Penggunaan mencit sebagai hewan laboratorium disebabkan beberapa sifat yang menguntungkan:

Mudah dipelihara dalam jumlah besar Menghasilkan banyak keturunan (6-15 ekor)

Masa reproduktif aktif yang relatif panjang (2-14 bulan)

Masa kehamilan yang relatif pendek

Berat badan mencit dewasa: Jantan : 20--40 g Betina : 18--35 g

Mencit dapat hidup dalam jangka waktu: 1 sampai 3 tahun Pubertas mencit jantan : Ditandai dengan turunnya testis ke kantung skrotum . Umur 40 hari
Mencit jantan mencapai kesuburan maks: Umur 100--300 hari

Mencit betina mempunyai siklus berahi Poliestrus, dengan tahapan sbb: - Diestrus - Proestrus - Estrus - Metestrus
Lama satu siklus: 4--5 hari Tahapan siklus yang sedang berlangsung dapat diketahui dengan: Metode Oles Vagina

Lama hidup Lama produksi ekonomis

1--2 tahun, bisa sampai 3 tahun 9 bulan

Lama bunting
Kawin sesudah beranak

19--21 hari
1 sampai 24 jam

Umur disapih
Umur dewasa

21 hari
35 hari

Umur dikawinkan
Siklus kelamin

8 minggu (jantan dan betina)

Poliestrus

Lama siklus estrus

Lama masa estrus

4--5 hari
12--14 jam

Perkawinan
Ovulasi Fertilisasi Cleavage, zigot blastula

Pada waktu estrus

Dekat akhir estrus, spontan 2 jam sesudah kawin 2,5--4 hari

Implantasi Berat dewasa

4--5 hari sesudah fertilisasi 20--40 g

Berat lahir
Jumlah anak

0,5--1,0 g
~6, dapat mencapai 15

Suhu (rektal)
Pernapasan Denyut jantung Tekanan darah

36--39o C
140--180/menit; anestesi 80/menit; stres 230/menit 600--650/menit; anestesi 350/menit; stres 750/menit 130--160 sistol, 102--110 diastol; anestesi 110 sitol, 80 diastol

Putting susu Plasenta

Uterus Perkawinan kelompok Kromosom Aktivitas

10 putting (3 pasang di daerah dada, 2 pasang di daerah perut) Diskoidal hemokorial

2 kornu, bermuara sebelum serviks 4 betina dengan 1 jantan 2n = 40 Nokturnal

Mencit laboratorium dapat ditempatkan dalam kandang yang tidak besar (sebesar kotak sepatu)
Kotak tersebut dapat terbuat dari: - Plastik - Aluminium - Baja tahan karat

Bahan harus tahan lama, dan tahan terhadap gigitan mencit.

Table 1.0.1 Minimum Space Requirements for Housing Laboratory Animals in Cagesa Weight Floor area/animal Cage heightb

Animal
Mouse

(g)
<10 10-15

(in2)
6.0 8.0

(cm2)
38.71 51.62

(in)
5 5

(cm)
12.70 12.70

15-25
>25

12.0
15.0

77.42
96.78

5
5

12.70
12.70

Rat

<100
100-200 200-300

17.0
23.0 29.0

109.68
148.40 187.11

7
7 7

17.78
17.78 17.78

300-400
400-500 >500

40.0
60.0 70.0

258.08
387.12 451.64

7
7 7

17.78
17.78 17.78

a Guidelines are derived from Guide for the Care and Use of Laboratory Animals (ILAR, 1985).b From the resting floor to the cage top From Current Protocols in Immunology Online

Minimizing Risk = Minimizing Exposure Procedures to Reduce Exposure

INDIVIDUALLY VENTILATED CAGES laminar air rack

From TENCIPLAST

From TENCIPLAST

Ventilation efficiency with low velocity Air Speed

Visibility

Easy access

From TENCIPLAST

Diuresis cage
From TENCIPLAST

Metabolic cage
From TENCIPLAST

 Lingkungan harus kering dan bersih Suhu sekitar 25oC Memberi ruangan yang cukup untuk bergerak bebas dalam berbagai posisi Harus dilengkapi dengan makanan dan minuman yang mudah dicapai oleh mencit. Kandang harus ditempatkan dalam rak yang cukup longgar

Kandang harus memiliki alas tidur yang bersih dan berkualitas baik

Table 1.0.2 Recommended Relative Humidity and Dry Bulb Temperature for Animals Housed in Cagesa
Dry bulb temperatureb
Animal
Relative humidity (%)

Mouse Rat

40-70 40-70

18-26 18-26

64.4-78.8 64.4-78.8

Hamster Rabbit
a Guidelines

40-70 40-60

18-26 16-21

64.4-78.8 60.8-69.8

from ILAR, 1985. b ILAR, 1965, 1977.

From Current Protocols in Immunology

 Harus tidak menarik mencit untuk dimakan

Tidak menarik binatang lain (Misal kutu) Harus dapat mengisap air
Tidak mengandung zat-zat yang dapat mengganggu penelitian

Alas tidur harus diganti minimal 2 kali dalam seminggu atau bila sudah tercium bau amonia
Alas tidur yang biasa digunakan:
Serbuk gergaji yang tidak terlalu berdebu Sekam padi yang tidak tercemar oleh kencing ataupun tinja mencit atau tikus liar

 Makanan berbentuk pelet dalam jumlah tanpa batas (ad libitum) Makanan ditempatkan dalam tutup kandang yang melekuk Seekor mencit dewasa makan sekitar 3-5 g/h
- Protein 20-25% - Lemak 10-12% - Karbohidrat 40-55% - Serat kasar ~ 4% - Abu 5-6% - Vitamin A 15.000-20.000 IU/kg - Vitamin D 5000 IU/kg - Alfa tokoferol 50 mg/kg - Asam linoleat 5-10 g/kg - Tiamin 15-20 mg/kg - Riboflain 8 mg/kg - Pantotenat 20 ng/kg - Vitamin B12 30 g/kg - Biotin 80-200 g/kg - Piridoksin 5 mg/kg - Inositol 10-1000 mg/kg - Kolin 20 g/kg

Kandungan bahan makanan:

Mouse handling and manual restraint. Apply slight, rearward traction on the tail (A). Grasp skin behind ears with thumb and index finger (B). Transfer the tail from the preferred hand to beneath the little finger of the hand holding the scruff of the neck (C).

From Current Protocols in Immunology

Placing a mouse on a cage lid and grasping the loose skin behind the ears by the thumb and forefinger

As soon as the mouses head is restrained, the mouse can be picked up and the tail secured within your ring finger and little finger

From Current Protocols in Immunology

A. Blood collection from tail vein B. Blood collection from orbital sinus C. Blood collection from cardiac puncture D. Blood collection from saphenous vein E. Intraperitoneal injection F. Subcutaneous injection G. Oral Feeding H. Sexing

 For collection of small amount of blood (Approximate 0.1 ml )

 75% alcohol cotton ball for surface disinfection Small plastic bottle with 1/2 cm diameter holes in both ends as mouse restrainer Scissors

Pipetteman and tips

A vial for blood collection

BLOOD COLLECTION FROM TAIL VEIN OF MOUSE AND RAT USING MICROHEMATOCRIT TUBE
Visualize a sampling site of the lateral tail vein at approximately midpoint on the length of the tail.

From Current Protocols in Immunology

BLOOD COLLECTION FROM TAIL VEIN OF MOUSE AND RAT USING MICROHEMATOCRIT TUBE

Collect blood from the hub of the needle with the microhemato crit tube.

From Current Protocols in Immunology

Push the mouse into the restrainer

Leave the tail of the mouse outside the cover of the restrainer

Amputate the tip of the mouse tail by scissors

Massage the tail and collect blood by pipetteman

 Should apply anesthetic before blood withdraw A convenience and easy apply method for blood collection in mouse

Collect amount up to 0.5 ml

75% alcohol cotton ball for surface disinfection Hypnorm for general anesthetic 27 G needle with 1 ml syringe for injection Glass capillary tube and vial for blood collection

Anesthetize a mouse by intraperitoneal injection of Hypnorm

BLOOD COLLECTION FROM ORBITAL SINUS OR PLEXUS OF MOUSE AND RAT

Introduce the end of the microhematocrit tube at the medial canthus of the orbit.

From Current Protocols in Immunology

BLOOD COLLECTION FROM ORBITAL SINUS OR PLEXUS OF MOUSE AND RAT

Slowly, and with axial rotation, advance the tip of the microhematocrit tube gently towards the rear of the socket until blood flows into the tube.

From Current Protocols in Immunology

BLOOD COLLECTION FROM ORBITAL SINUS OR PLEXUS OF MOUSE AND RAT Remove the microhematocrit tube from orbit and dab excess blood from the site with a gauze sponge or swab moistened in saline or PBS.
From Current Protocols in Immunology

Use a sharp end glass capillary tube to penetrate the orbital conjunctiva and rupture the orbital sinus

Collect blood with a vial

 For collect up to 1 ml of blood within a short period of time Must be performed under general anesthetic

75% alcohol cotton ball for surface disinfection Hypnorm used as anesthetic 27G needle with 1 ml syringe for injection 24G needle with 3 ml syringe for blood withdraw

Anesthetize a mouse by intraperitoneal injection of Hypnorm

Disinfect the thorax area with 75% alcohol cotton ball

Search for the maximum heart palpitation with your finger

Insert a 24G 1 needle through the thoracic wall at the point of maximum heart palpitation

Withdraw blood slowly by your right hand

 This method is used of multiple samples are taken in the course of a day

It can also be applied on rats, hamsters, gerbils and guineapigs

 75% alcohol cotton ball for surface disinfection 50 ml syringe tube with small holes at the end as restrainer a scalpel and shaver for remove of hair 24 G 1 needle for release of blood tips and pipetteman for blood collection

Placing a mouse on a cage lid and grasping the loose skin behind the ears with your thumb and forefinger

Place the mouse in the restainer

Pull out the leg and removed the hair by a assistant

Hair can also be shaved by using a small scalpel

Apply vaseline after disinfect the surface area to reduce clotting and coagulation during blood collection.

Use a 24 G 1 needle to puncture the vein and release blood from the saphenous vein

Use a Microvette or a pipetteman with tip to collect blood from the saphenous vein

Approximate 100 microliters can be collected

Flex the foot of the mouse to reduce the flow of blood back to the puncture site

A cotton ball is applied to the puncture site to stop further bleeding

 A common method of administering drugs to rodents

 75% alcohol cotton ball for surface disinfection 25G 1/2 needle with 1 ml syringe for injection

Place a mouse on a cage lid and grasping the loose skin behind the ears with your thumb and forefinger

As soon as the mouses head is restrained, the mouse can be picked up and the tail secured within your ring finger and little finger

The injection site should be in the lower left quadrant of the abdomen because vital organs are absent from this area. Only the tip of the needle should penetrate the abdominal wall to prevent injection into the intestine.

 The most common method for immunology studies

 75% alcohol cotton ball for surface disinfection 25G 1 needle with 1 ml syringe for injection

Pick up a nude mouse and spin its tail to put it in a faint condition

Grasp the loose skin on the back of the mouse from ears along the legs and restrain the legs with your ring finger and little finger

After disinfect the surface area, insert the needle in the lateral side of the abdominal wall and push upwards to the armpit of the mouse

A lump of injection substance can be seen through the skin after injection

INTRAVENOUS INJECTION OF MOUSE

1. Fill syringe with injectate and remove air bubbles.

Removal of air bubbles is critical to avoid air embolism.

2. Place mouse in a restrainer.

From Current Protocols in Immunology

INTRAVENOUS INJECTION OF MOUSE

3. Warm the tail with a heat lamp or by immersing in warm water to dilate vessels.
4. Swab the tail with 70% ethanol on a gauze sponge or swab.

From Current Protocols in Immunology

INTRAVENOUS INJECTION OF MOUSE

5. Immobilize the tail with gentle traction. 6. Visualize the lateral tail vein and insert needle parallel to the vein 2 to 4 mm into the lumen.

Keep the bevel of the needle facing up.

From Current Protocols in Immunology

INTRAVENOUS INJECTION OF MOUSE

7. Inject slowly.

No bleb should form if needle was properly located. If a bleb appears, indicating failure to cannulate the vein, additional attempts may be made proximally. Thus it is helpful to make the first attempt at injection as close to the tip of the tail as possible.

From Current Protocols in Immunology

INTRAVENOUS INJECTION OF MOUSE

8. Withdraw the needle and apply digital pressure if necessary to achieve hemostasis.

From Current Protocols in Immunology

 Gastric intubation ensures that all the material was administered Feeding amount limited to 1% of body weight

 A 18 G stainless steel, ball tipped needle a glove

Grasp the loose skin on the back of the mouse and restrain its tail with your ring finger and little finger. Then, introduce the feeding tube from the pharynx in to the esophagus when the mouse is in the act of swallowing.

Common complications associated with gastric intubation are damage to the esophagus and administration of substance into the trachea. Careful and gentle passage of the feeding needle will greatly reduce these possibilities.

The anatomy picture showed the position of the feeding needle tip inside the esophagus with the heart and sternum removed.

Sexing mice - The distance between the anal and genital orifices is greater in the male (left) compared to the female (right).

Male

Female

The top two mice are neonates and note that the anogenital distance is larger in the male than in the female neonates, the penis and vulva cannot be easily differentiated and so are referred to as a genital papilla. The bottom two animals are adults; genitalia are differentiated.

 Also, nipples become evident in females at about 10 days of age.

Male mice, like other rodent species, retain an open inguinal canal in adulthood. That is, the descended testicles communicate with the abdominal cavity. Depending on the position in which the mouse is held, the testicles may be retracted into the abdomen or descended into the scrotum.
Because of the open inguinal canal, castration of mice requires that the surgeon use caution when applying tension to the testicle. Too much tension can result in the intestines being pulled through the inguinal canal.

 Occasionally people experience difficulty in determining gender in mice at weaning age, although the anogenital distances are markedly different between males and females.

Newborns have a subtle difference in the anogenital distance, due to their small size and scale of the anatomical landmarks. In determining the gender of newborns, its best to examine several animals side by side to distinguish the males from the females. The difference becomes more apparent after a few days of age. Another landmark is the presence of nipples in the females from 10 days of age, which are absent in the male. Darker mice are more difficult to differentiate than light colored mice.

Newborn through Day 5 The images on the next slide show pups from 1 to 5 days of age. (The following slides show mouse pups from 6 to 15 days of age.)

Newborn, or 1 day old, mice are very red, helpless, and hairless. Because of their color and lack of hair, they are often referred to as pinkies.

Day 6 through 10

The next slide shows images of pups at ages of 6 through 10 days. The stages of fur growth are an important indicator of age during this period of time

Here are images of pups at ages of 6 through 10 days. The stages of fur growth are an important indicator of age during this period of time.

Day 11 through 28
The next slide shows images of pups from 11 days through 4 weeks of age.

This 18 day period is one of rapid growth and change as their eyes open and activity increases.

Carbon Dioxide Asphyxiation

60% to 100% CO2 (tank or house carbon dioxide) or dry ice Impervious container with raised floor and lid or CO2 chamber with lid

Carbon Dioxide Asphyxiation

Precharge the impervious container or the CO2 chamber with CO2 prior to introducing the animals.

If dry ice is used as the CO2 source, a raised floor is necessary to prevent animal contact with the dry ice. Placing warm water on the dry ice facilitates vaporization and filling of the container.

Carbon Dioxide Asphyxiation

Unconsciousness will occur within 30 sec, but animals should be left in the container for several minutes to ensure death.

4. Verify death by lack of cardiac pulse and fixed and dilated pupils prior to carcass disposal.

Carbon Dioxide Asphyxiation Cause perivascular edema in the lungs or alveolar hemorrhage. Neonates are resistant to the effects of high levels of CO2

Cervical Dislocation of Mouse

Institutional Animal Care and Use Committee policy.

Cervical Dislocation of Mouse

Remove the mouse from its cage by grasping the base of the tail. Place it on a smooth, hard surface without releasing the tail. Place the pencil or metal rod firmly behind the ears and across the neck. Pull the tail sharply to the rear while pressing down on the neck (to quickly dislocate the cervical vertebrae).