Pennsylvania Animal Diagnostic Laboratory System
PADLS User Guide - Contents Tests Toxicology
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9.8 Toxicology

Toxicology is performed only at NBC and PVL. Enclose a detailed history, including clinical signs and lesions, tentative diagnosis, a list of drugs administered, and the response to previous treatments. A complete history may enable us to decide which additional tests are warranted.

In situations involving small animals, reptiles, or birds, where the amount of sample indicated below may not be available, please submit as large a sample as possible. Each individual specimen must be in a separate, clean, clearly labeled container. Containers should be freezable, such as plastic bags. For liquids, use clean glass/plastic containers. Purity of sample is of extreme importance in toxicology.

Do not use any preservatives with the exception that EDTA or heparin should be used as anticoagulants for whole blood samples. Serum should be separated from the clot. Containers should be as full as possible, but allow for expansion if samples are to be frozen. If "source materials" are obtained, mail them in a separate package. Do not mail them in the same package as the tissue samples. Information regarding their availability, means of exposure, and an estimate of amount ingested should be included. Enclose the label or a list of all chemicals and concentrations on the label.

If a topical or injected toxicant is suspected, submit a large area of tissue from the suspected absorption site. Many poisons produce microscopic changes in tissues. Therefore, always submit sections of tissues in 10% formalin for histopathologic examination in addition to the fresh or frozen tissues required for chemical analysis.

If other analyses are desired or if you have questions, please call the laboratory. At present, samples are kept 6 months after analysis unless we are specifically advised to the contrary.

Test/ Agent	Site	Procedure	Species	Tissue/ Specimen	Container	Shipping	Turn- Around	Remarks
Aflatoxins see: mycotoxins	PVL	HPLC ELISA
Ammonia	PVL	Microdiffusion	Any	5 ml whole blood	Heparin vacutainer	Frozen in insulated container with gel packs	2 days	Since ammonia forms rapidly after withdrawal of blood, it is necessary to determine blood ammonia soon after withdrawal or freeze the blood immediately.
Anions: chloride, fluoride, nitrate, nitrite, phosphate, sulfate	NBC	Ion chromatography	Any	100 ml water 2-5 ml serum 1-2 ml ocular fluid	For water: use bottles provided by an extension agent, public health official, or an empty distilled water bottle.	Chilled/frozen in insulated container with gel packs	2 days	Nitrate and nitrite are the only anions performed at PVL.
Anticoagulant screen: brodifacoum, bromadiolone, chlorophacinone, coumafuryl, difenacoum, difethialone, diphacinone, pindone, valone, warfarin	NBC PVL	HPLC	Any	20 g liver, bait 2 ml whole blood	Whirl-pak or zip-lock plastic bag EDTA or heparin Vacutainer	Chilled/frozen in insulated container with gel packs Chilled (not frozen) in insulated container with gel packs	5 days	Stomach contents may not be useful due to length of time for clinical signs to develop after a single exposure. Literature reports indicate blood may contain higher concentrations than serum, although both have been used successfully.
Avicides Starlicide^® Avitrol^® 4- aminopyridine	NBC	GC/MS	Any	Crop contents; suspect bait	Whirl-pak or zip-lock plastic bag	Chilled/frozen in insulated container with gel packs	5 days
Bile Acids	NBC	Kit	Any	0.5 ml serum	Red top tube	Chilled/frozen in insulated container with gel packs	1 day	Collect blood after overnight fasting. Centrifuge samples promptly after clot has formed. Lipemic or icteric samples may give misleading results
Bromide (single anion)	NBC	Ion Chromatography	Any	1 ml serum	Take sample in red top vacutainer tube, remove serum from clot, and ship in clean Vacutainer tube or plastic vial	Chilled/frozen in insulated container with gel packs	2 days	Therapeutic monitoring for treatment with NaBr or KBr
Calcium	NBC PVL		Any	Any	Whirl-pak or zip-lock plastic bag	Chilled/frozen in insulated container with gel packs	2 days	This is a useful adjunctive test to confirm ethylene glycol toxicosis
Carbamate insecticides	NBC PVL	HPLC/MS	Any	Serum, liver, baits, stomach contents	See organic chemical screen	See organic chemical screen	5 days	Screens for carbamate pesticides
Cholinesterase	NBC PVL	Spectrophotometry	Any	1 ml whole blood	EDTA tube	Chilled	2-5 days	Cholinesterase values may vary among different brain regions; most laboratories use an entire half in order to assure consistency of results. Cholinesterase activity is often used to assess significance of exposure to organophosphate, and less reliably, carbamate pesticides.
				1 eye (cattle or sheep only)	Whirl-pak or zip-lock plastic bag	Chilled/frozen in insulated container with gel packs
				½ brain (right or left hemisphere) For small rodent or bird, submit entire brain	Same	Same
Clenbuterol	NBC	ELISA LC/MS	Any	Eyeball	Whirl-pak or zip-lock plastic bag	Chilled/frozen in insulated container with gel packs	7 days
				10 ml urine	Plastic container
				10 gm hair	Plastic bag
Convulsant screen: Bromethalin, Strychnine	PVL	TLC		Stomach contents Bait	Whirl-pak or zip-lock plastic bags	Chilled/frozen in container with gel packs.	5 days
Copper	NBC PVL		Any	Liver biopsy 100 mg wet weight fresh tissue	Whirl-pak or zip-lock plastic bags	Chilled/frozen in insulated container with gel packs	3 days	Useful for nutritional assessment of copper status. Copper levels in the blood have variable diagnostic reliability
				2 ml serum	Red top tube
				Plasma	Purple or green top tube
Corticosteroid Screen	NBC	LC/MS ELISA	Any	2 ml serum	Red top tube	Chilled/frozen in insulated container with gel packs	5 days
				10 ml urine	Plastic container
				5 gm liver	Whirl-pak or zip-lock plastic bags
Cyanide	NBC PVL	Test strip	Any	1-5 ml whole blood	EDTA tube	Chilled	2 days	Cyanide is volatilized by stomach acid, so stomach contents must be frozen as quickly as possible and shipped in an air-tight container.
				10 g stomach/rumen/crop contents	Air-tight container	Frozen in insulated container with gel packs
				200 gm forage or feed	Plastic bag	Same
Drug screen, prepurchase	NBC	ELISA GC/MS LC/MS	Equine	5 ml serum	Serum or red top tube	Chilled in insulated container with gel packs	5 days for screen; if positive on screen, confirmation will take 5 more days	Corticosteroid, benzodiazepin, promazine, tranquilizer, non-steroidal anti-inflammatories, procaine, lidocaine
Drug screen, prepurchase	NBC	ELISA GC/MS LC/MS	Equine	10-20 l urine	Plastic vial	Chilled in insulated container with gel packs
Ethylene glycol screen	NBC	GC	Any	10 gm kidney	Plastic vial Whirl-pak or zip-lock plastic bag	Chilled/frozen in insulated container with gel packs	3 days	Kidney calcium concentration is a useful confirmation in suspected ethylene glycol toxicosis Samples that test positive in our screening method will be forwarded to an outside laboratory for confirmation of the results
Feed microscopy	Referral		Any	1 lb feed	Plastic bag	Dry	Minimum of 2 weeks	Representative sampling is extremely important for all feed analyses
Fumonisins (see mycotoxin)	PVL	ELISA HPLC
Insecticide screen/Chlorinated Hydrocarbons	NBC	GC/ECD	Any	5 gm liver, bait, fat or brain	Whirl-pak or zip-lock plastic bag	Chilled/frozen in insulated container with gel packs	5-7 days
Ionophore screen: monensin, narasin, salinomycin, Lasalocid, laidlomycin, maduramicin	PVL	TLC	Any	400 g feed or GI contents	Plastic bag	Dry Chilled or frozen	5 days
	NBC	LC/MS	Any	400 g feed or GI contents	Plastic bag	Dry Chilled or frozen	5 days
Lead (blood)	PVL NBC	AA ICP/MS	Any	1 ml whole blood	EDTA tube preferred, heparin 2^nd best	Chilled (not frozen) in insulated container with gel packs	2 days
Lead [see Minerals]	PVL NBC	AA ICP/MS		Tissues	See mineral panel
Macrolide Endectocides: Ivermectin, moxidectin, doramectin, eprinomectin, selamectin	NBC	LC/MS	Any	Brain, liver, fat - 5 g (minimum)	Whirl-pak or zip-lock plastic bags	Chilled/frozen in container with gel packs	5 days	Call laboratory if specimen is not one of those listed.
Melamine and cyanuric acid	NBC	GC/MS		Feed	20 gm	Whirl-pak or zip-lock plastic bag	5-7 days
Mercury	PVL NBC	AA	Any	1 ml whole blood	EDTA tube preferred, heparin 2^nd best	Chilled (not frozen) in insulated container with gel packs	7 days
Mercury	PVL NBC	AA	Any	5 gm each liver and kidney 5 gm suspect material	Tissues in whirl-pak or plastic bag	Chilled (not frozen) in insulated container with gel packs	7 days
Minerals (heavy metal screen): arsenic, cadmium, lead, selenium, thallium	PVL NBC	AA ICP/MS	Any	5 g ea. liver, kidney 5 g stomach/ rumen/ crop contents 5 g suspect material 100 ml water 100 g feed 2-5 ml whole blood	All solid environmental samples: whirl-pak or zip-lock plastic bags (no metal contact) Water samples: consult laboratory Tissue samples: whirl-pak or zip-lock bag Whole blood: EDTA vacutainer preferred	Water, tissues, serum/ plasma/ urine, chilled/ frozen in insulated container with gel packs Whole blood chilled (not frozen) in insulated container with gel packs Other samples in sturdy shipping container.	3 days	PVL does only lead, arsenic and selenium
Mineral panel - complete: arsenic, cadmium, calcium, cobalt, copper, iron, lead, magnesium, manganese, molybdenum, phosphorus (total), potassium, selenium, sodium, thallium, zinc	NBC	ICP/MS	Any	100 g feed 5 g suspect material 100 ml water 2-10 ml bile 5 g stomach/ rumen/ crop contents 5 g ea. liver, kidney 2-5 ml urine 2 ml serum and 2 ml whole blood	All solid environmental samples: whirl-pak or zip-lock plastic bags (no metal contact). Water samples: consult laboratory Tissue samples: whirl-pak or zip-lock bag Serum/ plasma/ urine: royal blue top vacutainer tubes, plastic container	Water, tissues, serum, plasma, urine chilled in insulated container with gel packs.	3 days	The postmortem specimens of choice are liver and kidney. Liver is the sample of choice for evaluation of nutritionally important elements (e.g., Ca, Cu, Zn, Fe, Mg, Mn, Mo), whereas kidney is best for evaluation of toxic elements (e.g., As, Pb, Hg, Cr, Tl). It is suggested that both samples be analyzed for optimal evaluation of sudden death or unthriftiness where clinical and/or diagnostic findings are inconclusive. For live animals both serum and whole blood should be submitted Avoid contact with rubber if zinc is of interest Herd-based testing is available PVL does only As, Ca, Cu, Fe, Mg, K, Se, Na and Zn
Mineral panel-Nutritional: (Tissue or biopsy nutritional metal screen): calcium, cobalt, copper, iron, magnesium, manganese, molybdenum, selenium, zinc	PVL NBC	AA ICP/MS	Any	5 to 30 gm liver	Whirl-pak or zip-lock bags	Chilled/frozen in insulated container with gel packs	3 days	PVL does only calcium, copper, iron, magnesium, selenium and zinc
Mineral panel (biological fluid metal screen): calcium, copper, soluble iron, magnesium, total phosphorus, sodium, potassium, zinc	PVL NBC	AA ICP/MS	Any	3 ml serum (NBC) 5 ml serum (PVL) 1-2 ml ocular fluid	Take sample in royal blue top vacutainer tube, remove from clot, ship in clean royal blue top tube or plastic vial	Chilled/frozen in insulated container with gel packs	3 days	This test provides total phosphorus (Pi) and nonprecipitable iron, rather than the total phosphorus and total iron reported in the serum mineral analysis. The test thus eliminates the need for a separate Pi analysis and removes the interference of hemolyzed cells on serum Fe. Herd-based testing is available.
Monensin (see Ionophore screen)
Mycotoxins: aflatoxins, DON, DAS, fumonisins, ochratoxin, T-2 toxin, zearalenone	PVL	ELISA TLC/HPLC	Any	1 kg feed, silage	If dry, submit in plastic bag or carton; if high moisture, ship frozen	Insulated container with gel packs if high moisture and frozen	5 days	Representative sampling is extremely important for all feed analyses. Sample multiple locations in the feed.
NEFA's (nonesterified fatty acids)	NBC	Test Kit	Bovine	1 ml plasma	Green top or purple top plasma tube	Frozen in insulated container with freeze packs	3 days	Important: Contact the laboratory prior to drawing samples. Plasma-spin down and separate before freezing. Ship directly to the Toxicology Section by overnight express. Samples must remain frozen.
Nitrates/Nitrites (feed)	PVL	Spectrophotometry	Any	Feed, forage	If dry, submit in plastic bag or carton; if high moisture, ship frozen	Insulated container with gel packs if high moisture and frozen	3 days
Nitrates/Nitrites (feed)	NBC	Ion Chromatography	Any	Feed, forage			3 days
Nitrates/Nitrites (serum, ocular fluid)	PVL	Spectrophotometry	Any	5 ml serum, blood, or aqueous humor	Plastic tubes	Chilled/frozen in insulated container with gel packs	3 days	Blood from live animals with clinical signs should be examined for methemoglobin
Nitrates/Nitrites (serum, ocular fluid)	NBC	Ion Chromatography	Any	Whole eye	Plastic zip-lock or whirl-pak bags	Chilled/frozen in insulated container with gel packs	3 days
Nonsteroidal Antiinflammatory Screen	NBC	LC/MS	Any	2 ml serum	Red top tube	Chilled/frozen in insulated container with gel packs	5 days	Includes: flunixin, ibuprofen, indomethacin, ketoprofen, meloxicam, mefanamic acid, oxyphenylbutazone, phenylbutazone, salicylic acid, tolmetin, diclofenas
				5 gm liver	Whirl-pak bag
				10 ml urine	Plastic tube, RTT
Organic Chemical Screen	NBC	GC/MS	Any	10 ml serum, urine	Plastic vial	Chilled/frozen on gel packs	5 days minimum	This screen tests for a wide variety of pesticides, drugs, pollutants, and industrial chemicals For specific questions, please call the laboratory
				100 g rumen/stomach/crop contents, liver 10 g body fat 400 g suspect material	Whirl-pak or zip-lock plastic bag
				1 liter water	Distilled water bottle
PCBs	NBC	GC/ECD	Any	10 g tissues or biological samples, preferably liver and brain	Plastic or whirl-pak bag	Chilled/frozen in insulated container with gel packs	5 days	Quantitated Aroclor 1260
pH	PVL NBC	pH Meter	Water, biological fluids	100 ml water, GI contents	Clean plastic distilled water bottle		1 day
Plant identification	PVL NBC		Any	Whole plant, including roots, or rumen/GI contents; if weed in hay, submit representative sample of suspect plant material only	Plastic bag if fresh	Pressed or chilled if fresh	5-7 days	Contact laboratory before submittal. When submitting a fresh plant, roots should be wrapped in wet newspaper and the entire plant placed in a plastic bag. When chilled, the plant will arrive in good condition.
Selenium	PVL NBC	AA ICP/MS	Any	2 ml whole blood	Purple top tube	Chilled (not frozen)	7 days	Preferred sample is whole blood. Whole blood - send cold or frozen.
				2 ml serum	Royal blue serum or red top serum tube	Chilled/frozen in insulated container with gel packs		Serum - spin down and separate before freezing.
				5 g liver	Plastic bag	Chilled/ frozen in insulated container with gel packs
				5 ml milk	Plastic tube	Chilled/ frozen in insulated container with gel packs
				100 g feed (defined or undefined)	Plastic bag	Dry		Defined feed - please state the expected level of selenium (<10 ppm). Results are intended for clinical or diagnostic use only.
Tranquilizer Screen: xylazine, medetomadine, ketamine, diazepam, acepromazine, hydomorphone, midazolam, fluphemazine, detomidine	NBC	LC/MS	Any	2 ml serum	Red top tube	Chilled/frozen in insulated container with gel packs	3 days
	NBC	LC/MS	Any	5 gm liver	Plastic or whirl-pak bag	Chilled/frozen in insulated container with gel packs	3 days
Urea	PVL	Test Kit	Any	1 kg feed, rumen contents	Dry, in plastic bag or carton. If high moisture, ship frozen.	If high moisture, chilled/frozen in insulated container with gel packs	7 days
Vitamin E	PVL NBC	HPLC	Any	4 ml serum	Royal blue serum or red top serum tube	Chilled or frozen in insulated container with gel packs	7 days	Protect from light Serum-spin down and separate before freezing Herd-based testing is available
				5 g liver	Plastic bag	Fresh, frozen	7 days
				100 g feed	Plastic bag	Dry	14 days
Vomitoxin (deoxynivalenol, DON) (see mycotoxins)	PVL	ELISA TLC HPLC
Warfarin (see Anticoagulant screen)	PVL NBC	HPLC
Zearalenone (see mycotoxins)	PVL NBC	ELISA TLC HPLC
Zinc (serum) [see Minerals (serum)]	PVL NBC	AA ICP/MS	Any	5 ml serum (PVL) 1 ml serum (NBC)	Take sample in royal blue top vacutainer tube, remove from clot, ship in clean royal blue top tube or plastic vial	Chilled/frozen in insulated container with gel packs	2 days	Zinc will leach out from other rubber-stoppered tubes and invalidate the assay for zinc Please obtain the sample in a royal blue (trace metals) vacutainer tube (clot type), separate it from the clot, and transfer it to a clean royal blue top tube or plastic vial Hemolyzed samples should be avoided
Zinc phosphide	NBC	GC/MS	Any	Stomach content, bait, feed	Whirl-pak or zip-lock plastic bag	Chilled or frozen in insulated container with gel packs	5 days

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