INTRODUCTION

Bangladesh is agro based country. Its economy is mostly depends on agriculture (BBS 2002). Livestock plays an important role in nutrition - directly through the consumption of animal products by livestock owners and their families; and indirectly through the sale of animals and animal products as a source of income (FAO report ,1999).

The genetic composition of animals in the production system determines the response of the system to the different inputs. Often, local livestock breeds do not produce at a high level as they have been selected for survival under difficult conditions, including under-nutrition and exposure to various diseases. In contrast, highly productive breeds are more susceptible to disease, thereby increasing the need for animal health measures. These animals often require a high level of nutrition to gain the production benefits (FAO report, 1999).

Many animals receive maintenance or below maintenance levels of nutrition resulting in low levels of production. Increasing the quantity of feed by reducing the number of animals does not provide a solution as the nutritional value of the available feed is low. The addition of a small amount of higher quality feed can have a large effect on production in this situation. In the case of milk production, the use of cattle with a small body size would result in reduced maintenance requirements of the animal, thus enabling more efficient use of available feed for milk production (FAO report, 1999).

When animals are fed a low quality rations and are not protected from disease, genetic traits for survival are more important than those for production. With an increase in nutrition and health, large gains are made in productivity. Improvements in production traits only become important once certain conditions in health and nutrition are met and production levels reached. These conditions vary between species. Livestock health is a limiting factor to production (FAO report, 1999).

Physiological equilibrium is maintained mainly by the blood in the body (Geneser, 1986).but many physiological conditions may alter this equilibrium. When thorough history and physical examination fail to yield a diagnosis in difficult cases, many practitioners turn to blood samples for a complete blood count and chemistry panel, hoping these tests will identify the problem (Navarre Christine, 2007).

Normal blood work can rule out some diseases. And if there are abnormalities, they might aid in establishing a prognosis and/or developing a therapeutic plan, even if a specific diagnosis is lacking (Navarre Christine, 2007).

The importance of hematobiochemical indices in animal husbandry is well acknowledged. Metabolic disturbance usually by inappropriate feeding without manifestation of clinical symptoms are important in animal husbandry and may cause insufficiently developed breeding cattle (Radostits et al., 2003).

The changes in hematological constituents are important indicators of the physiological or pathological state of the animal (Ahmed Ijaz et al., 2003).Blood examination is also performed for screening procedure to asses general health (Gutienez et al., 1971; Jain, N.C; Peinado,V.I. et al., 1993).

The complete blood count (CBC) is an important  and powerful diagnostic tool as a component of a minimum database. It can be used to monitor  response to therapy, to gage the severity of an illness or as a starting point for formulating a list of differential diagnosis. Interpretation of the (CBC) can be broken down into three sections: evaluation  of the erythrocyte, leukocyte and platelets. Each of these parameters can be interpret individually: however , integration of the data is important for the highest diagnostic yield (Barger et al., 2003).

It is well known that variables such as breed, stage of growth, age, reproduction status and stage of lactation have an influence on many blood parameters (Doornenbal  et al., 1988).

Hematological values such as total red blood cell count (Koubkova et al., 2002) packed cell volume (El-Nouty et al., 1990), Mean corpuscular volume, Mean corpuscular hemoglobin and Mean corpuscular hemoglobin concentration and hemoglobin concentration (Kumar et al., 2000) and white blood cell (Gutienez-De Lar  et al., 1971), i.e lymphocyte and Monocyte are indicated adaptability to adverse environmental condition.  However, hematological values are used for indicating stress and welfare (Anderson et al., 1999). Determination of normal values for hematological and blood biochemical values are important for the clinical interpretation of laboratory data. These indices may vary depending on factors such as sex, age, weather, stress, season and physical exercise ( Kaneco et al.,1999).

Hence, the hematological values during different physiological situations should be known for the diagnosis of various pathological and metabolic disorders, which can adversely affect the productive and reproductive performance of cows, resulting in great economic losses to dairy farmers ( Pyne and Maria, 1981; Dutta et al., 1988 ).

There is less research work in Bangladesh on the hematological and biochemical profiling of cow and its calf.

Aim of the study:

1. To know the hematology and biochemical values in calf.
2. To compare the hematology and biochemical values of cow and calf.
3. To know the nutritional and disease status of animal.

MATERIALS and METHODS

Study Period

This study was conducted for a period of 17 days ( 26 , August-11 september,2009) in Metro Dairy Farm, Kulgaon, Hathazary, Chittagong.

Selection of Farm

Chittagong area is potential for dairy farm because of its high demand of  fluid milk, suitable weather, feeds and fodder availability, available veterinary facilities from Chittagong Veterinary and Animal Sciences University. So there is good communication with university and dairy farms as for giving various technical supports to the farms and for some research work also.  Metro Dairy Farm was selected for conducting the study because of its suitable location, large  population, satisfactory record  keeping system, proper feeding and management and also for their kind cooperation.

Study population

Study population was 30 cross bred multiparous (HF X Local,  Shahiwal X Local,  HF x Shahiwal X Local.) lactating cows and their 30 calves. Cows were in different age and production status. Calves  were also in different age and feeding stage.

Health status of animal

All animals involved in this study  were clinically healthy and for ensure this clinical history was reviewed with the farm manager with the  examination of physical condition.  Their physiological status of cows were non pregnant lactating cow . Calves were mostly lives on milk and some are grass also.

General Management System of METRO DAIRY  FARM

Housing System-The pattern of housing is both face -in and face-out system. There is different shed for different status of animal eg. milch cow shed, heifer shed, dry cow shed, pregnant cow shed and calf shed. In every shed there is individual cow distance, common allay, gutter etc. The floor is made of concrete. Surface of the floor is even and generally no bedding material is used. There is proper drainage facility and quick disposal of animal waste. Farm workers wash the floor three times daily and bath the cows once daily. There is both natural and artificial air  flow and available light in every shed.

Feeding System- There are separate mangers for each of the cattle. Both roughage and concentrates are offered to them. The farmer grows German grass besides the farm area and have own cattle feed mil. Available green fodder is supplied from own fodder land and Concentrate   from own feed mil called Metro Dairy Feed. Only straw is bought from other

Concentrate mixture contains- Rice polish,Wheat bran , Broken maize,Broken rice ,Tiloil cake,Mustard oil     cake, coconut oil cake,Pulse husk, Molasses, salt,Toxin binder etc.The feeding schedule-Concentrate and straw  is supplied at 10am, green fodder at 12am  again concentrate at 6.30pm and straw at 7.30pm.

Calf Management- The calves are weaned and usually fed by pale feeding. Colostrums feeding is practiced by nipple drinker. They supply drinking water from underground water source. They supply concentrate according to thumb rule (3 kg for maintainance+50% of the milk production).                         

Production Management--The farm produces about 310 lit liters in the morning and about 110  liters at afternoon. The average daily yield of the farm is about 420 liters. The farmer practice hand milking. Before milking milker's hands and teat dipping is practiced buy potassium per manganate PPM (0.1%). They also practice dry cow therapy for drying the cows.

Breeding management--Most of the milkers  detect heat of their cows during milking in the morning and AI usually done within 10-14hrs by AI technician. Usually AI is done 2 times  per conception for each of the cows . Sometimes natural service is practiced by the farmers.  Mostly they use HF semen.  Generally they keep  AI sheet as a breeding record which is provided by AI technician.  AI technician or Veterinarian diagnose the  pregnancy by rectal palpation.

Health Therapeutics and Preventive Management--There is a least prevalence of infectious diseases in the farm. When any symptom of sickness arises the farmer calls a veterinarian. Anthelmentic treatment and vaccination is regularly practiced by the farm owner.

Table: Vaccination program that follow in the farm:

Name of the vaccine

Frequency of use

Groups of animals administered

FMD  Vaccine

thrice/year

Cows, calves, heifers

BQ Vaccine

Twice/year

Cows, calves, heifers

Anthrax vaccine

once /year

Cows, calves, heifers

Table : Anthelmintic Treatment followed in the farm:

Name of Anthelmentics

Frequency of use

Dosage

Piper vet® powder(piperazine citrate)

Single dose at 1 month of age

5-10gm/40kg calf wt.

Antiworm bolus® (Levamisole Hydrochlodride +Trichlabendazole)

Every 3months alternate

1 bolus /60kg body wt

LT-vet®(Levamisole Hydrochlodride +Trichlabendazole)

Every 3months alternate

1 bolus /60kg body wt

Percentages of frequency of diseases of animals (last year):

Common diseases that occur in the farm are mastitis20%, milk fever30%, blot5%, metritis3%, abortion4%, arthritis15%, calf diarrhoea20%, Others 3%.

Table : Commonly used drugs in the farm:

Antibiotics

NSAIDs

Antihistaminics

Vitamine andMinerals

Hormonal drugs

Renamycin® Oxytetracycline)

Streptopen®

(Streptomycin + Penicillin)

S-P vet®

(Streptomycin + Penicillin)

Acigent® (Gentamycin )

Amoxyvet® ( Amoxycillin )

Diclovet® ( Diclofenac Sodium)

Ketovet® ( Ketoprofen)

Astavet® (Antihistamine)

Histavet®( Antihistamine)

Dellergen®(Antihistamine)

Calcivit plus®(Calcium preparation)

Acivit ADE® ( Vit A,D and E)

Sancal vet®(Calcium Preparation)

Oxcin® ( Oxytocin)

Record keeping :

Table : Individual Cow Records that are kept in the farm register book.

TAG NO

COW

I D

Age

(years)

Lac

tation no.

Milk Yield

(Lit/die)

TAG NO

COW

I D

Age

(years)

Lactation no.

Milk Yield

(Lit/die)

C-27

C1

6.5

4th

7

C-72

C16

5

2nd

8

C-08

C2

6

3rd

8

C-125

C17

7

3rd

5

C-16

C3

4

2nd

4

C-62

C18

4

1st

4

C-23

C4

4.5

2nd

5

C-76

C19

8

5th

3

C-36

C5

6

3rd

9

C-96

C20

7

3rd

6

C-48

C6

4

1st

7

C-90

C21

5

2nd

4

C-21

C7

6.5

3rd

4

C-115

C22

7.5

4th

6

C-61

C8

4

1st

6

C-85

C23

7

3rd

7

C-102

C9

6.5

4th

6

C-19

C24

7.5

4th

3

C-37

C10

4.5

2nd

8

C-27

C25

4

1st

7

C-43

C11

6

3rd

4

C-129

C26

7

4th

5

C-32

C12

7

4th

3

C-55

C27

6

3 rd

8

C-110

C13

5

2nd

7

C-41

C28

6.6

3rd

7

C-81

C14

7

3rd

5

C-79

C29

8

4th

9

C-66

C15

7

3 rd

8

C-93

C30

5

2nd

8

Sample preparation

Sample collection--8ml Blood sample was collected from jugular vein puncture with taking proper aseptic measure. 6 ml was kept in a vial containing anticoagulant (EDTA, 1 mg/ ml) and 2 ml was kept in syringe for smooth coagulation. Two ml anticoagulant mixed blood was centrifuged to collect the plasma.  Blood  samples were allowed to stand 2 hours at room temperature to allow proper clotting. All blood samples were collected between 10 am and noon in order to standardize time related variables which are known to influence certain blood components. 

Sample Transportation--Blood samples were transported to the lab within one hour keeping in a thermo flask with ice and then fresh blood was examined for TEC, DLC, Hb% and PCV%.                                      

Serum Sample Preservation :2 ml of coagulated blood was kept overnight in refrigerator for collecting the serum. The serum samples obtained after centrifugation were stored at 20C  in refrigerator for further biochemical test.

Sample examination:

Hematological Examination : Different hematological parameters were   studied according to the methods     described by Sastri ,1985 .The detail of the parameters studied as follows:For hemoglobin determinationfresh blood mixed with EDTA  was kept in the Wintrobe  tube and centrifuge at 3000 rpm for 30 minutes. Thus PCV was determined.For hemoglobin determinationN/10 HCl was taken in a graduated hemoglobinometer up to 10 mark and blood was mixed upto 20 cu mm mark. Then diluted the acid hematin using water and match the color thus hb% was determined. TEC determined from frehly collected blood  using hemocytometer .DLC determined by making of blood smear on the clear glass slide and then drying of the smear. Then blood smear was stained  by the Wright's stain and  dryed. Prepared smear was examined under microscope( 100x). Mean corpuscular volume, MCV=Hematocrit x   10/RBC count.Mean corpuscular hemoglobin,MCH =hemoglobin x 10/ BRC count. Mean corpuscular hemoglobin concentration, MCHC =hemoglobin x 100/  hematocrit value.

Biochemical Examination: Total Plasma protein, Plasma Albumin, Plasma Glucose ,Serum Calcium were determined by Automated Humulizer established  in physiology lab of Chittagong Veterinary and Animal Sciences University.

Statistical Analysis: The arithmetic means (±SD)  hematological parameters in different groups were calculated. Further analysis was done using analyses of variance (ANOVA) technique (Steel and  Torrie, 1984) and means were separated by Ducan's  multiple range test. The level of significance  was determined at P‹0.05. All  data are expressed throughout  as mean ± standard deviation.

RESULTS

Hematology

There is significant age related variation for most hematological and biochemical parameters .

Table 01: Hematological  parameters in crossbred  lactating cows and their calves.

COW

CALF

P-Value

Hematological parameters

Mean± sd

range

95% cl

Mean± sd

range

95% cl

Reference Value

TEC ( ×106/µl)

5.63 ± 0.73

4.6-7.0

6.37 ± 0.66

5.3-7.8

05.0-10.0

**

PCV                            %

27.97 ± 3.41

22-38

28.60 ± 4.047

21-37

24.0-46.0

NS

Hemoglobin conc.    (gm/dl)              

7.86 ± 0.679

6.5-9.2

8.28 ± 0.846

7.1-9.2

8.0-15.0

**

**= P<.05 (significant in 95% confidence interval), NS= Non significant

Table shows that TEC is higher in calf (6.37 ± 0.66 ×106/µl) than the cow (5.63 ± 0.73×106/µl) and the difference is significant.

Present study also shows that PCV in cow and calf is 27.97 ± 3.4% and 28.60 ± 4.047% resoectively. It indicate calf have somewhat higher PCV parcentage which is non significant.

At the same time Hb% also higher in calves (8.28 ± 0.846 ) than the cows (7.86 ± 0.679) and it is significant.

Table 02 : Hematological parameters ( RBC indices) in crossbred  lactating cows and their calves.

COW

CALF

P-Value

Hematological parameters

Mean± sd

range

95% cl

Mean± sd

range

95% cl

Reference Value

MCV                          (?t)

49.99 ± 5.16

37.68-60.37

44.62 ± 3.71

36.92-55.74

40.0-60.0

**

MCH                         (pg)

14.09 ± 1.47

11.83-16.96

15.15 ± 8.38

11.08-47.06

11.0-17.0

NS

MCHC                          %

28.08 ± 2.49

21.14-32.31

29.21 ± 2.56

23.82-35.71

30.0-36.0

NS

**= P<.05 (significant in 95% confidence interval), NS= Non significant

MCV from the above table shows that it is higher in cows (49.99 ± 5.16 ft) than the calf (44.62 ± 3.71) and the difference is significant. On the other hand MCH and MCHC both are higher in calves (15.15 ± 8.38 pg and 29.21 ± 2.56 % respectively) but are non significant

Table 03:  Hematological  parameters ( differential count) in crossbred  lactating cows and their calves.

COW

CALF

P-Value

Hematological parameters

Mean± sd

range

95% cl

Mean± sd

range

95% cl

Reference Value

Lumphocyte %                                      

61.46 ± 4.52

49-68

64.26 ± 5.91

55-72

45.0-75.0

**

Monocyte  %                                         

6.87 ± 2.37

3-17

9.26 ± 3.39

2-21

2.0-07.0

**

Neutrophil %                                          

21.10 ± 5.29

12-36

19.63 ± 4.75

6-27

15.0-47.0

NS

Eosinophil %         

10.07 ± 3.75

2-18

6.26 ± 2.93

2-14

0-20.0

**

Basophil %            

0.73 ± 0.86

0-4

0.26 ± 0.62

0-2

0-2.0

NS

**= P<.05 (significant in 95% confidence interval), NS= Non significant

Above table indicates that among the different leukocytes  Lymphocyte and Monocyte is higher in calves ( 64.26±5.91% and 9.26±3.39% respectively) and it is significant.

At the same time  Neutrophil and Basophil is somewhat higher in cows (21.10±5.29% and 0.73 ± 0.86 % respectively) but the difference is non significant.

With this, Eosinophil is higher (10.07 ± 3.7% )in cows and difference is significant.

Serum Biochemistry.

Table 04 :Biochemical parameters in crossbred  lactating cows and their calves.

COW

CALF

P-Value

Serum Parameters

Mean± sd

range

95% cl

Mean± sd

range

95% cl

Reference Value

Total protein (mg/dl)

7.70 ± 1.17

6.12-11.65

6.60 ± 0.81

4.98- 7.73

0.000

5.7-8.1

**

Albumin  (mg/dl)

3.54 ± 0.61

2.67-3.88

3.41 ±  0.57

1.9- 4.3

0.403

2.1-3.9

NS

Calcium

(mg/dl)

9.29 ± 2.06

7.30-16.90

10.45 ± 2.67

7.30- 16.90

0.066

8.0-12.5

NS

Glucose

(mg/dl)

43.81 ± 12.17

20.0-68.70

57.68 ± 2.67

31.7-90.8

0.000

35-55

**

**= P<.05 (significant in 95% confidence interval) NS= Non significant

Table shows that Glucose  level  in cow and calf is 43.81 ± 12.17(mg/dl) and 57.68 ± 2.67(mg/dl) respectively and this difference is significant.

At the same time Total protein level in cow and calf is 7.70 ± 1.17(mg/dl) and 6.60±.811(mg/dl) respectively which is also significant.

But here albumin in cow and calf is  3.54±.61(mg/dl) and  3.41±.57(mg/dl) respectively and this is non significant.

Another parameter calcium in cow and calf is 9.29±2.06(mg/dl) and 10.45±2.67(mg/dl) respectively which is also non significant difference.

DISCUSSION

Hematology :

PCV: The results of the present study shows that PCV  is higher in calf. This may be  due to higher RBC count  and higher MCV % in calf.

TEC: Here we find that TEC is higher in cows than cows .Hege C. Brun- Hansen et al., 1982 found that ,Compared with adult reference intervals, RBC count is higher in calves during the 6 month of life

Hb%: Hb concentration in calf  (8.28± gm/dl) is higher due to their higher RBC count and higher MCH%  and MCHC %.

MCV, MCH, MCHC: Here present study shows that  MCV is higher in calves which supports the findings of ( Hege C. Brun- Hansen et al., 1982). They found that ,Compared with adult reference intervals, the MCV  is lower in calves during the 6 month of life.  Present study also shows that MCH% and MCHC% slightly higher in calves and it is non significant. Previous study reveals that the MCH and MCHC values are affected by variation in Hb synthesis (Benjamin, 1978). The MCHC is a measure of the quantity of Hb in each RBC and also relates to the weight of Hb and volume of cell, the cows having higher Hb concentration showed higher MCH and MCHC values. Higher RBC count with lower Hb concentration may be due to increase in number of erythrocytes with decrease in their size (  Benjamin, 1978).

Differential Count: Lymphocyte and Monocyte  is significantly higher in calves.

Differences in neutrophil count in calf is somewhat lower than cow. Though it is  is non significant but it may be due to nutritional deficiency that occurs in starvation or anorexia are causes of neutropenia (Spivak et al., 1984).

Eosinophil is higher in cows and  it is significant. It may be due to their higher  parasitic load  than  the calves. Differences in Basophil % is not significant and it is normally 0-2% in the body.

Serum biochemistry:

Glucose : Glucose  level found in cow and calf were 43.81±12.17 mg/dl (2.56±mmol/lit) and 57.68±2.67 mg/dl (3.20±  mmol/lit) respectively and the difference is significant. This indicate milk feeding calf have higher glucose  percentages than lactating cow. This higher percentages in milk feeding calf may be due to their monogastric nature and their milk ingestion regularly.

Calcium : Calcium level found in cow and calf were 9.29±2.06 mg/dl (2.33±mmol/lit) and 10.45±2.67604 mg/dl (2.61±  mmol/lit) respectively. This indicate milk feeding calf have higher calcium percentages than lactating cow.  Lower  percentages in lactating cows may be due to calcium losses during milk production. This result agree with the Doornenbal  et al., 1988 .In Shorthorn cattle Doornenbal  et al., 1988 found that calcium and  Inorganic Phosphorus is higher in milk feeding calves and calcium and  Inorganic Phosphorus both generally decreased with increasing age  beyond one year of age. He sited that one of the main function of these elements is there involvement in skeletal growth in young animals. In older animal there is a  decreased need for calcium (Ca) and Pi for this purpose  and this is reflected in lower blood levels.

Total protein: Total protein level is higher in lactating cows than milk feeding calves and it is significant. This result agree with the Doornenbal  et al., 1988; Roussel  et al., 1982; Peterson et al., 1981. In shorthorn cattle Doornenbal et al., 1988 found except for the values at birth, total protein levels were lower ( p‹ 0.05) in young animals and higher in mature animals.  Roussel  et al., 1982, also found that in Jersey cows total protein  increased with age over a range of one to six years. In dairy cattle total protein levels were reported to be higher in dry cows ( Peterson  et al., 1981) . In animals there is a general modification in serum proteins with advancing age and in the very old; thus age is an important consideration in the interpretation of the serum proteins (Kaneoko et al.,1997).

Albumin: Albumin levels lower at birth  ( p‹ 0.05) and then increase, but fluctuate somewhat. There was no clear effect of age on albumin levels (Doornenbal  et al., 1988). Present study also agree with it because result shows that albumin in calf is somewhat higher ic adult than calf and the difference is insignificant.

Discrepancies in values for various hematological parameters between our findings and previous studies may be explained by differences in sampling interval, methods used, numbers of cows sampled, and/or degree of metabolic disturbances. Moreover, genetic differences between cows (Mallard et al., 1998) and subtropical conditions of the present study might have played a role for the differences with other studies.

LIMITATIONS OF THE STUDY

There were some limitations of this study. These are as follows:

1. Shortage of time period: This study was carried out only for a period of 30 days that was not sufficient.

3. I did not find any other such types of work done by other scientist in Bangladesh. So I could not compare my result with other scientist.

Conclusion.

Hematological and Biochemical values of crossbred lactating cows and their calves were determined, in Metro Dairy Farm, Hathazari , Chittagong from 09.08.09-08.09.09 for  comparing the values between cow and calf. The result found that there is some hematobiochemical difference between cow and calf but all parameters are not within the reference range indicated in case of Holstein Friesian.

Finally hematological and biochemical values were an efficient tool for evaluation of physiological status, metabolic disorders, management problems of the farm which have great relation to health status of the animal.

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