Professional Documents
Culture Documents
EM Cabana, DVSM, MVSt Professor, Veterinary Pathology College of Veterinary Science & Medicine Central Luzon State University Science City of Muoz, Nueva Ecija Philippines
All Rights Reserved 2008 First Published 2001 by the CLSU Alumni Association, Inc.
No part of the contents of this book may be reproduced or transmitted in any form or by any means without written permission from the author or publisher
Foreword
While conducting the lectures for undergraduate students in general veterinary pathology, my students and I noted the dearth of literatures dealing with procedures in performing necropsies. It appeared that similar to most skills that were transferred from generation to generation of veterinarians, necropsy techniques considerably varies, yet the object of the exercise remains the same, i.e., to systematically examine the animal carcass. While doing my postgraduate degree, I was tempted to ask my former professors an appropriate text for me to master the skills. They replied that there seems to be no one documenting the procedure prompted me to write one. This book therefore is an attempt to document the procedures. While I do not purport to be the original source of the routine necropsy techniques herein described, I attempted to include techniques for cosmetic necropsies. It happened to me one day that I was awestruck by the owner holding her dead pet and asking me if there was an alternative procedure that will not mutilate her pet dog. I get hold of her dead pet and made a deal that a cosmetic procedure will be done. My academic supervisor, seeing the results then told me to better document that one too. It is hoped that this book will be of value to both students and practicing veterinarians alike. The initiation to veterinary pathology seems to start at the necropsy rounds. Have our predecessors did not study the changes in organs and tissues through skillful necropsies, then I surmise that our knowledge about animal diseases could have been very, very limited indeed. Let the initiation then begin!
EM Cabana June 2001
Table of Contents
Chapter 1 Necropsy: An Introduction General Principles The Necropsy Record Describing Lesions Collection and submission of Specimens Chapter 2 Tissue Changes Normal Anatomical Structures Physiological Changes Senile Changes Agonal Changes Post Mortem Changes Factors influencing Rate of PM Autolysis Terms used to describe PM Changes PM Changes on Organs & Tissues Chapter 3 Routine Necropsy Dissection Stage Display Stage Examination Stage Chapter 4 Cosmetic Necropsy Preliminaries Necropsy Procedures Chapter 5 Avian Necropsy Physical examination Euthanasia Opening the Body Cavities Examination Stage 1 1 5 10 11 12 12 14 16 16 17 17 18 19 22 22 25 26 35 35 36 39 40 40 41 43
Necropsy: An introduction
Chapter
ecropsy may be defined as the systematic examination of an animal carcass aimed to search for lesions. It is an important diagnostic tool and supports other procedures performed in the diagnosis of disease cases in a herd or flock. The necropsy procedure employed by veterinary students and practitioner alike varies from examiner to examiner and from specimen to specimen. The conduct of a particular routine depends largely on the individual preferences of the examiner, the availability of the materials and equipment for the examination, the condition or state of the carcass, the extent of the examination required, and the mode of examination requested by the client or owner. It is often observed that necropsy done by the uninitiated or the untrained is characterized not by the voluminous information gathered that have little importance to the diagnosis of a particular case in question. It is the absence of information vital to the formulation of a diagnosis. Ill-performed necropsy thus confuses the understanding of a disease process. A working routine is desirable so that adequate information is gathered that will aid in the formulation of a diagnosis. One factor to consider in the formulation of a diagnosis is the accuracy of the data gathered. A systematic approach in necropsy is required to so that appropriate and adequate information be gathered during the examination. This manual will describe the standard procedures for the necropsy examination of domestic animals adaptable to most laboratories and in field conditions. It is hoped that it will help practicing veterinarians and students alike to adapt a working routine in performing necropsy.
GENERAL PRINCIPLES
IMPORTANCE OF NECROPSY
The examination of dead or terminally ill animals offers opportunities in studying the processes involved in disease situations. Various medical imaging techniques have evolved in recent years providing adequate information on the morphologic alterations of organs and tissues following disease. However, necropsy still provides a first hand look on what really happened along the course of the disease. In poorly understood disease situations, tissue alterations resulting from or as a reaction to the disease process may or may not be detected during clinical examination. Results of clinical
examination alone may not be sufficient to define the process involved. Thus, gross and microscopic examination of diseased organs and tissues may lend valuable information in understanding the pathogenesis of a disease. Morphological changes when correctly recorded and interpreted provide a basis for correlating functional changes seen in a particular disease process. Not all disease processes will show dramatic morphologic alterations in organs and tissues. Some clues may be derived however from necropsy examination that will provide valuable information in the recognition of such functional disturbances. The challenge is on the examiner to recognize these clues. While most students and veterinary practitioners would regard necropsy as purely of academic interest, the purpose of necropsy does not end on the recognition of the lesions alone. Skillfully performed necropsy with all the information gathered, accurately recorded and interpreted will provide valuable assistance in the formulation of animal health and production strategies aimed to prevent and control animal diseases in a herd or flock. The primary aims of necropsy are to uncover the cause of death of an animal by defining possible etiology and pathogenesis to arrive at a diagnosis. Yet, the usefulness of necropsy resides on the application of the information gathered in the formulation of appropriate treatment, control and disease prevention measures.
hours or during weekends, cut open the abdomen and remove the viscera, particularly the segments of the gastrointestinal tract and examine the parts right away. The other parts of the cadaver may then be saved in a refrigerator and examined the next instance or day, although some degree of post mortem changes must be anticipated. If necropsy will be delayed for some reason or another (example the cadaver will be shipped to a distant laboratory and will take considerable time before it reaches its destination), freeze the whole cadaver solid. Pack it in dry ice before shipping, observing the pertinent rules and regulation in the transport of suspected biological hazards. Never pack the cadaver with ordinary ice if it is anticipated that the specimen may not reach the laboratory within one or two hours. Even then, the whole specimen should be packed in a plastic bag and put enough ice packs above and below the cadaver in a Styrofoam container. Send the cadaver the soonest possible time, with the necessary information as will described later.
pumped. All dead animal cadavers should be considered possible sources of contamination that should be disposed responsibly.
BASIC EQUIPMENT
The choice of equipment for necropsy depends on the size of the animal, the type of examination requested (whether routine or cosmetic necropsy), and the individual preferences of the examiner. For most purposes, two sharp knives, a pair of scissors and forceps, a metal probe, and an ordinary mechanic hacksaw will be sufficient. Preferences for the size of the knives depend on the size of the cadaver, ease of handling and safety. A steel rod or sharpening stone to keep the knives sharp is essential. The forceps could be a lockable scissors type or a lifting forceps with rattoothed or serrated tips that grasp tissues without slipping. A mechanic hacksaw will prove useful for cutting bones and other hard structures. A metal probe made of stainless steel, copper or bronze, or an ordinary galvanized iron wire gauge 12 and about 10-12 inches long is useful in probing connections and patency of openings. Other useful equipment includes a small axe, mallet and chisel (for cutting bones especially in large animal cadaver), and an ordinary pruning shears (instead of a costotome) for cutting the rib cage, mandibular symphysis, and pelvic bones. Other tools or instruments may be included, as they prove useful and saves manual labor during the examination. Weighing scales and measuring instruments like a millimeter rule and graduated cylinders or measuring cups are essential for accurately recording dimensions and volumes. Two specimen bottles, one half-filled with 10% neutral buffered formalin are required for containing samples of tissues and body fluids for laboratory examination. Sterile swabs and petri dishes for the collection of samples for microbiological examination (if required) should be made available. Other materials that may be needed include disposable syringes and needles, glass slides, and ordinary fishing twine or thread for tying up hollow organs.
PROTECTIVE CLOTHING
The wearing of protective clothing is not meant to prevent soiling and/or preserve the appearance of what is worn by the examiner underneath. It should protect the examiner from contamination with blood, tissues and body fluids from the cadaver that are potential carrier of infectious particles. The recommended protective clothing should provide comfort to the examiner while not compromising protection from possible contamination. The wearing of cotton coverall, rubber boots, gloves, and butcher's plastic vest is recommended and provide ample protection from contamination. These articles must be washed clean and disinfected after each use. The common laboratory gown may be used. However, the hanging flaps may easily soak with the cadaver's blood and body fluids with the examiner not noticing it,
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and thus exposing the examiner to contaminants. Although lacking any protective clothing, the laboratory gown will be sufficient. A pair of ordinary garden latex gloves of appropriate size is useful for necropsy. Compared with the surgeon's latex glove, the latter are less expensive, more durable and provide equal protection. Necropsy without wearing any gloves is an open invitation for contamination. The gloves should fit the hands and fingers of the examiner without interfering with manual dexterity and causing numbness. Loose, also very tight fitting gloves may cause undue interference in handling organs and tissues, not to mention the awkward look and the uncomfortable feeling it may render to the examiner.
8) Examiner's Information - includes the name, qualifications and signature of the one who did the examination and formulated the diagnosis.
RECORDING INFORMATION
During necropsy, it is best to record all findings as the examination progresses. Yet, thorough examination of the animal cadaver will consume a considerable amount of time. An assistant that will record the findings as the examination progresses may not be always present. Thus, it is recommended that the findings be recorded immediately after the examination and before the cadaver is disposed. If an organ or systems have been inadvertently missed during the examination, or the examiner f r o one reason of another forgot some details of the lesions, the specimen may still be available for reexamination. In the laboratory, the use of a tape recording unit is ideal if many specimens will be examined in a single day. This unit is practical for a busy laboratory with large necropsy specimen accession. Models of tape recording units that are particularly suitable include those with a microphone that can be pinned to the lapel and with either a foot switch or a wireless microphone, or that of a voice-activated recording unit. Although this will involve additional investment, the accuracy in recording information may far outweigh the cost should necropsy accession be large enough. It is recommended to take at least a photograph of the animal and/or lesions noted in cases where legal proceedings are anticipated. In these cases, carefully record the location and appearance of identifying markings of the animal such brands, cowlicks, and ear notches. These data are particularly sought in establishing the identity of the animal particularly in insurance claims.
many interpretations and that one examiner may or may not agree with the interpretation of another examiner. This is particularly true if another person other than the one who examined the specimen will synthesize the findings and formulate the diagnosis. The finished report should be descriptive enough allowing other to clearly visualize what were observed during the examination to enable them to make their own interpretation and possible diagnosis.
DESCRIBING LESIONS
Recognition of lesions requires a sound background in anatomy and pathology. Veterinary students should be aware of the appearance of normal structures and the species difference within and between species. However, as one gain experience in examining various animal species, recognition of the normal structures and what form an abnormality in the structure (or the lesion) becomes routine. Students of necropsy may find it useful to pay a visit to the local abattoir to observe the appearance of organs and tissues of domestic animal species. However, one must be aware that specimens submitted for necropsy most often shows post mortem changes that may mask or alter the appearance of otherwise normal structures. The recognition of these changes requires knowledge of pathology and considerable experience. It is not uncommon for students and the uninitiated to have a problem in describing lesions. Although veterinary pathology is rich in descriptive terms that describe the forms and appearance of any given lesion, the use of adjectives from ordinary conversational language is desirable than having no descriptions at all. It is recommended to write plainly in the report that no remarkable alterations were noted in the organ/tissue examined should that be the case, than leaving it not mentioned in the report. Also, include in the report the organs not examined for one reason or another. For most purposes, the description of any given lesion should include the following information: A. Solid organs: 1) Organs/tissue involved 2) Position, relations, and involvement of adjacent structures; 3) Size and shape; 4) Weight; 5) Color (shade, tint, hues); 6) Appearance; 7) Consistency; 8) Texture or intact and cut surface; 9) Odor; B. Hollow Organs: 1) Organs involved 2) Appearance;
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3) Texture of intact and opened surfaces; 4) Contents, which should be qualified as to: a) Nature b) Volume/Amount c) Consistency d) Transparency e) Colour; f) Odour; In describing lesions, it is best to keep the descriptions in as few but very descriptive words as possible. Refrain from using verbose descriptions that tend to distract attention and present no clear meaning. Weights and measure should be recorded using the metric system of notation. The approximation of the weights and measure is desirable than comparing what was observed to common or ordinary articles. For example, it is far better to approximate the size of a tumor as "about 4-5 centimeter in diameter" than recording it as "about the size of a 25-cent coin in diameter." Although the latter may be considered more acceptable than having no description at all, its use in the necropsy report should be avoided.
It is the responsibility of the examiner to notify relevant government offices in cases where the specimen is collected from suspected cases of highly contagious, zoonotic or exotic disease. Adequately label specimens taken from such cases to warn others about the potential of spreading the infection or pose danger to the biological system, or to the courier of the specimen should improper handling occur. The full descriptions of the methods employed in specimen collection and preservation are beyond the scope of this work, and the readers are referred to the appropriate text in clinical pathology. However, a short guide will be discussed in the proceeding section.
cadaver. Hollow organs such as segments of the gastrointestinal tract are best handled by obtaining a loop tied at both ends and placed in a sterile petri dish.
Contact the toxicology laboratory where the samples will be sent to ensure that the right specimen and amount are collected and the adequate precaution on handling and preservation is observed.
ends and save all its contents. Scrapping deeply the mucosa of the affected intestinal segment and examining the scrapings as a wet smear may do the diagnosis of coccidial infection in poultry.
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Tissue Changes
Chapter
hanges in the appearance of the structure of organs and tissues make the recognition of lesions difficult for students of necropsy. Some are considered incidental findings during necropsy. These changes may be categorized into five major groups as follows:
1) Normal anatomical variation 2) Physiologic changes 3) Senile changes 4) Agonal changes 5) Post mortem changes. The findings of one or more of these changes are often difficult to interpret and correlated with other tissue changes and/or lesions that may be present. It is therefore essential to recognize these changes to prevent confusion. The following lists are not meant to be comprehensive.
5) Os cordis in cattle - consist of bones embedded in the myocardium at the base of the aortic valve. 6) Unguiculate papillae - these are keratinized papillae at esophageal groove of ruminants. 7) Torus pyloricus - this is a tongue- shaped epithelium covered bulge of tissue at gastro-duodenal junction in pigs and cattle. 8) Duodenal papillae - nodules in the mucosa of the proximal duodenum and these structures are present in most species. These are the sites where the bile duct and pancreatic ducts open into the duodenum. 9) Pigmentation of mesenteric lymph nodes and the presence of haemo- lymph nodes in ruminants. 10) Melanosis of organs - gray or black pigmentation of meninges, brain, parenchyma, kidney, adrenals, uterus, lungs, esophagus, oral cavity, gastric and intestinal mucosa, and intima of great vessels of the heart in most species. 11) Os penis of dogs - consist of bone at the corpus cavernosum of the penis. 12) Absence of seminal vesicle and bulbo-urethral glands in dogs. 13) Discrete fine nodules in the pancreas of cats - these are pacinian corpuscles normally found in the pancreas of this species. 14) Presence of caseous material in the prepuce of boars. 15) Presence of whitish plaques in the esophageal mucosa of animals that have not eaten for some time. 16) Conversion of red marrow into a gelatinous mass - seen in malnourished animals and is more pronounced in ruminants. 17) Hyperplasia of lymphoid follicle with formation of prominent nodules in pharynx and larynx of young horses. 18) Dilated lymphatics in epicardial surfaces of heart in horses. 19) Cysts in kidneys of pigs - maybe solitary or multiple, and most are congenital abnormalities of no clinical significance.
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PHYSIOLOGICAL CHANGES
The normal functions of the animal body usually lead to some alterations in the gross appearance of organs and tissues. This may be affected by a host of factors related to the state of nutrition, endocrine influence, circulatory status, and stage of production. Tissue changes under this category include the following: 1) Post-prandial physiologic hyperemia of stomach mucosa - occur as reddening of the stomach mucosa in most species. This change is more pronounced in the horse and pig. The visible absence of inflammatory exudates differentiates it from gastritis. 2) Copious amount of mucus in stomach mucosa in the horse and pig - this is due to continued secretion of mucus in the stomach even after a few minutes after death in these species. 3) Distended gall bladder in most species that have not eaten recently - the gall bladder contains watery-pigmented bile, and this finding is more pronounced in dogs and ruminants. 4) Pallor of the liver in pregnant and lactating animals, and this is particularly seen in ruminants. 5) Uterine mucosal changes in appearance, texture and contents - this is part of the normal cyclical activity of the uterus following pregnancy and parturition. If the animal has recently given birth, pink- colored sludge like material with no offensive odor may be seen contained in the uterus. 6) Atrophy of the prostate in male animals after castration - the prostate show great deduction in size and with shriveled connective tissue capsule. 7) Vascular congestion and hyperemia of the gastrointestinal tract - seen in animals that have taken a meal shortly before death. 8) Yellow precipitate in renal papillae of piglets, usually associated with dehydration and/or insufficient urine flow.
SENILE CHANGES
Changes in the structure and appearance of organs and tissues occur as the animal matures. These changes might be confused as lesions during necropsy. Senile tissue changes seen in domestic animals include the following:
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1) Tension lipidosis in the liver - seen most commonly in cattle and horses. Focal pallor of the liver parenchyma immediately next to the mesenteric attachment characterizes this. 2) Nodules in the liver of old dogs - these nodules are of variable size, and may be deep in the parenchyma or sub capsular in location. They are composed of hyperplastic liver parenchyma. 3) Fatty cyst in old cats - these are brownish nodular masses found on the sub capsular surfaces or deep into the parenchyma of the liver. 4) Cystic hyperplasia of gall bladder mucosa in dogs - this takes the appearance of fleshy folds in the mucosa with fluid-filled cysts. 5) Nodular hyperplasia of pancreas and adrenal glands - these structures should be differentiated from neoplastic nodules. 6) Multifocal pleural fibrosis in cattle and sheep - these are observed more commonly on the diaphragmatic surfaces of caudal lobes of the lungs. 7) Focal grittiness in trachea, bronchi and lung parenchyma of old dogs and cats. 8) Anthracosis in dogs and cats - frequently seen in those animals living in the city. The lung parenchyma may show fine dark coloration. The draining lymph nodes may also show the same changes. 9) Cholesteatoma (Cholesterol granulomas) in choroid plexus in the lateral ventricle of the brain in horses - these are discrete nodular masses that may be tan, sometimes mineralized gritty nodules. These are accumulations of cholesterol and macrophages on the distal tips of the choroid plexus. 10) Medullary calcification in the kidneys of dogs and cats - occur as white gritty radial streaks in the kidney medulla. Sometimes, they appear as whitish streaks that border the cortex and the medulla. 11) Par ovarian cysts in old dogs - these may be solitary or multiple and situated on the surface of the ovary and in the adjacent mesosalpinx or mesometrium. These structures may contain colorless fluid. 12) Prostatic changes in dogs - there may be enlargement, or hyperplastic nodules may be seen. Laminated and calcified masses deep within the parenchyma may be noted.
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13) Nodular endocardiosis in dogs - these changes occur in the atrio-ventricular valves and appear as shiny nodules at the edges of the valves. They may be multiple, and their significance should be assessed in terms of their effects on the patency of the affected valve. 14) Siderotic nodules and extra splenic tissues in dogs - Siderotic plaques are hard and gritty pale to yellowish nodules seen more commonly at the edges of the spleen. Extra splenic tissues occur as nodules of varying sizes scattered at the omentum near the spleen, and may represent previous t aumatic injury to the r spleen. 15) Osteoarthritis in dogs - this is common in large breeds of dog, and may take the form of erosion of the articular cartilages. Other findings include thickening of the joint capsule with deformation of the articular surfaces. 16) Conversion of red marrow into fat in most species. 17) Telangiectasis of liver in old debilitated cattle. 18) Small, atrophic lymph nodes in old sheep 19) Subendocardial materialization in ventricle or atria of adult horse. 20) Haemomelasma ilei in horses - consist of elevated subserosal plaques of variable size, shape and color on the antero- mesenteric surface of distal small intestine and colon. 21) Fibrous tags attached to the liver capsule, particularly on diaphragmatic surface in horses and buffaloes.
AGONAL CHANGES
Agonal changes are those tissue changes that occur immediately before death or following cessation of vital functions. While dying, some changes occur in tissues and include the following: 1) Vascular congestion of most organs - seen particularly in the lungs and pancreas. 2) Pulmonary emphysema in ruminants and pulmonary edema in dogs - the emphysema is due to labored breathing during the agonal period. The pulmonary edema is due to agonal impairment of the venous return of blood.
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3) Hemorrhages in the heart in most species - occur as streaks of hemorrhages and are more pronounced on the endocardial surfaces. 4) Adrenal congestion and hemorrhages - seen in most species and more commonly observed in cattle and horses. 5) Food materials in the lungs and airways - this is more commonly observed in ruminants and should be differentiated from ante mortem aspiration of foreign materials. 6) Congestion of the meningeal vessels in the brain of most species - may be related to redistribution of blood to vital organs.
autolysis is progressive with time, and the longer the time elapsed the greater the degree of post mortem changes in organs and tissues occur. 3) Cause and Mode of Death - Generalized infection such as septicaemic bacterial diseases hasten the appearance of post mortem changes. 4) Condition of the Animal before Death - The condition of the animal before death, particularly its nutritional status influences the rate of post mortem autolysis. Obese animals tend to show an accelerated appearance of post mortem changes. This condition is probably related to the insulating effect of the fat layer that retards cooling of the body after death. 5) Tissue-related Factors - The degrees of the expression of post mortem changes vary from tissues to tissues. The presence of bacterial flora, enzyme secretions, and the availability of moisture and substrates influences the rate of post mortem autolysis. SPECIAL TERMS USED TO DESCRIBE SPECIFIC POST MORTEM CHANGES 1) Rigor mortis - refers to the contraction and stiffening of muscles after death. Most literatures consider the fall in the availability of adenosine triphosphate (ATP) as the possible cause of terminal muscle fiber contraction after death. Other plausible explanation includes the influx of calcium ions after cessation of the sodium pump. Classically, rigor mortis begins from 1-6 hours after death and passes off in 24-48 hours. However, several factors influence the onset of rigor mortis and these include the following: a) Nutritional status of the animal b) Environmental and body temperature of the cadaver c) Cause of death Well-fed animals have large glycogen reserves, and may show a delay in the onset of rigor, while cachectic animals may develop rigor quickly. Animal cadavers that are exposed to hot temperatures may develop and passes off rigor mortis quickly. Those that died of septicaemic diseases may not develop rigor mortis at all. Once a part of the animal body that has passed into rigor is moved, that part will pass off rigor mortis. This has some significance in human medico-legal cases. 2) Algor mortis - Gradual cooling of the animal body after death, and is associated with a fall in ATP. 3) Livor mortis - The settling of blood to the down side of the animal body. Gravitational force causes this to happen. This gravitational settling of blood and
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body fluids results to intense reddish coloration of the organs and tissues at the down side of the cadaver. 4) Hemoglobin imbibition - Pinkish to reddish coloration imparted to tissues due to the lysis of red blood cells. This is most evident on the surfaces of large arteries and in outer surfaces of visceral organs. 5) Bile imbibition - Golden yellow coloration imparted on tissues following seepage of bile. Discoloration is most evident on the surfaces of organs in contact with the gall bladder, and on duodenal mucosa. 6) Pseudomelanosis - Greenish gray to dark coloration of tissues. This is due the action of bacteria to hemoglobin forming hydrogen sulphide. Pseudomelanosis suggests a more advanced stage of post mortem degradation of tissues. 7) Chicken Fat Clot - An old term referring to the gelatinous mass formed by separation and coagulation of plasma proteins from the component of blood. This is usually seen inside the major blood vessels and the heart. They sometimes give a cast of the ramifications of the vessels. To prevent misconceptions, the use of this term should be avoided. 8) Currant Jelly Clot - An old term applied to coagulated blood. The term becomes obsolete and should be avoided.
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The Pancreas
Post mortem change occur rapidly in the pancreas. It may be discolored red from hemoglobin imbibition, lose its lobular pattern, and become soft and translucent. The portion that is facing the gall bladder may show bile imbibition. In advanced stages of post mortem degradation, the pancreas turns into a sac-like structure containing red-tinged fluid. It may even disappear leaving a flimsy membranous structure.
The Kidneys
The kidneys may be discolored red from hemoglobin imbibition, and later discolored black (pseudomelanosis). The demarcation between cortex and medulla becomes not apparent. As time progresses, the organ becomes soft and gas bubbles form at the peri-renal tissues.
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The Brain
The brain, being located farther from the gut show delayed post mortem changes. Vacuolation of the brain parenchyma due to seepage of cerebrospinal fluid may be observed. With advanced stages of post mortem degradation, the brain turns into a soft mushy mass. Later, the brain liquefies and will ooze when the calvarium and meninges are opened.
Muscles
Discoloration of the muscles, fascia and tendon sheath occurs after death because of seepage of myoglobin. Intense coloration of the muscles may be seen at the down side of the animal. Air pockets demarcating muscle groups and fascial places occur in advanced stages of degradation.
The Eye
The cornea of the eyes turns opaque due to absorption of the aqueous humor. The globe may either protrude or collapse. When opened, detached retina may be noted.
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Routine Necropsy
Chapter
he routine procedures for the necropsy examination of domestic animals will be described first. To illustrate the technique, the dog will be used as the model for the methods employed. Before necropsy, evaluate the clinical history of the animal to have an idea of what organs or organ system should receive particular attention. Although the examination of all organs and systems is recommended, prior knowledge of the clinical history serve as a guide in knowing what to look for and what specimens should be collected to confirm the diagnosis. Specimens submitted for necropsy should be examined following a set of routine. Two methods are traditionally used in the necropsy examination of domestic animals: Routine necropsy, and cosmetic necropsy. Cosmetic necropsies are most often sought by the owner of companion animals, particularly dogs and cat owners and shall be dealt with in another section. This section shall deal with the routine necropsy procedures. The procedures for the routine necropsy examination of dead carcass can be conveniently divided into three major stages: 1) Dissection stage - consist of preliminary incisions to expose the various organs and body cavities for the examination. 2) Display stage - the stage where all organs are exposed but remained on their original position and relations. 3) Examination stage - the part of necropsy where specific organs and organ systems are systematically examined one after another.
DISSECTION STAGE
Position the specimen with its left side down, with the feet facing the examiner (Figure 1). Carefully examine the animal's exterior. Note the body openings for the presence of secretions/excretions, prolapse, and color of mucus membranes. Examine the hair coat, and note for the presence of ectoparasites, areas of alopecia, thickening of the skin, crust formations, tumour masses, and possible wounds. Penetrating
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wounds should be probed and the extent or depth noted. Palpate the continuity of bony structures and look for evidences of fractures and abnormal masses. Work with the right side of the specimen. The first incision is a straight line from the chin towards the ventral midline of the neck. Make an incision beginning from the chin and expose the mandibles and masseter muscles. Skin the neck and expose the underlying structures. Continue skinning backward to the flank of the right forelimb. Grasp and lift the right forelimb upward and cut all muscles between the subscapular area and the rib cage to free the limb. While doing this, locate, slice and examine the prescapular and axillary lymph glands. Examine the size and color and texture of the glands. After cutting all attachments of the forelimb, reflect the limb to the dorsum of the specimen (Figure 2). Hold the right hindlimb and cut the skin and underlying muscles of the hind flank. Expose the rim of the hip joint. With the aid of the tip of the knife, cut the round ligament to free the head of the femur and consequently the hindlimb. While doing this, take note of the articular surfaces of the acetabulum and the femur. Note also the amount, quality and quantity of the synovial fluid, and the appearance of the joint structures.
Figure 1. Position of the animal for necropsy. Arrows indicate the course of primary incisions.
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Figure 2. The specimen after skinning and cutting the attachments of both limbs. Broken lines indicate the cuts that should be made to open the thoracic and peritoneal cavities.
Reflect the freed hindlimb to the dorsum of the specimen. Continue skinning the ventral midline of the specimen from the incision made at the region of the rear flank and backward to the hind flank are. Reflect the skin at the dorsum of the specimen. While skinning the specimen, take note of the quality of the carcass in terms of the amount and appearance of the flesh. Note for any discoloration, bruises and prior bleeding points. The next step is to open the buccal cavity. To do this, cut deeply the submandibular muscles and underlying structures close to the inner rims of the mandible at both sides. With the aid of an ordinary pruning shear or costotome, split the mandibles at its symphysis. Alternatively, a hacksaw may be used to do this. Grasp the tongue and pull it backward. Cut all muscular attachments to free the tongue. Severe the hyoid bones at the articulation of the great and the small cornu. Free the tongue by cutting all structures behind the tonsillar tissues. Examine the palate, pharyngeal mucosa, and tonsillar tissues. Drag the tongue backward and dissect the trachea and oesophagus cutting all attachments up to the thoracic inlet. Leave the freed tongue, oesophagus and trachea still attached at the thoracic inlet. The abdominal cavity is then opened. Palpate the free edges of the last rib and make a shallow incision sufficient to cut the abdominal muscles and peritoneum at this region while not cutting deeper structures. Lift the opening and continue cutting the abdominal wall from the dorsum and into the area of the xiphoid cartilage of the sternum. Continue cutting the abdominal wall at its dorsal and caudal boundaries down
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to the inguinal region. While this is done, note for the presence of ascitic fluid. Save as much fluid as possible for measurement of volume. Be careful not to cut the intestinal segments while doing this maneuver. After exposing the abdominal organs, leave the omentum that covers the intestinal loops for a while to avoid drying of the exposed segments. Position the knife at the angle formed by sternal part of the diaphragm and the xiphoid cartilage of the sternum. Cut the sternal part of the diaphragm and note the presence or absence of negative pressure within the thoracic cavity (Figure 3). The presence of a negative pressure is suggested by the backward displacement of the diaphragm. Continue cutting the costal part of the diaphragm close to the inner rims of the ribs. The right side of the rib cage is then severed from its attachment to the sternum. In young subjects, cutting the costo-chondral articulation with the aid of a knife can easily do this. Old subjects may require the use of ordinary pruning shears or costotome. Cut the costo-chondral articulation from the last articulation and towards the first rib. Be careful not to severe the tongue, trachea and oesophagus lying freely at this region. Detach the wall of the rib cage by cutting the neck of the ribs and associated intercostal muscles to expose the thoracic organs. The pelvic cavity is then opened. This is done by sawing the ilium close to the rim of the acetabulum at both sides (Figure 4). Then, saw the ischium from behind at both sides. Remove the sawed portion of the pelvic bone by cutting all underlying attachments to expose the pelvic cavity and its contents. At this point, the buccal, thoracic, abdominal and pelvic cavities and their contents are exposed for detailed examination.
DISPLAY STAGE
The display stage is that part necropsy examination when all the organs are exposed for close inspection. This stage is the best time to examine the whole carcass where all organs are exposed and remains untouched. Examine the exposed organs and note their relations, position, and external appearance. Lift carefully the organs for a much-detailed examination of the whole structure. Exercise extra care so as not to unnecessarily displace the organs at this stage. If frank or clotted blood is present in any of the body cavities, carefully look for possible bleeding points. Should this be noted in the abdominal cavity, take particular attention to the surfaces of the liver and look for small fissures and cracks on the surface. In most traumatic conditions (e.g., vehicular accidents involving small animals), fissures and cracks on the liver parenchyma may be subtle or not readily apparent. Unfortunately, handling may easily produce the same and thus information concerning this condition is easily lost or overlooked. Manipulation and unintentional cutting of blood vessels may leak blood into the body cavity and make the appreciation
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of internal bleeding difficult. However, antemortem bleeding into the body cavity usually has accentuated lymphatic vessels filled with blood. Evidence of gastrointestinal accidents (torsion, volvulus, intussusception, strangulation, rent and tears in the omentum and mesenteries) is best examined at the display stage. Excessive manipulation of organs could easily dislocate relationship of organs, making the examination and recognition of strangulation, volvulus and intususception difficult. Similarly, small holes in the omentum or mesentry caused by abdominal accidents most often are overlooked, easily lost, or inadvertently produced. Should specimens for laboratory examination be required, the display stage is the best time to collect the required samples. Tissue blocks intended for histopathological studies should be collected at this stage. This is recommended since excessive handling of organs and tissues during the examination stage will most often produce artefacts in tissue sections. After carefully observing the organs at this point, the detailed examination of any organ or organ system follows.
EXAMINATION STAGE
Examination of the Abdominal Organs
The abdominal organs, particularly the gastrointestinal tract takes priority in the examination of organs. Their content of enzymes and bacterial flora render these organs to undergo rapid post mortem autolysis. It is desirable to remove the entire alimentary tract from the rest of the carcass. This will minimise soiling of the carcass with spilling digesta from the opened alimentary tract segment. Remove the whole segment of the gastrointestinal tract. To do this, grasp the large intestine and cut its mesenteric attachment. Cut the portion of the large intestine as it enters the pelvic cavity. While holding the cut segment of the large intestine, severe the mesenteric attachment of the large and small intestine. This will free the whole segment of the gastrointestinal tract. Grasp the whole structure, including the stomach and spleen and expose the duodenal loop where the bile duct opens. Gently press the gall bladder and note if the bile would flow freely from the bile duct towards the duodenal loop. A small cut at the segment of the duodenum where the bile duct opens may be required to determine this condition. Then, grasp the body of the stomach and gently press the cardia and the oesophagus close to the oesophageal hiatus of the diaphragm. Free the stomach the segments of the intestines by cutting the oesophagus. Remove the spleen by cutting the omental attachment from the body of the stomach. Place the whole gastrointestinal segments on one side of the table. With the aid of a pair of scissors, the stomach is first opened by cutting from the cardia and down at the greater curvature towards the pylorus. Spread open the stomach and gently
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remove its contents. Examine the contents and qualify and the thickness of the walls. Note for the presence of ulcers, evidence of calcification, strictures, perforations, foreign bodies, and exudates. Free the entire length of the intestine by cutting close to the mesenteric attachment segment by segment. While freeing the duodenum, exercise care not to damage the pancreas which should be examined at this stage. Note for nodule formations, colour, and texture of the organ. Examine also the adjacent adipose tissues and look for evidence of necrosis of fat. Cut open the intestine as the segment is freed from its attachment. Some workers suggested that the whole segment of the gastrointestinal tract be opened. While this is ideal, the time involved in opening the whole segment may be staggering. For an average sized animal, this may reach to four meters and consequently require much time. Also, the subdivisions of the intestinal tract may not be appreciated if the whole segment is freed and lying as a straight tubular structure on the table. Moreover, accidental cutting of segments may leave the examiner losing track of the continuity of the structure. As an alternative, the intestines may be examined leaving their mesenteric attachment intact. Locate and open a representative segment of the duodenum, jejumum, ileum, caecum and colon. Examine the contents, the appearance of the mucosal surfaces, thickness of the walls, presence of ulcers and strictures, foreign bodies, and exudates. Qualify the color, odour, and consistency of the contents. Samples taken from the segments of the gastrointestinal tract intended for laboratory examination should be evaluated as to its relative merits. Considerations should include the time elapsed since death of the animal.
palpation. Look for changes in colour and consistency of individual lobes, collapsed or dilated lobes, and for the presence of abnormal tissue masses. Areas of consolidations should be characterised as to location and degree of involvement of lung parenchyma and its distribution. An apical distribution is most often an indication of bronchopneumonia.
Rotate the heart until the left auricle faces the examiner. M ake a horizontal cut at the left auricle extending to its extremities. Remove any clotted blood and note the patency of the bicuspid valve. Open the left ventricle by positioning the knife vertically into the chamber wall. Cut open the wall down to the apex. Examine closely the bicuspid valve and the papillary muscles. After doing this, position the knife into the opening of the aorta which is partly covered by the bicuspid valve. Cut open the valve and the aorta. When all the chambers have been Figure 5. Opening the heart while still opened, compare the thickness of the attached to thoracic viscera muscles and valves. For the heart valves, note its thickness, presence of strictures and/or local tissue masses, and torn moderator bands. Examine the endocardial surfaces and note for areas of haemorrhages, egenerations, fibrosis, anomalous openings among others. Then, examine the lumen of great vessels and note for evidence of regurgitation of blood. At this point of the examination, go over the rest of the carcass and cut open the entire length of the aorta down to the iliac bifurcation. Look for possible areas of thrombosis.
For male animals, slice and examine the prostate and cut open the urethra up to the penis. Look for possible small calculi that usually lodge at the urethral flexure. In female animals, free the vulva and the vagina from their attachments in the pelvic cavity. Cut them open and examine.Lymph nodes are examined whenever they are encountered during dissection. In cases of suspected malignancy, it is imperative to examine the regional lymph node and look for evidence of metastases.
Hold the head firmly with the aboral surface (occipital bones) slightly tilted and facing the examiner. Grasp the whole head with one hand pressing the thumb against the nasopharyngeal openingAnchor the index finger on the orbital rim and support the nasal region with the rest of the fingers. Press firmly the head on one corner of the table. Carefully saw the condyloid fossa just above the occipital condyles. Continue the
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cut slightly oblique and forward cutting the junction of the squamous temporal and occipital bones, and up to the supraorbital process of the frontal bones. Take care when cutting the lateral part of the temporal bone since at this part the bones are relatively thin and lie close to the brain. After completing cuts at the right side of the head, rotate the head to cut the bones at the other side. Grasp the head with the thumb anchored on the orbital rim and the index finger pressed against the nasopharyngeal opening. Support the head using the rest of the fingers grasping the nasal region. Make a similar cut until both the cuts at both side of the head meet at the median plane. Place the head on the table and hold it firmly with one hand. Make a diagonal cut continuing the cuts made at either side, and saw off about quarter thick of the head. This completes the cuts to open the calvarium.
Hold the head slightly tilted with the foramen magnum facing the examiner. To remove the sawed portion of the calvarium, pry it open by sticking the knife or a flat instrument onto the sawed portion where the occipital and temporal bones meet forming a ridge. This part of the skull is relatively thick. Twist the knife and force open the sawed bone. If the cuts made were deep enough, this should pry open the calvarium without much difficulty. Once the sawed portion is lifted and removed, cut the meninges covering the brain. The brain is now exposed for examination. Before removing the whole brain from the cavity, examine the surface of the brain and look for evidence of oedema such as cerebellar coning and flattening of the gyri and sulci. Remove the brain by inverting the head with the palm supporting the falling brain. With the aid of a pair of scissors, cut all the cranial nerves and attachments until the whole brain drops slowly to the palm of the hand holding the head. The whole brain is best fixed in ten-times the brain volume of 10% formalin solution overnight before examination. This will harden the brain and makes it amenable to slicing and manipulation. Detailed examination of the brain requires making serial sections no thicker than 0.5 cm. In looking for possible lesions in the brain, the idea of duplication in the appearance of both sides of the brain should be considered. Any alteration on the appearance of one side compared to that of the other side may be a suspect for possible lesion, provided that cuts were made perpendicularly. When slicing the brain, note any difference on the diameter of the ventricles and any abnormal tissue masses. Save the slices of the brain in fresh formalin solution. For routine histopathological evaluation of the brain, at least six sections are required composed of the following transverse sections: 1) Root of spinal cord 2) Mid cerebellum
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3) Cerebrum at either side (2 sections) including the hippocampus 4) Brain stem taken at the level of the pons 5) Cerebral peduncle
Examination of the Spinal Cord Clinical history of sudden and/or progressive paralysis warrants the examination of the spinal cord, nerve plexus, and associated ganglia. Remove the entire spinal column by sawing off all attachments of the ribs at either side and muscles. Cut the spine from the rest of the carcass at the articulation of the last lumbar and first sacral vertebra. Remove as much muscles as possible. Note for evidence of fracture and/or dislocations. The spinal cord should be removed by opening the spinal column. Two approaches may be used to do this. First, the spinal column may be split Figure 11. Diagram of the vertebra showing the sites (dorsal through the aid of a saw cutting arches and/or splitting of the vertebra) where the cuts should be made. about one third of the vertebra, and second, by removal of the dorsal arches. Splitting the vertebral column enables the examination of the intervertebral discs and vertebral canal. To do this, hold the spinal column with one hand pressing it on the far end of the table. With one or three vertebral bodies extended beyond the edge of the table, carefully saw the vertebral body longitudinally. Keep the blade of the saw positioned medially to the spinous process cutting the dorsal arches and the vertebral body at one side. Complete the cuts until the whole length of the spinal column is split open adjusting the Figure 12. Illustration showing the the sawed segment of the spinal column by splitting extended portion one or two the verterbra.
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vertebral bodies at a time as cuts are made. While sawing, be careful not to damage the enclosed spinal cord. Once the spinal column is split opened, mark the specific regions (cervical, thoracic, and lumbar regions) using pins or by loosely tying pieces of twine at each division. Lift the cord by grasping the spinal meninges and severe all attachments. The second alternate method employs the removal of the dorsal arches. This may be accomplished using a chisel cutting the arch of each vertebral body. For large animals however, it may be convenient to detach first the individual vertebral bodies and saw off the dorsal arches. The spinal cord, like the brain is best examined after fixation in 10% formalin solution. After fixation, remove the meningeal covering. Hold the cord vertically on one hand. Gently palpate the cord passing it between the thumb and the index finger. Note for pits and depressions, and difference in texture. Examine also the vertebral canal and note for narrowing of the lumen, evidence of fractures, and character of the intervertebral discs. Evidence of disc degeneration includes dryness and changes in the colour of the disc (from yellowish to greenish) with or without apparent protrusion into the spinal canal.
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34
Cosmetic Necropsy
Chapter
he bonds between owners and their pets have progressed to a point that pet animals are no longer considered as mere pets but members of the household. This is particularly true for dogs and cats that have been elevated to a certain position in the society where their principal role is not just to provide companionship, but as part of the family. Society has changed and currently recognizes the rights of these animals. Thus, in cases of unexpected death, the dead animal body is accorded equal respect. Accordingly, it is not uncommon to find pet owners objecting in part to have their pet be a subject for necropsy examination. While it may be argued that pet owners do not object to the aim of the necropsy per se, what might be intolerable are the procedures employed in routine necropsy. As evident by now, routine necropsy accompanies dismemberment of body parts eventually turning the dead carcass into one heap of dissected bones and flesh. This obviously would be revolting to some pet owners who would most likely want to have their pet a decent burial in one piece. Despite the development of techniques of post mortem examination in human medicine, the autopsy procedure itself being cosmetic in nature, description of the same technique in animals remains to be developed. While veterinary students are taught the procedures for routine necropsy, the incorporation of a suitable technique that will not hurt the sensibilities of pet owners received little attention. Thus, when confronted with such a case, students and practitioners alike hardly had the interest to pursue the necropsy examination. Cosmetic necropsy, as the term suggests employs methods aimed at limiting the procedures that will disfigure the carcass while maximizing the information gained during necropsy. As such, cosmetic necropsy requires carefully planned incisions and the methods for the restoration of the specimen into its original appearance after the examination. The proceeding sections attempt to define the basic procedure for cosmetic necropsy of domestic animals, with the dog as the animal model.
PRELIMINARIES
As in routine necropsy, the exterior of the animal should be carefully examined. Evaluate the clinical history and decide which organs of organ systems must be
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examined. Evaluate the conditions by which such organs will be made accessible considering the limitations of the cosmetic necropsy procedures. Should examination of the same will produce unsightly incisions and cuts, it is best to explore the alternatives and discuss these with the pet owner. If the intentions are discussed exhaustively with the pet owner describing the benefits of doing such an examination for the formulation of a diagnosis, consent may be sought with little difficulty. Under no circumstances that extra incisions and maneuvers be made without the owner's consent. Even then, the standard cuts that will be mentioned in the proceeding sections should be cleared with the owner to prevent possible misunderstanding and unfavorable outcomes.
NECROPSY PROCEDURES
The rule of thumb in cosmetic necropsy is to limit the number and the length of incisions to as few as possible. In planning the incisions, a few things must be considered: 1) The organs or systems that must be examined; 2) The site for incisions that will not disfigure the specimen, and with due considerations that the incisions would not be so obvious once sutured back to place; 3) The length of the incision considering the accessibility of the organs; and 4) The time that will be involved in the examination.
The best approach is to position the animal lying on its back (dorsal recumbency). The head of the carcass should face the left side of the examiner. Sandbags or other objects placed at either side of the thorax would help to hold the carcass in place. Using a sharp knife, make a straight longitudinal incision on the skin only from the xiphoid cartilage of the sternum and to the midabdomen. Dissect the skin towards the side taking care not to damage the hide,
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Figure 13. Illustration showing the lines of incision for cosmetic necropsy.
and expose the underlying tissues. Open the abdominal wall by cutting open the abdominal muscle at the exposed site. Note for the presence of fluid and try to assess the arrangements of organs. Retract both sides of the opened abdominal wall and reach for the diaphragm. From this small opening, puncture the diaphragm and note for the presence of a negative pressure. Open the thoracic cavity by cutting the diaphragm close to the inner walls of the rib cage. Grasp the thoracic organs and free them from their attachments. Drag the thoracic organs en masse and reach for the esophagus and trachea at the thoracic inlet. Cut free these structures and set them aside for examination later. From the same opening at the abdomen, grasp the visceral organs out of the cavity and severe all attachments. Remove first the liver, and then the stomach with the spleen. Drag the segments of the intestine and cut the mesenteries close to the intestinal wall segment by segment. Remove the intestine and set them aside for examination. Reach for the kidneys at both sides and severe them from their attachments. Palpate the urinary bladder, and if required, reach for it and severe its attachments. After these, examine the visceral organs as with routine necropsy. After removal of most organs, it is recommended to drain the carcass of blood and other body fluids. If needed, the body cavities may be washed with saline or tap water. Examine the body cavities and allow the carcass to drain. The examination of the brain should only be done if clinical history suggests neurological disturbance. To remove the brain from the bony calvarium, make a midline incision from Figure 14. Illustration showing the between the eyes and towards the procedures for opening the brain level of the first cervical vertebra (atlas). Dissect the skin and reflect all muscles covering the calvarium, cutting them towards the side. The vault of the cranium is best opened using a mallet and a sharp chisel. With the skin and muscles reflected at either side, cut the bone starting from the frontal region and around the exposed portion of the vault of the cranium. Take care not to damage the enclosed brain. After removing the vault of the cranium, cut the meninges
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to expose the brain. Severe the spinal cord and lift the brain carefully. Cut all the nerves at the vase. Tilt the head upward and backward to simplify removal of the brain from the cranial cavity.
POST EXAMINATION
One basic requirement of cosmetic necropsy is to restore the animal carcass back to its original state as much as possible. Sutures should close all incisions, and soiling of the hide by blood and body fluids should be cleaned. After examination of organs and before suturing back the incisions, the organs examined may be returned to the body cavities to give form to the carcass. Alternatively, the spaces may be stuffed with old newspapers that while giving forms to the hollow cavities, it also absorbs much of the remaining blood and body fluids. In closing the incisions made a continuous suture on the skin using heavy twine and surgical needle will be sufficient. In restoring the form of the head following removal of the brain, replace the chiseled vault of the cranium. Suture back the reflected skin at this site. Neat sutures and arranging the hair or fur will sufficiently hide many sutures. Other organs and body parts m be examined when deemed required, planning all ay incisions and suturing them back is required. While cosmetic necropsy provides some ways by which dead animal carcasses may be examined without hurting the sensibilities of the owner, some organs or body parts (example, the spinal cord) may be difficult to reach using this method. As was previously discussed, it is best to discuss with the owner all the necessary procedures that will be done in the carcass. Doing so assure confidence and by that maximizes the information gained during necropsy examination.
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Avian Necropsy
Chapter
ecropsy examination of the fowl present special problems. First, diseased or dead birds submitted for necropsy examination most often come from a large flock. Thus, there are no guarantees that the bird samples submitted represent the true picture of the health status of the flock. Isolated cases of diseases are therefore possible, and information gained following necropsy may thus be difficult to interpret for the rest of the flock. The basic question that must be answered is whether necropsy examination will shed light on the disease process affecting a significant proportion of the flock. Therefore, for necropsy examination to be meaningful, careful sampling of the flock is essential, and that samples submitted for necropsy should be those representing a wide spectrum of clinical signs observed on the flock. It is best to submit birds that are showing the earliest signs of the disease in question, at its worse state, and include also those dead birds that succumbed from the disease in question. Second, avian species tend to have a limited repertoire of signs following disease, such that critical history taking is essential. Moreover, it is generally recognised that most avian diseases are multi-factorial in aetiology. Information on the type of housing, ventilation, diet, husbandry practices, sources of stocks, number of birds affected, the stage of production, and the rapidity by which the disease spread of in terms of days elapsed following the appearance of the clinical signs should be obtained to properly evaluation the situation. Third, diagnosis of most avian diseases requires ancillary laboratory tests apart from necropsy. In most cases, there are no substantial gross changes that may be considered diagnostic, although clues may be derived from these gross changes on the best laboratory test that must be employed. Fourth, it must be remembered that species difference exists, and that the examiner should be aware of these differences between and within species of birds. Many veterinarians are caught off guard whenever species other than domestic chicken is presented for necropsy. The fact that birds has feathers lend themselves not amenable to physical examination, the feathers causing much difficulty for the examiner. Consider the number of birds submitted for necropsy for a given disease in question, this may require a considerable amount of time. Yet, physical examination is an essential prelude to necropsy. The proceeding discussions of the techniques for avian necropsy will use the domestic chicken as the model. The same technique may be easily adapted to the necropsy examination of most avian species.
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PHYSICAL EXAMINATION
Birds submitted alive for necropsy should first be examined prior to euthanasia. A systematic approach in the physical examination of the samples should be done. Ruffling of the feathers to view the skin from the head and proceeding caudally is best. For live birds, take particular attention to the body conformation and the behavior of the birds bearing in mind what is supposed to be normal for the species, breed and age. Allow the birds to rest for a while if they were transported. If blood samples may be required, it should be taken at this stage.
Figure 15. Cervical disarticulation in domestic fowl. Note the positioning of the thumb and the rest of the fingers. Inset: Direction of pull.
Take note of any swelling on the infraorbital sinuses, discharges on the nostrils, beak deformation, feathering, distention of the crop, the appearance of the foot pads and cloacal discharges. With the aid of a magnifying glass, ruffle the feathers under the wings and around the cloaca and examine for the presence of ectoparasites. The skin should be examined for the presence of lumps, bruises, wounds and other abnormalities. In interpreting cloacal discharges, it must be remembered that the cloaca or vent is the common opening for both the alimentary and uro-genital system. Soiling of the feathers around the vent need not indicate gastrointestinal pathology. The discharges should be qualified as to nature, colour, consistency and odor.
EUTHANASIA
Most avian species can be humanely killed by atlanto-occipital disarticulation. This technique however requires practice and care must be taken not to break the skin or totally obliterate the head from the neck makes a lot of mess. To do this, hold both legs and the wings with one hand. Grasp the head and neck with other hand. Secure the head between the thumb and the pointing finger. Position the thumb at the base of the neck and the pointing finger at the base of the lower beak. Hold the bird firmly and with a sudden jerk, twist the hand holding the head to disarticulate the neck.
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Large birds may react violently to the procedure such that adequate restraint is essential. Alternatively, chemical means of euthanasia may be done. For large birds, intraperitoneal or intravenous administration of barbiturates may be used. Small birds may be euthanased using inhalant agents such as chloroform or ether. Soak a piece of cotton in any of these agents and put it on a container with a lid. Place the bird on the container and close the lid.
Figure 16. Skinning the bird. The skin is manually dissected from the abdomen to the neck region. Note the line of incision to open the abdominal cavity
the breast and cut the set of ribs at both sides. While doing this, examine the paired anterior and posterior thoracic air sacs. Free the breast from attachments of the liver and heart. Using bone cutter or a large heavy-duty scissors cut the pectoral muscle and clavicle at both sides to free the breast. Care must be exercised not to unnecessarily severe the trachea, syrinx, brachial vessels, or cut open the crop. Remove the freed breast. Next, position the bird so that the feet are facing the left side of the examiner and the head of the bird facing the examiner's left side. Open the mouth and insert the point of the scissors at the commissure of the beak. Cut the commissure of the beak and Figure 17. Schematic diagram of the relative location of the air sacs in domestic fowl. Ce- cervical; Cl- clavicular; continue the cut towards the throat. Locate the opening of the Ax- axillary; At- anterior thoracic; Pt- posterior thoracic; oesophagus and cut through Ab- abdominal air sacs. towards the crop. Examine the contents of the crop and the appearance of the mucosa. Do the same for the trachea and cut through towards the syrinx. While doing this, examine the thymii, thyroids and parathyroids which are located at the lower neck region and at the thoracic inlet. Reposition the bird so that the feet are facing the examiner.
DISPLAY STAGE
Examine the organs in situ after completing the preliminary cuts to expose the visceral organs. As in mammalian necropsy, the arrangements of the organs, presence of fluids in body cavities, and evaluation of the appearance of most organs are done at the display stage. Also, samples for laboratory examination may be obtained at this stage. The relative size of the organs can be gauged at this stage, but the normal conformation for the species, breed and age should be known. Since most students of necropsy encounter difficulties identifying some structures in avian species, a short review follows. In birds, there is no structure homologous to the mammalian diaphragm. Thus, there is no distinct separation between thoracic and abdominal cavity except for two thin membranes, the pulmonary aponeurosis and oblique septum. The lungs are small
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and are attached to the ribs. The liver is usually large and bi-lobed. A gall bladder may be present in some species (e.g., chicken, duck and goose) or may be absent (e.g., pigeons). The gizzard is usually reflected to the right side of the specimen. An ovoid spleen is positioned near the proventriculus. Segments of the intestines, caecum, and colon can be observed in the intestinal mass reflected with the gizzard. The duodenum forms a loop that accommodates the pancreas. From the duodenum, there is no clear demarcation into jejunum and ileum. A paired caeca is present, and a short large intestine is not clearly delineated into colon and rectum. This enters into the formation of the cloaca. The kidneys are lobulate and closely attached to the sub-lumbar region. The ureters open into the cloaca. A urinary bladder is absent in avian species. Located close to the cloaca is the bursa of Fabricius. A left ovary and oviduct are always found at the left side, the right structures being undeveloped. There are no lymph nodes in birds. The long bones usually contain air spaces with minimal amount of marrow at the extremities.
EXAMINATION STAGE
The visceral organs in avian species may be examined in situ. If required, the liver, spleen, gizzard, proventriculus, and the whole intestinal tract may be detached to facilitate the examination of other organs. Grasp the proventriculus and cut the oesophagus. Lift the gizzard, the proventriculus, liver and the whole intestinal tract and severe their attachments. Cut the large intestine a short distance from the cloacal opening. Lay the severed organs at the table. Examine the liver and spleen. Open the proventriculus and gizzard longitudinally and note the contents and the mucosal surfaces. Peel the keratinised layer on the mucosal surface of the gizzard. Examine the pancreas and cut open the duodenal loop. Cut open the other parts of the intestine, preferably the whole segments. Note the colour and consistency of the contents, colour and appearance of the mucosa. Open the caecal junction and the caecum at either side. Examine the colour, consistency and appearance of the mucosa and its contents. Open the trachea down to the syrinx. Closely examine the mucosa of the trachea by sliding the blunt portion of the knife or scissors at the opposite side. Incise the lungs and note for any exudate and tissue masses. Remove the heart and make transverse section through it. Examine the kidneys, adrenals, ovaries/testes, and oviduct. Cut open the cloaca and examine its mucosa, contents and discharges. Locate the bursa of Fabricius and cut it open.
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Dissect out the muscles at the posterior portion of the legs to expose the sciatic nerves. Do the same for the other leg and compare the size of the nerve. Note for any haemorrhage on adjacent tissues. Examine also the brachial plexus by cutting the muscles between the scapula and the vertebral column. Note for any enlargements of the nerves. Examine the muscles, joints and tendons of the legs and wings. Remove the head from the rest of the carcass. Cut away the skin at the cranium. Using bone cutters or heavy duty scissors, cut the bones just behind the orbits and extend the cut caudally to the foramen magnum. Remove the cranial vault to expose the meninges and the brain. As in mammalian necropsy, fix the brain in 10% formalin solution prior to slicing. To remove the spinal cord, it is best to cut the dorsal arches of the spinal column, taking care not to damage the cord. Alternatively, severe the spinal column segment and fix them in 10% formalin prior to dissecting out the spinal cord.
Figure 18. Display stage in avian necropsy
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