In: Veterinary Toxicology, V. Beasley (Ed.

)
Publisher: International Veterinary Information Service (www.ivis.org), Ithaca, New York, USA.

Methemoglobin Producers (9-Aug-1999)

V. Beasley
Department of Veterinary Biosciences, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Urbana,
IL, USA.

Chapter Sections
Acetaminophen
Naphtalene
Local Anesthetics
Nitrate - Nitrite
Chlorates

Acetaminophen

Major Species Usual Time of Onset Usual Duration (if survives) Full Table for
Methemoglobin
Cats, dogs 4 - 24 hours Days; highly lethal Producers

Sources

z Many "aspirin-free" pain relievers contain acetaminophen, e.g., Allerest, Anacin-3, Comtrex, Datril, Dristan-AF,
Excedrin, Nyquil, Sinarest, Sine-Aid, Sine-Off, Sinutab, Tylenol, Vanquish, many others.
z Some analgesics contain phenacetin, which is rapidly metabolized to acetaminophen.

Absorption, Distribution, Metabolism and Excretion (ADME)

Rapidly absorbed from the gastrointestinal tract. Most is absorbed within 60 minutes. Previous ingestion of a high
carbohydrate diet may delay gastric emptying so that peak plasma levels may occur as late as 4 hours after ingestion.

Mechanism of Action

z Two major conjugation pathways are used by most species for metabolism. Both involve P-450 metabolism, and for a
major fraction of the compound, this is followed by glucuronidation or sulfation.
z Reactive metabolite from P-450 system can be conjugated by 1 of these 2 means or:
z Be conjugated with glutathione - probably becomes toxicologically inert.

z Bind to cellular macromolecules - causing cell dysfunction and/or cell death.

Major pathways of acetaminophen biotransformation. Adenosine 3'-phosphate 5'-phosphosulfate (PAPS), uridine
diphosphoglucuronic acid (UDP-glucuronic acid), and uridine diphosphoglucuronosyltransferase (UDP-
glucuronosyltransferase).

z Man, dogs, and cats have low ability for sulfation.

z Cats lack the specific glucuronyl transferase required to conjugate acetaminophen.
z Reactive metabolites form.

z Endogenous hepatic glutathione is quickly exhausted.

z When glutathione levels drop to 20 - 30% of normal, metabolites bind to cells causing cell death.

Species Susceptibility

z Dogs and man.

z Tolerate normal therapeutic dosages.

z Usually no clinical signs are observed until dosage exceeds 100 mg/kg.

z Signs are mainly referable to acute hepatic damage.

z Three of four dogs exposed to acetaminophen at 200 mg/kg developed clinical methemoglobinemia.

z Cats.
z Acetaminophen-Containing Products Should Never Be Administered To Cats!!

z Lower dosages produce toxicosis, especially if the dose is repeated. Signs are largely referable to

methemoglobinemia. Cats also develop liver lesions when poisoned by acetaminophen.

z An oral dose of 60 mg/kg produced 21.7% methemoglobin 4 hours after administration.

z Problems may be noted in cats exposed to as low as 40 mg/kg.

Signs and Lesions

z Cats.
z Cyanosis.
z Methemoglobinemia. Specific quantitation of the percent hemoglobin is preferred. A screening assay can be
performed by putting a drop of blood on a piece of absorbent white paper. A chocolate brown color generally
indicates at least 15% methemoglobin.
z Dyspnea.

z Severe depression/weakness.

z Anorexia.

z Vomiting.

z Facial and/or paw edema (mechanism not understood).

z Dogs and man.

z Acute centrilobular hepatic necrosis usually 24 - 36 hours postingestion. Manifestations may include anorexia,

vomiting, abdominal pain, shock, etc.

Treatment

z Goals:
z Prevent absorption from the gastrointestinal tract with an emetic, activated charcoal, and a cathartic (because
of rapid absorption, such measures are likely to be of benefit only for very recent ingestions, i.e., < 2 hours).
z Do not use an emetic if animal is hypoxic.

z Intercept reactive metabolites and promote their excretion.

z Convert methemoglobin (reduce it) back to hemoglobin (in cats).

z Support cardiovascular/respiratory system function.

z Activated charcoal and a cathartic (if appropriate as mentioned under goals above and under acetylcysteine

below).
z Activated charcoal may possibly absorb and inactivate orally administered N-acetylcysteine (this is an area of
controversy): for now, do not give together.
z N-acetylcysteine is highly effective as a therapy for acetaminophen toxicosis.

z N-acetylcysteine provides sulfhydryl groups, directly binds to acetaminophen metabolites to enhance

elimination, and serves as a glutathione precursor.

z For severely poisoned animals, initially give 280 mg/kg PO or IV in 5% dextrose (for milder cases, you can

start with 140 mg/kg). If the IV route is used, the dose is given slowly over a period of 15 - 20 minutes. The
IV route should be chosen if an emetic is to be used or if activated charcoal is still present in the upper
gastrointestinal tract. For IV use, it is preferred to filter the acetylcysteine solution through a 0.2 micron filter.
Most pharmacies will be able to provide a suitable bacteriologically filtered solution.
z Follow with 70 mg/kg PO QID for a total of 6 doses after the loading dose (monitor as withdraw to assure that

this course of therapy is long enough).

z For most cases of acetaminophen toxicosis, acetylcysteine is best given solely by the oral route.

z More of acetylcysteine will reach liver if administered as oral dose.

z Sources:
z Generic acetylcysteine of 98% purity from chemical company - 14% suspension in distilled water.

z Mucomyst ® (Mead, Johnson, and Co.).

z Sterile 10% and 20% solutions.

z As an alternative to acetylcysteine, sodium sulfate at 50 mg/kg has been evaluated after being given every 4 hours for
a total of 3 doses (given IV as a 1.6% solution). Although significant benefit was apparent and the sodium sulfate was
essentially equivalent to 3 doses of acetylcysteine (140 mg/kg, 70 mg/kg, 70 mg/kg), hepatic lesions were noted in
both antidote treated groups. These results may suggest that treatments for longer periods of time (see above) are
warranted.
z Ascorbic acid (20 mg/kg) up to QID - oral or parenteral.
z Empirically recommended in an effort to convert methemoglobin to oxyhemoglobin.

z Ascorbic acid is safe for cats (at doses recommended for acidification of the urine).

z Supportive fluid therapy.

z If severe cyanosis is present, supply oxygen.
z Cage rest.
 z Steroids and antihistamines are contraindicated!!

N-acetylcysteine sources

1. Fisher Scientific Company.*

1600 Glenlake Road, Itasca, IL 60143.
*An account is required for purchasing. Application required for first time orders.
Cost: 25 grams (powder concentrate), 100 grams
Observe precautions for storage, handling, and administration.
2. University of Illinois, Veterinary Medicine Pharmacy. 20% solution, in stock
4 ml vials
10 ml vials
3. Redbook prices. Mead-Johnson, Mucomyst ®.
10%
4 cc each x 12 vials
10 cc each x 3 vials
30 cc each x 3 vials
20%
4 cc each x 12 vials
10 cc each x 3 vials
30 cc each x 3 vials
4. Local hospital or pharmacy.
Prices will vary
Most practical and fastest to obtain
20% solution may be most cost-effective.
Carle Pharmacy, Urbana:
20% - 10 ml x 3 vials/pack
10% - 10 ml x 3 vials/pack

Case Example:

a. A 5-kg cat, presented with severe complications:

Loading dose (maximum at 280 mg/kg) 1400 mg
Maintenance dose (350 mg x 6 doses) 2100 mg
Total needed 3500 mg

b. Using:
10% solution, 10 ml vial
requires 3.5 vials (4 vials)

20% solutions, 10 ml vial

requires 1.5 vials (2 vials)
 Weight-Dosage Conversion Chart N-Acetylcysteine (Mucomyst ®)

Maintenance Dose

Loading Dose
Total
Animal Weight % Solution Total % Solution Needed
mg mg/dose mg (6 mg
kg (lb) 10 20 doses) 10 20

1 (2.2) 140 1.4 ml 0.7 ml 70 420 0.7 ml 0.35 ml 560

2 ( 4.4) 280 2.8 ml 1.4 ml 140 840 1.4 ml 0.7 ml 1120

3 ( 6.6) 420 4.2 ml 2.1 ml 210 1260 2.1 ml 1.05 ml 1680

4 ( 8.0) 560 5.6 ml 2.8 ml 280 1680 2.8 ml 1.4 ml 2240

5 ( 11.0) 700 7.0 ml 3.5 ml 350 2100 3.5 ml 1.75 ml 2800

6 ( 13.2) 840 8.4 ml 4.2 ml 420 2520 4.2 ml 2.1 ml 3360

7 ( 15.4) 980 9.8 ml 4.9 ml 490 2940 4.9 ml 2.45 ml 3920

8 ( 17.6) 1120 11.2 ml 5.6 ml 560 3360 5.6 ml 2.8 ml 4480

9 ( 19.8) 1260 12.6 ml 6.3 ml 630 3780 6.3 ml 3.15 ml 5040

10 ( 22.0) 1400 14.0 ml 7.0 ml 700 4200 7.0 ml 3.5 ml 5600

20 ( 44.0) 2800 28.0 ml 14.0 ml 1400 8400 14.0 ml 7.0 ml 11200

30 ( 66.0) 4200 42.0 ml 21.0 ml 2100 12600 21.0 ml 10.50 ml 16800

40 ( 88.0) 5600 56.0 ml 28.0 ml 2800 16800 28.0 ml 14.00 ml 22400

50 (110.0) 7000 70.0 ml 35.0 ml 3500 21000 35.0 ml 17.50 ml 28000

1. Mucomyst ® (N-acetylcysteine), Mead-Johnson.

2. Available Forms:
- in packs of 3 - 10 ml vials
10% solution
20% solution
3. Each 10 ml vials contains:
10% solution - 1000 mg N-acetylcysteine (100 mg/ml)
20% solution - 2000 mg N-acetylcysteine (200 mg/ml)
4. Loading and maintenance doses determined from protocol, i.e., loading dose of 140 mg/kg (280 mg/kg in severe
clinical situations), followed by maintenance dose of 70 mg/kg every 6 hours for a total of 36 hours (7 treatments).
Note - Do not administer N-acetylcysteine orally with activated charcoal.
Pathways in the reduction of methemoglobin.
Abbreviations: Met HB = methemoglobin; Hb = hemoglobin; NADH-MR = NADH-dependent methemoglobin reductase;
PG = phosphogluconate; DPG = diphosphoglycerate; GSSG = oxidized glutathione; GSH = reduced glutathione; GR =
glutathione reductase; DHA = dehydroascorbic acid.

Naphthalene

Major Species Usual Time of Onset Usual Duration (if survives) Full Table for
Methemoglobin
Dogs, cats Hours Days; often lethal Producers

Sources

z Naphthalene is present in "old-fashioned" mothballs. It is the most common active agent in mothballs. Naphthalene is
also present in some types of mothflakes or crystals; but in these products, paradichlorobenzene may be used more
often.
z Example - Enoz Old Fashioned mothballs contain 2.7 grams of naphthalene per mothball.
z Naphthalene mothballs are approximately 2 times as toxic as paradichlorobenzene mothballs, the only other
commonly encountered type.

Naphthalene
Toxicity

z Naphthalene appears to be quite toxic to cats.

z Less than one naphthalene mothball may induce toxicosis in a child.
z Acute hemolytic anemia has been reported in dogs at 411 mg/kg or greater.

Signs and Clinical Pathology

z GI irritation, vomiting may account for the most common complaints.

z Methemoglobinemia.
z Heinz body anemia associated with hemolysis; accounts for majority of serious effects.
z Hemoglobinuria.
z CNS abnormalities (possible seizures) are infrequently seen.
z Rarely, hepatitis occurs 3 - 5 days postingestion.

Treatment

z Emesis.
z Activated charcoal.
z Saline cathartic.
z Control methemoglobinemia.
z In cats and usually in dogs, use ascorbic acid at 20 mg/kg.

z In other species use methylene blue.

z Low doses of methylene blue (1.5 mg/kg) have recently been reported to be beneficial to cats with nitrite-

induced methemoglobinemia. Methylene blue given in the absence of methemoglobinemia can cause Heinz
body formation.
z For animals with hemolytic crisis, fluids and bicarbonate (2 - 3 mEq/kg slow IV added to fluids over a period of
hours), are suggested to minimize precipitation of hemoglobin in the kidney and associated nephrosis.
z For anemic animals, a blood transfusion may be of value.
z Control seizures.
z Supportive care.

Paradichlorobenzene Naphthalene
Surface appearance wet, oily dry
In ethanol rapidly soluble slowly soluble
In 60ºC water soluble insoluble
Reaction with chloroform, aluminum chloride, intense blue
no reaction
anhydrous powder color
Reaction with copper wire in a flame bright green flame no color
Immersion in a saturated solution of table salt sink float

z Simple specific gravity method to differentiate type of moth balls. Test for sink vs. float: start with 4 ounces of tepid
water. Add 3 heaping tbsp of table salt. Stir vigorously until salt will not further dissolve.
z Naphthalene mothballs - Sink in water, but float in saturated salt solution

z Paradichlorozene mothballs - Sink in water, and sink in salt solution.

z Apparently this is a very reliable test.

Local Anesthetics

Specific Agents Major Species Usual Time of Onset Usual Duration (if survives) Full Table for
Methemoglobin
Local Anesthetics Most species Hours Hours to a day; often lethal Producers
Resorcinol in cats Mostly cats Hours Days; often lethal

Sources

z Local anesthetic preparations including lidocaine, benzocaine, and tetracaine are present in a variety of preparations
for topical use. These include mucosal sprays, suppositories, nonprescription cremes, teething gels, and aerosols.
z They are used to facilitate endotracheal intubation, especially in the feline animal in an effort to control laryngeal
spasm.
z Xylocaine ® contains lidocaine.
z Cetacaine ® contains benzocaine and has been recommended for use as a laryngeal or pharyngeal spray to control
pain or gagging. Some veterinarians have suggested its use locally in the urethra in attempts to relieve feline urologic
syndrome. These uses are not without risk.
z Lanacane ® cream and Bicozene ® contain 6% benzocaine and resorcinol, and Vigisil ® contains 5% benzocaine and
resorcinol. These products are available in 1 oz (30 g) or 2 oz (60 g) tubes.

Lidocaine

Susceptible Species

z Humans, dogs, and cats have all been poisoned by local anesthetics.
z Cats are well recognized as the domestic species having hemoglobin most susceptible to the action of oxidizing
agents, including aromatic amines.

Absorption, Distribution, Metabolism and Excretion (ADME) and Mechanism of Action

z Local anesthetics or, more likely, their metabolites cause oxidation of hemoglobin to produce methemoglobin and
cause Heinz body formation.
z Local anesthetics contain a common aromatic group connected via an intermediate chain to an amino group by either
an amide or an ester linkage. Of the local anesthetics which have been implicated in the formation of methemoglobin,
lidocaine and prilocaine contain amide linkages. These bonds are metabolized slowly by liver microsomal enzymes,
resulting in intermediates which are believed to act as oxidants.
z In cats, an injection of prilocaine or o-toluidine, its proposed metabolite, induced approximately equal levels of
methemoglobinemia. In the same study, however, both lidocaine and its proposed metabolite, 2,6-xylidine, failed to
 produce methemoglobinemia in cats, suggesting an alternate mode of metabolism or, at least, lesser susceptibility to
this local anesthetic in the cat.
z Benzocaine contains an ester bond and is metabolized by esterases in the plasma. However, the specific metabolite
responsible for its effect on hemoglobin is unknown.
z Conjugation of local anesthetics is a necessary step prior to their elimination in the urine. The deficiency of
glucuronyl transferase in cats may contribute to their susceptibility to benzocaine and other oxidant drugs.
z Benzocaine is one of the least water-soluble local anesthetics, a factor which reportedly slows its absorption. The
Cetacaine ® preparation, however, contains preservatives which are reportedly irritating to the mucous membranes.
This irritation may, along with any pre-existent laryngeal lesions, increase absorption.
z Whether instillation into the trachea increases the fraction absorbed, as compared to oral exposure, remains to be
experimentally documented.
z Additional properties of local anesthetics which are less often associated with clinical toxicoses include:
z Cardiac effects: interference with intraventricular conduction, myocardial contractility, and electrical

excitability, resulting in arrhythmias. Hypotension and cardiac arrest can occur.

z Direct vasodilation may result in hypotension.

z At low doses local anesthetics are anticonvulsants, and at high doses they are convulsants.

z Respiratory effects: depression and apnea can also occur.

z Nervous effects: may include seizures, tremors, agitation, lethargy, and coma.

z Acidosis may also be observed.

Toxicity

z Similar metered sprays of Cetacaine ® (benzocaine) or Xylocaine ® Spray (lidocaine) were administered to cats.
When cats were treated with a one-second spray of Cetacaine ® (8.1 mg benzocaine), the animals consistently
experienced significant methemoglobinemia (up to 30%), while a similar exposure to Xylocaine ® Spray (10 mg
lidocaine) had no such effect.
z In spite of the absence of an effect in this study, it has been noted that lidocaine-metered sprays, intended for human
patients, do deliver a greater and, therefore, excessive dose for cats.
z An appropriate method of laryngeal treatment would be the use of a 1 - 2% solution on a cotton swab, applied directly
to the laryngeal folds.
z Cats intravenously dosed with benzocaine at up to a total dose of 41 mg (2.1 mg/kg) developed methemoglobinemia
up to a high value of approximately 22%.
z Dogs are sometimes clinically poisoned by ingestion of ointments or cremes containing local anesthetics.
z Methemoglobinemia has been experimentally produced (at up to 14%) in dogs intubated with endotracheal tubes to
which a gel containing 20% benzocaine was applied.

Signs

z In experimentally treated cats, methemoglobinemia developed within 5 - 10 minutes of a one-second laryngeal spray
of Cetacaine ® (benzocaine) and peaked at 20 - 30 minutes after application. Two of the 7 treated cats developed
clinical signs of cyanosis.
z Elevated values of methemoglobin persisted 30 minutes to 3 hours.
z Had greater degrees of exposure been tested, more severe effects would have been likely.
z Cyanosis, rapid, shallow respiration, and prolonged sedation may also occur.
z It was reported that the peripheral veins of Cetacaine ®-treated cats were difficult to palpate and visualize following
treatment.

Diagnosis

z Normal methemoglobin is reported as approximately 1% in cats.

z Diagnoses are based upon the presence of significant methemoglobinemia and/or a Heinz body anemia, a history of
excessive exposure to a local anesthetic, and absence of exposure to other oxidants (especially acetaminophen in the
cat).
Treatment

z For very recent oral exposure, in patients in which contraindications do not exist, an emetic may be used, and in any
case, activated charcoal and a saline cathartic are suggested.
z Diazepam is suggested for seizures in dogs. Phenobarbital is suggested if diazepam fails to control the seizures.
Phenobarbital is suggested for other species if seizures occur. If phenobarbital fails, pentobarbital is suggested.
z For acidosis, sodium bicarbonate in fluids is suggested.
z Forced diuresis has been shown to be of benefit for some local anesthetics but may not be of benefit for others.

For Methemoglobinemia

z Ascorbic acid:
z Ascorbic acid (vitamin C) has been suggested to correct methemoglobinemia at a dose of 20 mg/kg. However,

controlled studies of its effectiveness remain to be conducted.

z Methylene blue:
z It is best to avoid the use of methylene blue in the dog unless absolutely necessary (i.e., if vitamin C fails to

provide sufficient benefit.

z When given to the dog, the recommended dose of methylene blue is 4 mg/kg. There is little available data on

which to base this recommendation, and it is reported that a dose of methylene blue at 10 - 15 mg/kg can cause
acute intoxication and death in dogs due to hemolytic anemia with Heinz body formation.
z Low doses of methylene blue (1.5 mg/kg) have recently been reported to be beneficial to cats with nitrite-

induced methemoglobinemia. Methylene blue given in the absence of methemoglobinemia can cause Heinz
body formation.
z Oxygen:
z Oxygen therapy may be of limited benefit after the administration of agents intended to reduce

methemoglobin back to hemoglobin.

Nitrate - Nitrite

Usual Time of Usual Duration (if

Specific Agents Major Species
Onset survives)
Nitrate and nitrite Ruminants, occasionally Minutes to hours Hours to 2 days; often
other species lethal
Nitrate-accumulating plants, Ruminants, occasionally Hours to days Hours to 2 days; often
e.g.: other species lethal
Sorghum (Sorghum), Sudan (See Cyanogenic Glycoside-Containing Plants)
grass
Corn, especially green chop
(Zea mays)
Oats (Avena sativa) Full Table for
Methemoglobin
Wheat (Triticale) -
Producers
Beets (Beta) (See Toxicants that Affect the Kidneys)
Kochia (See Poisonous Plants that Affect the Liver)
Locoweeds (organic nitrates) (See Toxicants with Mixed Effects on the CNS)
Fiddleneck (Amsinckia) (See Poisonous Plants that Affect the Liver)
Fescue (Festuca) (See Toxicants that Affect Peripheral Circulation) (Methemoglobinemia
is rare)
Pigweed (Amaranthus) (See Plants that Affect the Kidneys) (Methemoglobinemia is rare)
Lamb's quarters (See Plants that Affect the Gastrointestinal Tract) (Toxicosis is rare)
(Chenopodium)
Many others -
Sources

Nitrate-Accumulating Plants
Sudan Grass (Sorghum spp.)* Sweet Clover (Melilotus)
Johnson Grass (Sorghum spp.)* Flax, linseed (Linum)
Sorghum (Sorghum spp.)* Pigweed (Amaranthus)
Fireweed (Kochia)* Lambsquarters(Chenopodium)
Beets (Beta)* Canada Thistle (Cirsium)
Rape (Brassica)* Jimson weed (Datura)
Corn (Zea)* Wild Sunflower (Helianthus)
Rye (Secale cereale) Cheeseweed (Malva)
Wheat (Triticum) Smartweed (Polygonum)
Oats (Arena) Dock (Rumex)
Fescue (Festuca) Nightshades (Solanum)
Alfalfa (Medicaqo) Soybeans (Glycine)
* Indicates plants of greatest risk.

z Factors Increasing Nitrate Concentrations in Plants

z Species of plant (nitrate accumulators).

z Heavy fertilization.

z Factors that tend to reduce plant nitrate reductase activity):

z Drought or rainy period after drought.

z Decreased light (cloudy weather, early in the day before exposure to much sunshine). For these

reasons, it is wise to cut hay or green chop relatively late on a sunny day to minimize concentrations of
nitrate.
z Decreased temperature.

z Nitrates in Water
z Run-off from fertilized fields.

z Run-off from decaying manure in barnlots.

z Silage juices.

z Nitrates or nitrites may leach through soil to deep and especially shallow, poorly cased wells.

z Concentrations are less likely to be excessive in ponds with abundant plants or algae. Algae blooms decrease

nitrates which decreases the risk of nitrate poisoning for exposed animals but (blue-green) algae is sometimes
a worse hazard.
z Concentrations may be highest after high surface run-off (heavy rains, spring snow melts), especially when

recently applied fertilizer washes into small ponds or very small streams.
z Fertilizers
z Some nitrogen fertilizers are high in ammonium nitrate.

z Occasional sources for large animals (sodium, potassium, or ammonium nitrate).

z May be eaten directly from bags, spilled, unevenly spread, or washed into water (see above).

z Fertilizer is the primary source of nitrate for small animals.

z Some nitrogen fertilizers are high in urea.

z Time
z Concentrations in plants are generally highest just prior to flowering and rapidly drop off after pollenation

occurs and grains or fruits set on.

z There is evidence of postharvest reduction of nitrate to nitrite in feedstuffs such as moist hay and silage, a

factor which could increase toxicity. Conversely, with completion of the ensiling process, nitrate-containing
feeds often become somewhat less hazardous with conversion of the nitrite to ammonia and dissipation.
z Drying
z Most nitrate is retained when plants are air-dried at ambient temperatures (e.g., hays). The nitrate content of

hay may even increase during storage due to moisture loss.

z Acid soils.

z Low molybdenum, sulfur, phosphorus, manganese, or magnesium in soils.

z Application of phenoxy (2,4-D like) herbicides.

z Nitrates accumulate in the vegetative tissues of the plant, not in the fruits or grain. Concentrations are usually
 greatest in the stalk and less in the leaves.

Mechanisms of Action

z Nitrite is the actual toxic compound.

z Ruminants are much more susceptible to nitrates than monogastric animals.
z Rumen microorganisms reduce nitrate (NO3) ion to nitrite (NO2) ion.
z Nitrite oxidizes hemoglobin to methemoglobin.
z 30 - 40% methemoglobin causes mild clinical signs.

z 75 - 90% methemoglobin causes severe clinical signs to death.

z Fetuses and neonates are more susceptible than are the older young or the adults.
z The small quantities of methemoglobin formed during (normal) metabolism of physiologic amounts of nitrites or
other oxidizing agents are converted to hemoglobin by an enzyme in erythrocytes, methemoglobin reductase. When
the formation of methemoglobin exceeds the turnover rate of methemoglobin reductase, abnormally high
concentrations of methemoglobin will occur.
z Vitamin A deficiency may be an effect of nitrate toxicosis and contribute to the occurrence of abortion (but this is
only a theory, it has not been substantiated).

Toxicity

z Acute nitrate poisoning is most likely to occur when forage nitrates exceed 10,000 ppm (1%) (dry weight basis) or
when water contains 1500 ppm (0.15%) nitrate. Since the effects of feed and water nitrate are additive, both should be
tested when the probability of toxicosis is being evaluated.
z Lethal doses of nitrate and nitrite are included in the following table:

Source Species Lethal Dose in mg/kg of body weight

Nitrate

Adult humans 80 - 130

Cattle 330 - 616

Sheep 308

Sodium Nitrite

Adult humans 110 - 165

Rabbits 80 - 90

Pigs 90

Sheep 170

z The amount of preformed nitrite influences toxicity, especially in monogastric species.

z The susceptibility of an animal to nitrate toxicosis is dependent upon several factors including: 1) rate of intake, 2)
variations in gastrointestinal nitrate reduction (adaptation), 3) diet (nutrition), 4) metabolic state of the animal as
influenced by age, pregnancy, and disease.
z Nitrite reduction in the gastrointestinal tract is attributed solely to action of microorganisms. In ruminants, this
reduction primarily occurs in the rumen; and nitrate, nitrite, and ammonium ions are absorbed directly into the blood.
z The inclusion of readily fermentable carbohydrates in the diet has been shown to protect ruminants against nitrate
toxicosis.
z Pregnant ruminants should not be fed over 0.2% nitrates.
z In monogastric species, with the exception of the human infant, the stomach contains relatively few microorganisms
due to the presence of hydrochloric acid. Monogastric species are, therefore, relatively resistant to toxicosis due to
nitrates but are susceptible to poisonings by preformed nitrites.
 z Human infants are more susceptible to nitrates and nitrites than adults. This appears to be due to differences in the
gastric flora as a result of lower gastric acidity and the comparative ease with which hemoglobin of the fetus is
converted to methemoglobin.
z Young rats and pregnant rats have also shown an increased sensitivity to nitrites.

Nitrate and Nitrite Expressions and Conversion Factors for Converting from One Way of Expressing Nitrate to Another

Form B
Form A
N NO2 NO3 KNO3 NaNo3
Nitrate-Nitrogen (N) 1.0 3.3 4.4 7.2 6.1

Nitrite-Nitrogen (N) 1.0 3.3 4.4 7.2 6.2

Nitrate (NO3) 0.23 0.74 1.0 1.63 1.37

Nitrite (NO2) 0.3 1.0 1.34 2.2 1.86

Potassium Nitrate (KNO3) 0.14 0.64 0.61 1.0 0.84

Sodium Nitrate (NaNO3) 0.16 0.54 0.72 1.2 1.0

To convert Form as Expression A to the equivalent amount of Form B, multiply A by the appropriate conversion factor.
(Nitrate as Form A x Conversion Factor = nitrate as Form B.)

Feed Concentration as Nitrate Ion (Dry

Estimated Toxicological Significance for Cattle
Matter Basis)
0.0 - 0.44% Considered safe to feed under all conditions.
Safe for nonpregnant animals under all conditions. Limit pregnant animals to 50% of
0.44 - 0.66%
the total dry matter in the ration
0.66 - 0.88% Limit to 50% of the total dry matter in the ration
Limit to 3 5 - 40% of the total dry matter in the ration. Feeds with over 0.88% nitrate
0.88 - 1.54%
ion should not be used for pregnant animals.
1.54 - 1.76% Limit to 25% of the total dry matter in the ration. Do not use for pregnant animals.

Over - 1.76% Feeds with over 1.76% nitrate ion may be toxic. Do not feed.
Source - Pennsylvania State University

Absorption, Distribution, Metabolism and Excretion (ADME)

z Both nitrate and nitrite are passively absorbed from the gastrointestinal tract and reach their highest concentrations 5 -
6 hours following their introduction into the rumen.
z In addition to conversion of nitrate to nitrite in the rumen, nitrate appears to be converted to nitrite by liver enzymes
following absorption, but the rate of the hepatic reduction must be slow since no significant methemoglobinemia
occurs following the intravenous administration of nitrates.
z Most nitrate is excreted by the kidney, but some is recycled back into the gastrointestinal tract by salivary and
gastrointestinal tract secretion.

Signs

z Acute Syndrome (onset within 1/2 - 4 hours after feeding).

z Direct gastrointestinal irritation - vomiting, salivation, diarrhea.

z Dyspnea.

z Tremors.
 z Ataxia.
z Rapid, weak heart beat.
z Terminal convulsions.

z Death in 6 - 24 hours. The number of deaths is increased when animals are stressed due to increased oxygen

requirements.
z Chronic Syndrome.
Note - These are the results of field observations. Chronic toxicosis has not been experimentally reproduced. Other,
unrecognized factors may be involved.
z Abortions - "Lowland abortion syndrome". Some authors report that abortions may occur most often at 3 - 5

days after an acute toxicosis.

z Poor growth and feed efficiency.

z Decreased milk production.

z Infertility.

z Goiter, primarily in sheep (interference with iodine metabolism).

z Increased susceptibility to infections.

Lesions

z "Chocolate-brown" blood.
z Brownish cast to all tissues.
z Agonal changes.
z Mucous membranes cyanotic.
z Congestion of ruminal or abomasal mucosae.
z Placental necrosis suggested by some authors.

Diagnosis

z Appropriate clinical signs and lesions.

z Laboratory analysis of nitrate and/or nitrite of: blood, serum, forages, feed, water, rumen contents, urine (NO3 stable
in urine for around 48 hours) and aqueous humor. Aqueous humor of aborted fetuses may also reveal elevated nitrate
concentrations. Except for dry feeds, dry forages, and blood, specimens should be collected as rapidly as possible and
shipped to the laboratory in a frozen state.
z Blood samples for methemoglobin analysis must be collected within 2 hours of death and rapidly frozen or preferably
preserved in phosphate buffer to stabilize the methemoglobin (See Osweiler G. Clinical and Diagnostic Veterinary
Toxicology, 3rd ed. Kendall/Hunt Publishing Company, 1984.).
z All samples should be shipped with as much air removed as possible.
z Results are reported in various ways (usually as NO3, nitrate ion).
z Serum NO3 of greater than 20 micrograms (µg) per ml is diagnostic.
z Fetal aqueous humor nitrate concentrations above 50 ppm have been deemed consistent with nitrate toxicosis and 30 -
50 ppm are considered suspect; thus when such levels are encountered diets and water also should be checked for
nitrate.
z Field Analysis:
z Diphenylamine test for nitrates.

z Alpha-naphthylamine test for nitrites.

A Qualitative Field Test for Detection of Nitrate or Nitrite

Nitrate Screening Diphenylamine Test

1. Add 0.5 g diphenylamine in 20 ml of water.
2. Add sulfuric acid QS to 100 ml. Cool the resulting solution and store in a brown bottle.
3. Mix equal parts of the stock solution and 80% sulfuric acid.
 A Qualitative Field Test for Detection of Nitrate or Nitrite

Nitrate Screening Diphenylamine Test

4. Test suspect material by dropping 1 drop of reagent on the cut surface of the plant.
5. A color change from green to blue is a positive reaction indicating more than 2% nitrate.
Nitrite Screening Test
1. Reagents needed are a) sulfanilic acid (0.5 g in 150 ml of 20% glacial acetic acid) and b) alphanaphthylamine hydrochloride
prepared by dissolving (with heat) 0.2 g of the salt in 20% glacial acetic acid.
2. Add 2 ml of unknown to a test tube. Then add 2 ml of sulfanilic acid, followed by 2 ml alphanaphthylamine. A pink to red color is
positive for nitrites. The solutions applied to cut surfaces of plants in the same order will give a pink to red reaction if positive

Treatment

z Don't stress any more than necessary.

z Methylene blue.
z 4 (dogs) to 15 (cattle) mg/kg IV in a 1% solution. Treatment may need to be repeated.

z In preliminary studies, methylene blue has been used successfully and safely to treat nitrite toxicosis in cats. A

single dose of 1.5 mg/kg IV can reverse methemoglobinemia much more rapidly than ascorbic acid or N-
acetylcysteine without causing anemia. Multiple doses or a single dose in the absence of nitrite toxicosis,
however, may cause Heinz body hemolytic anemia.
z Avoid overtreatment since excessive doses of methylene blue may increase methemoglobin formation.

z Dosage depends on severity.

z Function of methylene blue:

z Sources of methylene blue and diphenylamine: Sigma Chemical Co., Food and Drug Division, P.O. Box

14508, St. Louis, MO 63178, USA.

Blood Physical
Toxicant Mechanism Treatment
Color Characteristic
Nitrate (Nitrite) Brown Methemoglobin Methylene blue

Sodium Chlorate Brown Methemoglobin Methylene blue

Silo Gases (Nitrous
Dioxide, Nitric Oxide)
Cyanide
Carbon Dioxide
Carbon Monoxide
Slight Irritates deep portions of lungs; Methylene blue; Ca
Heavier than air
brown slight methemoglobin gluconate
Cherry
Blocks cytochrome oxidases Nitrite-thiosulfate
red
Dark Displaces oxygen Oxygen; fresh air Heavier than air
Bright Reduces ability of hemoglobin to Fresh air; oxygen + 5% CO2,
Lighter than air
red carry O2 (stable) thionine solution
Prevention

Forages

z Cut forage late in the day on a warm, sunny day.

z Analyze forage for nitrate before feeding suspect material.
z A diphenylamine-based test for screening is available.

z Wait as long as possible before harvesting; nitrates decrease with maturity of plant.
z Wait at least 1 week prior to harvest following a rain which ended a drought.
z Harvest corn for silage rather than green-chop. Ensiling will decrease nitrates by 30 - 50% after 1 - 2 months.
z For each of the following, cut higher on the stalk: corn for silage or green chop, or forage for hay.
z Feed a grain supplement to increase the conversion of nitrates to protein.
z Dilute with feed known to be low in nitrate.
z If hay must be salvaged:
z Grind hay and determine nitrate concentration.

z Mix with good hay until concentration is no more than 0.5% nitrate.

z Supplement diet with vitamins A, D, and E and iodized salt.

z Animals should be well fed before their first access to suspect feed to decrease the initial amount consumed as a
bolus.

Water

z Nitrate additions to ground water can be reduced by limiting nitrogen application, in the form of fertilizer and animal
waste to amounts that can be taken up by plants. Soil type influences percolation and time of year is a factor in plant
uptake.
z Wide green bands with abundant native plants tend to reduce runoff of nitrate into streams, ponds, and lakes.
z Nitrate can be reduced in water with ion exchange and reverse osmosis although an economic assessment is often
necessary to determine whether this is a practical alternative.
Johnsongrass. Sorghum halepense (L.) Pers. 1, stout stem base, roots, and a young rhizome; 2, panicle; 3, group of spikelets;
4, section of stem showing base of leaf; 5, seed. Perennial, reproducing by large rhizomes and seeds. Root system freely
branching, fibrous. Rhizomes stout, creeping, with purple spots, usually with scales at the nodes. Stems erect, stout, from 11/2
to 6 feet (0.45 to 1.8 m) or more tall. Leaves alternate, simple, smooth, 6 to 20 inches (15 to 50 cm) long, about 1/2 to 11/2
inches (1.3 to 3.8 cm) wide. Panicles large, purplish, hairy. Seed nearly _ inch (3 mm) long, oval, reddish-brown, marked
with fine lines on surface, bearing a conspicuous awn easily broken off. Found especially on rich soil. Troublesome in corn
and soybeans on overflow bottoms. Difficult to control or eradicate. Listed as noxious weed in states where found.

Images

z Johnson grass (Sorghum halapense). Knight A.P. and Walter R.G. (Eds.). A Guide to Plant Poisoning of Animals in
North America. Ithaca: International Veterinary Information Service (www.ivis.org), 2003. - To view this image in
full size go to the IVIS website at www.ivis.org . -
z Sorghum spp.. Source: Cornell University, Poisonous Plants Informational Database
(www.ansci.cornell.edu/plants/index.html). - To view this image in full size go to the IVIS website at www.ivis.org . -

z Johnson grass, Sorghum halapense - Google Image Search. - To view this image in full size go to the IVIS website at
www.ivis.org . -
Chlorates

Major Species Usual Time of Onset Usual Duration (if survives) Full Table for
Methemoglobin
All species Hours Hours to days Producers

Source

z Chlorate salts (usually sodium or potassium chlorate used as herbicides and defoliants, more commonly used in the
past). Looks like salt, therefore can be accidentally mixed in feed.
z Careless disposal of chlorates.
z Mistaken use of chlorates for sodium chloride in animal feeds.
z Improper application of the dry salt with clumps of chlorate being left on ground.
z An average book of 20 matches contains 200 mg of potassium chlorate.

Toxicity

z According to Osweiler et al. (1985), the LD for most species is 0.5 - 1 g/kg; but for chlorate salts, Gosselin et al. lists
the toxicity rating as 4 which translates into a potentially lethal dose of 50 - 500 mg/kg. Monogastrics and ruminants
are of similar sensitivity.
z Chlorate decomposes in approximately 7 days in warm, moist soil.

Mechanism of Action

z Powerful oxidizing agent which converts hemoglobin (Fe II) to methemoglobin (Fe III).
z Chlorate is not inactivated in the reaction and continues to form methemoglobin until excreted - may also result in
hemolysis.
z Chlorate is an irritant of the gastrointestinal tract.

Signs

z Onset often occurs within 1 hour. Since chlorates don't require reduction before they are active, the onset of action is
faster than that observed in nitrates. This is the reason that monogastrics are just as sensitive as ruminants.
z Ataxia.
z Prostration.
z Hypersalivation.
z Purgation.
z Hematuria and hemoglobinuria.
z Cyanosis and dyspnea.
z Chocolate-brown blood
z Methemoglobin.

Lesions

Dark brown tissues and blood, congested organs, blood-stained urine, erosions in stomach (abomasum) and
duodenum.

Diagnosis

z Methemoglobin and history of chlorate exposure; chlorate determinations in plasma, urine, or aqueous humor.
z Differentials:
 z Nitrate/nitrite toxicosis.
z Acetaminophen toxicosis in SA.
z Local anesthetic toxicosis.

Treatment

z Gastric lavage.
z Intravenous administration of methylene blue at 4 mg/kg in dogs, 10 - 15 mg/kg in cattle using a 2 - 4% solution; may
have to retreat - treat until blood no longer turns dark on standing. Low doses of methylene blue (1.5 mg/kg) have
recently been reported to be beneficial to cats with nitrite-induced methemoglobinemia. Methylene blue given in the
absence of methemoglobinemia can cause Heinz body formation.
z Vitamin C may be given to dogs and cats at 20 mg/kg. Low doses of methylene blue (1.5 mg/kg) have recently been
reported to be beneficial to cats with nitrite-induced methemoglobinemia. Methylene blue given in the absence of
methemoglobinemia cause Heinz body formation.

Prevention

z Remove animals from area treated with chlorates.

z If not feasible, give animals adequate salt intake at least 1 week prior to application and apply chlorates in form of a
spray.
z Safety cannot be assured by this approach.
z Properly dispose of unused material.
 References

Acetaminophen

1. Rumbeiha WK, Oehme FW. Methylene blue can be used to treat methemoglobinemia in cats without inducing heinz
body hemolytic anemia. Vet Hum Toxicol 1992; 34:120-122.
2. Savides MC, Oehme FW, Leipold HW. Effects of various antidotal treatments on acetaminophen toxicosis and
biotransformation in cats. Am J Vet Res 1985; 46:1485-1489.

Naphthalene

1. Fukuda T, Koyama K, Yamashita M, Koichi N, Takeda M. Differentiation of naphthalene and paradichlorobenzene

mothballs based on their difference in specific gravity. Vet Hum Toxicol 1991; 33:313-314.
2. Hayes WJ. Pesticides Studied in Man. Baltimore: Williams and Wilkins, 1982; 136.
3. Rumbeiha WK, Oehme FW. Methylene blue can be used to treat methemoglobinemia in cats without inducing heinz
body hemolytic anemia. Vet Hum Toxicol 1992; 34:120-122.

Local Anesthetics

1. Rumbeiha WK, Oehme FW. Methylene blue can be used to treat methemoglobinemia in cats without inducing heinz
body hemolytic anemia. Vet Hum Toxicol 1992; 34:2.

Nitrate - Nitrite

1. Osweiler G. Clinical and Diagnostic Veterinary Toxicology, 3rd ed. Dubuque: Kendall/Hunt Publishing Company,
1984; 460-466.
2. Howard JL, ed. Current Veterinary Therapy: Food Animal Practice. WB Saunders Co, 1981; 443-435.
3. Rumbeiha WK, Oehme FW. Methylene blue can be used to treat methemoglobinemia in cats without inducing heinz
body hemolytic anemia. Vet Hum Toxicol 1992; 34:2.

Chlorates

1. Osweiler, et al. Clinical and Diagnostic Veterinary Toxicology, 3rd ed. Dubuque: Kendall/Hunt Publishing Company,
1984.
2. Rumbeiha WK, Oehme FW. Methylene blue can be used to treat methemoglobinemia in cats without inducing heinz
body hemolytic anemia. Vet Hum Toxicol 1992; 34:2.

All rights reserved. This document is available on-line at www.ivis.org. Document No. A2651.0899.