grinding sound(43). Furthermore,
increase the representation of the vital sign with an increase in body temperature,
respiratory and heart rate as well as increase the capillary refill time which show the
acute form of the disease will also be mentioned by (1,37). It has been documented
that the capillary refilling time is a fast technique apply to check some clinical sign
troubles such as systemic dehydration, peripheral vascular disease, and shock as well
as hypothermia, therefore, the prolonged time of the refill time of the blood vessels
might indicate the low amount of blood flow which will reach to tissues (19).
The results of the current study show no significant changes in values of total
erythrocyte count and hemoglobin concentration, However, PCV was found high, the
same results were also demonstrated by (33,44, 45,46) which might be referred to the
excessive loss of body fluid and dehydration and the haemoconcentration mechanism
of the blood, which lead to decrease plasma volume in diarrheic buffalo calve breeds.
It has been documented that, The primary result of BVD infections is a decrease in
immune system ability and strength due to leukocytopenia (immune-suppression) due
to suppression of the defence cells like macrophages, nutrophiles and lymphocytes,
which are the first to respond to infection (47,48). The results of the current study
show a significant leukopaenia and Lymphopenia in BVD of buffalo local calve
breeds which agreed with the same data reported by (37) who refers to the low
number of both B- and T-lymphocytes in peripheral blood as a consistent finding in
acute BVD infection. Furthermore, (49) added that, previous studies indicate that the
causative agent was localized in the lymph nodes, Peyer's patches, enterocytes,
spleen, thymus, tonsils and liver causing destruction of lymphocytes in those tissues.
The Infection due to BVD could result in mild, less or severe decrease Lymphopenia,
which always has a good associated with the infection and the lesions of the
lymphoid tissues. Further, it was proven that, during BVD infection, Cytotoxic Tlymphocytes
will be more influenced than the helper one with less or with no any
325
Bas.J.Vet.Res.Vol.19, No.2, 2020.
harm on circulating γ/δ T-cells. The CD4+ depletion increases the period of virus
shedding (50). Therefore, BVD infection could increases the susceptibility to another
secondary microbial infections because the lymphocytes from diseased animals will
has impaired memory responses to BVDV and other antigens (42,51).
A significance difference has been indicated in clotting factor indices of
diseased buffalo calve breeds and controls in the current study, Since, it was shown
that, thrombocytopenia might occurs in a regular pattern when BVD was acute,
although, the low values of thrombocytes will not always reflected a severe
hemorrhages (52,53), It was also indicated by the results of the present study. The
exact reason of decreased platelets count might unclear completely, however, the
damage of megakaryocytes and the reduction of thrombocytes production by
megakaryocytes as well as the increased consumption of the platelet cells in the
periphery, and functional defects of the platelets have all been indicated as such a
contributing factors (54). Moreover, (55), added that the development of
thrombocytopenia is directly related to the infection of bone marrow with BVD and
that in the bone marrow, BVD can be detected in all cellular elements including
megakaryocytes. On the other hand, Bleeding tendency or hemorrhagic diathesis
might only occur when thrombocytes have reached a very low number (56,57), Which
was also detected in the current study, reflecting the intense of the clinical signs in
BVD diseased buffalo calve breeds.
In specific situations , some virulent strains of BVDV might result in high
mortalities at the beginning of the disease. As, the bone marrow tissue could be
affected in late stages than other lymphohematopoietic tissues, and thrombocytopenia
develops after infection of the bone marrow, Therefore, the animal might die before
hemorrhagic diathesis will confirm, this could clarify the difference in frequency of
bleeding indicated in the fields and the experimental cases of severe acute disease
(58). The present study indicates a significantly high value of clotting factor indices in
diseased buffalo calve breeds than in controls, which also agreed by(45,57). BVD in
buffalo calves alters the coagulation system which might enhance the development of
disseminated intravascular coagulation(DIC). The most common coagulation
abnormality in diseased animals is a status of Hyper-coagulation associated with the
(DIC) with a severity counting on the duration and virulence of the infectious agent
(59).
326
Bas.J.Vet.Res.Vol.19, No.2, 2020.
Different significant changes have been detected in biochemical analysis of
diseased buffalo calve breeds compared with controls. As, the results show significant
high values of the Aspartate and Alanine aminotransferase, Alkaline phosphatize as
well as the blood urea nitrogen which, could reflect the harmful effects of the skeletal
and cardiac muscles, hepatic as well as renal tissues. High values of the Aspartate and
Alanine aminotransferase was estimated in the diseased buffalo calves with acute
BVD. Both the aminotransferase could be an indicator of hepatic tissue injuries,
However, it could also originate from other tissues such as skeletal and cardiac
muscles. Both of those enzymes could be liberated and found during the pathological
conditions. These findings agree with the same evidence of the literature (33, 60).
Moreover, ALP was also used as an index of a hepatic injury, However, It could also
be useful in Skelton diseases. Since, The enzyme was found in the intestinal, hepatic,
and renal tissues beside bones and will increase with its level when those tissues were
already damaged (61). An increased level of BUN may indicate indirect damage of
renal tissue, and the presence of globins catabolites, however, Heart failure and
dehydration will also elevate the BUN level, which is indicated in the deceased
buffalo calves of the current study (62).
Significant slight hypoproteinemia was indicated in diseased buffalo calve
breeds compared with controls. Same results were also obtained by (44, 46), who
concluded that the relative difference and reduction in total protein level might be due
to, starvation, malabsorption, and diarrhoea, (digestive disturbances) destruction of
proteins due to fever, However, it could be attributed to stress which might affect the
hepatic parenchyma causing hepatic depletion resulting in less protein synthesis.
The present study shows a significant difference in the acute phase response. Since
high values of haptoglobin were indicated in diseased buffalo calves than controls.
This might reflect the unspecific and complex innate reaction that occurs quickly after
tissue injuries, As, the pro-inflammatory cytokines could be released initially at the
site of an insult and will be responsible for the starting of local and systemic defences
(63). It has been shown that the major mediators of acute-phase protein synthesis in
the liver, are, Tumor necrosis factor-alpha (TNF-α), inflammatory cytokines, the
interleukin-6, and interleukin-1-beta. (64). On the other hand, where is knows that
The main and important functions of acute-phase protein and response are an
enhancement of phagocytosis, activation of the immune system, clearance of the
product of inflammations? Nevertheless, It has been thought that the acute phase
327
Bas.J.Vet.Res.Vol.19, No.2, 2020.
protein response is more active and sensitive than leukocyte count as a marker of
inflammations, and are more stable than cellular components, Moreover, they also
confirmed that the acute phase response has a faster response rather than alterations in
leukocyte count in situations where new leukocytes must be generated by the bone
marrow(65).
Hypo-fibrinogenemia and prolonged clotting time which indicated in the
diseased buffalo calve breeds of the present study might suggest the prevalence of
petechial and ecchymotic hemorrhages which detected on the ocular mucous
membranes, However, the thrombocytopenia might also play a good role because of
the depression of the activities, of the bone marrow, enlargement of spleen and
sequestration of the thrombocytes, which reflected due to disturbed homeostasis and
could terminate with infarction due to micro-thrombosis of an important tissue such as
the pulmonary, the brain a well as the intestine (53,55).
Results of the present study show different macro and micro pathological
changes of the dead carcasses which were also mentioned by (5,66,67), who
confirmed that BVD could affect all body tissues and caused harmful changes when
occurring in different ages, however, in young ages, different degrees of enteritis
including the hemorrhagic type was predominant some time. Which is associated with
sloughing of the epithelial lining of the villi of the small intestine, as well as the
hyperplasia of goblet cells, Furthermore, with infiltration of different inflammatory
cells in the intestinal mucosa, and congestion of blood vessels which indicated in the
buffalo calve breeds of the current study.
دراسة سریریة وتشخیصیة للإسھال البقری الحموی فی عجول الجاموس لمحافظة البصرة ،
العراق
کمال الدین مھلھل السعد , حسین عبد الکریم عبد الواحد
فرع الطب الباطنی والوقائی ، کلیة الطب البیطری ، جامعة البصرة
الخلاصة
تم الاشتباه بإصابة عجول جاموس محلیة فی محافظة البصرة بمرض الاسھال البقری الحموی. أذ تم
فحص ( ٩٨٠ ) عجل جاموس بعمر اقل من سنھ واحدة ومن کلا الجنسین ،تاکد اصابة( ١٦٨ ) منھم بفحص
تفاعل البلمرة المتسلسل کما استخدم خمس وعشرون من عجول الجاموس المحلیة السلیمة سریریاً کمجموعة
سیطرة .اظھرت العجول المریضة علامات سریریة مختلفة تعود لمرض الاسھال البقری الحموی مع حدوث
ارتفاع معنوی فی معدلات درجات حرارة الجسم ، معدلات التنفس ، ضربات القلب وسرعة رجوع الدم فی
328
Bas.J.Vet.Res.Vol.19, No.2, 2020.
الاوعیة الدمویة فی العجول المریضة بالمقارنة مع مجموعة السیطرة .کما اظھرت النتائج حدوث زیادة معنویة
فی معدل حجم خلایا الدم المرصوصة فی عجول الجاموس المصابة بالمقارنة مع الصحیحة فی حین انخفضت
معنویا معدلات العدد الکلی لخلایا الدم البیض بسبب انخفاض معدلات الخلایا اللمفیة فی العجول المریضة
بالمقارنة مع العجول السلیمة . وقد اوضحت نتائج معاملات تخثر الدم حدوث انخفاض معنوی فی العدد الکلی
للصفیحات الدمویة فی حین ارتفعت معاملات حجم الصفیحات وسرعة انتشارھا ، معدل تخثر الدم ، معدل
الخثرین وحرک الخثرین الجزیئی فی العجول المریضة بالمقارنة مع مجموعة السیطرة . کما تبین ایضا حدوث
ارتفاع معنوی فی خمائرالاسبارتیت ناقلة الامین و الالنینن ناقلة الامنین والفوسفاتاز القاعدی ویوریا نتروجین
الدم فی العجول المریضة بالمقارنة مع السلیمة وعل العکس من ذلک انخفضت معنویا معدلات البروتین الکلی
فی العجول المصابة بالمرض بالمقارنة مع مجموعة السیطرة . بینت نتائج الدراسة وجود ارتفاع معنوی
ملحوظ فی معدلات بروتینات الطور الحاد حیث ارتفعت معنویا معدلات الھابتوکلوبین فی العجول المصابة
بالإسھال البقری الحموی بالمقارنة مع مجموعة السیطرة بینما انخفضت معدلات منشیء اللیفین فی العجول
المریضة بالمقارنة مع العجول السلیمة . تبین من الفحص العیانی للجثث النافقة بسبب المرض وجود احتقان
الاوعیة الدمویة المعوی الشدید المترافق مع التھاب الامعاء النزفی الکدمی فضلا عن تضخم متعدد وملحوظ
للغدد اللمفیة المساریقیة على طول اجزاء الامعاء الدقیقة والغلیظة مع وجود مود برازیة عجینیة فضلا عن
ضمور واضح للزغابات المعویة وتقشر البطانة الظھاریة لزغابات الامعاء الدقیقة وکذلک تضخم الخلایا
الکأسیة وانسلال الخلایا الالتھابیة مع احتقان واضح للاوعیة الادمویة . استنتج من خلال ھذة الدراسة ان
لمرض الاسھال البقری الحموی تأثیر ضار وسلبی على المجترات المصابة والتی فی الغالب ینتھی بھلاکھا
لذلک فأن اتخاذ تدابیر الوقایة اللازمة والصحیحة ھو الخیار النھائی والمناسب للسیطرة والقضاء على المرض
بشکل نھائی .
REFERENCES
1-Constable, P.D., Hinchcliffe, K.W., Done, SH., Grunberg, W.(2017). Veterinary
medicine: A textbook of the diseases of cattle, sheep, goats and horses. 11th ed.
Philadelphia: WB Saunders Co.
2-Liu, L.H., Xia, H.Y., Wahlberg, N., Belak, S. and Baule, C.(2009). Phylogeny,
classification and evolutionary insights into pestiviruses. Virol.385:351-357.
3-Yazici, Z., Serdar, M.S., Gumusova, S.O. and Albayrak, H. (2012). Molecular
diagnosis and seroepidemiology of pestiviruses in sheep. Vet. Arhiv. 82: 35-45.
4-Ridpath, J.F. (2015).Emerging pestiviruses infecting domestic and wildlife hosts.
Anim. Health Res. Rev.16:55–59.
5-Bachofen, C ., Braun, U ., Hilbe, M ., Ehrensperger, F ., Stalder, H., Peterhans,
E.(2010). Clinical appearance and pathology of cattle persistently infected with
329
Bas.J.Vet.Res.Vol.19, No.2, 2020.
bovine viral diarrhea virus of different genetic subgroups. Vet Microbiol. 141(3):
258–267.
6-Lanyon, S.R., Hill, F.I., Reichel, M.P., Brownlie, J. (2014). Bovine viral diarrhoea:
Pathogenesis and diagnosis. Vet. J. 199:201-209.
7-Firat, I.,Bozkurt, H.H., Turan, N. and Bagcigil, F. (2002). Distribution of bovine
viral diarrhoea virus (BVDV) in the genital system tissues of cattle. Vet. Arhiv.
72:35−48.
8-Hilbe, M., Stalder, H., Peterhans, E., Haessig, M., Nussbaumer, M. and Egli, C.
(2007).Comparison of five diagnostic methods for detecting bovine viral diarrhea
virus infection in calves. J. Vet. Diag.Invest.19:28−34.
9-Ezanno, P., Fourichon, C. and Seegers, H.(2008).Influence of herd structure and type
of virus introduction on the spread of bovine viral diarrhoea virus (BVDV) within
a dairy herd. Vet. Res., 39(5):39.
10-Garoussi, M.T., Haghparast, A.R. and Rafati, M.S. (2011). The prevalence of
bovine viral diarrhea virus in persistently infected cows in industrial dairy herds in
suburb of Mashhad-Iran. Iran. J. Vet. Med.5: 198-203.
11-Hessman, B.E., Fulton, R.W., Sjeklocha, D.B., Murphy, T.A., Ridpath, J.F. and
Payton, M.E. (2009). Evaluation of economic effects and the health and
performance of the general cattle population after exposure to cattle persistently
infected with bovine viral diarrhoea virus in a starter feedlot. Am. J. Vet.
Res.70:73−85.
12-Everman, J.F. and Ridpath, J.E . (2002). Clinical and epidemiologic observations of
bovine viral diarrhea virus in the northwestern United States. Vet. Microbiol., 89:
129-139.
13-Kaiser, V., Nebel, L., Schüpbach-Regula, G. R., Zanoni, G. and Schweizer, M.
(2017). Influence of border disease virus (BDV) on serological surveillance within
the bovine virus diarrhea (BVD) eradication program in Switzerland. Vet.
Res.13:21.
14-Zajac, A.M., and Conboy, G.A. (2012). Vet. Clin. Parasite., 8th ed, (p 3-39). UK:
Wiley-Blackwell.
15-Weiss, DJ., Wardrop, KJ. (2010).Schalm's Veterinary Hematology,6th ed(Ames,
Wiley-182 Blackwell).
16-Bush, B. M. (1975). Veterinary Laboratory Manual, The Gresham Press, London.
330
Bas.J.Vet.Res.Vol.19, No.2, 2020.
17-Maxie .M.G .(2016). Pathology of domestic animals Sixth Ed (Vol. 2,3). Academic
press. Elsever.
18-Leech,N.,Barrett,K., Morgan,G.A. (2013).SPSS for intermediate statistics :Use and
interpretation. Routledge.
19-Smith, BP.(2004). Large animal internal medicine, 4th ed., New York, Mosby.
20-Almeida, L.L., Miranda, I.C.S., Hein, H.E., Neto, W.S., Costa, E.F., Marks, F.S.,
Rodenbusch, C.R., Canal, C.W. and Corbellini, L.G. (2013). Herd-level risk
factors for bovine viral diarrhea virus infection in dairy herds from Southern
Brazil. Res. Vet. Sci.95: 901-907.
21-Al-Ajeeli, K. S. A. and Hasan, A. S. h.( 2011). Detection of Bovine Viral Diarrhea
Virus by Conventional RT-PCR: A comparative Study. Al-Anbar J. Vet. Sci. 4
(2):121-128.
22-Hasan, S. D. and Alsaad, K.M. (2018a). Evaluation of clinical, hematological, blood
coagulation and some biochemical parameter changes in clinically infected cattle
with bovine viral diarrhea. IOSR Journal of Agriculture and Veterinary Science
(IOSR-JAVS) Volume 11, Issue 3 Ver. II .64-70
22-Hasan, S. D and Alsaad, K.M.(2018b). Bovine Viral Diarrhea And Persistently
Infection Of cattle At Nineveh Province, Iraq. Bas.J.Vet.Res. 17(2).14-32.
23-Safarpoor, D. F. and Haghighi, N. (2012). Detection of bovine viral diarrhea virus in
bovine and buffalo milk thorough conventional and real-time reverse transcriptase
polymerase chain reaction. Res. Opin. Anim. Vet. Sci. 2: 263-267.
24-Yilmaz, V.(2016).Prevalence of antibodies to Bovine Viral Diarrhea Virus (BVDV) in
blood and milk serum in dairy cattle in Kars district of Turkey., Indian J. Anim.
Res. 50 (5) : 811-815.
25-Soltan, M. A., Wilkes, R. P., Elsheery, M. N., Elhaig, M. M., Riley, M. C. and
Kennedy, M. A .(2015). Circulation of bovine viral diarrhea virus – 1 (BVDV-1)
in dairy cattle and buffalo farms in Ismailia Province, Egypt. J . Inf. Dev. Ctri.,
9(12):1331-1337.
26-Thabti, F., Kassimi, L. B., M’zah, A., Ben Romdane, S., Russo BEN Said, P. M.S.,
Hammami, S. and Pepin, M. (2005). First detection and genetic characterization
of bovine viral diarrhea viruses (BVDV) types 1and 2 in Tunisia. Rev. Med. Vet.,
156(8-9):419-422.
27-Wernicki, A. R., Urban-Chmiel, D.,Stęgierska1, Ł., Adaszek, M.,Kalinowski, A.
and Puchalski, M.(2015). Detection of the bovine viral diarrhoea virus (BVDV)
331
Bas.J.Vet.Res.Vol.19, No.2, 2020.
in young beef cattle in eastern and southeastern regions of Poland. Polish J. Vet.
Sci.,18(1):141–146.
28-Deregt, D., Carman, P.S., Clark, R.M. Burton, K.M.,Olson, W.O. and Gilbert,
S.A. (2002). A comparison of PCR with and without RNA extraction and virus
isolation for detection of BVD virus in young calves. J .Vet. Diagn.
Invest.14(5):433-437.
29-Stevenson, Mark .(2005). An Introduction to Veterinary Epidemiology. EpiCentre,
IVABS. Massey University. New Zealand
30-Farhad, S. D.(2011). Prevalence study of Bovine viral diarrhea virus by evaluation of
antigen capture ELISA and RT-PCR assay inBovine, Ovine, Caprine, Buffalo and
Camel aborted fetuses in Iran. AMB Express.,1(32):1-6.
31-Graham, D. A.,Clegg, T. A., Lynch, M. and More, S. J. (2013). Herd-level factors
associated with the presence of bovine viral diarrhea virus in herds participating in
the voluntary phase of the Irish national eradication program. Prev. Vet. Med.
112:99–108.
32-Grooms, D., Baker, J.C., Ames, T.R. (2002). Diseases caused by bovine virus
diarrhea virus. In: LargeAnimal Internal Medicine, 3rd Ed. Ed. Smith BP,pp: 707–
714. Mosby, St. Louis, MO.
33-Hasan, S. D and Alsaad, K.M.(2018b). Evaluation of clinical, hematological, blood
coagulation and some biochemical parameter changes in clinically infected cattle
with bovine viral diarrhea. IOSR-JAVS. 11(3)Ver. II .64-70.
34-Zhong, F., Li, N., Huang, X., Guo, Y., Chen, H., Wang, X., Shi, C. and Zhang, X.
(2011).Genetic typing and epidemiologic observation of bovine viral diarrhea
virus in Western China. Virus Gen. 42: 204-207.
35-Van Vuuren, M. (2006).A review of bovine viral diarrhea virus infection in livestock
in Southern Africa. Perspectives in Agriculture. Vet. Sci. Nutrit. Natural Resou.
1:9.
36-Alsaad, K. M., Al-Obaidi, Q. T. and Hassan ,S. D.(2012a). Detection of bovine
viral diarrhea virus antibodies in cows and buffaloes milk in Mosul, Iraq. Res.
Opin. Anim. Vet. Sci.,2(3):158-160.
37-Brodersen, B. W. (2014). Bovine Viral Diarrhea Virus Infections: Manifestations of
Infection and Recent Advances in Understanding Pathogenesis and Control.
Vet.Pathol.51(2): 453-464.
332
Bas.J.Vet.Res.Vol.19, No.2, 2020.
38-Bazargani, T. T., Hemmatzadeh, F., Nadjafi, J. and Sadeghi, N. A. (2008). BVDV
induced gastro-neuropathy outbreak in a feedlot calves around Tehran (Iran). Iran.
J. Vet. Res.9(3):271-276.
39-Talafha, A,; Hirche, S.; Ababneh, M.; Al-Majali, A. and Ababneh, M. .(2009).
Prevalence and risk factors associated with bovine viral diarrhea virus infection in
dairy herds in Jordan. Trop. Anim. Health Prod., 41: 499-506.
40-Tao, J., Liao, J.H., Wang, Y., Zhang, X.J., Wang, J.Y. and Zhu, G.Q.(2013).
Bovine viral diarrhea virus (BVDV) infections in pigs. Vet. Microbiol. 165:185-
189.
41-Liebler-Tenorio, E.M., Ridpath, J.F,, Neill, J.D (2003a). Distribution of viral
antigen and development of lesions after experimental infection of calves with
aBVDV 2 strain of low virulence. J. Vet. Diagn. Invest. 15:221–232.
42-Liebler-Tenorio, E.M., Ridpath, J.F., Neill, J.D (2003b). Lesions and tissue
distribution of viral antigen in severe acute versus subclinical acute infection with
BVDV2. Biologicals 31:119–122.
43-Brock, K.V. (2004). The many faces of bovine viral diarrhea virus. Vet. Clin. North
Am. Food Anim. Prac. 20:1-3.
44-Fernández-Sirera, L., Mentaberre, G., López-Olvera, J. R., Cuenca, R., Lavín, S.
and Marco, I.(2011). Hematology and serum chemistry of Pyrenean chamois
(Rupicapra pyrenaica) naturally infected with a border disease virus. Res. Vet.
Sci. 90(3): 463-467.
45-Oguzhan, A., Sibel, Y. and Oya, B.(2014). Changes in Hematological Parameters in
Cattle Infected with Bovine Viral Diarrhea Virus. Acta Sci. Vet. 42: 1173.
46-Galbat, S. A., El-Shemy, A. and Keshta. H. G.(2015). Clinical, Hematological and
some Biochemical alterations in calves during diarrhea. Int. J. Adv. Res., 3(12):
191 – 196.
47-Bruschke, C., Weerdmeester, K., van Oirschot, J. and van Rijn, P.
(1998).Distribution of bovine virus diarrhoea virus in tissues and white blood cells
of cattle during acute infection.Vet. Microbiol. 64:23–32.
48-Goens, S.D.(2002). The evolution of bovine viral diarrhea: A review. Can. Vet.
J.43:946–954.
49-Pedrera, M., Gómez-Villamandos, J.C., Risalde, M.A., Molina, V. and Sanchez-
Cordon, P.J.(2012b). Characterization of apoptosis pathways (intrinsic and
333
Bas.J.Vet.Res.Vol.19, No.2, 2020.
extrinsic) in lymphoid tissues of calves inoculated with non-cytopathic bovine
viral diarrhoea virus genotype 1. J. Comp. Patho.,146: 30–39.
50-Ellis, J. A., W. C. Davis, E. L. Belden & D. L. Pratt. (1988). Flow cytofluorimetric
analysis of lymphocyte subset alterations in cattle infected with bovine viral
diarrhea virus.Vet. Pathol. 25, 231–236.
51-Ridpath, J.F., Bendfeldt, S., Neill, J.D., and Liebler-Tenorio, E.(2006).
Lymphocytopathogenic activity in vitro correlates with high virulence in vivo for
BVDV type 2 strains: Criteria for a third biotype of BVDV. Virus Res.,118(1-
2):62-69.
52-Corapi, W., French, T. and Dubovi, E.(1989).Severe thrombocytopenia in young
calves experimentally infected with noncytopathic bovine viral diarrhea virus. J.
Virol.,63:3934–3943.
53-Radwińska, j. (2010). Effect of the BVD-MD virus on coagulation And fibrinolytic
systems in dairy cows. Bull .Vet. Isnt. Pulawy. 54:293-298.
54-Walz, P., Steficek, B. and Baker, J.(1999a). Effect of experimentally induced type II
bovine viral diarrhea virus infection on platelet function in calves. Am. J. Vet.
Res.60:1396–1401.
55-Walz, P. H., Bell, T. G., Grooms, D. L., Kaiser, L., Maes, R. K. & Baker, J. C.
(2001).Platelet aggregation responses and virus isolation from platelets in calves
experimentally infected with type 1 or type II bovine viral diarrhea virus. Can. J.
Ve. Res. 65: 241–247.
56-Rebar, A.H., Mas Williams, P.S., Feldman, B.F., Metzger, F.L., Pollock,
R.V.,Roch,J.( 2005). Platlets: Overview ,Morphology,Quantity ,Platelets function
disorders. Int. Vet. Inf.21:805-825.
57-Alsaad, KM., Al-Obaidi, QT. And Hassan, SD. (2012b). Cinical, haematological
and coagulation studies of bovine viral diarrhoea in local Iraqi calves ,Bulg.
J.Vet.Med.15: (1) 44-50.
58-Walz, P. H., Bell, T. G. , Steficek, B. A. , Kaiser, L. , Maes, R. and Baker, J.
(1999b). Experimental model of type II bovine viral diarrhea virus-induced
thrombocytopenia in neonatal calves. J. Vet. Diag. Invest.11:505–514.
59-Bick, RL.( 2003). Disseminated intravascular coagulation: Current concepts of
etiology, pathophysiology, diagnosis and treatment. Hematol. Oncol. Clin. North.
Am.17:149.
334
Bas.J.Vet.Res.Vol.19, No.2, 2020.
60-Fiore, F., Cubeddu, G.M., Lai, M.G. and Pintori, G. (2006). Clinical considerations of
bovine viral diarrhea (BVD). XIV Cong. Inte.Fe. Me. S. P. Rum, Lugo (Spain).
61-Kaneko,J.J., Harvey, J.W., Bruss.M.L.(2008). Clinical biochemistry of domestic
animals.6th ed. Elsevier
62-Stockham, S.L., Scott, M.A.(2008). Fundamentals of Veterinary Clinical Pathology,
2nd ed. Ames, IA: Blackwell.
63-Cary, C., Zaias, J. and Altman, N.H. (2009). Acute Phase Response in Animals:A
Review. Cattle. Vet. Rec. 162, 514-517.
64-Jain, S., Gautam, V., and Naseem, S. (2011). Acute-phase proteins: As diagnostic tool.
J. Pharm. Bioallied.Sci. 3(1): 118–127.
65-Tothova, C., Nagy, O., Kovac, G.(2014). Acute phase proteins and their use inthe
diagnosis of diseases in ruminants: a review. Veterinarni Medicina,59.(4): 163–
180.
66-Acland, S. T. (2001), Tissue distribution of bovine viral diarrhea virus antigens in
persistently infected cattle. J. Vet. Sci. 2:81–84.
.67-Duncan, C. G ., Ridpath, J. F ., Palmer, M ., Spraker, T.(2008). Histopathologic and
Immunohistochemical Findings in Two White-Tailed Deer Fawns Persistently
Infected with Bovine Viral Diarrhea Virus. Vet. Diag. Investigate.
