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Instructions
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Techniques of Experimentation.
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The purpose of this chapter is to select and summarize
Instructions the available information in an attempt to emphasize two
Introduction
major concepts:

Resources
Tech of Exp (1) The technique employed in animal experimentation is
often the critical factor in determining the success or
failure of a research protocol, and ;
Videos
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(2) a mastery of selected techniques is extremely useful
to the veterinary clinician in performing diagnostic and
Other
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Digestive System Technique

Digestive System Technique

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1. Oral Examination
Instructions
Introduction 2. Oral Administration of Substances

3. Tooth Extraction
Resources
Tech of Exp 4. Pulpectomy and Pulpotomy

5. Bile Duct Manipulations

Videos
GI
6. Partial Hepatectomy and Liver Biopsy

7. Pancreatic Exocrine and Endocrine Studies

Other
Urinary
8. Intestinal Cannulation
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9. Intestinal Loop Isolation

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 1. Oral Examination

ORAL EXAMINATION
Tubular Device For Intraoral Examination Of
Rodents

A tubular device overcomes the difficulties created by the

large diastema and small oral cavity of rodents.

Proved to be reliable for intraoral examination of mice, rats,

hamsters, and guinea pigs.

The device consists of a tube with a conical end, a base,

and a movable stem.

The rodent is anaesthetized lightly with ether and inserted

quickly into the tube so that the head extends beyond the
opening at the conical end, where cheeks, upper and lower
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incisors are secured to permit illumination and direct visual

or microscopic examination of the palate, molars or gingiva.
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 1. Oral Examination

ORAL EXAMINATION
3.5v Bivalve Nasal Speculum Certain species have unique anatomical adaptations
The bivalve nasal speculum from Welch Allyn is
that are utilized in research investigations.
useful for oral exams. A more sophisticated
technique for examination of the oral cavity is Example:
dental endoscopy
• Hamster cheek pouch.
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 2. Oral Administration Of Substances

ORAL ADMINISTRATION OF SUBSTANCES

Catheters and Needle

Used for gavage in small rodents.

Durable, stainless steel, ball-tipped needles ,
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Gastric intubation
and semirigid disposable needles are used
more frequently
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Ensures that all the material is administered

 2. Oral Administration Of Substances

ORAL ADMINISTRATION OF SUBSTANCES

Gastric Intubation

❑ minimize trauma to oropharyngeal tissues

❑ make inadvertent endotracheal passage less
likely.
❑ safer for the animal, and a recommended animal
welfare refinement.
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 2. Oral Administration Of Substances
VIDEO

ORAL ADMINISTRATION OF SUBSTANCES

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 2. Oral Administration Of Substances

ORAL ADMINISTRATION OF SUBSTANCES

Common Complications Associated With Gastric Intubation

❑ Aspiration of solution into lungs

▪ Due to: tube length being too short from not pre‐measuring to the correct length
resulting in accidental administration into trachea or regurgitation.

▪ Response: Place the animal in a cage, administer oxygen if available and

monitor breathing closely.
▪ If the solution was aqueous, it may recover.
▪ If the substance was oily or viscous, the animal should be euthanized. Monitor
closely and if any signs of respiratory distress continue after 10 minutes, the
animal should be humanely euthanized
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 2. Oral Administration Of Substances

ORAL ADMINISTRATION OF SUBSTANCES

Common Complications Associated With Gastric Intubation

❑ Perforation of the esophagus, trachea or lungs

▪ Due to: improper restraint and excessive struggling by the animal or forceful
▪ insertion of gavage tube
▪ Response: Humane euthanasia

❑ Chewing off a piece of the tubes (plastic tubes only)

▪ Response: If this happens, keep the rat restrained and use atraumatic forceps to
▪ attempt to grasp the portion of the tube that is in the esophagus and gently
remove it.
▪ If swallowed, monitor animal closely over the next few days to ensure it is
behaving normally and is not losing weight and that the piece has not resulted in
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a perforation or blockage. Look for it passing in the feces. Contact the facility
veterinarian if animal is not doing well
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 2. Oral Administration Of Substances

ORAL ADMINISTRATION OF SUBSTANCES

Common Complications Associated With Gastric Intubation

❑ Damage to the oral cavity

▪ Due to: improper restraint and excessive struggling by the animal or forceful
insertion of gavage tube.

▪ Response: If any blood is seen in the animal’s mouth or on the gavage tube,
monitor closely to ensure bleeding does not persist.
▪ Re‐check animal’s mouth after a few minutes. If any bleeding is seen,
consult the facility Veterinarian since the animal may need to be humanely
euthanized.
▪ If no blood is seen, check the animal again at the end of the day to ensure
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they have recovered.

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 2. Oral Administration Of Substances

ORAL ADMINISTRATION OF SUBSTANCES

Common Complications Associated With Gastric Intubation

❑ Esophagitis (inflammation of the esophagus)

▪ Due to: irritation of the esophagus by the substance administered or the gavage
tube (e.g. rough surfaces on a plastic tube, animal struggles during gavage, or
tube advanced too forcefully)

▪ Response: Consult the facility Veterinarian for treatment options

❑ Other traumatic injuries related to improper animal restraint

▪ Response: Dependent on severity of symptoms: humane euthanasia or
consultation with facility Veterinarian for treatment options.
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 3. Tooth Extraction

TOOTH EXTRACTION
A tooth extraction is a dental procedure during
which the tooth is completely removed from its
socket.

PURPOSE:
● Severe tooth decay (cavities).
● A fractured tooth.
● An impacted tooth.
● Crowded teeth.
● Severe gum disease.
● Tooth luxation or other dental injuries.
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 3. Tooth Extraction

TOOTH EXTRACTION
The technique of tooth extraction using the dental elevator involves:
1. Inserting the dental elevator between the alveolar bone and the tooth;
2. Applying pressure on the elevator, using a rotating wrist action while
directing it toward the apex of the root on all sides; and
3. Attempting to pull the tooth only after all periodontal ligaments have
been severed.

Dental procedures may be adapted from those described from dogs and cats.

Canine tooth extraction and reduction in nonhuman primates appears less

widely used than in the past, and policy statements by the AVMA and the U.S.
Department of Agriculture (USDA) are in opposition to the practice if used for
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nonmedical reason
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 3. Tooth Extraction
VIDEO

TOOTH EXTRACTION
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 3. Tooth Extraction

PULPECTOMY AND PULPOTOMY

❑ Popularized as alternatives to canine

tooth extraction.
❑ These procedures have been used in
veterinary medicine for treating tooth
fractures and pulp infections and for
disarming dangerous animals.
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 3. Pulpectomy And Pulpotomy

PULPOTOMIES PULPECTOMIES

opening of the root canal and the refer to the total removal of the pulpal
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partial removal of the pulpal tissue. tissue.

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 3. Pulpectomy And Pulpotomy

PULPOTOMY VIDEO

PROCEDURE
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 3. Pulpectomy And Pulpotomy

PULPECTOMY
VIDEO
PROCEDURE
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 3. Pulpectomy And Pulpotomy

PULPECTOMY/PULPOTOMY PROCEDURE

❑ Anesthetizing the animal or a nonhuman primate

❑ Placie on a surgery table in dorsal recumbency.
❑ Rubber dams are used on all teeth to prevent contamination of the root canal
with microorganisms from the oral cavity.
❑ Each canine tooth is cut off level with adjacent teeth, using tapering crosscut
bars or damascus separating disks (attached to a dental drill.
❑ Broaches are used to remove all pulpal tissue from the root canal.
❑ The root canal is irrigated with sterile saline.
❑ Paper points are used to dry the canal and check for the presence of blood
▪ If bleeding occur, the canal must be explored, so that all pulpal tissue is
removed.
❑ Other techniques have employed epinephrine flushes or electrocautery to
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stop the bleeding and dry the root canal.

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 3. Pulpectomy And Pulpotomy

PULPECTOMY/PULPOTOMY PROCEDURE

❑ Root canal paste formulas include:

▪ an RC-2W mixture
▪ the commercial product Hypo-Cal which is calcium hydroxide in a
calibrated disposable syringe with a special applicator needle.
❑ After the canal has been filled, excess paste is removed, and an
amalgam plugger is used to firmly pack the hardening paste into the filled
canal.
❑ A silver alloy amalgam filling is used to permanently seal the filled root
canal.
❑ Excess alloy is removed, and the tooth is sculptured smooth.
❑ Some have advocated a more permanent technique involving the
placement of a permanent pulp cap Although this technique is more
costly and time-consuming, it does give the investigator an opportunity to
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easily convert troublesome temporary caps to more durable crowns at a

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later date. Pulpotomy using formocresol and electrosurgery

 3. Pulpectomy And Pulpotomy

Other Nonhuman Primates

❑ In baboons and macaques

❑ A technique termed Submucosal Vital Root Retention,

involving the amputation of the crown of the canine tooth
below the margin of the gum, has been described by
Schofield et al., 1991.

❑ In this technique, the gum flap is repositioned over the

remaining tooth, with retention of nerve function and
minimization of subsequent problems such as toothroot
abscesses and devitalization of the tooth
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 4. Bile Duct Manipulation

BILE DUCT MANIPULATIONS

Bile Anatomy & Functions

Bile is a fluid produced in the liver and stored in the
gallbladder.
Function: Digestion of fats and fat-soluble vitamins.
The primary role of bile is to emulsify (blend) lipids
(fats) prior to digestion

Bile Duct Manipulations/Exploration

Is a procedure that is performed to see if anything,
such as a stone, is blocking the flow of bile from the
liver and gallbladder to the intestine.
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 4. Bile Duct Manipulation

BILE DUCT MANIPULATIONS

Simple, Short-term Method For Collecting Bile
1. Direct Surgical Cannulation Of The Gallbladder
❑ Laparoscopic cholecystectomy
❑ Endoscopic Retrograde Cholangiopancreatography (ERCP)
▪ Use in larger species, such as the dog, pig, and nonhuman primate.
▪ It is done with placement of an 11-mm duodenoscope to collect bile and
evaluate the biliary and pancreatic ductal system.
▪ Bile can be aspirated by cholecystocentesis under ultrasound guidance
▪ Permits reliable and safe catheterization of the gallbladder without the
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need for endoscopic sphincterotomy

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2. Ligation Of The Common Bile Duct

 4. Bile Duct Manipulation

BILE DUCT MANIPULATIONS

Complete biliary retention
❑ can be accomplished easily by ligation of the common bile duct.
❑ This technique results in obstructive jaundice.
❑ Partial obstruction, to mimic disease conditions can be produced in the rat by ligation
of the bile duct over a 0.5-mm rod, followed by removal of the rod
▪ bile duct stricture
▪ Cholelithiasis
▪ biliary tumors
▪ extrinsic compression of the bile duct

Complete biliary retention without obstruction

❑ can be accomplished by implanting a catheter between the common bile duct and
the venous return
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❑ This technique allows for studies of complete bile recirculation without the pathologic
effects of complete obstruction
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 4. Bile Duct Manipulation

VIDEO
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 4. Bile Duct Manipulation

VIDEO
BILE DUCT MANIPULATIONS
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 5. Partial Hepatectomy And Liver Biopsy

PARTIAL HEPATECTOMY AND LIVER BIOPSY

❑ Liver has a considerable capacity to regenerate after surgical or
toxic insult.
❑ Partial Hepatectomy (PH)
▪ Is performed to get sufficient tissue for certain types of studies,
to remove an abnormal portion of the liver which may contain a
tumor, infection, or some unknown disease.
▪ Served as a valuable experimental tool to study liver
regeneration and function
▪ A comprehensive review of rodent hepatectomy models
identifies the advantages and disadvantages of these various
techniques
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▪ Using this technique, 5–10% of the original liver is needed for

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survival.
 5. Partial Hepatectomy And Liver Biopsy

PARTIAL HEPATECTOMY AND LIVER BIOPSY

❑ Liver biopsy
▪ The most straightforward method for performing liver biopsies
a. transcutaneous skin puncture
b. percutaneous liver biopsies

Open Liver Biopsy

▪ for larger tissue samples
▪ Advantage: Larger samples of biopsy material are provided and
hemostasis of the severed liver lobe is excellent

Laparoscopic Biopsies Of The Liver

▪ involves inserting a laparoscope, a thin tube with a tiny video camera
attached, through a small incision to look inside the body to view the
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surface of organs
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 5. Partial Hepatectomy And Liver Biopsy

VIDEO
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 4. Bile Duct Manipulation

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

The pancreas performs two essential

purposes.
❑ “Exocrine Function” produces and
secretes digestive enzymes

❑ “Endocrine Function” produces

hormones that control healthy levels
of blood sugar (glucose) in the
bloodstream.
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 4. Bile Duct Manipulation

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

Techniques
❑ Surgical creation of pancreatic fistulas

❑ Duodenal fistulas, or duodenal

pouches.
❑ Catheterization of the pancreatic duct

▪ A study employing catheterization

of a surgically created pancreatic
fistula allowed studies to be done
in conscious, freely mobile rats.
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 6. Pancreatic Exocrine And Endocrine Studies

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

Models Of Pancreatic Endocrine Function Largely

Involve Study Of The Ff:
❑ Diabetes Mellitus (DM)

▪ Type 1 DM.

❑ Non-obese-diabetic (Nod) - Mouse

❑ Biobreeding (Bb) – Rat / Or Can Be Induced

Induced Models Usually Involve The

❑ Chemical Ablation Of The Pancreatic Islets

▪ Using Alloxan Or Streptozotocin (STZ)

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▪ Surgical Removal Of The Pancreas.

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 6. Pancreatic Exocrine And Endocrine Studies

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

Chemical Induction Techniques

❑ based upon STZ’s specific β-cell toxicity

❑ Chemical ablation is effective in
▪ Mice
▪ Rats
▪ Rabbit
▪ nonhuman primates
▪ Dogs
▪ pigs
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 6. Pancreatic Exocrine And Endocrine Studies

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

Chemical Induction Techniques
❑ Cats -resistant to the diabetogenic effects of
both alloxan and STZ
▪ surgical removal, or a combination of
surgical and chemical techniques
❑ In rats and swine
▪ Coadministration of Nicotinamide and STZ
can create a stable state of moderate post-
prandial or fasting hyperglycemia
❑ Alloxan - used, alone and in combination with
STZ, in creating chemically induced models of
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diabetes
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 6. Pancreatic Exocrine And Endocrine Studies

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

Pancreatectomy Models
▪ Provide insights into DM caused by Total
Pancreatectomy
▪ Islet Auto Transplantation
▪ Allotransplantation
▪ Immunosuppression protocols bioengineered
pancreas
▪ Cell regeneration

❑ Total pancreatectomy in dogs and swine is well described

❑ In laboratory rodents, surgical removal of the pancreas is
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very difficult because of its diffuse anatomy

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 6. Pancreatic Exocrine And Endocrine Studies

PANCREATIC EXOCRINE AND ENDOCRINE STUDIES

❑ Microsurgical anastomosis of the extirpated pancreas to recipient sub-
renal aorta and porto-portal venous drainage are most physiologic.
❑ Exocrine pancreatic drainage can be accomplished by diversion into the
recipient intestine, urinary tract, or peritoneal cavity.
❑ All techniques have potential shortcomings.
▪ Whatever technique is used, sophisticated microsurgical skills,
close attention to reducing ischemia time in the graft, and prevention
of hemorrhage and hypothermia in the recipient are required to
improve graft survival and minimize postsurgical complications
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6. Total Pancreatectomy

VIDEO
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 7. Intestinal Cannulation

INTESTINAL CANNULATION
❑ There are many designs and materials used for insertion of rigid or
flexible cannulas into alimentary tract fistulas of experimental
animals.
▪ The choice of cannula depends on several factors, such as:

▪ size of the lumen of the organ to be fistulated,

▪ particle size and viscosity of the material to be sampled or

infused
▪ site of exteriorization of the cannula,

❑ For the case of reentrant fistulas

▪ volume and consistency of ingesta expected to flow through the
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cannulas to avoid producing excessive resistance to flow

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 7. Intestinal Cannulation

INTESTINAL CANNULATION

❑ An intestinal re-entrant cannulation

technique was devised which did not
necessitate gut transection.
❑ The technique provides for the total
collection and return of digesta, but
restores a comparatively uninterrupted
flow of digesta between collection
periods.
❑ The technique involves the displacement
of a short section of intestine to a
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subcutaneous site between two simple

T-shaped cannulae.
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 7. Intestinal Cannulation

INTESTINAL CANNULATION

Cannula Designs and Materials

▪ flexible plastic cannulas made of

▪ polyvinyl chloride [PVC]

▪ tygon, may be preferable to rigid cannulas in
▪ polyethylene some cases
▪ silicone

❑ Design modifications in the catheter tips have improved patency of

intestinal catheters and includes formation of a slit or burp valve that
allows infusion of materials while preventing ingesta from occluding
the catheter.
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 7. Intestinal Cannulation

INTESTINAL CANNULATION

❑ These devices can be modified for longer

patency by adding in barrel plugs to prevent
ingesta from occluding the cannula.
❑ The cannulas have also been used as access
for repeated endoscopy and biopsy collections
❑ Vascular access ports may be used for
delivering drugs and solutions to various
segments of the gastrointestinal tract without
the problems associated with externalized
catheters or cannulas
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 8. Intestinal Loop Isolation

INTESTINAL LOOP ISOLATION

❑ Isolated Loops Of Small Intestine Are Referred To As:
1. Thiry fistula (one end exteriorized)

2. Thiry–Vella loop fistula (both ends exteriorized)

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 8. Intestinal Loop Isolation

INTESTINAL LOOP ISOLATION

❑ The crucial stage in preparing such a segment

is the method of anastomosis used on the
intestinal wall.
❑ For most species, the preferred technique is
the use of a simple, interrupted, approximating
suture pattern that limits intestinal tissue
trauma and minimizes luminal narrowing.
❑ Maintaining vascular viability is critical and
tension should not be placed on the segment
that could occlude the vasculature.
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 8. Intestinal Loop Isolation

INTESTINAL LOOP ISOLATION

RITARD (Reversible Intestinal Tie–adult Rabbit
Diarrhea)
❑ model allows temporary occlusion of a section of
bowel for bacterial colonization studies in the
rabbit.
❑ This technique utilizes sterile umbilical tape tied
in such a way that the ligature can be loosened
and removed by gentle traction on the ends that
are exteriorized through the incision site
❑ have been utilized in vaccine development for
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prevention of Vibrio cholera using this technique

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Urinary System Technique

Urinary Sustem Technique

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Method
Instructions
Introduction 1. Urine collection
2. Exteriorization of the ureters
Resources
Tech of Exp 3. Implantation of the ureters into the
intestine.
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1. Reconstructive surgery of the ureters
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4. Denervation and decapsulation of the kidney
 1. Urine Collection

Urine Collection
Many abnormalities of the urinary system can be diagnosed from the
signalment, history and physical examination findings, serum chemistry
profile, urinalysis, and aerobic bacterial urine culture.

Urine may be collected by one of five (5) methods:

(1) Collection During Spontaneous Micturition
(2) Manual Compression Of The Urinary Bladder
(3) Catheterization
(4) Cystocentesis
Use Of Metabolic Cages
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(5)
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 1. Urine Collection

Urine Collection

Collection During Spontaneous Micturition

Micturition / Urination
❑ process of expelling urine from the bladder
❑ Involves the passive storage and the active voiding of
urine.
❑ Spontaneous Micturition patterns can be used to
evaluate how changes in environment, age, and
genetic background impact urinary function.
❑ Manually restrained mice often urinate
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spontaneously; 50–500μl can be collected in a 2-min

period.
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 1. Urine Collection

Urine Collection
Manual Compression Of The Urinary Bladder

❑ The stream of urine flowing, is being

produced by manually compressing the
bladder.
❑ Complete emptying is impeded by
contraction of the striated urethral
sphincter (urethralis muscle) in this
patient.
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 1. Urine Collection

Urine Collection
Manual Compression Of The Urinary Bladder

Manual urine expression may be best

accomplished in an anesthetized rabbit.
The full bladder can be palpated and
pressure applied in a firm, steady manner.
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 1. Urine Collection

Urine Collection
Catheterization

ADVANTAGE DISADVANTAGES
❑ Distended urinary bladder ❑ Urinary bladder may be traumatized
compressed at convenience of ❑ Infected urine may be force into
collector ureters, kidney and prostate
❑ Technique cannot be used
immediately post cystotomy and
may be uncomfortable in other post-
operative laparotomy patients.
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 1. Urine Collection

Urine Collection
Catheterization
❑ A very narrow sterile catheter is
passed up the lower urinary
passage/urethra into the bladder.
❑ less invasive and is a good option
when a voluntary sample is not
available, especially in male dogs.
❑ Catheterization causes mild
irritation to the urethra, and may
carry bacteria from the urethra
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into the bladder

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 1. Urine Collection

Urine Collection
Catheterization

❑ Catheterization of the urethra can be accomplished most readily in male

rabbits.
❑ A subcutaneous dose of acepromazine at 0.8mg/kg will tranquilize the animal
and encourage protrusion of the penis.
❑ The buck is supported in a sitting position, tilted slightly back; a seated handler
can restrain the animal held in the handler’s lap with one hand holding the
scruff and the other restraining the rear legs
❑ A sterile, flexible, lubricated catheter, size 9 French is passed into the urethra
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in a downward direction for about 2 cm.

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 1. Urine Collection

Urine Collection
Catheterization

❑ The catheter and penis are manually depressed, and the catheter tip is then
gently advanced into the bladder.

❑ To catheterize a doe, the sedated rabbit is placed in ventral recumbency. The

catheter is first directed vertically in the caudal part of the vagina. The
catheter is then brought to a horizontal position to enter the urethral opening
in the ventral vagina. A cupped hand can be used to apply abdominal
pressure to express urine through the catheter.
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 1. Urine Collection

Urine Collection
Catheterization

ADVANTAGE DISADVANTAGES
❑ Bladder does not have to distended ❑ Risk for iatrogenic infection, blood
ad cellular contaminants
❑ Risk for trauma or perforation of the
urethra or urinary bladder
❑ Not possible if there is urethral
obstruction
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 1. Urine Collection

Urine Collection
Catheterization

Diagnostic catheterization is indicated for:

1. collecting bladder urine for urinalysis or bacterial culture
2. studying renal function
3. instilling contrast media for radiography
4. evaluating the urethral lumen for strictures and/or obstruction
5. surgically repairing the urethra and surrounding structures.
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 1. Urine Collection

Urine Collection
(1) Cystocentesis

❑ Sterile needle and syringe are

used to collect urine from the
bladder.
❑ The needle is passed through
the abdominal wall into a full
bladder and urine is withdrawn
directly into the sterile syringe.
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 1. Urine Collection

Urine Collection
(1) Cystocentesis

❑ performed on an anesthetized rabbit, placed in dorsal

recumbency.
❑ The full bladder is palpated and pressed between the
fingers and thumb, up against the abdominal wall.
❑ A 20-gauge needle is inserted directly through the skin
into the bladder.
❑ A 30–60-ml syringe is used to aspirate the urine.
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 1. Urine Collection

Urine Collection
(1) Cystocentesis
DISADVANTAGES
ADVANTAGE ❑ slightly more invasive than other methods
❑ Less risk for iatrogenic infection ❑ Must have sufficient amount of urine in
and contaminants bladder to perform
❑ Localizes source of cells and or ❑ Risk for microscopic hematuria
bacteria
❑ Inadvertent intestinal sampling
❑ Urinary bladder tear, leakage of urine into
abdomen in animal with severe bladder
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distention due to urethral obstruction.

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 1. Urine Collection

Urine Collection
Use Of Metabolic Cages

❑ designed to allow measurement of fluid

intake, and to separate and collect feces
and urine for numerous qualitative and
quantitative determinations.
❑ permits observation of the animal, feces,
and urine at all times, is readily cleaned
or sterilized, and is easily fashioned from
common laboratory equipment.
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 1. Urine Collection

Urine Collection
Use Of Metabolic Cages

Metabolic cages designed to collect urine separately from feces are

commercially available. If urine samples free from cage contamination are
needed, then urine can be collected by manual expression of the bladder,
cystocentesis, or urethral catheterization
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 1. Urine Collection

Urine Collection

❑ With spontaneous collection and following manual expression,

▪ first part of the urine stream should not be used for urinalysis or
bacterial culture, because it may contain debris, bacteria, or exudate
flushed from the urethra or genital tract.
❑ In male rats, which are difficult to catheterize, urine may be collected by
retraction of the prepuce and attachment of silicone tubing over the
glans penis
❑ In the female dog, visualization of the external urethral meatus aids in the
placement of the catheter.
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 1. Urine Collection

Urine Collection

❑ Regardless of the specific procedure employed, a meticulous aseptic and

gentle atraumatic technique should be used for catheterization of all
species
❑ Conscious patients should be restrained by an assistant in order to
minimize contamination of the catheter as well as trauma to the urethra.
❑ Verification of this position may be accomplished by injection of a known
quantity of air through the catheter
▪ inability to remove all of the air indicates improper positioning of the
catheter.
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▪ Proper positioning of the catheter facilitates removal of all of the

urine from the bladder.
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 1. Urine Collection

Urine Collection

❑ Potential complications associated with catheterization in all species

include hematuria and infection
❑ Procedures that may be used to reduce the incidence of infection
following catheterization include:
1. strict adherence to principles of asepsis
2. administration of oral or parenteral antibiotics
3. use of catheters impregnated with antibacterial agents,
4. irrigation of the bladder with antibacterial solutions, such as
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neomycin or furacin.
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 1. Urine Collection

Urine Collection

❑ Use Of Catheters In The Tethered, Freely Moving Animal

▪ Rabbitat: the ureters are very friable and subject to intraluminal
bleeding.
o Reduced urine output during anesthesia, combined with bleeding
tendencies, will often result in occlusion of the catheter with
blood clots.
o In order to compensate for this problem, designed a double-
lumen catheter.
o Irrigation of blood from the catheter is accomplished by perfusion
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of heparinized saline through a small inner catheters.

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 1. Urine Collection

Urine Collection

❑ Urine collection in neonatal (12-hour- to 10-day-old) rabbits can be

accomplished by stroking the abdomen.
❑ The kit is held gently but firmly on its back in one hand, while the other
hand strokes from the stomach area to just beyond the bladder, with
most pressure in the bladder area.
❑ After 10 days of age, muscular control of the bladder and increased size
of the kit makes it more difficult to use this method, and a longer period
of stroking is needed.
❑ Up to 5ml of urine can be collected in this manner
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 Resources

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1. Collection Of Pharyngeal Fluids
Instructions
Introduction 2. Tracheobronchial Washings

Resources
Tech of Exp
3. Endotracheal Inoculation
4. Intubation
Videos
GI
5. Tracheal Pouch Formation
6. Tracheostomy
Other
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7. Bronchoscopy
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Groups
Respi.. 8. Bronchopulmonary Lavage
 1. Collection of Pharyngeal Samples

Collection of Pharyngeal Samples

❑ A straight tube formed from a 1-ml tuberculin

syringe is used as a speculum for the
passage of sterile swabs into the pharynx.
❑ The transparent syringe barrel allows for easy
visualization of the larynx.
❑ Commercial sources for sheathed applicator
swabs are also available technique can be
performed in several species with materials
readily available in the laboratory and at a
much lower unit cost.
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 1. Collection of Pharyngeal Samples

Collection of Pharyngeal Samples

❑ Oral and pharyngeal samples are routinely

collected for
❑ polymerase chain reaction (PCR) and reverse
transcription (RT)-PCR for detection of
various diseases
❑ BVD in cattle
❑ West Nile virus in tree squirrels
❑ rabies in bats and dogs
❑ Pneumocystis in rats
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 1. Collection of Pharyngeal Samples

Collection of Pharyngeal Samples

❑ Oral and pharyngeal samples are routinely

collected for
❑ polymerase chain reaction (PCR) and
reverse transcription (RT)-PCR for detection
of various diseases
▪ BVD in cattle
▪ West Nile virus in tree squirrels
▪ rabies in bats and dogs
▪ Pneumocystis in rats
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❑ Oral swabs have also been used for

genotyping transgenic mice using PCR
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 2. Endotracheal Intubation

Endotracheal Intubation

❑ A patent airway is essential in

reducing mortality during and after
surgical procedures

❑ Endotracheal intubation is a medical

procedure in which a tube is placed
into the windpipe (trachea) through
the mouth or nose. In most
emergency situations, it is placed
through the mouth
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 2. Endotracheal Intubation

Endotracheal Intubation

INDICATION
❑ Endotracheal intubation is done to:
▪ Keep the airway open in order to give oxygen, medicine, or anesthesia.
▪ Support breathing in certain illnesses, such as pneumonia, emphysema,
heart failure, collapsed lung or severe trauma.
▪ Remove blockages from the airway.
▪ Allow the provider to get a better view of the upper airway.
▪ Protect the lungs in people who are unable to protect their airway and are at
risk for breathing in fluid (aspiration). This includes people with certain types
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of strokes, overdoses, or massive bleeding from the esophagus or stomach.

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 2. Endotracheal Intubation

Endotracheal Intubation

CONTRAINDICATION
❑ Patients whose respiratory status might improve with less invasive methods
should be tried on modalities such as non-invasive positive pressure ventilation or
other modes of oxygenation.
❑ Severe orofacial trauma can obstruct oropharyngeal intubation due to significant
bleeding or disruption of the facial and upper airway anatomy.
❑ Cervical spine manipulation during intubation can be harmful to patients with
spine injury and immobility
❑ No absolute contraindications to intubation, and the decision to place a definitive
airway should take into consideration each patient’s unique clinical condition.
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 2. Endotracheal Intubation

Endotracheal Intubation
CONSIDERATIONS FOR ENDOTRACHEAL INTUBATION

Lubricate the cuff

Select the right size
The length of tube required can be The cuff should be lubricated with a
Before to consider intubation, it
estimated by measuring it against water-soluble lubricant prior to use.
need to test the cuffs – even for
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the dog. The tip should lie at the To facilitate intubation & to reduces
brand new tubes.
point of the shoulder and the trauma to the tissues and allows a
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connector at the incisor arcade. better cuff seal.

 2. Endotracheal Intubation

Endotracheal Intubation
CONSIDERATIONS FOR ENDOTRACHEAL INTUBATION

Anesthetic Depth Secure the Tube

Use Visual techniques require the use of Securing the ET tube to minimise movement is
laryngoscopes to visualize and reached a suitable essential. This is achieved by placing a ‘tie’ around
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depth of anesthesia before you intubate, usually the body of the ET tube, being careful not to
when they have lost the gag reflex to ensure an obstruct the cuff inflation tube, and then securing it
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atraumatic intubation. to the patient.

 2. Endotracheal Intubation

Endotracheal Intubation
❑ Method and Techniques of Endotracheal Intubation
▪ Visualization of the vocal cords with a laryngoscope or video
laryngoscope
▪ Blind Nasotracheal Technique
- insertion is aided by use of breathe sounds.
▪ Fiberoptic Bronchoscope
- insertion of the tube into the glottis is aided by visualization of the
glottic opening and passage of the endotracheal tube over the
fiberscope
❑ Confirmation of correct tube placement in both techniques is by use of
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capnography and auscultation of the chest for breathe sounds.

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 3. Tracheobronchial Washing and Inoculation

Tracheobronchial Washing and Inoculation

❑ Tracheobronchial lavage or wash differs from a bronchoaveolar lavage

based on the anatomical structure being evaluated

▪ Tracheobronchial Lavage - assesses the upper airway exudate and

cellular population,

▪ Bronchoaveolar Lavage - concentrates on the lower airway

structures.

❑ Pediatric Laryngoscope - permits easy visualization of the larynx and

passage of the tubes.
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 3. Tracheobronchial Washing and Inoculation

Tracheobronchial Washing and Inoculation

TRACHEOBRONCHIAL WASHINGS

▪ Obtained using a 3 French feeding tube, 40.6 cm long.

▪ A 5-ml syringe containing sterile saline is then attached to the feeding

tube and used to infuse saline into the trachea.

▪ Saline is cleared from the tube using 2–3ml of air.

▪ Aspirating the 5-ml syringe will generally yield 1–2ml of foamy

tracheobronchial fluid
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 3. Tracheobronchial Washing and Inoculation

Tracheobronchial Washing and Inoculation

TRANSTRACHEAL ASPIRATION TECHNIQUE

▪ involves passing a needle between the tracheal rings or cricothyroid

membrane and into the tracheal lumen.

▪ A catheter is then directed through the needle and advanced to a

bronchus. Sterile saline (10–15ml for a macaque) is infused and
aspirated through the catheter

▪ It circumvents the potential problems of pharyngeal and laryngeal

contamination
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 4. Tracheal Pouch Formation

Tracheobronchial Washing and Inoculation

TRACHEOBRONCHIAL ASPIRATION TECHNIQUE

❑ EQUIPMENT
▪ required for intratracheal inoculation in small rodents includes two
pairs of small curved forceps
▪ a 100-μl microsyringe
▪ a 38 × 1mm diameter needle with a blunt tip bent at a 30° angle
10mm from the end
▪ laryngoscope made from a disposable polypropylene
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micropipette tip
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 4. Tracheal Pouch Formation

Tracheal Pouch Formation

❑ Involves resecting a segment of

cervical trachea, with blood and nerve
supply intact, and relocating this
closed segment subcutaneously for
ease in sampling.
❑ A more simplified technique for
creating a tracheal pouch has been
described in the ferret.
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 4. Tracheal Pouch Formation

Tracheal Pouch Formation

❑ Is used to study:
▪ nonadrenergic
▪ non-cholinergic inhibitory (NANCI)
▪ neurotransmission in guinea pigs
▪ evaluation of mucolytic drops in miniature
▪ vasoactive intestinal peptide as a neurotransmitter of nonadrenergic
inhibition of guinea pigs
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 5. Tracheostomy

Tracheostomy
❑ This technique does necessitate daily
management of the stoma and tube
❑ A permanent tracheostomy has been
developed for dogs that eliminates the need
for the tracheostomy tube
❑ The procedure consists of dissecting
portions of the cartilaginous rings free from
the underlying tracheal mucosa, cutting
through the mucosa, and suturing the
mucosa to the skin.
❑ This procedure results in a permanent,
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maintenance-free, mucocutaneous stoma

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 6. Bronchoscopy

Bronchoscopy

❑ FFB is a safe procedure, which has a high

diagnostic yield of respiratory disease.

❑ Its ability to assist in establishing the

diagnosis has been improved with the
presence of rapid technology revolutions.

❑ It can be used therapeutically in cases of

foreign body aspiration, ablation or debulking
of endobronchial masses, airway stenosis,
and lung lavage
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 6. Bronchoscopy

Bronchoscopy

❑ Plays a significant role in diagnosing various

situations such as:
▪ patients with hemoptysis
▪ chest infection
▪ parenchymal lung disease
▪ lung nodules or masses
▪ persistent lung infiltrates
▪ mediastinal lymphadenopathy
▪ suspected lung transplant rejection
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 6. Bronchoscopy

Bronchoscopy

❑ Flexible fiberoptic bronchoscopy

▪ an invasive procedure that has been used for
a long time for diagnostic and therapeutic
purposes.
▪ It contains a light source, fiber optics, and a
camera that allows direct visualization of the
upper and lower airways.
▪ Can be applied to most species including
rats, ferrets, and others.
▪ technique has been utilized as a diagnostic
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and therapeutic tool in primate medicine

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 7. Bronchoalveolar Lavage

Bronchoalveolar Lavage

❑ used as a therapeutic procedure in

humans and as a means of recovering
cells, surfactant, and inhaled
particulates from the lungs of animals
❑ Highlights of the technique described
by Singletary can be applied to rabbits,
dogs, swine, and other larger species.
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 7. Bronchoalveolar Lavage

Bronchoalveolar Lavage

1. The subject is deeply anesthetized and can be placed in a prone

position
2. A mouth speculum can be placed behind the incisor teeth and an
appropriately sized flexible endoscope or bronchoscope is advanced
beyond the epiglottis and into the trachea
3. A saline-filled syringe can be attached to the working channel of the
endoscope for instillation of lavage fluid.
4. The volume of saline to be used for each wash is determined by
individual pressure–volume measurements prior to lavage.
5. In dogs and swine, the volume may range from 50 to 300ml and larger
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animals (horses, sheep, etc.) may require larger volumes.

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 7. Bronchoalveolar Lavage

Bronchoalveolar Lavage

6. Lavage is accomplished by
hyperventilating the subject, instilling the
calculated volume of warmed normal
saline, and immediately withdrawing the
saline until a slight resistance is felt on the
plunger.
7. Another syringe and the tracheal aspiration
tube are used to clear residual fluid from
the catheter, and the animal is then
ventilated until effective spontaneous
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breathing is reestablished.
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 7. Bronchoalveolar Lavage

Bronchoalveolar Lavage
8. For more specific localization:
▪ the bronchoscope can be wedged into a
subsegmental bronchus to create a seal, which
then allows fluid to be instilled and aspirated from
the area of interest.
▪ The total volume of fluid should be reduced with
this procedure to prevent peripheral lung
damage.
▪ A total of three to four wash sequences will
provide adequate sampling, taking care to drain
as much fluid as possible from the lungs between
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washes
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 REFERENCES

REFERENCES
● https://citeseerx.ist.psu.edu
/viewdoc/download?doi=10.1.1.
487.5728&rep=rep1&type=pdf

● https://animalcare.ubc.ca/sit
es/default/files/documents/TE
CH%2009b%20-
%20Oral%20Dosing%20%28Gavage%
29%20in%20Adult%20Rats%20Apri
l%202021%20Final.pdf
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