Wirelab textbook pdf download

Gram Staining: A. Add crystal violet stain over the fixed smear and let it stand for 30 to 60 seconds. Wash off the stain with tap water. Add the iodine solution on the smear and let it stand for 30 to 60 seconds.

Pour off the iodine solution and rinse the slide with running water. Add a few drops of decolorizer so the solution trickles down the slide. Rinse it off with water after 5 seconds. Counter stain with Safranin solution for 40 to 60 seconds. Wash off the solution with water. Air dry the smear, and examine it under microscope, x10 and then x The method of collection should always be noted on the urinalysis requisition at the time of analysis.

Free catch voided sample Free catch samples are the easiest to collect and are often brought in by owners. They are an adequate sample for routine urinalysis, but are not appropriate for bacterial culturing. Owners can be instructed on how to collect samples at home in a manner that still maintains adequate quality control.

A midstream, first-morning collection will provide the best sample for analysis. Like free catch samples, this collection method provides an adequate sample for routine urinalysis, but is not appropriate for bacterial culturing. Catheterization Catheterization involves inserting a rubber catheter into the urethra and feeding it up to the bladder of the patient to collect a sample.

A sterile syringe is attached to the end of the catheter and urine is drawn out. This procedure varies slightly depending upon species and sex. In any case, sterility is important. Sterile catheters, sterile lubricant, and sterile gloves should be worn to prevent infection.

Samples collected by this method in a sterile manner may be suitable for bacterial culturing. Cystocentesis Cystocentesis is another method for collecting a sterile urine sample, and therefore is appropriate for bacterial culture. The procedure should only be performed on quiet, easily restrained animals. Sedation may be required to achieve this. Typically, the patient is restrained in dorsal recumbence and the bladder is palpated.

An ultrasound may also be used to visualize the bladder and help guide the needle. Sample preservation Fresh urine should be examined within 20 minutes of collection. If that is not possible, then a method of preservation should be considered.

Before choosing a preservative, consider the following: 1 The method of sample collection used 2 The analyses to be performed 3 The biochemical method that will be used 4 The time delay between urine collection and analysis. Before analyzing the sample, be sure to allow it to return to room temperature first.

This is important for the enzymatic reactions that occur during the chemical examination of urine. Generally, enzymatic reactions occur at a slower rate when the sample is at a lower temperature. This results in false readings. Lowering the temperature may also encourage crystal formation. As the sample returns to room temperature, these crystals will dissolve. The accuracy of specific gravity SG readings on refractometers can also be compromised when analyzing a sample that is below room temperature.

Freezing Preserving urine by freezing will inhibit bacterial growth, thus avoiding changes caused by the actions of microbes. Freezing is also adequate for most chemical tests and preserves the mineral content of urine.

Freezing, however, is an unsuitable method of preservation for sediment and hormonal analyses due to the resulting cellular and protein damage. Toluene Toluene is an organic solvent that can be used to preserve a urine sample.

Toluene floats on the surface of the sample, preventing the loss of acetone and contamination. Two milliliters of toluene will be adequate for mL of urine.

Due to the fact that toluene is a solvent, care should be taken in choosing an appropriate container in which to preserve the urine. Formalin prevents microbial growth, therefore preserving the cellular integrity of various structures within the sample. Formalin is a reducing agent, so it does interfere with the chemical analysis of substances that use this principle in their enzymatic reactions. One example is glucose determination using reagent strips.

The use of enough formalin can result in a false negative. Boric acid 0. Boric acid is an appropriate preservative for sediment analysis and, with the exception of pH, chemical analysis as well. Commercial preservation tablets Commercial preservation tablets are designed to inhibit microbes and control the pH of a urine sample, usually for a period of up to 5 days. They also provide good preservation of urine sediment, such as leukocytes, erythrocytes, casts, and crystals.

Physical examination of urine The physical examination of urine consists of assessing the volume, color, turbidity, odor, and SG of a urine sample. Volume An accurate representation of urine volume should be assessed over a period of 24 hours.

All voided urine is collected and measured to determine if the quantity produced is appropriate. Color Normal urine is described as being straw colored. The pigment of the urine often correlates with the concentration of the sample, but this should still be confirmed with a SG reading. To avoid discrepancies among technicians, use standard color terms when recording your findings. Common urine colors and associated causes.

Straw ……………Normal Colorless-pale ………. Dilute urine Deep yellow ………….. Concentrated urine; bilirubinuria Orange-red …………Hematuria; hemoglobinuria Red-brown ……….. Myoglobinuria Milky white…………..

An increase in turbidity in the urine can be caused by cellular elements, crystals, microorganisms, or mucous.

Items listed above that can cause an increase in turbidity will settle to the bottom of a standing sample. Again, it is important to use standard terminology to minimize discrepancies among technicians: Strong odor: The urine of intact male animals, especially cats and goats, is often quite strong and should be described as such.

Ammonia: Ammonia-scented scented urine can be associated with cystitis. Some species of bacteria metabolize urea in the urine to ammonia, producing a distinguishing odor. The presence of bacteria should be confirmed during the the sediment examination or with a urine culture. Putrid: Putrid- or foul- smelling urine is associated with the degradation of protein. This can occur during infections, and should be confirmed with further testing.

Fruity: Increased ketones present in tthe he urine give off a fruity odor. These are multi-parameter strips that contain a number of pads, each designed to test for a specific urine constituent.

There are individual tests available that can be used as well. Whichever method is chosen, it is important to review the package insert and make note of the testing time required and any possible false positives or false negatives that can occur.

Testing for chemical composition of the urine. Glucose The term used to describe glucose in the urine is glucosuria. The reagent strip method for detecting glucose is specific for glucose, meaning no other sugar will cause a positive result. False negatives or decreased values can be seen in urine samples preserved with formalin.

Clinitest tablets are also available to detect glucose in the urine. These tablets will detect any sugar or reducing substance in the urine, and therefore are not specific to glucose. Bilirubin is unstable in sunlight or artificial light and should therefore be tested immediately. Delaying testing will result in false negative values. Ictotest tablets are available to test for the presence of bilirubin in a urine sample.

Ictotest tablets are more sensitive than reagent strips and are able to detect smaller quantities of bilirubin. The presence of ketone bodies in the urine, or ketonuria, is often accompanied by a fruity odor. In addition to the reagent strips, individual tablet tests called Acetest are available. Acetest relies on the same testing principle as the reagent strips, but is more sensitive. Blood Reagent strips have the ability to detect intact RBC hematuria , free hemoglobin hemoglobinuria , and myoglobin myoglobinuria in a urine sample.

The test is quite sensitive, and positive samples may appear normal in color during the physical exam. Hematest tablets that were originally developed for detecting blood in feces can be used in urine samples; however, they are less sensitive than reagent strips.

To avoid false negatives, be sure to mix and resuspend the sample before testing. Another source of a false positive is contamination with oxidizing disinfectants. Results are read at 0. Care should be taken when using the reagent stick as not to contaminate the pH pad with the acidic buffer contained within the neighboring protein pad. Protein When protein is present in the urine, the term proteinuria is used. Reagent strips are not specific for any particular protein. They primarily detect albumin and are less sensitive for globulins and mucoproteins.

False negatives may occur due to the low concentration of protein contained within the sample. The concentration may fall below the sensitivity level of the reagent strip. For the detection of microalbuminuria, certain in-house enzyme-linked immunosorbent assay ELISA test kits are able to detect the trace amounts of albumin that would be missed by the reagent strip method.

Urobilinogen When testing for urobilinogen, the same light-sensitivity precautions used for bilirubin should be taken. Other false negatives can result from the use of formalin as preservative Nitrite Nitrites are products formed from nitrates by the actions of certain species of bacteria Because of this, the presence of nitrites can suggest a bacterial infection, but the absence of nitrites should not rule it out.

The reagent strips are specific for nitrites only, but there are several factors to consider when recording nitrite levels as negative. First, only certain species of bacteria will convert nitrate to nitrite. Second, it takes time for this to occur. A short urine retention time within the bladder may not be adequate for this conversion to take place.

The reagent strip method for detecting leukocytes relies on the detection of a leukocyte esterase enzyme. Unfortunately, this test is insensitive in dogs, and almost always gives a positive result for feline samples, regardless of the presence of leukocytes. The presence or absence of leukocytes should always be confirmed with a sediment examination. A sediment exam not only allows the technician to visualize structures such as cells, crystals, and casts, but it also serves to confirm suspected findings during the physical and chemical examinations previously conducted.

The SG of the sample itself can have an effect on the appearance of some structures, specifically cally leukocytes and erythrocytes. This is due to the process of osmosis. Before examining the sediment, we must first concentrate the sample. This is done by centrifuging the urine. After centrifugation, the urine will be separated into two portions: sediment the supernatant and the sediment. Sediment stain Sediment stain can be a useful tool in the examination of urine.

Many microscopic elements are translucent, and stain can provide some color or contrast to make visualization easier. If using stain, it is still a good practice to prepare one sample without stain and one with stain, then examine and compare both. Stain can bring with it some undesirable findings as well. Older stain can precipitate, causing clumps which could obscure or interfere with your readings.

Stain can also harbor bacteria and fungus, resulting in a possible false positive for bacteria in your microscopic examination. Comparing a stained sample with an unstained sample can help in eliminating these false positives.

To maintain good quality control, check your stain regularly. Scan the area for bacteria and stain precipitate. If these contaminants are found, discard the stain. Sediment stain is relatively inexpensive and should be routinely replaced on a monthly basis. Procedure for microscopic examination of urine 1 Pour 10mL of the urine sample into a labeled conical centrifuge tube.

If 10mL is not available, note the volume used on your requisition. Tip: To verify the presence of cellular components, make a blood smear with the sediment. Brucellosis Brucellosis is a disease caused by a group of bacteria from the genus Brucella. These bacteria can infect both humans and animals.

Brucellosis is often spread when people eat contaminated food, which can include raw meat and unpasteurized milk. The bacteria can also be spread through the air or contact with an open wound. Symptoms of Brucellosis The symptoms of brucellosis in humans are similar to having the flu. The symptoms may include: appetite loss, back pain, chills, lethargy, headaches, pain in the abdomen, pain in the joints, fever that comes and goes, weight loss.

Diagnosing Brucellosis The Veterinarian doctor may test The patient for brucellosis symptoms, Later on Testing may include: 1. Although the overall sensitivity reported for RBT varies widely, with the use of good quality antigens made by experienced or reference laboratories, the sensitivity of RBT can increased.

Procedure Whole blood, Plasma and Serum 1. Remove the test kit from the foil pouch, and place on it a flat and dry place. Add 4 drops of assay dilute in to the sample hole S vertically. Start the timer the sample will flow across the result window, if it does not appear after 1 minute add one more drop of assay diluents.

For the test result you will see the purple band in the result window of the kit. Do not interpret after 20 minutes. Interpretation of the test result 1. Color band will appear in the left section of the result window to show that the test is working properly, this is a band is the control line C 2.

The right section of the result window indicates the test result. Bovine Tuberculosis Bovine Tuberculosis, or TB, is an infectious bacterial disease caused by Mycobacterium bovis, which most commonly affects the lungs.

Laboratory diagnosis 1. Intradermal tuberculin test 2. Serology Test Anigen-Antiboday Test 3. Antigen-antibody reactions are visualized on the lines using anti-human antibody bound to substances. These technologies are very attractive candidates for the simple, accurate, and ideally inexpensive diagnosis of TB.

Procedure whole blood specimen 7. Dispense 3 drop of the whole blood diluents in to the test tube for the whole blood dilution. Add 1 drop of a whole blood sample with a disposable dropper and mix them for 1 minute. Dispense 3 drops of the developing buffer in to the developing hole. Do not interpret after 30 minutes. Serum and plasma specimen 1. Remove the test kit from the foil pouch and place it on a flat and dry surface.

For the result you will see the purple band in the result window of the kit. Interpret test result at 20 minutes, Do not interpret after 30 minutes. Interpretation of the test result 3. Color band will appear in the left section of the result window to show that the test is working properly, this is a band is the control line C 4.

Invalid: If the control line purple color band is Not visible within the result window after performing the test, the result considered Invalid. Spread the specimen evenly over the central area of the slide using a continuous rotational movement. Place slides on dryer with smeared surface upwards, and air dry for about 30 minutes.

Heat fix dried smear 4. Cover the smear will carbol fuchsin stain 5. Heat the smear until vapour just begins to rise. Do not overheat. Allow the heated stain to remain on the slide for 5 minutes. Wash off the stain with clean water. Wash well with clean water 9. Wash off stain with clean water Wipe the back of the slide clean, and place it in a draining rack for smear to air dry. Examine the smear microscopically, using the x oil immersion objective and scan the smear systematically.

The disease affects cattle, swine, sheep, goats and other cloven-hoofed ruminants. Intensively reared animals are more susceptible to the disease than traditional breeds.

The disease is rarely fatal in adult animals, but there is often high mortality in young animals due to myocarditis or, when the dam is infected by the disease, lack of milk. FMD is characterised by fever and blister-like sores on the tongue and lips, in the mouth, on the teats and between the hooves. The disease causes severe production losses, and while the majority of affected animals recover, the disease often leaves them weakened and debilitated.

The organism which causes FMD is an aphthovirus of the family Picornaviridae. Each strain requires a specific vaccine to provide immunity to a vaccinated animal. However, FMD cannot be differentiated clinically from other vesicular diseases, such as swine vesicular disease, vesicular stomatitis and vesicular exanthema.

Laboratory Diagnosis 1. Virus Isolation 2. Procedure whole blood specimen 1. Add 3 drops of the developing buffer in to the developing hole 4. Interpretation of the test result 5. Color band will appear in the left section of the result window to show that the test is working properly, this is a band is the control line C 6.

Trypanosomes found in mammals including humans are blood and sometimes tissue parasites of the order Kinetoplastida, family of the Trypanosomatidae, genus Trypanosoma, principally transmitted by biting insects, in which most of them undergo a biological cycle. Animal Trypanosomoses are nowadays a permanent constraint for livestock in Africa, Asia, and Latin America, but their geographical distribution is still evolving. The main African pathogenic trypanosomes belong to three subgenera of the salivarian section, namely, Nannomonas Trypanosoma congolense , Duttonella Trypanosoma vivax , and Trypanozoon Trypanosoma brucei group.

These parasites are mostly transmitted cyclically by the tsetse fly in which the procyclic forms undergo a cycle of transformations and multiplications leading to infective metacyclic forms, which may be inoculated by the tsetse flies with its saliva into a new host. However, in some instances, in addition to the movements of their hosts, the geographical distribution of trypanosomiasis does not fit that of the tsetse fly, due to several other ways of transmission.

Amongst them, while direct vertical, oral, sexual, and iatrogenic transmission may have an occasional impact, the most important alternative way is mechanical transmission by biting insects.

This way of transmission does not involve a specific biological relation between parasite and vector. These tests directly measure the interaction between antigen and antibody.

Antibody against trypanosomal antigen is used to coat ELISA plates and any antigen present in test sera binds. Although numerous nematodes infect humans, six spend the majority of their lifecycle in the bowel lumen and are classified as intestinal nematodes: Ascaris lumbricoides; Trichuris trichiura whipworm ; Ancylostoma duodenale and Necator americanus the two human hookworms ;Enterobius vermicularis pinworm ; and Strongyloides stercoralis.

Ascaris, Trichuris, and hookworms are commonly grouped together as the soil-transmitted helminths STHs due to a shared aspect of their life cycles. Nematode eggs should be examined using flotation technique.

Procedure 1. Collect fecal with the green part of the fecalyzer 2. Set in the white part and fill half way with the floatation solution 3. Mix thoroughly 4. Fill up the fecalyzer until a meniscus forms 5.

Place a coverslip on top and set the timer for mins any longer than that and crystals will start to form 6. Set coverslip on microscope slide, read under 4xx for roundworms and tapeworms and 10x- 40x for protozoa. Cestode tapeworms Tapeworm infection is caused by ingesting food or water contaminated with tapeworm eggs or larvae. If you ingest certain tapeworm eggs, they can migrate outside your intestines and form larval cysts in body tissues and organs invasive infection.

If you ingest tapeworm larvae, however, they develop into adult tapeworms in your intestines intestinal infection. An adult tapeworm consists of a head, neck and chain of segments called proglottids. When you have an intestinal tapeworm infection, the tapeworm head adheres to the intestinal wall, and the proglottids grow and produce eggs. Adult tapeworms can live for up to 30 years in a host.

Intestinal tapeworm infections are usually mild, with only one or two adult tapeworms. But invasive larval infections can cause serious complications The intestinal cestodes e.

The intestinal forms of these helminths can be asymptomatic or can mimic some symptoms of intestinal nematode infection e. Passage of tapeworm proglottids in the stool can occur, although these appear macroscopically different from nematodes. Ascaris is the only intestinal nematode large enough to potentially be confused with intestinal cestodes, but Ascaris is non-segmented and tapeworms are segmented. Signs and symptoms of intestinal infection include: Nausea, Weakness, Loss of appetite, Abdominal pain, Diarrhea, Dizziness, Salt craving, Weight loss and inadequate absorption of nutrients from food.

Tape-worm Laboratory diagnosis Tapeworm infection in animals is not particularly difficult to diagnose. This procedure involves processing a fresh stool sample and examining the end-product under a microscope for tapeworm eggs, which are quite large and usually are readily distinguishable from the eggs of other intestinal parasites. Trematodes flukes Trematodes flukes have small flat leaf-like bodies with oral and ventral suckers and a blind sac-like gut.

They do not have a body cavity acoelomate and are dorsoventrally flattened with bilateral symmetry. They exhibit elaborate gliding or creeping motion over substrates using compact 3-D arrays of muscles. Most species are hermaphroditic individuals with male and female reproductive systems although some blood flukes form separate male and female adults. Signs and symptoms of intestinal infection include: Abdominal pain, Nausea, diarrhea, and vomiting, Weight loss and Fever Laboratory diagnosis The sedimentation technique is a qualitative method for detecting trematode eggs in faeces.

Weigh out estimate 2 or 5 grams of feces. Mix with 10ml of sugar solution. Place tube into the centrifuge. Fill tube with sugar solution to a slight positive meniscus and cover with a coverslip. There should be a small bubble under the coverslip if correct amount of flotation solution was added.

Centrifuge at rpm for 5 minutes. Make sure the centrifuge is balanced. Let stand for 10 minutes. Remove coverslip from tube and place on slide labeled with the animal name or number. Examine entire coverslip at 10X.

Use 40X to identify parasites or eggs. Liver-fluke Mastitis The Etiology micro-organisms of mastitis are Streptococcus agalactiae, Streptococcus uberis, and Micrococcus pyogenes. These organisms usually cause a chronic mastitis and a loss in milk yield with or without the appearance of clinical symptoms.

The reason is the electronic devices divert your attention and also cause strains while reading eBooks. Electrical wiring is an electrical installation of cabling and associated devices such as switches, distribution boards, sockets, and light fittings in a structure.

Wiring is subject to safety standards for design and installation. Allowable wire and cable types and sizes are specified according to the circuit operating voltage and electric current capability, with further restrictions on the environmental conditions, such as ambient temperature range, moisture levels, and exposure to sunlight and chemicals.

Wiring safety codes vary by locality, country or region. The International Electrotechnical Commission IEC is attempting to harmonise wiring standards amongst member countries, but significant variations in design and installation requirements still exist.

LearnEngineering team try to Helping the students and others who cannot afford buying books is our aim. For any quarries, Disclaimer are requested to kindly contact us , We assured you we will do our best. Thank you. Please Note : This list is not the final book list. Electric Wiring: Domestic By J. Coker and W. Mullin and Phil Simmons. Del Vecchio Free Download.