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Toxicology of selected pesticides, drugs, and chemicals. Anticoagulant, cholecalciferol, and bromethalin-based rodenticides.

The control of rodent pests is a continuing goal of mankind. To this end, a multitude of rodenticides have been produced, each designed to kill rodents by exerting their toxic effects on various body systems. As examples, veterinarians have had to manage companion animal poisonings due to anticoagulant, sodium fluoroacetate (compound 1080), thallium, barium carbonate, and zinc phosphide-based rodenticides. Many of these rodenticides were introduced because of their anticipated safety in relation to nontarget species; unfortunately, this has not been the case. Veterinarians must attempt to identify the specific rodenticide involved in poisoning cases. Therapeutic success in these poisonings is often more dependent upon symptomatic and supportive care rather than the use of antidotal therapy.

Aniline Compounds

Difethialone (LM-2219): a new anticoagulant rodenticide for use against warfarin-resistant and -susceptible strains of Rattus norvegicus and Mus musculus.

Data on the efficacy of difethialone, a new anticoagulant rodenticide derived from 4-hydroxybenzothiopyranone against rats and mice, are reported. After one day of feeding with 25 mg/kg of bait, 100 and 85%, respectively, male and female warfarin-susceptible Rattus norvegicus were killed. With the resistant strain, mortality was 90 and 94% for males and females, respectively. 100 and 96% of warfarin-sensitive males and females, respectively, were killed after 3 days of feeding with the same bait. Those of the resistant strain were respectively, 100 and 93%. In Mus musculus, after one day of feeding, mortality was 97, 94, 95 and 93% for males and females of susceptible and resistant strains, respectively, and 100, 97, 100 and 91% after 3 days of feeding. No significant difference in bait intake was observed between 25 mg/kg difethialone bait and a control bait. Compared to other anticoagulant rodenticides, these results show that difethialone is a promising rodenticide.

4-Hydroxycoumarins

Toxicity of a vitamin D3 rodenticide to dogs.

As a follow-up to an investigation of 2 dogs that died as a result of apparent toxicosis attributable to a cholecalciferol-containing rodenticide, we tested the toxicity of this product in dogs. Two groups of 2 dogs each were fed amounts of rodenticide that provided 20 and 10 mg of cholecalciferol/kg of body weight (approx one fourth and one eighth of the published LD50, respectively). All dogs developed hypercalcemia and hyperphosphatemia and then died. Major lesions were gastrointestinal hemorrhage, myocardial necrosis, and mineralization of vascular walls. Our data indicate that cholecalciferol-containing rodenticides pose a much greater hazard to dogs than was previously believed.

Animals

Laboratory evaluation of pyriminyl used as a rodenticide against the lesser bandicoot rat, Bandicota bengalensis.

The properties of pyriminyl (N-3-pyridylmethyl-N'-p-nitrophenyl urea) as a rodenticide against the lesser bandicoot rat (Bandicota bengalensis) in Rangoon, Burma, were investigated in the laboratory. The acute LD 50 and LD 95 dose of orally administered pyriminyl for B. bengalensis were found to be 6.7 mg/kg and 23.0 mg/kg of body weight respectively. When caged bandicoots were given a choice between plain and poisoned baits, the optimum rodenticidal concentration in the bait was found to be 0.25-0.5%. Symptoms of pyriminyl poisoning appear from 1 to 4 h after feeding starts, giving individual animals time to consume from 2 to over 30 LD 50 doses of 0.5% pyriminyl before feeding stops. Deaths occurred from 4 to 96 h after either oral dosing or free-choice feeding. There appeared to be no significant aversion to the poison at 0.25% or 0.5% concentration in foods. The potential hazards and use of pyriminyl as a field bait against populations of B. bengalensis are discussed.

Administration, Oral

Laboratory evaluation of gophacide as a rodenticide for use against Rattus norvegicus and Mus musculus.

Laboratory tests were carried out to assess the efficacy of gophacide as a rodenticide against the Norway rat (Rattus norvegicus) and the house mouse (Mus musculus). Results of feeding tests with wild animals suggest that the compound would be more useful against mice than rats, and that 0.3% would be a near optimal concentration for field trials for both species. The hazards of using gophacide as a rodenticide are discussed.

Amidines

Laboratory evaluation of difenacoum as a rodenticide.

The efficacy of difenacoum as a new anticoagulant rodenticide was evaluated by blood coagulation studies and laboratory feeding tests using warfarin-resistant and non-resistant common rats (Rattus norvegicus), ship rats (R. rattus) and house mice (Mus musculus). Prothrombin assays indicated that the compound had as marked an activity with warfarin-resistant common rats as coumatetralyl had with non-resistant animals. Feeding tests confirmed that 0-005% would be a near-optimal concentration for field use, although there was some evidence of unpalatability. Results with ship rats and house mice were less favourable. Trials with enclosed colonies of warfarin-resistant mice confirmed the laboratory finding that although difenacoum was more effective than all other currently used anticoagulants, it was unlikely to give complete control. It is concluded that difenacoum is a valuable new rodenticide, especiaaly for controlling warfarin-resistant common rats.

4-Hydroxycoumarins

Multicomponent determination of 4-hydroxycoumarin anticoagulant rodenticides in blood serum by liquid chromatography with fluorescence detection.

A sensitive liquid chromatographic method was developed for the analysis of 4-hydroxycoumarin anticoagulant rodenticides in blood serum. The method can simultaneously measure the serum levels of five anticoagulant rodenticides: brodifacoum, bromadiolone, coumatetralyl, difenacoum, and warfarin. Serum proteins are precipitated with acetonitrile and the supernatant is mixed with ethyl ether. The organic phase is separated, evaporated to dryness, and the residue subjected to chromatographic analysis. The anticoagulants are separated by reversed-phase gradient chromatography with fluorescence detection at an excitation wavelength of 318 nm and emission wavelength of 390 nm. Extraction efficiencies of 68.1 to 98.2% were obtained. The within-run precision (CV) ranged from 2.19 to 3.79% and the between-run precision (CV) from 3.72 to 9.57%. The anticoagulants can be quantitated at serum levels of 10 to 20 ng/mL.

4-Hydroxycoumarins

Case studies on second-generation anticoagulant rodenticide toxicities in nontarget species.

Specimens from 10 cases of second-generation anticoagulant rodenticide poisoning in dogs and cats were submitted to the Texas Veterinary Medical Diagnostic Laboratory during 1986 and 1987. The clinical signs most frequently observed were lethargy, dyspnea, and ventral hematomas; common necropsy findings included hemoperitoneum, hemothorax, and pulmonary hemorrhage. In the instances when histopathological examination of the tissue was done, it supported a diagnosis of coagulopathy. The presence of anticoagulants in serum or liver was confirmed by high pressure liquid chromatography, gas chromatography/mass spectrometry, or a combination of the two. Five cases of brodifacoum poisoning, 2 of bromadiolone, and 3 of diphacinone toxicity were verified. Concentrations of these rodenticides ranged from approximately 0.001 to 12 ppm.

Animals

Trials of the anticoagulant rodenticide WBA 8119 against confined colonies of warfarin-resistant house mice (Mus musculus L.).

The efficacy of the newly developed anticoagulant rodenticide WBA 8119 was evaluated against the house mouse (Mus musculus L.) using individual and family groups of warfarin-resistant animals. WBA 8119 at 0-002%, 0-005% and 0-01% in pinhead oatmeal bait gave complete kills of mice in 'no-choice' feeding tests carried out in cages and small pens. In replicated 21-day treatments on families of mice confined in larger pens conditioned to feeding on plain foods, the overall mortalities obtained using the three formulated poison baits were 71/72, 62/63 and 57/57 respectively. The results of the WBA 8119 toxicity tests are considered in relation to previous findings on other anticoagulant rodenticides, particularly difenacoum. In equivalents tests, WBA 8119 performed better than difenacoum. The data thus suport the laboratory findings that WBA 8119 is the most active anticoagulant so far tested for the control of warfarin-resistant house mice.

4-Hydroxycoumarins

Trials of the anticoagulants rodenticide WBA 8119 against confined colonies of warfarin-resistant house mice (Mus musculus L.).

The efficacy of the newly developed anticoagulant rodenticide WBA 8119 was evaluated against the house mouse (Mus musculus L.) using individual and family groups of warfarin-resistant animals. WBA 8119 at 0-002%, 0-005% and 0-01% in pinhead oatmeal bait gave complete kills of mice in 'no-choice' feeding tests carried out in cages and small pens. In replicated 21-day treatments on families of mice confined in larger pens and conditioned to feeding on plain foods, the overall mortalities obtained using the three formulated poison baits were 71/72, 62/63 and 57/57 respectively. The results of the WBA 8119 toxicity tests are considered in relation to previous findings on other anticoagulant rodenticides, particularly difenacoum. In equivalent tests, WBA 8119 performed better than difenacoum. The data thus support the laboratory findings that WBA 8119 is the most active anticoagulant so far tested for the control of warfarin-resistant house mice.

4-Hydroxycoumarins

Hypercalcemia secondary to cholecalciferol rodenticide toxicosis in two dogs.

Hypercalcemia secondary to cholecalciferol rodenticide toxicosis was identified in two dogs. The first dog died shortly after admission. The second dog responded to treatment with sodium chloride solution, prednisolone, furosemide, and calcitonin. Treatment was needed for a longer period than anticipated and the serum calcium concentration did not stabilize for approximately one month. Although not conclusively demonstrated, calcitonin was considered the cause of severe anorexia. This new class of rodenticides has great toxic potential for dogs, and it is recommended that serum calcium concentration be carefully monitored as treatment for hypercalcemia is gradually withdrawn.

Animals

[Toxicity to rodenticides and its treatment].

A case is presented of exposure to a rodenticide which caused a severe disorder of hemostasis, including macroscopic hematuria, several months after the patient was exposed to it. Response to treatment with vitamin K and barbiturates came only after 70 days. The cause-and-effect relationship between exposure to the rodenticide and the disorder in hemostasis is based on the clinical course and on the late response to treatment.

Adult

Hypercalcemia associated with rodenticide poisoning in three cats.

Hypercalcemia (12.0 to 18.3 mg/dl) was detected in 3 cats that had eaten a rodenticide that contained cholecalciferol. Clinical signs included lethargy, anorexia, vomiting, and polydipsia. Treatment with furosemide and fluids administered IV resulted in normalization of the serum calcium concentration and in remission of the clinical signs in 2 cats. One cat with a serum calcium concentration of 18.3 mg/dl did not have clinical signs, was not treated, and was reportedly normal 9 months after initial examination. We attributed the uniformly favorable outcome of exposure to the rodenticide in these cats to the small quantity of the toxin ingested.

Animals

Kinetics of bromadiolone, anticoagulant rodenticide, in the Norway rat (Rattus norvegicus).

Rats (Rattus norvegicus) dosed orally with the rodenticide bromadiolone (0.8 and 3 mg/kg) were sacrificed in groups of 4 rats at various times up to 97 hours after administration. Bromadiolone was assayed in plasma, liver and kidney by an HPLC method. The compound disappeared slowly from the organism with a half-life of 25.7 hours for the 0.8 mg/kg dose and 57.5 hours for the 3 mg/kg. Concentrations in liver were rapidly established and were 14- to 46-fold higher than plasma concentrations. 97 hours after 3 mg/kg dose, liver concentrations were about 1.5 micrograms/g. Bromadiolone levels in kidney were slightly higher than those observed in plasma, with a longer half-life.

4-Hydroxycoumarins

The toxicity and mechanism of action of bromethalin: a new single-feeding rodenticide.

Bromethalin is a new rodenticide for the control of commensal rodents. Doses in excess of the LD50 (2 mg/kg in rats) will cause death within 8-12 hr and it is preceded by one to three episodes of clonic convulsions with death usually due to respiratory arrest. Multiple low doses or sublethal intoxication yields hind leg weakness and loss of tactile sensation in rodents. Histopathology of the brain and spinal cord of these animals revealed a spongy degeneration of the white matter which was shown upon ultramicroscopic examination to be intramyelenic edema. No inflammation or cellular destruction of neuronal tissue was noted. LD50 values ranged from 1.8 mg/kg in the cat to approximately 13 mg/kg in rabbits. The only apparent nonsusceptible species was the guinea pig which could tolerate doses in excess of 1000 mg/kg without effect. Identification of the desmethyl metabolite was demonstrated in the blood and liver of treated animals by comparison of chromatographic retention times to that of a reference standard, but direct mass spectral identification was unsuccessful in part due to the low dose which could be administered. Therefore, the metabolism of bromethalin was studied by indirect means. Animals were pretreated with three inducers of microsomal drug metabolism: phenobarbital, 3-methylcholanthrene (3MC), and Aroclor 1254 (Aroclor) and one inhibitor, SKF-525A. Pretreated mice or rats were given an LD50 dose of bromethalin or the desmethyl analog and the percentage of surviving animals was determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Aniline Compounds

Determination of diastereoisomers of bromadiolone, an anticoagulant rodenticide, in animal tissues by high-performance liquid chromatography.

Two components isolated by semi-preparative normal phase high-performance liquid chromatography (HPLC) of bromadiolone reference material were tentatively identified as diastereoisomeric forms. Examination by mass spectroscopy confirmed this identification and supporting evidence was provided by identical UV fluorescence characteristics. The separated isomers were used to examine the chromatographic properties of bromadiolone in ion-pair, ion-suppression and weak ion-exchange HPLC modes. Conditions suitable for the analytical determination of the individual diastereoisomers were established for each mode. The influence of mobile phase pH on the resolution of coumarin-based rodenticides by weak ion-exchange HPLC on an aminopropyl-bonded phase was studied. Clean-up techniques for the determination of residues of bromadiolone in animal tissue extracts were compared. A combined gel permeation and adsorption chromatographic procedure was preferred for sensitive assay; it permitted the use of either fluorescence or UV detection. The lower practical limit of determination of each isomer in animal tissues was 0.005 mg kg-1 using UV detection and 0.0005 mg kg-1 using fluorescence detection.

4-Hydroxycoumarins

Diagnosis and therapy of anticoagulant rodenticide intoxications.

The mechanism of toxicity and agents of anticoagulant rodenticides are discussed. The diagnosis of anticoagulant poisoning is outlined and discussed by applying clinical, laboratory, and therapeutic response measures as a means to confirm poisoning. Additional therapeutic concerns and newly developed diagnostic tests are discussed. Application of the therapeutic and diagnostic measures provides a successful plan to manage anticoagulant poisonings.

Animals

Laboratory test of seven rodenticides for the control of Mastomys natalensis.

Laboratory feeding tests were carried out to assess the efficacy of seven rodenticides against Mastomys natalensis. The poisons (warfarin, coumatetralyl, difenacoum, brodifacoum, bromadiolone, calciferol and zinc phosphide) were all toxic at the concentrations normally used against Rattus norvegicus (Berk.), although several were unpalatable. Trials are now needed to demonstrate the relative efficacy of these poisons in the field, but it is likely that, given suitable bait formulations, they would all be useful as practical control agents.

4-Hydroxycoumarins