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Pakistan Vet. J., 25(4): 2005
CLINICAL, HAEMATOLOGICAL AND SERUM MACRO MINERAL
CONTENTS IN BUFFALOES WITH GENITAL PROLAPSE
S. Ahmed, I. Ahmad, L. A. Lodhi, N. Ahmad and H. A. Samad
Department of Animal Reproduction, University of Agriculture, Faisalabad, Pakistan
ABSTRACT
In the present study, 20 buffaloes suffering from genital prolapse and 10 normal pregnant (control
group) buffaloes were used to investigate clinical and haematological changes and serum macro mineral
contents. Blood samples of all animals were collected aseptically in two portions i.e. with and without
anticoagulant. Haematological parameters were determined from samples containing anticoagulant. Serum
was separated from the other portion for determination of calcium, phosphorus and magnesium
concentrations with spectrophotmetric kits. The results showed that there was a drastic decrease in serum
calcium (6.42 ± 1.05 v 10.96 ± 0.95 mg/dl), phosphorus (2.90 ± 0.85 v 5.50 ± 1.61 mg/dl) and magnesium
(1.50 ± 0.53 v 2.40 ± 0.53 mg/dl) levels in prolapsed animals as compared to the controls (P<0.01). There
was also a significant decrease in PCV, Hb concentration, lymphocytes and monocytes, while an increase
in ESR, WBC counts and neutrophils was observed in prolapsed animals as compared to controls.
However, there was no difference in haematological and serum macro mineral contents between vaginal
prolapsed and uterine prolapsed buffaloes. It was concluded that deficiency of calcium, phosphorus or
magnesium might be possible causes of genital prolapse in these buffaloes.
Key words: Haematology, serum biochemistry, genital prolapse, buffaloes.
INTRODUCTION
Livestock play an important role in the economy of
Pakistan by providing most essential items of human
consumption like milk, meat, skins and wool. Nili-Ravi
buffaloes occupy a key position in the rural economy of
Pakistan, particularly for the landless farmers or those
with smallholdings. Due to its importance in the small
and landless farmer’s economy, it is often called as the
“Black gold of Pakistan”. It has also been suggested
that due to its large size, emphasis on selection of
buffaloes for meat production can yield high returns
(Cady et al., 1983).
Genital prolapse is a common obstetrical problem,
which adversely affects productive and reproductive
performance of buffaloes by affecting postpartum
return to oestrus, conception rate and calving interval.
Samad et al. (1987) reported the incidence of genital
prolapse as 42.9% among various obstetrical problems
in buffaloes. The maximum incidence of prepartum
vaginal prolapse was recorded in May, while
postpartum uterine prolapse showed the highest
incidence during July and September.
However, there is little information in the literature
on haematological parameters and serum macro mineral
levels in buffaloes with genital prolapse. Therefore, the
present project was planned to investigate clinical,
haematological and serum macro mineral contents in
prolapsed animals and to compare them with normal
pregnant animals of the same species.
MATERIALS AND METHODS
Experimental animals
The present study was conducted on buffaloes
brought to the clinic of the Department of Animal
Reproduction, University of Agriculture, Faisalabad or
maintained in and around Faisalabad city. For this
purpose, a total of 30 animals were selected, out of
which 20 were suffering from genital prolapse (10 with
vaginal and other 10 with uterine prolapse), while the
remaining 10 were normal pregnant (control). Complete
history regarding the clinical status of the disease was
obtained.
Blood collection and processing
About 20 ml of blood was collected in 2 parts, one
with and other without anticoagulant, from each animal
aseptically in clean, sterilized test tubes by jugular
vene-puncture method. The blood samples containing
anticoagulant were used for haematological parameters
including red blood cells (RBC) count, packed cell
volume (PCV), haemoglobin (Hb) concentration, mean
corpuscular volume (MCV), mean corpuscular
haemoglobin (MCH), mean corpuscular haemoglobin
concentration (MCHC), erythrocyte sedimentation rate
(ESR), white blood cell (WBC) count and differential
leucocytic counts. Serum was separated from blood
samples without anticoagulent and stored at -20°C until
analysed for the calcium, phosphorus and magenisum
contents.
167
Pakistan Vet. J., 25(4): 2005
168
Serum calcium concentrations were estimated
using the calcium kit (Human, Germany, Cat No.
10011). Serum phosphorus contents were determined
using photometric UV Test kit (Cat No. 10027). Serum
magnesium levels were studied by using the
magnesium kit. (Crescent, Cat. No. CE 502).
Statistical analysis
Mean values (± SE) of various parameters for control
buffaloes and those suffering from vaginal prolapse
including (vagino-cervical) and uterine prolapse, were
computed. In order to ascertain magnitude of variation in
various parameters between respective groups, the data
were subjected to unpaired T-test, using computer
programme M-STAT C (Freed, 1987).
RESULTS AND DISCUSSION
Haematology
Mean (± SE) value of red blood cell (106/ul) count
in prolapsed animals (6.31 ± 0.69) was higher than the
controls (6.10 ± 0.43), the difference was statistically
non significant (Table 1). Similar results were reported
by Tarjinder and Singh (1993), who observed RBC
count of 6.5 ± 0.2 x 106/ul in prolapsed animals.
A highly significant (P<0.01) decrease in PCV in
prolapsed animals as compared to the control group was
observed (Table 1). These results are in agreement with
those of Kinney (1967), who stated that the decrease in
PCV in prolapsed animals might be due to possible
release of antidiuretic hormone as a result of stress.
According to the Agarawal (1987), a significant
decrease in PCV due to anorexia and toxaemia was
observed which may hold true for the present study
also. However, no difference was observed in PCV
when the vaignal prolapsed animals were compared
with uterine prolapsed animals (Table 2).
In the present study, a significant (P<0.01)
decrease in haemoglobin concentration was found in
prolapsed animals as compared to the normal control
group (Table1). Tarjinder and Singh (1993) reported
similar results in buffaloes. Decrease in haemoglobin
concentration in prolapsed animals may be due to loss
of body fluid (Maxwell and Kleeman, 1972) or
reduction in the size of erythrocyte rather than number
of RBCs (Benjamin, 1978). This may be considered a
possible explanation of decrease in haemoglobin
concentration observed in prolapsed buffaloes in the
present study. However, no difference was observed in
haemoglobin concentration between vaginal prolapse
and uterine prolapse groups (Table 2). There was a
significant decrease (P<0.01) in MCH in buffaloes
suffering from genital prolapse as compared to controls.
However, there was no difference in MCV and MCHC
between animals of the two groups (Table 1). Similarly,
no difference was found in MCH, MCV and MCHC in
buffaloes with vaginal or uterine prolapse (Table 2).
Table 2: Haematological parameters in buffaloes
with vaginal and uterine prolapse (mean
± SE)
Parameters Vaginal
prolapse Uterine
prolapse
RBC (106/ul) 6.26 ± 0.76 6.42 ± 0.58
PCV (%) 35.54 ± 2.26 36.00 ± 2.24
Haemoglobin
(gm/dl) 11.59 ± 0.98 10.66 ± 1.33
MCV (fl) 64.82 ± 6.73 56.20 ± 5.33
MCH (pg) 18.75 ± 2.68 16.60 ± 2.63
MCHC (gm/dl) 32.73 ± 3.31 29.90 ± 4.94
ESR (mm/hr) 36.34 ± 6.06 36.39 ± 4.07
WBC (103/ul) 12.83 ± 0.75 12.80 ± 1.65
Values for each parameter differ non-significantly
between the two groups.
Table 1: Hematological parameters in prolapsed
and control groups of buffaloes (mean ±
SE)
Parameter Prolapsed
group Control
group T-value
RBC (106/ul) 6.31 ± 0.69 6.10 ± 0.43 0.89 NS
PCV (%) 35.70 ± 2.20 42.10 ± 2.23 7.47**
Hb (gm/dl) 11.26 ± 1.17 14.05 ± 0.84 6.69**
MCV (fi) 81.30 ± 10.98 69.39 ± 6.98 0.49 NS
MCH (pg) 18.00 ± 2.78 23.04 ± 2.32 4.91**
MCHC
(gm/dl) 31.74 ± 4.07 33.29 ± 2.32 1.11 NS
ESR (mm/hr) 36.36 ± 5.33 31.00 ± 1.70 4.10**
WBC (103/ul) 12.82 ± 1.10 6.63 ± 0.56 7.47**
** = Highly significant (P<0.01), NS = Non
significant.
A significant (P<0.01) increase in erythrocyte
sedimentation rate (ESR) was observed in prolapsed
animals as compared to controls in the present study
(Table 1), but no difference was observed between
vaginal and uterine prolapsed animals (Table 2).
Similar results were found by Tarjinder and Singh
(1993). According to Jain (1986), ESR increases in
inflammatory conditions and in acute generalized
infection, in addition to change in concentration of
various proteins in blood. However, according to
Benjamin (1978), change in ESR is a nonspecific
reaction. The ESR in bovine is so variable that a normal
value for use as a diagnostic tool for a specific
condition cannot be estimated.
Higher WBCs counts (Table 1) in prolapsed
group (P<0.01) compared to the control animals as
observed in the present study are similar to those
recorded by Verma et al. (1989), who stated that
increase in WBC could be a result of higher level of
cortisol due to stress in affected buffaloes. Singh et al.
(1992) observed a progressive increase in WBC count
with advancement of normal pregnancy, while Sindhu
(1975) stated that this increase was due to stress. Mohy
et al. (1985) reported that increase in WBC count in
prolapsed animals was due to release of
adenocorticotrophic hormone from the adrenal gland.
Pakistan Vet. J., 25(4): 2005
169
According to Benjamin (1978), the degree of
leukocytosis depends upon several factors such as
nature of the causative agent, severity of the infection,
resistance of the animal and localization of the
inflammatory response. When vaginal prolapsed
buffaloes were compared with uterine prolapsed
animals, no difference was observed in WBC count
between both the groups (Table 2). This may be due to
the fact that degree of leukocytosis increases with the
advancement of pregnancy, as both the conditions occur
approximately at the same stage of parturition, hence no
difference was observed.
In the present study, a significant increase in
neutrophils (P<0.01), while a significant decrease in
lymphocytes (P<0.05) and monocytes (P<0.01), was
recorded in prolapsed animals as compared to control
animals (Table 3). These findings agree with the results
of Tarjinder and Singh (1993), who also reported
neutrophilia in prolapsed animals. This may be due to
increased level of cortisol due to stress (Verma et al.,
1989), or inflammatory process in the genital prolapse
(Malik et al., 1990). However, neutrophilia has also
been reported during excitement, exercise and
adrenaline release (Sastry, 1989). Basophils and
eosinophils remained unchanged in animals of
prolapsed and control groups (Table 3). Similarly, no
difference in differential leukocytic count was observed
between vaginal prolapsed and uterine prolapsed
animals (Table 4). Prepartum vaginal and postpartum
uterine prolapse both usually cause the same degree of
inflammation in the genital tract, which may be a
possible reason for similar differential leukocytic
counts between animals of the two groups in this study.
Serum macro minerals
Calcium
Hypocalcemia was observed in prolapsed group as
compared to the control group (P<0.01) in the present
study (Table 5). Marques et al. (1996) and Salmanoglu
and Salmanoglu (1998) found similar results in their
studies, while the results of Paul et al. (2000) are
contrary to the present findings. Risco et al. (1984)
reported that hypocalcemia resulted in loss of muscular
contractions and ultimately uterine prolapse in
buffaloes. Goff et al. (1990) reported that cortisol might
cause immuno-suppression, hypocalcaemia and loss of
muscular contractions.
When comparison between vaginal prolapsed and
uterine prolapsed groups was carried out, no significant
difference was observed in serum calcium levels
between the two groups (Table 6). This may be due to
the reason that same factors like hypocalcaemia,
inflammation and stress are responsible for both
conditions.
Table 5: Some biochemical parameters (mg/dl) in
prolapsed and control buffaloes (mean ±
SE)
Parameters Prolapsed
group Control
group T-value
Calcium 6.42 ± 1.05 10.96 ± 0.95 11.55**
Phosphorus 2.90 ± 0.85 5.50 ± 1.61 4.79**
Magnesium 1.50 ± 0.53 2.40 ± 0.53 4.38**
* =Highly significant (P<0.01), NS = Non significant.
Table 6: Mean values (± SE) of minerals in
buffaloes with vaginal and uterine
prolapse (mg/dl)
Parameters Vaginal
prolapse Uterine
prolapse
Calcium 6.48 ± 1.04 6.30 ± 1.12
Phosphorus 3.05 ± 0.90 2.61 ± 0.74
Magnesium 1.52 ± 0.61 1.46 ± 0.40
Ca : P 2.12 : 1 2.41 : 1
Values for each parameter differ non-significantly
between the two groups.
Table 3: Differential leukocyte counts (%) in
prolapsed and control buffaloes
(mean ± SE)
Parameter Prolapsed
group Control
group T-value
Neutrophils 37.50 ± 4.95 31.20 ± 3.46 1.28**
Lymphocytes 50.15 ± 4.31 54.20 ± 4.57 2.29*
Monocytes 4.20 ± 4.58 9.20 ± 1.55 4.41**
Eosinophils 3.70 ± 1.84 4.60 ± 1.78 3.60 NS
Basophils 1.00 ± 1.30 0.80 ± 0.92 0.64 NS
* = Significant (P<0.05), ** = Highly significant (P<0.01)
NS =Non significant.
Table 4: Differential leukocytic counts (%) in
buffaloes with vaginal and uterine
prolapse
Parameters Vaginal
prolapse Uterine
prolapse
Neutrophils 36.00 ± 4.67 40.29 ± 4.46
Lymphocytes 50.77 ± 4.42 49.00 ± 4.16
Monocytes 7.92 ± 1.80 7.42 ± 2.94
Eosinophils 4.08 ± 2.00 3.00 ± 1.41
Basophils 1.38 ± 1.50 0.57 ± 0.54
Values for each parameter differ non-significantly
between the two groups.
Phosphorus
There was a significant (P<0.01) decrease in serum
phosphorus level in prolapsed animals as compared to
the control group (Table 5) in the present study. These
results are in agreement with those of Marques et al.
(1996). Pathak and Janakiraman (1987) reported
calcitonin and progesterone insufficiency as a cause of
hypophosphoremia in cattle. Between vaginal and
uterine prolapsed groups of animals, no difference was
found in serum phosphorus levels (Table 6).
Magnesium
In the present study, a significant (P<0.01)
decrease in serum magnesium level in prolapsed
animals was recorded as compared to the control
animals (Table 5). These results are in agreement with
the study of Marques et al. (1996). However, no
Pakistan Vet. J., 25(4): 2005
170
difference was observed in serum magnesium levels
between vaginal prolapsed and uterine prolapsed groups
of animals (Table 6). These results do not match with
those of Paul et al. (2000). The variation in these results
might be due to differences in diet fed to animals used
in these two studies.
Based on the results of the present study, it may be
concluded that deficiency of calcium, phosphorus and
magnesium might be the possible factor that lead to
prolapse of genital tract in buffaloes included in the
present study.
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