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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↗

Profiling, mimicking and masking the flavor of a selected rodenticide.

In Experiment 1, rats drank strychnine solution followed by an injection of LiCl. Generalization of learned strychnine avoidance to 4 non-toxic flavors was then assessed. Additional conditioning and generalization trials followed until 24 flavors had been presented. In Experiment 2, rats were conditioned to avoid individual flavors, or flavor mixtures concocted on the basis of avoidance generalization observed in Experiment 1. Tests followed for generalization of learned avoidance from the simple flavors to the mixtures, from the mixtures to the simple flavors, and from either to strychnine. In Experiment 3, two concentrations of NaCl were mixed with strychnine or one of the flavors (SOA) used in the previous experiments. These stimuli, as well as SOA alone, strychnine alone, and each of the NaCl concentrations, were presented to rats during conditioning. Generalization followed, as in the previous experiments. In Experiment 1, strychnine avoidance generalized to 'bitter' flavors (ps less than 0.01). In Experiment 2, avoidance of flavor mixtures generalized more strongly to strychnine than did learned avoidance of simple flavors (ps less than 0.01). In Experiment 3, NaCl masked or otherwise suppressed the 'bitter' flavor of strychnine or SOA insofar as no groups conditioned with a 'bitter'-salt mixture generalized avoidance to 'bitter' alone (ps less than 0.01). Rats are therefore capable of recognizing the flavor components of strychnine. Moreover, when these components are mixed in proportion to the degree of generalized avoidance, a mimic (either in terms of flavor characteristics or perceived intensity) of strychnine is obtained. Avoidance learning appears useful in the development of rodenticide baits and pre-bait formulations.

Animals↗

Effects of the rodenticide Vacor on cultured rat pancreatic beta cells.

Human ingestion of the rodenticide Vacor has been implicated in the onset of diabetes mellitus. We report here studies of the effects of Vacor on cultured rat pancreatic beta cells and fibroblasts to determine if this agent preferentially intoxicates insulin-producing beta cells. Cytotoxicity in beta cells was monitored both by phase contrast microscopy and the release of insulin into the culture medium. Changes in cellular protein were correlated with morphologic observations to determine fibroblast viability. Beta cells were 10-fold more sensitive to the toxic effects of Vacor than were fibroblasts. Vacor toxicity was reduced by treatment with nicotinamide but was unchanged by 3-O-methyl glucose. Both of these agents reduced the toxic effects of the known beta cell poison streptozotocin. The present studies indicate that Vacor possesses a toxic affinity for pancreatic beta cells and that its mechanism of toxicity may be similar but not identical to that of streptozotocin.

Animals↗

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↗

Wartime rat control, rodent ecology, and the rise and fall of chemical rodenticides.

The story of how World War II stimulated the development of DDT, and the ensuing postwar dependence on such chemical insecticides, is well known. However, less recognition has been given to the wartime efforts to synthesize new rodenticides to fight rat-borne epidemics. Baltimore, Maryland served as the site for field tests of the powerful new compound alpha naphthyl thiourea (ANTU) from 1942-1946. This experimental campaign sparked debates over the efficacy of controlling rats via chemical warfare instead of environmental sanitation, which led to the ironic conclusion that urban rat control demanded an ecological, rather than technological, approach.

Animals↗

Lethal paradoxical cerebral vein thrombosis due to suspicious anticoagulant rodenticide intoxication with chlorophacinone.

Superwarfarin exposure is a growing health problem, described in many countries. The authors report a case of suspicious chlorophacinone poisoning with a problematic diagnosis. They review the literature and discuss particularities of anticoagulant rodenticide intoxication, as well as the apparent contradiction between anticoagulant intoxication and lethal thrombosis.

Adult↗

Determination of chlorophacinone and diphacinone in commercial rodenticides by liquid chromatography-UV detection and liquid chromatography-electrospray ionization mass spectrometry.

This work describes a simple method for the determination and mass-spectrometric confirmation of the indanediones in commercial rodenticides. The sample is sonicated in methanol containing 2% formic acid and analyzed by liquid chromatography-UV detection. Once retention time and UV-Vis spectrum provide tentative identification, mass-spectrometric confirmation is obtained by analyzing a second aliquot by LC and electrospray ionization mass spectrometry (LC-ESI-MS). The extensive fragmentation of the indanedione molecule under MS/MS conditions provides sufficient structural information for positive identification on analyte levels as low as 20 ng on column.

Chromatography, Liquid↗

The multi-residue determination of coumarin-based anticoagulant rodenticides in animal materials by high-performance liquid chromatography.

The rodenticides brodifacoum, difenacoum, coumatetralyl and warfarin are determined in animal relicta by high-performance exclusion chromatography on porous silica. The first three compounds are not separated, but are subsequently differentiated by adsorption or reversed-phase high-performance liquid chromatography of the appropriate eluate fraction collected from the exclusion column. The method is rapid, and clean-up (on Sep-Pak silica cartridges) is simple. Mean recoveries from spiked substrates were generally above 80% at levels of 0.1-1.0 mg/kg. Routine limits of determination are about 0.05-0.1 mg/kg for warfarin and about 0.02 mg/kg for the other compounds. If analysis for warfarin is not required, the latter limit can be lowered to about 1 microgram/kg by a slight modification to the clean-up step.

4-Hydroxycoumarins↗

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↗

A scheme for the placement of rodenticide baits for rat eradication on confinement livestock farms.

We investigated whether the allocation of rodenticide baiting points to specific structural elements would result in complete rat eradication on livestock farms, as opposed to assigning the baiting points only to places where there were obvious signs of rat activity. The goal was to establish an effective rodent-control program that is easy for untrained persons to conduct.Rat-control strategies were examined on 25 farms in Velen (Muensterland), Germany, where an average of 20% of trapped rats were resistant for bromadiolone according to a blood-clotting response (BCR) test. All farms were investigated for signs of rat activity prior to and after the control measure. Differences in the percentage level of farmer compliance in setting up the baiting points as prescribed were analysed for each type of baiting point and in total, and were compared between the group of farms which achieved complete rat eradication and those which did not. Farms achieving complete eradication had an average of 81% compliance with prescribed control plans, whereas a significantly lower compliance level of only 51% was recorded on farms that did not achieve eradication. A >/=75% level of implementation of the control plan always resulted in complete control success. The new method of bait-point allocation was incorporated into a self-explanatory computer program, which was verified to be effective during a rat-control campaign in the restricted area after an outbreak of classical swine fever near Soltau in northern Germany, in July 2001. This program, which is available on the Internet, enables the creation of individualised rat-control plans, including complete documentation of the control measure.

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↗

Spatial and temporal analysis of second-generation anticoagulant rodenticide residues in polecats (Mustela putorius) from throughout their range in Britain, 1992-1999.

Polecats (Mustela putorius) in Britain are currently expanding their range eastwards from Wales to reoccupy central and eastern areas of England. Second-generation anticoagulant rodenticides (SGARs), to which polecats are exposed by eating contaminated prey, are used more extensively in these central and eastern regions, leading to fears of increased exposure, and possible resultant mortality. We measured bromadiolone, difenacoum, flocoumafen and brodifacoum concentrations in the livers of 50 polecats from areas that included newly recolonised habitats and found that at least one SGAR was detected in the livers of 13 out of 37 (35.1%) male and 5 out of 13 (38.5%) female polecats. Difenacoum and bromadiolone were detected most frequently. We then combined these data with measurements on another 50 individuals from earlier studies to create a dataset for 100 polecats collected throughout the 1990s from across the whole of their current range. Using this dataset, we determined if there was any evidence that contamination in polecats had increased during the 1990s and whether animals from England were more contaminated than those from Wales, as might be expected given regional differences in the patterns of SGAR use. Overall, 31 of the 100 polecats analysed to date contained SGAR residues. The incidence was a little higher (40%) in animals that died between January and June and this probably better reflects the overall proportion of animals that are sub-lethally exposed. There was no statistically significant change during the 1990s in the proportion of polecats exposed to SGARs nor any evidence that greater use of SGARs in England resulted in more contamination of polecats. Contrary to expectation, the proportion of animals that contained difenacoum was marginally higher in Wales than elsewhere.

4-Hydroxycoumarins↗

Immunotoxicity of epicutaneously applied anticoagulant rodenticide warfarin: evaluation by contact hypersensitivity to DNCB in rats.

The immunotoxicity of epicutaneously administered anticoagulant rodenticide warfarin (WF) was examined in this work by using experimental contact hypersensitivity (CHS) reaction to hapten dinitrochlorobenzene (DNCB). WF (0.05 and 0.5 mg/kg) administration 24 h before the induction of CHS does not change expression of CHS evaluated by ear swelling assay. Regional draining lymph node response during sensitization phase was characterized by decreased cellularity but increased spontaneous and IL-2 stimulated proliferation of draining lymph node cells (DLC). No changes in IL-2 production and in numbers of CD25(+) cells were noted and even decreased proliferative index (ratio of IL-2 stimulated to unstimulated DLC proliferation) was detected. Increase in granulocyte activity (MTT reduction and adhesion to plastic) was noted following application of WF solely with further increase following subsequent application of DNCB, when granulocyte activation (NBT reduction) was noted also. Access of WF into general circulation might be responsible for observed changes, what was supported by ex vivo changes in DLC and granulocyte functions assessed before initiation of sensitization and by in vitro effect of exogenous WF as well. Differential effects of WF on lymphocytes and granulocytes noted in this study highlight the need for simultaneous testing of both cell type activity what might constitute a more integrated approach in immunotoxicity studies.

Administration, Topical↗

Use of an ion-pairing reagent for high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry determination of anionic anticoagulant rodenticides in body fluids.

The on-line combination of high-performance liquid chromatography with mass spectrometry (HPLC-MS) has become a powerful tool for trace analysis thanks to the developments in interface techniques. However, non-volatile salts such as ion-pairing reagents are considered to be incompatible with HPLC-MS systems; they cause drops in analyte signals because of contamination of mass analyzers and also because of blocking of the capillary transferring ions from atmospheric pressure to the vacuum manifold. In this work, a new type of ion-pairing reagent, di-n-butylammonium acetate (DBA), was evaluated for use in HPLC-MS. DBA did not cause these problems to HPLC-MS systems; a possible explanation might be that DBA decomposed to volatile compounds under APCI conditions. In addition, DBA was very useful for obtaining sharp peaks, which resulted in high sensitivity. With this ion-pairing reagent, we developed a procedure for the measurement of five (including internal standard) anticoagulant rodenticides in whole blood and urine samples by SIM detection of [M-H]- ions. Calibration range, recoveries and precision of the method were examined; detection limits as low as 1-5 ng/ml blood sample or 0.5-2.5 ng/ml urine sample were achieved.

Animals↗

QSAR study on competition binding of rodenticides (PATs) to H(1) receptor in rat and guinea pig brain.

A quantitative structure-activity relationship (QSAR) study on binding activity for a series of rodenticides (PAT analogues) to the [(3)H]-mepyramine-labeled H(1) receptor in rat and guinea pig brain is attempted topologically. From the pool of molecular descriptors we, observed that the most significant descriptor is the molecular redundancy index (MRI). Multiple regression analysis gave excellent results with MRI upon introduction of some dummy parameters (indicator parameters). Predictive ability of the proposed models is discussed using cross-validation parameters.

Aniline Compounds↗

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↗

The susceptibility of Bandicota bengalensis from Rangoon, Burma to several anticoagulant rodenticides.

The baseline susceptibility of the lesser bandicoot rat, Bandicota bengalensis, from Rangoon, Burma, to five anticoagulant rodenticides was established with no-choice feeding in the laboratory. The susceptibility of lesser bandicoots to the several poisons (brodifacoum, difenacoum, diphacinone, coumatetralyl, and warfarin) was such that they were offered at a 0.001% concentration. B. bengalensis was most susceptible to brodifacoum, and in descending order, difenacoum, coumatetralyl, diphacinone and warfarin. In comparison with Rattus norvegicus on warfarin at 0.005%, B. bengalensis proved more susceptible. Feeding tests at 0.005% concentration indicated that a 1-day feeding on brodifacoum and difenacoum would result in complete mortality, whereas coumatetralyl and warfarin would require 4 days feeding to a 100% kill. Brodifacoum and difenacoum are recommended at 0.002-0.005% bait concentrations and coumatetralyl at 0.005--0.01% concentrations for the control of B. bengalensis in the field in Rangoon. The use of any anticoagulant material in rat control should be alternated with acute toxicants to retard the possible development of anticoagulant resistance.

4-Hydroxycoumarins↗