HEMATOLOGY I

By :

Name : Fatahalani RizkikaStudent ID : B1K014017Entourage : VIIGroup : 2Assistant : Lucky Pratama Suharto

PRACTICUM REPORT OF ANIMAL PHISIOLOGY I

MINISTRY OF RESEARCH, TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITYFACULTY OF BIOLOGY

PURWOKERTO

2015

I. INTRODUCTION

1.1. Background

Blood was the body fluid that flows in the veins and circulates to whole

body. Blood in generally consist of these elements and cellular matrixes fluid

called plasma. Blood consist of plasma and components of cellular, plasma is

liquid that contain ions and organic molecules such as proteins and electrolytes,

nutrient, waste materials, substance dissolved and materials dissolved (Prosser

and Brown, 1961).

Blood was formed from the cells are free and matrix (plasma). Various

vertebrate animals both living in water and on land would be considered to have

composition that almost the same as that consists of 52% balance plasma-colored,

also the red blood cells (erythrocytes), cells would have had to white blood cells

(leukocyte) and the cells of the small namely platelets or thrombocytes a crucial

part of the coagulation of blood. Based on components immense can functions of

blood, which as oxygen carrier from the lung to tissue, the result digestivus

absorbed, place minerals dissolved, waste product, secretion, especially enzymes

and the antibodies (Yuwono, 2001).

Blood was considered as special tissue circulation. The flow of blood in

the body to ensure environment that is still, so that all of the cells and tissues to be

able to perform its function. Chemical Composition blood is very complex

because of the blood brought great nutrient, waste product and organic ion, so it

possible to sustain coordination and integration metabolism in several animals on

high-level (Dukes, 1995).

Hematology is branches of science health who studied blood, blood

forming organs and disease, hematology used as guide severity disease. Changes

hematology and blood chemical with both qualitative and quantitative will be able

to define animal health. Cells and blood plasma has role that physiological

couples important in diagnosis, prognosis and disease therapy (Soetrisno, 1987) .

In practicum hematology this time using blood samples blood of chicken

(Gallus domestica), mouse (Mus muscullus) and fish (Oreochromis niloticus)

because it is one of the representative samples from aves, pisces (constellation)

and mammals that have blood that is quite a lot easier to get, observed, and make

them affordable .

1.2. Objective

The objective practical class this time is to provide skills to students about

taking blood animals, know the difference between blood cells form in many

animals, and how to make calculations red blood cells, white blood cell, and

levels of hemoglobin blood animals.

II. MATERIAL AND METHOD

2.1. Material

The tools that used in practicum is haemometer, haemositometer, tube

Sahli, pipette capillaries, light microscope, and cover glass, a syringe, the cup of

petri, batang stirring spoon or rod, pipette thoma erythrocytes, pipette thoma

leukocyte and hand counter.

The materials used in practicum is blood of mouse (Mus muscullus),

chicken (Gallus gallus ), Nila fish ( Oreochromis niloticus ), solution hayem, turk

solution, 0.1 N HCl and EDTA solution.

2.2. Method

A. Count The Total of Leukocyte

1. Blood animals taken with micropipet that was named EDTA to

dilution shows the number 1, and then the tip of mikropipet cleaned

with tissue.

2. Turk solution taken on tube reaction by using micropipet to number

11.

3. Micropipet held on both tip with his mother fingers and index finger

and reshuffled.

4. A few drops solution in micropipet used for the calculations that

dropped in inner chambers count haemositometer.

5. The room count viewed under microscope and counted leukocyte

found in the square booth (big square).

6. Number of leukocyte per mm 3 counted.

B. Count The Number of Erythrocytes

1. Blood animals taken with micropipet that was named EDTA to

dilution shows the number 1, and then the micropipet cleaned with

tissue.

2. Solution hayem taken on tube reaction by using micropipet to number

11.

3. Micropipet held on both ends with his mother fingers and index finger

and reshuffled.

4. A few drops solution in micropipet used for the calculations that

dropped in the inner chambers count haemositometer.

5. The room count viewed under microscope and counted leukocyte

found in the square.

6. Number of erythrocytes per mm 3 counted.

C. Measurements levels of Hemoglobin Using methods Sahli

1. Tube sahli inside given blood animals to show the number 10.

2. Tube sahli that contains a blood test animals dropped with aquades

until the solution stained in tubes in accordance with the indicators tube

sahli.

3. After solution stained in tubes and comparator sahli as seen on a

smaller scale, then reduced by blood in tubes before given akuades.

4. Write the results that need.

D. Hematocrite value measurement

1. Blood samples are taken by capillary pippete.

2. Centrifuge in 1200 rpm until 3 minutes.

3. Check the hematocryte value with hematocryte reader.

III. RESULT AND DISCUSSION

3.1. Result

Table 1. Observations The count Hematology

Group Animal Test Eritrocyte

(cell/ mm3)

Leukocyte

(cell/mm3)

Hematocryte

(%)

Hb

(gr/dl)

1 Fish 805.000 164.576 20 6,8

2 Chicken 1.340.000 218.950 20 5

3 Fish 2.545.000 87.275 22 5

4 Mice 14.340.000 10.325 66 40

5 Fish 1.008.889 185.252 17 6,4

Table 2. Blood Glucose Level

Group Blood Glucose Level (mg/dl)

1

2

3

4

5

122

67

49

62

73

Count Eritrocyte Count Leukocyte

Box 1 = 56 Box 1 = 2848

Box 2 = 55 Box 2 = 1792

Box 3 = 59 Box 3 = 1798

Box 4 = 52 Box 4 = 2320

Box 5 = 46 Box 5 = -

268 x 5000 = 1.340.000 cell/mm3 218.950 cell/mm3

Hematocryte Value = 20 %

Hemoglobin = 5 gr/dl

3.2. Discussion

Taking blood in the fish , mice, probandus and chicken can be do with

injected in the under wings part. In the Fish taking blood will do with injected in

the veins caudal, heart, and their gills. In mice taking blood will be do with

injected in the eyes and cut in the tail, cutting lambs in mouse is the way that

efficient to get blood whereas the Chicken., taking blood be do with injected in

the part wing at Jugular venous. Probandus taking blood injected in one of the

finger (Soetrisno, 1987).

Hematology often used to detect physiological changes that follow

conditions under pressure from different. Hematology can be treated as important

index to health status. The most common hematological variable measure the

pressure includes title nobility white blood cells and hemoglobin content, red,

hematocrit values and index red blood cells. Parameter fish hematology often

defined as index of their health status. When the values parameter hemotology

obtained under abnormalities must be possible to monitor changes in their

physical and chemical property water (Al-Attar, 2005).

The measurement method hematology animals on the measurement of

leukocyte, erythrocytes, the value hematocryt and measuring levels of hemoglobin

blood animals. Leukocyte is white blood cells that played an important role in

defense and improvements to the body organisms which functions as its main

phagocytosis, producing antibodies and eating seeds diseases that have entered in

the body, According to Hoffbrand (1987) , said that only 1 percent (in mammals)

to 10 percent (in the fish) leukocyte in blood stream function smashed the objects

foreign matter through the phagocytosis process, when initiate many leukocyte

that led dead, this number is not always remained. The live leukocyte has

variation, begin from few hours to granulocytes, until monthly to manosit and

even yearly to lymposit. In the bloodstream most white blood cells are non-

functional and without being transported to the network when necessary only

(Frandson, 1992). The total of leukocyte influenced by gender, age activities and

environmental conditions (Lagler et al .,1977).

The tools that used are haemometer that serves to measure the levels of

haemoglobin; the haemositometer function in the calculation of the number of

leukocytes and erythrocytes; sahli tube serves as diluent tube in measuring the

levels of hemoglobin; thoma serves as pipette pumps blood and hayem solution or

turk with a certain volume; light microscope observations using the function

haemositometer; spuit injection serves to take the blood test animals; handcounter

functions to count the number of erythrocytes or leukocytes are observed under a

microscope; and the cups functions to the container with a solution of EDTA

blood dilution. Solution-a solution that is used i.e. a solution of turk as a diluent

solution hayem, leukocytes as a diluent solution of HCl, erythrocytes function

change of blood acid hernitin, and EDTA (Ethylen Diamin Tetra Acetic Acid)

serves as anticoagulants or substance causes the blood not frozen (Hoffbrand,

1987).

Blood is the body fluid that flows in the veins and it circulates to the

whole body. Blood in generally consist of these elements and cellular matrixs

fluid called plasma. As consisting blood plasma and components of the cellular,

plasma is balance that contain ions, organic molecules such as on proteins and

electrolytes, nutrient, waste materials, substance dissolved and materials dissolved

(Prosser and Brown, 1961). Blood cells can be divided into erythrocytes (red-

blood cell), leukocyte (white blood cells) and platelets (blood chip) (Bevelander

et al ., 1988).

The basic component of the blood that major blood plasma and blood cells

composed of erythrocytes, leukocytes, and trombosyte. Blood plasma is the liquid

component that contains ions and organic molecules which include proteins,

electrolytes, waste materials, substances and control of dissolved substances. Part

of the blood plasma which has important functions is the serum. Serum is blood

plasma issued or separated fibrinogen by way of turning the blood in sentrifuge.

Serums look very clear and contains antibodies. This antibody serves to destroy

foreign proteins into the body. Incoming foreign proteins into the body are called

antigens. While the cell is made up of discrete cells that have specific forms and

different functions, while the components of plasma fibrinogen in addition there

are also inorganic ions (Alamanda, 2006). 

Functions of blood is as transport (food, oxygen, carbon dioxide, waste

and water), termoregulasi (thermostat in the body), immunology (contains

antibodies body), and homeostasis (adjusting the substance, pH regulator). Red

blood cells like discs generally small bikonkaf, sunken on both sides. Red blood

cell structure consists of wrapper or stroma, contains the hemoglobin.

Erythrocytes function as oxygen carrier. Hemoglobin in the erythrocytes possible

appearance ability to carry the oxygen, and to cause the color red in the blood.

Hemoglobin join with oxygen that there are in their lungs formed

oxysihemoglobin, and then would release oxygen to tissues and cells in the body

(Pearce, 1989).

Hematrocryte value is the ratio between eritrocyte cell volume with blood

plasma. Decreasing the level hematrocyte can as indication low protein (Coles,

1974). While Kadar glukosa dalam tubuh makhluk hidup dapat digunakan

untuk memprediksi metabolisme yang mungkin terjadi dalam sel dengan

kandungan gula yang tersedia. Jika kandungan 1 glukosa dalam tubuh sangat

berlebih maka glukosa tersebut akan mengalami reaksi katabolisme secara

enzimatik untuk menghasilkan energi. Namun jika kandungan glukosa

tersebut dibawah batas minimum, maka asam piruvat yang dihasilkan dari proses

katabolisme bisa mengalami proses enzimatik secara anabolisme melalui

gluconeogenesis untuk mensintesis glukosa dan memenuhi kadar normal glukosa

dalam darah (Oslon, 1973).

Berdasarkan hasil pengamatan, jumlah eritrosit dari sampel darah

ikan nila (Oreochromis niloticus) kelompok 1 adalah 805.000 sel/mm3, jumlah

eritrosit darah ayam (Gallus gallus) kelompok 2 adalah 1.340.000 sel/mm3 dan

kelompok 3 ikan nila (Oreochromis niloticus) adalah adalah 2.545.000 sel/mm3,

dan jumlah eritrosit pada sampel darah mencit (Mus muscullus) kelompok 4

adalah 14.340.000 sel/mm3 dan kelompok 5 pada ikan nila (Oreochromis

niloticus adalah 1.008.889 sel/mm3. jumlah leukocyte dari sampel darah ikan nila

(Oreochromis niloticus) kelompok 1 adalah 164.576 sel/mm3, jumlah eritrosit

darah ayam (Gallus gallus) kelompok 2 adalah 218.950 sel/mm3 dan kelompok 3

ikan nila (Oreochromis niloticus) adalah adalah 87.275 sel/mm3, dan jumlah

eritrosit pada sampel darah mencit (Mus muscullus) kelompok 4 adalah 10.325

sel/mm3 dan kelompok 5 pada ikan nila (Oreochromis niloticus adalah 18.525

sel/mm3 . Jumlah leukosit normal pada mencit (Mus muscullus) adalah 4.000-

11.000 sel/mm3 dan jumlah eritrosit pada mencit (Mus muscullus) adalah 4 juta

sel/mm3. Jumlah leukosit normal pada ikan nila (Oreochromis niloticus) adalah

20.000-150.000 sel/mm3 dan jumlah eritrosit pada mencit (Mus muscullus) adalah

50.000-3 juta sel/mm3.

Hasil pengamatan jika dilihat dari banyaknya eritrosit dan leukosit yang

diperoleh, terdapat ketidaksesuaian dengan pendapat Frandson (1992). Hal ini

disebabkan karena keterbatasan ketelitian penglihatan dalam menghitung jumlah

leukosit dan eritrosit didalam mikroskop cahaya serta faktor – faktor internal

maupun eksternal. Faktor – faktor tersebut seperti stres, kualitas air, polusi,

malnutrisi, jenis kelamin, kondisi tubuh, umur, variasi harian dan penyakit-

penyakit. Ikan dapat beradaptasi pada kondisi lingkungan yang buruk dengan

merubah fisiologi aktivitas mereka, namun hasil pengamatan sesuai pula dengan

pendapat Bevelander (1979) yang menyatakan bahwa besarnya jumlah leukosit

selalu dipengaruhi oleh jumlah eritrosit dan jumlah leukosit selalu lebih rendah

daripada jumlah eritosit. Jumlah eritrosit lebih banyak dibandingkan dengan

jumlah leukosit karena fungsi eritrosit sebagai transport oksigen sedangkan

leukosit berfungsi memelihara dan menjaga tubuh dari serangan penyakit.

Berdasarkan pengamatan kadar hemoglobin dari sampel darah ikan

kelompok 1 yaitu 6,8 gr/dl, kelompok 2 dari sampel darah ayam yaitu 5 gr/dl,

sedangkan pada kelompok 3 pada ikan yaitu 5 gr/dl kadar haemoglobin pada

kelompok 4 pada mencit didapatkan yaitu 40 gr/dl dan kelompok 5 pada fish

adalah 6,4 gr/dl. Menurut Hoffbrand (1987) yang menyatakan bahwa kadar

hemoglobin ayam yang sehat yaitu 5,92 g/dl – 6,33 g/dl dan hemoglobin ikan lele

yang sehat sekitar antara 6,46 – 7,93 g/dl. Kadar Hb pada ikan dipengaruhi oleh

afinitas darah pada hewan.Ikan budidaya dengan kondisi tidak sakit dapat ditandai

dengan rendahnya nilai eritrosit, hematokrit dan hemoglobin (Hb), jika kondisi

kurang sehat maka banyak diproduksi sel darah putih dan lender (Oslon, 1973).

Berdasarkan pengamatan nilai hematocryte dari sampel ikan pada

kelompok 1 adalah 20 %. kelompok 2 sampel darah pada ayam didiapatkan 20%,

dan pada kelompok 3 pada ikan menghasilkan nilai hematocryte sebanyak 22%.

Kemudian pada mencit kelompok 4 yaitu 66%, dan kelompok 5 pada ikan yaitu

17 %. Nilai hematocrit normal antara 30 – 45 % (Oslon, 1973). Berdasarkan

pengamatan tingkat kadar glukosa pada probandus masing-masing kelompok.

Pada kelompok 1 adalah 122 mg/dl, kelompok 2 67 mg/dl. Kadar glukosa

probandus pada kelompok 3 dan 4 adalah 49 mg/dl dan 62 mgl/dl, kelompok 5

mendapatkan kadar glukosa sebanyak 73 mg/dl. Menurut Hoffbrand (1987) kadar

glukosa normal seseorang pada pagi hari setelah semalaman berpuasa seharusnya

70-100 mg/dl. Kadar gula darah biasanya kurang dari 120-140 mg/dl pada 2 jam

setelah makan atau minum cairan yang mengandun gula maupun karbohidrat.

Berdasarkan data praktikum tersebut terdapat perbedaan antara jumlah

eritrosit dan leukosit serta kadar hemoglobin dalam darah. Hal tersebut

dipengaruhi oleh dipengaruhi oleh jenis kelamin, umur, aktivitas dan kondisi

lingkungan sedangkan jumlah eritrosit dalam darah dipengaruhi oleh jenis

kelamin, umur, variasi harian, ketinggian tempat dan tekanan emosional. Jumlah

leukosit jauh lebih kecil dibawah eritrosit dan bervariasi tergantung dari spesies

atau jenis hewannya. Leukosit berperan penting dalam pertahanan seluler dan

humoral organisme terhadap organ-organ asing. Bila tersuspensi dalam sirkulasi

darah mereka berbentuk steris, tetapi mampu bersifat amoboid. Melalui proses

diapedesis leukosit dapat meninggalkan kapiler dengan menerobos antara sel-sel

endotel dan menembus ke dalam jaringan ikat ( Hadikastowo, 1982).

Sedangkan factor yang mempengaruhi nilai hematocryte adalah radius

sentrifuge, kecepatan sentrifuge dan lama pemusingan. Kadar gula darah di

pengaruhi oleh makanan dan minuman yaitu semakin banyak kandungan

karbohidrat dalam makanan, glukosa darah juga semakin tinggi, yang di

konsumsi, aktifitas fisik menyebabkan shift volume Antara compartment di dalam

pembuluh darah dan interstitial, kehilangan cairan karena berkeringat, dan

perubahan kadar hormonr. Akibatnya akan terjadi perbedaan besar Antara kadar

glukosa di arteri dan vena (Evans, 1988).

IV. KESIMPULAN

Berdasarkan praktikum tersebut dapat disimpulkan bahwa :

1. Cara pengambilan darah hewan uji yaitu jika pada ayam pengambilan

darahnya melalui vena jugularis yang terletak pada sayapnya. Darah

mencit cara pengambilannya dengan memotong ekor mencit tersebut.

Darah ikan cara pengambilannya bisa dari pangkal ekor, vena caudalis,

atau dari jantungnya. Cara pengambilan darah probandus dengan cara di

suntik pada salah satu jari.

2. Erythrocytes form in vertebrates except Mammals are oval office is not

and leukocyte is and is motil.

3. jumlah eritrosit dari sampel darah ikan nila (Oreochromis niloticus)

kelompok 1 adalah 805.000 sel/mm3, jumlah eritrosit darah ayam (Gallus

gallus) kelompok 2 adalah 1.340.000 sel/mm3 dan kelompok 3 ikan nila

(Oreochromis niloticus) adalah adalah 2.545.000 sel/mm3, dan jumlah

eritrosit pada sampel darah mencit (Mus muscullus) kelompok 4 adalah

14.340.000 sel/mm3 dan kelompok 5 pada ikan nila (Oreochromis niloti-

cus adalah 1.008.889 sel/mm3.

4. Jumlah leukocyte dari sampel darah ikan nila (Oreochromis niloticus)

kelompok 1 adalah 164.576 sel/mm3, jumlah eritrosit darah ayam (Gallus

gallus) kelompok 2 adalah 218.950 sel/mm3 dan kelompok 3 ikan nila

(Oreochromis niloticus) adalah adalah 87.275 sel/mm3, dan jumlah

eritrosit pada sampel darah mencit (Mus muscullus) kelompok 4 adalah

10.325 sel/mm3 dan kelompok 5 pada ikan nila (Oreochromis niloticus

adalah 18.525 sel/mm3

5. Berdasarkan pengamatan nilai hematocryte dari sampel ikan pada kelom-

pok 1 adalah 20 %. kelompok 2 sampel darah pada ayam didiapatkan 20%,

dan pada kelompok 3 pada ikan menghasilkan nilai hematocryte sebanyak

22%. Kemudian pada mencit kelompok 4 yaitu 66%, dan kelompok 5 pada

ikan yaitu 17 %

6. Berdasarkan pengamatan tingkat kadar glukosa pada probandus masing-

masing kelompok. Pada kelompok 1 adalah 122 mg/dl, kelompok 2 67

mg/dl. Kadar glukosa probandus pada kelompok 3 dan 4 adalah 49 mg/dl

dan 62 mgl/dl, kelompok 5 mendapatkan kadar glukosa sebanyak 73 mg/

dl.

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