Clinical Biochemistry in

Toxicology
• The availability of accurate historical information including a list of the animal species
affected, clinical signs, toxins suspected, potential route of exposure, vehicle, relative
amount, and timing of exposure is often a limiting factor in the diagnosis of toxic
disease.

• In the absence of such detailed history, the identification of target organs using clinical
biochemistry may help clinicians to create a list of potential toxins retrospectively.
 HEPATOTOXICITY
The susceptibility of the liver to toxic insult is in part a consequence of its location
between the digestive tract and the rest of the body and the central role it plays in
biotransformation and disposition of xenobiotics.

Extrahepatic metabolism of toxins by mixed function oxidases may affect the

target organ and potential hepatotoxicity of a given xenobiotic.

Lipophilic compounds tend to be more hepatotoxic than hydrophilic ones because

the latter are eliminated by the kidney.

Many toxins are hepatotoxic and nephrotoxic, however, and most toxins have
multiple target organs.
Biotransformation

Potentially toxic xenobiotic Relatively harmless

Metabolic
Detoxification activation

Inactive metabolite Reactive intermediate

Converting lipophilic to water
soluble compounds
Lipophilic
Xenobiotic (non-polar)
Phase I - Activation

Reactive intermediate

Phase II - Conjugation

Conjugate
Water soluble
Excretion (polar)
Phase I

• introduction of functional group

• hydrophilicity increases slightly

• may inactivate or activate original compound
• major player is CYP or mixed function oxygenase (MFO)
system in conjunction with NAD(P)H
• location of reactions is smooth endoplasmic reticulum
Phase II

• conjugation with endogenous molecules

(GSH, glycine, cystein, glucuronic acid)

• hydrophilicity increases substantially

• neutralization of active metabolic intermediates
• facilitation of elimination
• location of reactions is cytoplasm
Phase I reactions
 Oxidation
 Hydroxylation (addition of -OH group)
 N- and O- Dealkylation (removal of -CH side chains)
 Deamination (removal of -NH side chains) O epoxide
 Epoxidation (formation of epoxides) C C
 Oxygen addition (sulfoxidation, N-oxidation)
 Hydrogen removal

 Reduction
 Hydrogen addition (unsaturated bonds to saturated)
 Donor molecules include GSH, FAD, NAD(P)H
 Oxygen removal
C O
 Hydrolysis
 Splitting of C-N-C (amide) and C-O-C (ester) bonds
Biotransformation
• Activation of xenobiotics is a key element
(e.g. benzene, vinyl chloride)
• Reactive intermediates include epoxides and free
radical species (unpaired electrons) that are short-lived
and hence highly reactive
• Protection is provided by
• endogenous antioxidant substances, e.g. GSH
• vitamins C and E
• antioxidant enzymes, SOD, GPX, CAT in coupled reactions
• Antioxidant molecules are oxidized in the process but
have the capacity to regenerate the reduced form from
the oxidized - NAD(P)H is a key player
Metabolic enzymes
1. Microsomal:
1. CYP450 monooxygenases
2. Flavin monooxygenase
2. Non-microsomal
1. Alcohol dehydrogenase
2. Aldehyde dehydrogenase
3. Monoamine and diamine oxidases
3. Both
1. Esterases and Amidases
2. Prostaglandin synthase
3. Peroxidases
Phase II reactions
• Glycoside conjugation - glucuronidation
• Sulfate - sulfation
• Glutathione (GSH)
• Methylation
• Acylation
• Acetylation
• Amino acid conjugation
• Deacetylation
• Phosphate conjugation
 NEPHROTOXICITY
• Acute nephrotoxicity may initially induce polyuria that is followed by oliguria or
anuria.
• Nephrotoxins affecting approximately 66% of the nephrons will result in inability to
concentrate urine to a specific gravity greater than 1.030 in the dog, 1.035 in the cat,
and 1.025 in the horse and cow.
• Chronic toxicity may result in isosthenuria (constant urine osmolality in the range of
glomerular filtrate, 1.008 to 1.012).
• Azotemia, elevation in blood urea nitrogen (BUN) or creatinine, may
occur as prerenal, renal, or postrenal.
• Elevations of BUN and creatinine are not proportional in renal disease of
ruminants because of reutilization of urea by the rumen.
• Creatinine concentration is not affected significantly by diet, pro- tein
catabolism, or urinary flow.
 TOXINS AFFECTING SKELETAL AND CARDIAC MUSCLE
• The clinical signs of weakness, dysmetria, and incoordination suggest not only the possibility of
neurological disease but also skeletal muscular or cardiovascular disease.
• Acute toxicity of skeletal or cardiac muscle can be detected by elevations in serum creatine kinase
(CK).This dimeric enzyme catalyzes the reversible reaction, phosphocreatine + ADP  creatine +
ATP, and has three isoenzyme types: CK1, CK2, and CK3.

• CK1 is found in brain, peripheral nerves, cerebro-spinal fluid, and viscera, but it is not found in
serum during neurological disease. CK2 is found in cardiac muscle and minute amounts in skeletal
muscle. CK3 is found in cardiac and skeletal muscle.
• CK elevations maximize within 6 to 12 h and return to normal within 24 to 48h. Continuing necrosis
can result in persistent elevation.
• Cardiac troponin T and I have been demonstrated to be sensitive and specific biomarkers of cardiac
injury in dogs and laboratory animals
• LDH5 is the principal isoenzyme in skeletal muscle and erythrocytes.
• Necrosis of skeletal muscle may result in release of myoglobin and potassium resulting in
myoglobinemia and hyperkalemia.
TOXINS AFFECTING THE GASTROINTESTINAL TRACT
• Vomiting and diarrhea may produce dehydration that results in mild to moderate elevations
of BUN, plasma protein, packed cell volume (PCV), and urine specific gravity.
• Metabolic alkalosis (increased pH, normal or increased PCO 2, increased HCO3-and
HCO3-/H2CO3) and hypochloridemia may result from chloride loss associated with vomiting.
• Metabolic acidosis (decreased pH, normal or decreased PCO 2, decreased HCO3- and
HCO3-/H2CO3) may result from secretory loss of bicarbonate in diarrhea.
• In the absence of evidence of malnutrition or hepatic or renal disease, chronic gastrointestinal
malabsorption or protein loss should be considered as a potential cause of hypo-proteinemia.
• The seleniferous plants, which may be associated with acute gastroenteritis in herbivores.
• Fusarium spp. and the macrocyclic trichothecene produced by Stachybotrys alternans are
highly irritant and produce acute ulceration and hemorrhage of the gastrointestinal tract.
• estrogenic mycotoxin, zearalenone, may induce rectal prolapse in pigs in addition to affecting
reproductive performance.
•
 TOXINS AFFECTING THE NERVOUS SYSTEM
• Many acute and chronic neurotoxins produce illness or death without
alterations detectable by routine clinical biochemistry performed on blood or
serum.
• The cerebrospinal fluid (CSF) is usually normal in neurotoxicity; however, mild
elevations in protein and leukocyte count may occur with lead poisoning
• Accumulations of endogenous toxins secondary to hepatotoxicity and
nephrotoxicity may produce neurological dysfunction.
• Increased urinary excretion of delta-aminolevulinic acid is a potential indicator
of lead intoxication.
• Organo- phosphates induce cholinesterase inhibition, which can be detected
as reduction of plasma or whole blood cholinesterase activity.
• Thiaminases that may induce polioencephalomalacia in herbivores are present
in Equisetum arvense and Pteridium aquilinum.
 TOXINS AFFECTING THE LUNG AND RESPIRATORY TRACT
• Disease affecting the respiratory tract is often clinically apparent on the basis of dyspnea. Dyspnea
in veterinary medicine is more often the result of pneumonia rather than intoxication. However,
when body temperature is normal, the possibility of pulmonary edema induced by toxins affecting
the lung and respiratory tract or cardiovascular system should be considered.
• Serum Angiotensin converting enzyme (ACE) activity is altered in chronic and acute pulmonary
disease .
• Because CO2 is approximately 20 times more diffusible than O2, early pulmonary edema typically
results in decreased PaO2.
• Severe pulmonary edema may result in elevated PaCO2 (hypercapnia and respiratory acidosis)
• Cattle are susceptible to several toxins that induce acute pulmonary edema including ingestion of
perilla ketone in Perilla frutescens, 4-ipomeanol in sweet potatoes (Ipomoea batatas) infected with
Fusarium solani
• Organochlorines and organobromines include chlorinated naphthalenes, polychlorinated biphenyls
(PCBs), polybrominated biphenyls (PBBs), and dibenzofurans. These industrial toxins are cumulative
and result in hypovitaminosis A that is associated with squamous metaplasia of columnar
epithelium of the respiratory tract and hyper-keratosis.