This is a viral disease of mice that affects multiple organs. The objective of this book is to concisely present information with respect to appropriate use of experimental rodents in research. Cage Density: 5 in²/mouse Bleed Route: Cardiac puncture Analyzing Equipment: Drew Scientific HemaVet Charles River Technical Data 2010: Spontaneous Neoplastic Lesions in the Crl:CD-1® (ICR) Mouse in Control Groups from 18-Month to Two-Year Studies 2007: Reproductive and Behavioral Evaluations in Crl:CD-1® (ICR) Mice 2000: Spontaneous Neoplastic Lesions in the Crl:CD-1® (ICR) BR Mouse … .. There have been recent outbreaks at several facilities in United States associated with the injection of mouse sera or tissue culture material containing mouse sera into mice. Because of the thermoregulatory function of the tail no more than the distal 3 mm should be taken at a time. A 3-CC syringe with 25-gauge needle (Becton Dickinson and Co., Franklin Lakes, NJ) was pre-coated with heparin lock flush … Ovulation occurs at the end of metestrus. Procedures performed on the animal should be clearly indicated. 0000007537 00000 n 0000022714 00000 n The inguinal canal remains patent throughout life. Cardiac puncture Up to 1 ml of blood can be obtained from the heart of a deeply anesthetized mouse in a terminal procedure. 0000000016 00000 n Found inside – Page 847The control group giogenesis in a mouse model of urethane - induced primary lung received the control diet . ... from blood collected by cardiac studies have established that the pure compound is stable for many puncture and placed into ... 0000003604 00000 n Vena cava and abdominal aortaBlood can be obtained from the posterior vena cava or abdominal aorta in a deeply anesthetized mouse following laparotomy. The goal is to prevent introduction of a disease into a colony rather than to treat animals after disease outbreak. 3. Submandibular vein bleed in mice. After swabbing the lower right quadrant with alcohol, a fine gauge needle is introduced slowly through the skin, subcutaneous tissue and abdominal wall. Generally opioids e.g. It appears to be of no sequelae for the health of the mice. While mice are timid, they may bite. Download PDF. iii. A butterfly catheter with only about 5 mm of tubing attached to it (rest cut off) may be used instead of a needle and syringe. All physiological measurements described below were carried out in the last 2 … For repeat bleeds at shorter intervals, a maximum of 1% of an animal's circulating blood volume can be removed every 24 hours. Murine pharmacokinetics (PK) represents the absorption, distribution, metabolism, and elimination of drugs from the body, which helps to guide clinical studies, ultimately resulting in more effective drug treatment. Use of cervical dislocation or decapitation in absence of deep anesthesia must be scientifically justified. If the protocol involves more than one species, blood collection procedures must be addressed for each species. Protocol. Up to 10% of the circulating blood volume may be taken on a single occasion from a normal healthy animal on an adequate plane of nutrition with minimal adverse effect. Knockout mice has a protocol yields a surgical scissor and try to the same. a. Rodents (Mouse and Rat) i. Males have a greater (1.5-2 times) anogenital distance than females as well as a larger genital papilla. 1. Diseases of mice are usually handled as a herd (colony) health problem rather than on an individual animal basis. 0000069292 00000 n Outbreaks result in widespread deaths in neonates and occasionally weanlings, with or without diarrhea. In ventilated cages air is forced into the cage at up to 60 air changes per hour. 2. We are interested in inflammation and oxidative stress in the heart. 0000014922 00000 n Gentle pressure over the puncture site or relaxation of the restrainer's grip is usually sufficient to stop the blood flow. The following volumes can be injected into mice safely (based on 25 g mouse): 2-3 ml subcutaneously, 0.05-0.1 ml intramuscularly (0.03 ml per site), 0.50 ml intravenously, 0.1-0.3 ml into the stomach and 2-3 ml intraperitonealy. Mice have a pair of incisors and three pairs upper and lower of molars. Blood was serially diluted and cultured on a tryptic soy blood agar plate at 37 °C for 48 h (n=10 mice). The mouse is probably the most genetically and biologically characterized mammal in the world. Because of potential damage to the heart, blood collection by cardiac puncture is usually permitted only as a terminal procedure. They are born hairless and their eyes open after 10-12 days. They then underwent laparotomy and a 22 G needle attached to a 1 mL syringe was used to puncture through the diaphragm into the heart and 800 µL of blood was collected, … Blood samples were collected by cardiac puncture for clinical biochemistry analysis. 0000010748 00000 n Individual mice can be identified using ear punches, ear tags, tattoos, fur dyes, indelible mark on tail or microchips. PROTOCOL FOR CARDIAC PUNCTURE[10,11] Requirements include animal, anesthetic agent, towel, cotton, 19 to 25G needle with 1 to 5 ml syringe, surgical blade, tube (internal diameter of 0.1 to 0.3 mm) for thoracotomy, plastic disposable bag and blood sample collection tubes. 300-500ul) Done under anesthesia Generally terminal e.g. Mice were first used for the study of respiration in the 17th century. Males have a greater (1.5-2 times) anogenital distance than females as well as a larger genital papilla. Mums with newborns will likewise aggressively defend their pups and territories. acetaminophen, ketoprofen, caprofen, ibuprofen are used to relieve pain. Successfully reported this slideshow. Preferred sites for blood collection from mice and rats include tail veins, saphenous veins (hind limb), or, with proper training, the facial vein and jugular vein (competency certification by OLAC required prior to … Cardiac puncture is a suitable technique to obtain a single, large, good quality sample from a euthanised mouse or a mouse under deep terminal anaesthesia if coagulation parameters, a separate arterial or venous sample or cardiac histology are not required. Cardiac puncture should not be used if the peritoneum needs to be lavaged to harvest cells, as this technique can cause blood to escape into the peritoneal cavity. One person is required to take the blood sample. Parasuraman S, Raveendran R, Kesavan R (2010). This behavior is especially evident among females and is more common in some strains than others. 0000003805 00000 n I have a quite dismal success rate with the cardiac puncture technique and I wonder if anyone here can provide some tips on how to do it. Avoid contaminating urine with faeces. 1 Weigh the adult mouse to the nearest 0.1 gram. 2 Administer anesthetic. 3 Place the injected mouse in a heated cage for 5-10 minutes. 4 Assess responses to tail/toe pinches and the intactness of the ocular reflex. 5 Secure the mouse in the supine position (lying on the back with face upward) by gently. Most of the common laboratory strains were developed in the early 1900s. Make a cut through the skin at the side of the thorax into the angle of the forelimb (axilla) to expose the axillary vessels. Sodium pentobarbital can be used, but it has a narrow safety margin and is associated with a prolonged recovery period. Found inside – Page 52New Insights and Experimental Protocols Mojca Božič-Mijovski ... Up to 1 mL of mouse blood was drawn by cardiac puncture of deeply anesthetized fasting mice in a terminal procedure and collected in plastic tubes containing trisodium ... Materials to be administered to mice can be given orally e.g. 3. The amount of secretions increases during stress and appears as 'red crusts' around the eyes and nostrils. Ectromelia causes high mortality and drastic measures including depopulation are usually taken to eliminate the disease. A. There are some suggestions that it may be a cooperative and/or learnt behavior. Found inside – Page 112... was collected by cardiac puncture from each mouse and placed in an micro-tube tube containing 10μl of Heparin. ... The mouse blood was chemically deoxygenated following a protocol developed by Briley-Sæbø and Bjørnerud [42]; 1.5 mg ... There should be no movement of the head during the procedure. Position a microhematocrit tube along the inner corner of the eye (medial canthus) beside the eyeball. At day 33 post challenge the mice are sacrificed with the caecum collected and assessed for the presence or absence of worms. Species. SexingSex is determined using the anogenital distance. Some basic science research has been conducted to begin to understand the implications of variant HFE genotype on cancer, but the story is far from complete. It also includes brief technical reports … This hierarchical system creates a situation whereby the dominant animals often barber (chew off the hair) those of lower rank. Translate PDF. A fine gauge needle should be used for this procedure. Disassemble a 3 cc syringe.Thread a loop of OO gut suture material through the syringe. All individuals performing euthanasia must be properly trained. Serum was collected by centrifugation at 1500×g, 4°C for 15 The expression of miR-193-3p, STAT3 and HMGB1 in the myocardial tissue of septic mice were detected. Blood should flow freely, if, the tube is properly inserted. This page contains links to older films and slide shows produced at the Norwegian School of Veterinary Science, and links to newer resources produced by international colleagues. Tissue Harvest Protocol. Procedures performed on the animal should be clearly indicated. There is no charge. To register: Click here: Training Form. Could monochromatic X-rays revolutionize breast imaging? One hour later, the abdomen was reopened and blood was drawn by means of a terminal cardiac puncture. Cardiac puncture allowed the collection of appro-priate amounts of blood from a single animal. The footpad of each animal was pinched a few times to ensure that all physical sensa-tion had been lost. PneumocystisThis is a fungal disease affecting a wide range of laboratory animals and humans. This is a terminal procedure. Hello fellow lab rats. 1. In addition blood is collected by cardiac puncture and serum isolated for confirmation of productive infection. Cages with filter tops may have a slightly higher temperature, relative humidity, carbon dioxide and ammonia than the room air. A variety of restraint devices are available to assist in handling mice. 0000004639 00000 n The tail vein in a mouse cannot be used to collect small amounts of blood. To overcome this limitation, we sought to develop a simplified method in the mouse that uses ultrasound-guided injection of potassium chloride directly into the heart. C. The lateral saphenous vein is not accessible in the mouse. 8ai) (p = 0.5217 by paired t test) while in TBI mice, plasma GFAP concentrations from saphenous blood were 163.4 ± 1.8% of that from cardiac puncture (Fig. Cardiac puncture (Terminal Procedure) The technique for cardiac puncture from the hamster is identical to that described for the mouse. Cardiac puncture (terminal procedure) Tail snip for genetic analysis; Euthanasia with CO2/O2; All research personnel must be listed on an approved Animal Protocol to participate in this class. <> 3 0 obj cMyC has three distinct isoforms encoded by different … Molars are permanently rooted while the incisors have an open root and grow continuously. Wipe the shaved area with alcohol or sterile lubricate gel and use a 25-gauge needle to puncture the vein (Vein is highlighted by the dark line in the picture below). Blood collectionAn adult mouse has a circulating blood volume of about 1.5-2.5 ml (6-8% of the body weight), however in older and obese animals this may be lower. Cardiac puncture: Following collection of the TT sample, the mice were immediately anaesthetized by intraperitoneal injection with ketamine (100 mg/kg body weight) and xylazine (5 mg/kg body weight). Table 1: Mouse Site Mice were first used for the study of respiration in the 17th century. Collection from the auricular vein or artery of the rabbit is also relatively unstressful to the animal. Babies are born either head or tail first (breech). Place mouse in isofluorane chamber and lightly anesthetize the animal. c. Wash the tail with 70% ethanol. However, in what appears to be the first description of the cardiac puncture procedure using a needle, tubing, and suction using the mouth, Hicks and Little describe drawing 0.5–1.0 mL blood samples from laboratory-raised house mice (Mus musculus L.): “The animals were lightly anaesthetized, fastened on their backs to a small board and swabbed with 70 per cent … BarberingMice are communal animals with a social hierarchical system. If the animal struggles or appears to be in respiratory difficulty withdraw the tube and begin all over again. Usually 100% of the animals are infected. Sudden cardiac arrest mice have secondary organ damage, including 28 significant kidney injury, but no clear evidence of neurologic dysfunction. It is important that barbering is differentiated from other causes of hair loss or skin problems e.g. For mice and rats, the circulating blood volume is approximately 55-70 ml/kg of body weight (1,2). Here, the basic approaches for analyzing hematopoiesis in mice are described. This protocol was repeated with a second batch of mice with the histological data being pooled for both groups. For mice in this group, the above-described procedures [see High- intensity/short-duration (10 days) protocol], differing only in exercise intensity and duration (3.53 m/min, 60 min/day, 5 days/wk for a total of 6 wk), were followed. Separating the dominant mouse often leads to rise of another dominant mouse. MOUSE INTRACARDIAC PUNCTURE PROCEDURE 2.1 Anesthetize mouse (according to approved protocol) 2.2 Place mouse in dorsal recumbency 2.3 Prep area just below sternum with alcohol swab. My protocol to get PBMCs from mouse blood is a bit crude but simple. *Cardiac collection subject to stipulations outlined in this guideline and in the animal use . Mouse Blood Draw: Cardiac Puncture. Ketamine and xylazine is a common injectable anesthetic combination. Place a needle into the loop and pull the loop tight against the tip of the syringe.Tie the long ends of the suture material together. Septic cardiac dysfunction is the main cause of death in septic patients. Found inside – Page 31Clinical protocol and patients' characteristics are described in the Supplemental Information (Table S1). ... Mouse whole blood was drawn in anesthetized mice by cardiac puncture using heparin as anticoagulant. Cardiac puncture Up to 1 ml of blood can be obtained from the heart of a deeply anesthetized mouse in a terminal procedure. The development of cardiac dysfunction in sepsis results in an increase of mortality. Cardiac puncture in the mouse through the anterior thoracic aperture, 1979 An adult mouse has a circulating blood volume of about 1.5-2.5 ml (6-8% of the body weight), however in older and obese animals this may be lower. Found inside – Page 33427.2.4.1 Blood Collection Mouse Blood 1. To perform cardiac puncture, euthanize mice using carbon dioxide. Then access the ventricle with an 18 gage needle and aspirate 400–500 μL blood into a 2 mL syringe. Clerical errors can prove fatal. The new WHO guidelines provide recommended steps for safe phlebotomy and reiterate accepted principles for drawing, collecting blood and transporting blood to laboratories/blood banks. Conspicuous bilateral rows of nipples are visible in females at about 9 days of age. Bleeding can be stopped by applying gentle finger pressure. The disease produces massive die off in adult mice and amputation of the limbs (ectromelia) in surviving animals. 4. <>>> This behavior is especially evident among females and is more common in some strains than others. mites, fungi or bacteria by a veterinarian. This paper. Rotate the tube gently as you proceed. 0000121526 00000 n check your protocol, the size of the mouse, the frequency and interval of collections whether or not replacement fluids are given whether it is a terminal blood collection. The protocols offered in this unit describe collection of blood from the orbital sinus or plexus of the mouse, rat, or hamster. Found inside – Page 452Tail vein venipuncture in mice: A scalpel blade and a 25–30 gauge needle are required. The mouse is restrained ... Cardiac puncture represents an accepted method of blood collection from mice when more than a few drops are required. For a detailed discussion of pain relief in mice refer to module 2. The animal must be Drugs can be administered in water, in jello, as oral drops, by gavage or injected. Pain reliefFor a detailed discussion of pain relief in mice refer to module 2. The disease has a severe negative impact in research involving immunocompromised animals, pulmonary function and immunology. Absence of testicles is not a useful criterion for sexing since the testis is retractable into the open inguinal canal throughout life. 0000006776 00000 n ReproductionMice have two distinct cervices and uterine bodies. The laboratory mouse, Mus musculus, belongs to the order Rodentia and family Muridae. Blood Collection • Large volumes (e.g. Open the chest widely, taking care not to puncture the lungs. At the end of the collection apply pressure to the cut end with a gauze bandage and ensure that blood has completely stopped flowing before returning the mouse to the cage. Turn on isoflurane chamber to 5. second group of mice was examined. Mice were monitored using a clinical severity score every 12 hours, as previously reported. 1 0 obj Mouse rooms are usually maintained at 30-70% relative humidity and a temperature of 18-26ºC (64-79ºF) with at least 10 room air changes per hour. 0000008381 00000 n This hierarchical system creates a situation whereby the dominant animals often barber (chew off the hair) those of lower rank.
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