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Field investi afions on Lameness Due To Chronic Hypophosphatemia in it gyptian Buffaloes: Clinical Hematological and
Biochemical Studies with Trials of Treatment
Abd EI-Raof, F.M.
Department of Animal Medicine, Fac. Vet. Med., Benha University
ABSTRACT
This study was conducted from April 2003 to May 2004, at Tukh area, Kalubia
Governorate. Diseased buffaloes suffered from severe emaciation and pronounced
lameness. Also there was decrease of milk yield and depraved appetite. Blood picture
showed a significant decrease in Hb, PCV, RBCS, while WBCS, MCV and MCH showed
significant increase. Also, there was significant increase in lymphocytes. On the other
hand, Neutrophils, Basophils, Monocytes did not change significantly. Biochemical
analysis for serum from cases with chronic hypophosphatemia indicated a significant
decrease in serum inorganic phosphorus, sodium, potassium, iron, total iron binding
capacity, copper and glucose. While serum calcium, magnesium and zinc did not change
signif- cantly. There was a significant decrease in serum total proteins and albumin. Also
serum enzyme activities showed a significant increase in serum ALP, AST and LDH,
while ALT did not change significantly. Also serum urea and creatinine increased
significantly. These Animals were successfully treated with Toldimfos 20%
(Tonopltosphan) with vitamin D (Devarol). Also, anti-inflammatory as declofenac acid
2.5% was injected to control locomotor disturbance.
INTRODUCTION
Phosphorus has been reported to have some known functions in the body than other
mineral elements. It is responsible for rigidity of bones and many metabolic processes as
well as it is essential in the structural of phospholipids, phosphoproteins, ATP, enzymes
and chemical components of the cells (Swenson, 1984). The phosphorus status of cattle
has been evaluated frequently with blood serum inorganic phosphorus concentration,
which is however subjected to the influence of recent phosphorus intake. Serum
phosphorus is also influenced by phosphorus mobilization from bone. Bone contains from
80 to 85% of the total phosphorus in the body of mammals. Most of the phosphorus in
bone exists in the form of calciumphosphate. Resorption and formation of bone can occur
simultaneously, but net change in bone phosphorus storage in dairy cows may vary with
stage of lactation. Cows undergo a net loss of both Ca and Phosphorus from bone to help
in supply of these elements during early lactation. This is reversed in later lactation (Wu
and Satter, 2000). However, if phosphorus intake is too low over a prolonged period of
time, resulting in severe loss of phosphorus from bone. The bone will eventually become
weak and porous and could be deformed or broken if stress occurs on bone (Sinner,
1980). Hypophosphatemia is primarily affecting highly producing cattle and usually
associated with prolonged feeding on barseem which is low in phosphorus content (4hd
E1-fatif and
154
A£ivad, 1963). Hypopirespiiatcmia may be classified into acute as a result of influx of
phosphorus into cells or bone and chronic as a result of decreased phosphorus absorption
and may cause muscle weakness and osteomalacia (Marvel! and Kleeman, 1990). Signs of
chronic hypophosphatemia are weakness, pica, lameness, and infertility (Radostitis et a!,.
2000).
Blood picture in chronic hypophosphatemia revealed a significant decrease of RBCs,
Hb concentration and PCV%. While, there was significant increase of WBCs, MCV and
MCH (Selina et at, 1998 and Rezk, 2005). Serum biochemical alterations in chronic
hypophosphatemia included significant decrease of serum phosphorus, sodium,
potassium, glucose, iron, total iron binding capacity, and copper. Also, there was
significant decrease of serum total proteins and albumin. While, there was significant
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increase of serum ALP, AST, LDH activities, urea and creatinine, (Pandy and Misra,
1987, Radostits et a!., 2000, Kaneko et al., 1997, Mousa, 1998, Selim et al., 1998 and
Rezk, 2005;.
Treatment of chronic hypophosphatemia was based on serum biochemical analysis so,
there was good response to administration of phosphate and glucose. Also, administration
of anti-inflammatory is recommended to overcome locomotor disturbance (Selim et at,
1998 and Rezk", 2005).
The objective of this study was to obtain some information about the clinical pattern of
chronic hypophosphatemia among Egyptian bufiaiocs as well as hematological and biochemical changes with trials of treatmen),Abd 1;!-Pao
MATERIALS AND METHODS This study has been carried out on twenty two
Egyptian buffaloes. Twelve of them (8 lactating and 4 pregnant) were examined during
the period from April 2003 to May 2004 at the area of Tukh, Kalubia Governorate. These
animals suffered from pronounced lameness, pica, severe emaciation and normal body
temperature. Barseem was the sole feed for these animals. Another ten clinically healthy
buffaloes from the same locality were fed on concentrate mixture and roughage were
served as a control group. Clinical examinations were carried out according to Kelly
(1984). Two blood samples were collected from jugular vein from diseased and
apparently healthy buffaloes at initial clinical examination and 10 days and 30 days posttreatment according to Kelly (1984). The first sample was taken with anticoagulant
(EDTA) for blood picture examination, while the other was taken without anticoagulant,
left to clot and centrifuged, then the serum was obtained. Some biochemical analyses were
carried on these serum samples._Blood picture including erythrocytic count (RI3Cs),
white blood cells count (WBCs), differential leucocytic count, hemoglobin concentration
(Hb), packed cell volume (PCV), mean corpuscular volume (MCV). mean corpuscular
hemoglobin (MCH) were estimated according to Coles (1986). Commercially available
diagnostic kits were used for coloritnetric determination of serum inorganic phosphorus
(Marinai and Prox, 1973,), calcium (Glinder and King, 1972), magnesium (Bauer, 1982),
iron and total iron binding capacity (TIBC), (Tietz, 1976), sodium and potassium, (Henry
et a1., 1974), glucose (Trinder, 1969), total prothins, albumin, globulin and A/G ratio chewing of inanimate. Moreover the (Alper, 1974),
alkaline nhosphatase, milk yield decreased. The history in(Roy, 1970), Alaaine aminotransferase and Aspartate aminotransferase, (Reitinan and
Frankel, 1957) lactate dehydrogenasc (Cabuud, 1958), urea (Patton and Crouch, 1977)
and creatininc, (Thomas ,1992). Copper and zinc were estimated by using 5 PG atomic
absorption spectrophotometer according to Willis (1960). Diseased cases were treated
with toldimfos 20% (Tonophosphan from intervet Co.) was injected intramuscularly as 25 CC for successive days. In the same time 60 ,,successiv
phosphate were administered orally once daily. Vitamin D (Devarol from Memphis Co.)
was injected intramuscularly (11,000 [U/kg B.W) (Radostitis et at, 2000). Dextrose 25%
was injected intravenously and oral drench of glycerine (110 ml twice daily for 2 days) for
treatment of concurrent ketosis. To control locomotor distur bance, anti-inflammatory
drug as Deciofenac acid 2.5% (Declofianle from Arabco Co.) 4 C.0/100 kg B.wt. was
injected intramuscularly for 5 successive days. On the other hand wheat bran,
concentrates and wheat straw replaced barseem feeding (Radostitis at al., 2000). Data
were statistically analysed according to Tanrhane and Dunlop (2000).
RESULTS AND DISCUSSION
Twelve buffaloes (8 lactating and 4 pregnant) were suffered from severe locomotor
disturbances as there were stiffness in gait and inability to move, (Fig.l & Fig.2). The
lameness shifted from leg to leg and there was crackling sound during walking. Some
animals preferred to lie down for long periods. The appetite decreased and depraved as
there was licking anddicated exclusive feeding on barseem for the last five months
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without supplementation of any source of pllosphonrs in the ration. The results were
similar to the same mentioned by .Smith (1996), Mousa (1998) and Radostitis et al.
(2000). Pregnancy, lactation and heavily feeding on barseem were the main risk factors in
the present study Nagpal at a!. (1968). Locomotor disturbance and muscle weakness
recorded with these cases are usually related to the reduction of 2, 3- diphosphoglyceratc
(2. 3-DPG) in association with deficiency of phosphorus as erythrocytes with low 2, 3DPG concentration have increased binding affinity for oxygen and reduced delivery of
oxygen to peripheral tissues (Kaneko at al., 1997). The pulse and respiratory rates
showed significant increase (70 t 0.55 / min and 32 ± 0.55 / min respectively Table, 1).
This finding is simulating that reported by Selim at at. (1998) and Radoslitis at at
(2000). This acceleration of pulse and respiratory rates may be due to anaemic anoxia
(Radostils at al., 2000). However, the body temperature was within normal value. This
observation was previously reported by Sclinr at a!. (1998). Ruminal movement
decreased significantly (2 f 0.07/2 min Table,2). An increased susceptibility to bloat has
been postulated as an effect of phosphorus deficiency (Radostitis at at., 2000). Values of
pulse and respiratory rates and rurninal movements returned to the normal limits after
treatment.
Regarding, blood picture (Table 2), hemoglobin concentration (Hb%), packed cell
volume (PCV%) and total erythrocytic count (RBCs) showed significant decrease (3.94 ±
0.34%) (27.40 ± 1.23%) and (3.87 ± 0.41 106 / cui mn) respectively. These results were
similar to the those recorded with Selim et al. (1998). The decrease in RBCs, Hb and PCV
might be due to inadequate phosphorus in the plasma and subsequent subsequently there
was a decrease of ATP synthesis (Wang et al., 1985). ATP is essential for maintenance of
shape and deform-ability of RBCs (Kaneko et al., 1997). The reduction of ATP caused
cells to become spherical with increased osmotic fragility and shortening of life span
(Ogawa et at, 1989). Reduction of ATP leading to depletion of 2, 3DPG which is essential
for RBCs integrity (Smith, 1996). Also, phosphate is necessary for the erythrocytes glycolytic pathway (Kaneko et at, 1997). Barseem contains high amount of saponins that
produce hemolytic anemia (Loxey, 1983 and F,1-Bagoury, 1987). The total leucocytic
count showed significant increase (8.052±0.13 x 10 ca/nun, Table,2). Concerning the
values of Neutrophils, Eosinophils, Basophils and Monocytes, there were non significant
changes 52.20 ± 1.1%, 5.6 ± 0.25%, 0.00 ± 0.00% and 6.2 ± 0.45% respectively (Table,
2). On the other hand, lymphocytes increased significantly 36 ± 1.22% (Table, 2). This
increase of total Icucocytic count may be related to some degrees of inflammation
recorded in joints (Radostitis et aL, 2000). The mean corpuscular henroglobin and the
mean corpuscular volume increased significantly 28.952 ± 1.17 and 88.378 ± 3.48,
respectively (Table, 2). These results were nearly similar to those recorded by Selim et at
(1998). The hematological values returned almost to the normal limits after treatment.
Biochemical analysis for serum of diseased animals indicated a significant decrease in
inorganic phosohoi-us 3.36 t 0.17 mg /dl (Table, 3), sodium 117 ± 6.71 mEq/L (Table, 3),
po tassium 3.56 ± 0.55 mEq/L (Table, 3), iron 149 ± 8.94 ug/dl (Table, 3), tota: iron
binding capacity 477.6 ± 11.84 ug/dl (Table 3), copper 58.28 ± 3.13 ug/dL (Table, 3) and
glucose 16.22 ± 1.09 gm/dL (Table, 3). These results were nearly similar to those
recorded by Seffner (1972), Forar et al. (1982), Abdou et at ((986), Ismail and Hussin
(1988), Yates (1990), Hegiazi et at (1993), Mousa (1998), Selin: et at (1998) and Mata
and Bliardwaj (2000). The decrease in the serum inorganic phosphorus wa greatly
attributed to the disturbance in calcium: phosphorus ratio in diet and blood due to
prolonged feeding on barseem which is rich in calcium 0.33% and poor in phosphorus
0.06 (Ca: Ph 5.3 : 1) Morrison (1956). The decrease of iron may be due to the decrease of
serum copper level which is essential for iron absorption and metabolism through
activation of ferroxidase enzyme (Abdel-Maksoud and Abdel-Raof, 1998). The decrease in
the serum copper might be due to the poor nutrition which is accompanied by decrease of
appetite and gradual starvation of the diseased animals through which the stimulation of
gonadotrophins was inadequate as the copper is considered as gonadotrophic hormone
index. Estrogen induces the synthesis of ceruoplasmin copper containing a2 globulin and
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initiats the absorption and metabolism of copper (Henley and Judith, 1995). Ilypoglycemia in chronic hypohosphatemia may be due to anorexia or may he due to some
interference in carbohydrate metabolism or due to less glyconegensis (Selim et at., 1998).
The serum calcium level showed non-significant change 10.9 ± 0.19 mg/dl (Table, 3).
This result is nearly similar to that recorded by Monsa (1998). Also, serum magnesium
level showed non significant change 2.682 ± 0.21 mg/dl (Table, 3). This result is
simulating that recorded. by Mousa (1998) and Selim eta!. (1998).
The serum total proteins and albumin showed decreased significantly 7.239 ± 0.08
gm/dl and 2.60 ± 0.07 gm/dl respectively (Table, 4). While total globulin had nonsignificant change (4.639 ± 0.04 gmldl, Table, 4). On the other hand A/G ratio decreased
significantly (0.56 t 0.04, Table, 4). These results are nearly similar to those mentioned
by Pandey and Misra (1987), Abdel-Maksoud and Abdel-Raof (1998) and Selim et at
(1998). The changed serum biochemical parameters almost returned to the normal limits
after treatment.
Regarding serum enzymes activities, the serum alkaline phosphatasc values increased
significantly 76.42 ± 2.32 U/L (Table, 4). This result is simulating that reported by Abdou
et at (1986) and Mousa (1998). Serum aspartate aminotransferase (AST) increased
significantly 85.07 ± 4.22 U/L (Table, 4). This increase of AST may be regarded to
locomotor disturbance, (Kaneko el at, 1997 and Selim et at, 1998). Also, serum alanine
aminotransferase did not significantly changed 26.87 ± 1.44 IU/L (Table, 4). This result
coincids with that reported by Se//m et at (1998). Also, serum lactate dehydrogenase
activity in-.creased significantly 586 =„7.6 U/[. (Table, 4). These finding are in agreement
with those reported by Mac/son and Neilson (1944) and Pandey and Misra (1987), The
in-crease of lactate dchyciogenase may be related to the advanced lameness and
locomotor disturbance. Concerning the kidney function parameters, serum urea and
creatinine increased sipziificantly 58.52 ± 2.55 fug/dl and 1.998 ± 0.02 mg/dl respectively
(Table, 4). These results are nearly similar to those recorded by Mousa (1998) and Selim
et at (1998). The high level of urea may be due to the result of kidney dysfunction as
there were degenerative changes in tubular epithelium (Kurundkar ct a1., 1981).
There was a good response to the program of treatment and this result similar to the
same record obtained by Radostitis el at (2000) and Sells. et at (1998).
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