Iophenoxic Acid Research Articles

Competitive Displacement Restores the Hyperpolarized 15N NMR Signal in Blood Plasma.

Hyperpolarized (HP) NMR can improve the sensitivity of conventional NMR experiments by several orders of magnitude, thereby making it feasible to detect the signal of low sensitivity nuclei such as 13C and 15N nuclei in vivo. Hyperpolarized substrates are usually administered by direct injection into the bloodstream, and interaction with serum albumin can cause rapid decay of the hyperpolarized signal due to the shortening of the spin-lattice (T1) relaxation time. Here we report that the 15N T1 of 15N labeled, partially deuterated tris(2-pyridylmethyl)amine decreases dramatically upon binding to albumin to such an extent that no HP-15 signal could be detected. We also demonstrate that the signal could be restored using a competitive displacer, iophenoxic acid, which binds stronger to albumin than tris(2-pyridylmethyl)amine. The methodology presented here eliminates the undesirable effect of albumin binding and should widen the range of hyperpolarized probes for in vivo studies.

Uptake of baits by wild badgers: Influences of deployment method, badger age and activity patterns on potential delivery of an oral vaccine

In parts of the United Kingdom and Ireland, the European badger is a wildlife host for Mycobacterium bovis (the causative agent of bovine tuberculosis). Badger vaccination is one management option for reducing disease spread. Vaccination is currently achieved by parenteral vaccination of captured badgers, but an oral vaccine delivered in a bait may provide an additional approach in the future. We conducted two field experiments in wild badger populations to identify factors that influence uptake (% of individuals with evidence of bait consumption) of candidate oral vaccine baits. In both instances, baits containing the biomarker iophenoxic acid (as a proxy for the vaccine) were fed at burrows (setts) associated with badger social groups (study A = 48 groups, study B = 40 groups). Badgers were captured following a period of bait deployment to quantify uptake in relation to age, sex and social group. In addition, groups were allocated different treatments and the bait deployment protocol was varied to identify effects on uptake. Study A tested the effects of season, bait type, bait placement and packaging, while study B investigated the effects of bait quantity and badger activity levels. Overall bait uptake was low (Study A = 24 %, Study B = 37 %) but this varied among treatment groups (range 0–58 %). In both studies, bait uptake was significantly higher in cubs than in adults. Uptake was substantially higher where baits were placed directly into sett entrances (rather than under tiles near setts), and by badgers caught at main setts rather than at outlier setts. Season, bait type and packaging did not influence uptake, while increasing the quantity of bait available increased uptake by cubs but not by adults. Levels of badger activity at setts varied over time (suggesting potential disturbance), but were positively associated with levels of bait uptake.

Placebo Oral Rabies Vaccine Bait Uptake by Small Indian Mongooses (Herpestes auropunctatus) in Southwestern Puerto Rico

The small Indian mongoose (Herpestes auropunctatus) is a rabies reservoir in areas of the Caribbean including Puerto Rico, but no rabies vaccination program targeting this host exists. We used two derivatives of iophenoxic acid (IPA) to evaluate placebo oral rabies vaccine bait uptake by mongooses in southwestern Puerto Rico. We hand-distributed baits at an application rate of 200 baits/km2 at three, 400 ha, sites during autumn 2016 and spring 2017. Each site contained 90-100 cage traps in a 100 ha central trapping area. We used ethyl-IPA as a biological marker during the autumn and methyl-IPA during the spring. We live captured mongooses for 10 consecutive days, beginning 1 wk following bait application. We obtained a serum sample from captured mongooses and analyzed the sera for ethyl- and methyl-IPA by liquid chromatography-mass spectrometry. During autumn 2016, 63% (55/87) mongooses sampled were positive for ethyl-IPA. In spring 2017, 69% (85/123) of mongooses were positive for methyl-IPA. Pooling seasons, accounting for recaptures between years, and disregarding marker type, 74% (133/179) unique mongooses were positive for IPA biomarker, indicating bait consumption during either the autumn, spring, or both trials. We conclude that distributing baits at an application rate of 200 baits/km2 is sufficient to reach over 60% of the target mongoose population in dry forest habitats of Puerto Rico.

Bait uptake by wild badgers and its implications for oral vaccination against tuberculosis.

The deployment of baits containing vaccines or toxins has been used successfully in the management of wildlife populations, including for disease control. Optimisation of deployment strategies seeks to maximise uptake by the targeted population whilst ensuring cost-effectiveness. Tuberculosis (TB) caused by infection with Mycobacterium bovis affects a broad range of mammalian hosts across the globe, including cattle, wildlife and humans. The control of TB in cattle in the UK and Republic of Ireland is hampered by persistent infection in European badgers (Meles meles). The present study aimed to determine the best strategy for maximising uptake of an oral vaccine by wild badgers, using a surrogate novel bait deployed at 40 badger social groups. Baits contained a blood-borne biomarker (Iophenoxic Acid, IPA) in order to measure consumption in badgers subsequently cage trapped at targeted setts. Evidence for the consumption of bait was found in 83% (199/240) of captured badgers. The probability that badgers had consumed at least one bait (IPA >10 μg ml-1) was significantly higher following deployment in spring than in summer. Lower uptake amongst social groups where more badgers were captured, suggested competition for baits. The probability of bait consumption was significantly higher at groups where main and outlier setts were provided with baits than at those where outliers were present but not baited. Badgers captured 10–14 days post bait feeding had significantly higher levels of bait uptake compared to those caught 24–28 days later. Uptake rates did not vary significantly in relation to badger age and whether bait was placed above ground or down setts. This study suggests that high levels of bait uptake can be achieved in wild badger populations and identifies factors influencing the potential success of different deployment strategies. The implications for the development of an oral badger vaccine are discussed.

Testing of a palatable bait and compatible vaccine carrier for the oral vaccination of European badgers (Meles meles) against tuberculosis

The oral vaccination of wild badgers (Meles meles) with live Bacillus Calmette–Guérin (BCG) is one of the tools being considered for the control of bovine tuberculosis (caused by Mycobacterium bovis) in the UK. The design of a product for oral vaccination requires that numerous, and often competing, conditions are met. These include the need for a highly palatable, but physically stable bait that will meet regulatory requirements, and one which is also compatible with the vaccine formulation; in this case live BCG. In collaboration with two commercial bait companies we have developed a highly attractive and palatable bait recipe designed specifically for European badgers (Meles meles) that meets these requirements. The palatability of different batches of bait was evaluated against a standardised palatable control bait using captive badgers. The physical properties of the bait are described e.g. firmness and colour. The microbial load in the bait was assessed against European and US Pharmacopoeias. The bait was combined with an edible vaccine carrier made of hydrogenated peanut oil in which BCG vaccine was stable during bait manufacture and cold storage, demonstrating <0.5 log10 reduction in titre after 117weeks’ storage at −20°C. BCG stability in bait was also evaluated at +4°C and under simulated environmental conditions (20°C, 98% Relative Humidity; RH). Finally, iophenoxic acid biomarkers were utilised as a surrogate for the BCG vaccine, to test variants of the vaccine-bait design for their ability to deliver biomarker to the gastrointestinal tract of individual animals. These data provide the first detailed description of a bait-vaccine delivery system developed specifically for the oral vaccination of badgers against Mycobacterium bovis using live BCG.

Feasibility and efficacy of oral rabies vaccine SAG2 in endangered Ethiopian wolves

Diseases are a major cause of population declines in endangered populations of several canid species. Parenteral vaccination efforts to protect Ethiopian wolves (Canis simensis) from rabies have targeted the domestic dog reservoir, or the wolves themselves in response to confirmed outbreaks. Oral vaccination offers a more cost-efficient, safe and proactive approach to protect Ethiopian wolves and other threatened canids from rabies. Field trials of the oral vaccine Rabigen® SAG2Dog were undertaken in the Bale Mountains of southeastern Ethiopia. Four different bait types and three delivery methods were tested in twelve Ethiopian wolf packs, and the oral vaccine (using the preferred bait) was trialled in three packs. Vaccine uptake and immunization rates were measured through direct observations and in live-trapped animals through the assessment of biomarker levels and serological status. Commercial baits were never taken by wolves; goat meat baits had the highest uptake, compared to rodent and intestine baits. Targeted delivery from horseback and nocturnal delivery within a pack’s territory performed favourably compared to random bait distribution. Bait uptake by non-target species was lowest during the nocturnal blind distribution. Of 21 wolves trapped after vaccination, 14 were positive for the biomarker iophenoxic acid (i.e. ingested the bait and most likely pierced the sachet with the vaccine). Of these, 86% (n=12/14) had levels considered sufficient to provide protective immunity to wildlife (⩾0.20IU/ml), and 50% (n=7/14) demonstrated antibody titres above the universally recognised threshold (⩾0.5IU/ml) -the baseline average was 0.09IU/ml (n=12 wolves). All but one of the wolves vaccinated in 2014 were alive 14months later. Our trials confirm the potential for SAG2, delivered in a goat meat bait, to effectively protect Ethiopian wolves against rabies, supporting the initiative for a more efficient and proactive approach to manage and eventually eliminate rabies in Ethiopian wolf populations.

Selective analysis of human serum albumin based on SEC-ICP-MS after labelling with iophenoxic acid.

Human serum albumin (HSA) is the most abundant protein in the human plasma. HSA has several physiological roles in the human body, including storage and transport. Owing to the predominance of albumin in plasma, HSA is often involved in the protein binding of drugs. The aim of this work was to develop a selective, quantitative method for determining albumin in plasma with the purpose of clarifying the fate of metal-based drugs in biological systems. The method can also be applied for determination of urine albumin, which is of relevance in diagnostics of kidney disease. A selective method for quantification of HSA based on labelling the protein with iophenoxic acid (IPA) was developed. Samples were subjected to size exclusion chromatography (SEC) and detection by inductively coupled plasma mass spectrometry (ICP-MS) monitoring iodine and platinum. The iodine signal for the HSA-IPA complex showed linearity in the range 1 to 250 mg L(-1). The precision was 3.7% and the accuracy 100.7% determined by analysis of a certified HSA reference material. The limit of detection (LOD) and limit of quantification (LOQ) were 0.23 and 9.79 mg L(-1), respectively. The method was applied for analysis of HSA in human plasma and urine samples and for studying the binding of cisplatin to proteins in the human plasma.

Iophenoxic Acid and Rhodamine B as Biomarkers of Bovine Tuberculosis Vaccine Bait Uptake by White-tailed Deer

Bovine tuberculosis (bTB), caused by the bacteria Mycobacterium bovis, exists in free-ranging white-tailed deer in portions of northeastern Lower Michigan where deer herds serve as reservoirs and sources for reinfection of livestock herds. Density reduction and vaccination of reservoir host populations could be used together to reduce prevalence or eliminate the disease. Voluntary oral uptake of vaccine by deer is the most feasible mode of delivery. High probability of eliminating bTB would depend, in part, on a high proportion of deer being vaccinated. Chemical biomarkers could be used to estimate the proportion of a deer population consuming baits. Three analogs of iophenoxic acid were evaluated relative to their pharmacokinetic profile in blood-serum, and presence of rhodamine B was evaluated in facial whiskers using captive white-tailed deer.

Iophenoxic acid derivatives as markers of oral baits to wildlife

The bait-marker iophenoxic acid (IPA) and its derivatives are increasingly used for evaluating and optimizing the cost-effectiveness of baiting campaigns on wildlife, particularly on game species such as the wild boar. We aimed to determine whether concentrations of the three main IPA derivatives ethyl, methyl and propyl-IPA measured on thoracic liquid extracts (TLE) of hunted wild boars may be representative of two exposure doses, 40 and 200 mg, from 20 to 217 days after ingestion. Then we developed a method of detection of the three IPA derivatives by LC/ESI-MS-MS in muscle and liver to evaluate the suitability of these two other tissues for monitoring the marked bait consumption and for measuring available residues in the meat of marked animals. Three semi-captive wild boars received 40 mg of each IPA derivative, three received 200 mg, and three, as controls, did not receive IPA. Blood serum was sampled 20, 197 or 217 days after IPA exposure according to animals and to the derivative. Wild boars were shot by gun after the different times of serum sampling times, and TLE, muscle and liver were sampled. Our results suggest that TLE is not a relevant tissue for quantitatively expressing IPA exposure. Due to interference, no analytical method was validated on TLE containing digestive material. On the other hand, quantifications in the muscle and particularly in the liver could discriminate wild boars that had ingested the two IPA doses from 20 days until 7 months after exposure, especially for the two long term markers ethyl and propyl-IPA. So IPA quantifications in the liver sampled on hunted animals appear to be a reliable tool for monitoring bait consumption in the field at a large scale. Nevertheless, whatever the ingested dose, ethyl- and propyl-IPA concentrations measured in the muscle and the liver of tested animals until 217 days after exposure, remained higher than 0.01 mg/kg, the Maximal Residue Limit (MRL) is recommended for molecules for which no toxicological data are available. Based on the range of IPA residues available in these two tissues, implications for humans consuming marked animals are discussed.

Uptake of buried baits by badgers: Implications for rabies control in Great Britain and the delivery of an oral TB vaccine

AbstractThe British Government's contingency plan for the control of wildlife rabies is based on oral vaccination around a focal outbreak, with the option of the targeted lethal control of foxes (Vulpes vulpes) and badgers (Meles meles) using buried poisoned bait. We investigated uptake by badgers of nonpoison meat‐based baits buried along transects for 10 consecutive days in 1995. The original aim was to evaluate the likely effectiveness of this contingency option. However, the results are also pertinent to the development of an oral tuberculosis (TB) vaccine baiting strategy for badgers, for which research has primarily been carried out since this study was completed. Bait disappearance steadily increased over time, reaching an asymptote after 7 days. By incorporating iophenoxic acid biomarkers into baits, we showed that about 51% of subsequently captured badgers had consumed bait, the majority of which did so during the first 7 days. Bait uptake was significantly higher among adults. Rates of uptake by badgers of baits buried along transects could be increased by shortening the prebaiting period or extending the baiting period. However, considerably higher levels of uptake by adults and cubs are likely to be achieved by targeting bait deployment at badger setts, and this is now included in Britain's rabies contingency plan. Although meat‐based baits cannot be used for TB control, the relevance of these results to the delivery of an oral TB vaccine bait to badgers is discussed. © 2012 Crown copyright

Iophenoxic acid as a bait marker for wild mammals: efficacy and safety considerations

Abstract Bait delivery of vaccines, toxicants or contraceptives to target wild mammals is an essential component of management strategies worldwide. Marking agents are required to enable the identification of individuals that consume the baits in order to evaluate and optimize the cost‐efficacy of baiting campaigns. Ethyl‐iophenoxic acid (Et‐IPA) is an organic iodine‐containing compound that has been used increasingly since the 1980s as a bait marker. It binds to proteins in animal blood and therefore can be detected indirectly by measuring plasma or serum iodine concentration or directly by measuring plasma or serum Et‐IPA concentration with liquid chromatography. Methyl‐iophenoxic acid and propyl‐iophenoxic acid can also be used to provide quantitative information on temporal or spatial patterns of bait consumption in a range of mammalian species. We review the suitability of Et‐IPA and its analogues as bait markers in mammals. The highly variable persistence of analogues and in species highlights the need for calibration testing of each compound as a marker for each species and for each proposed use before starting a bait delivery trial. After absorption, the fate of IPAs (by metabolism, distribution and excretion) is variable, but most are bound to plasma proteins. Marking efficacy is therefore high. The implications of exposure to IPAs for predators and humans were analyzed to evaluate its safety for delivery to wild mammals. Although it is highly unlikely that secondary exposure through ingestion could involve sufficiently large quantities to produce adverse effects, further studies are necessary to assess long‐term effects after chronic exposure to these compounds. IPAs can mark animal serum for long periods of time, and improvements in detection methods for them are currently being developed, so that they are a good choice for oral baiting field experiments with mammals.

Crystallographic analysis reveals the structural basis of the high-affinity binding of iophenoxic acid to human serum albumin

BackgroundIophenoxic acid is an iodinated radiocontrast agent that was withdrawn from clinical use because of its exceptionally long half-life in the body, which was due in part to its high-affinity binding to human serum albumin (HSA). It was replaced by Iopanoic acid, which has an amino rather than a hydroxyl group at position 3 on the iodinated benzyl ring and, as a result, binds to albumin with lower affinity and is excreted more rapidly from the body. To understand how iophenoxic acid binds so tightly to albumin, we wanted to examine the structural basis of its interaction with HSA.ResultsWe have determined the co-crystal structure of HSA in complex with iophenoxic acid at 2.75 Å resolution, revealing a total of four binding sites, two of which - in drugs sites 1 and 2 on the protein - are likely to be occupied at clinical doses. High-affinity binding of iophenoxic acid occurs at drug site 1. The structure reveals that polar and apolar groups on the compound are involved in its interactions with drug site 1. In particular, the 3-hydroxyl group makes three hydrogen bonds with the side-chains of Tyr 150 and Arg 257. The mode of binding to drug site 2 is similar except for the absence of a binding partner for the hydroxyl group on the benzyl ring of the compound.ConclusionsThe HSA-iophenoxic acid structure indicates that high-affinity binding to drug site 1 is likely to be due to extensive desolvation of the compound, coupled with the ability of the binding pocket to provide a full set of salt-bridging or hydrogen bonding partners for its polar groups. Consistent with this interpretation, the structure also suggests that the lower-affinity binding of iopanoic acid arises because replacement of the 3-hydroxyl by an amino group eliminates hydrogen bonding to Arg 257. This finding underscores the importance of polar interactions in high-affinity binding to albumin.

Specificity and success of oral-bait delivery to Eurasian wild boar in Mediterranean woodland habitats

Eurasian wild boar (Sus scrofa) is an important reservoir host for pathogens affecting humans and domestic animals. The eradication of these diseases may require the development of control strategies that reduce pathogen transmission between wildlife and domestic animals. Baiting for oral vaccine delivery is often considered for wildlife disease control. The effective and efficient field vaccination of wildlife requires species-specific baits as delivery vehicles for oral vaccines and designing appropriate baiting strategies. The objective of this study was to determine the proportion of young and adult wild boar and non-target animals that consumed baits containing a chemical marker, iophenoxic acid (IPA), in delivery trials conducted in summer in four different sites in the Mediterranean region of Spain where wild boar are abundant. The proportion of wild boar showing IPA markers in serum in autumn ranged from 11.5% to 56.4%. When attending to age classes, 12.6% to 72.7% of young individuals presented IPA. The results evidenced that the percent of wild boar that ingested the baits varied among study sites and age classes. Placing baits inside selective cages (for juveniles) and under heavy pavel stones (for adults) contributed to improve age specificity in bait consumption. We suggest ways for improving the age specificity of bait delivery systems used for young and adult wild boar.

Analysis by LC/ESI-MS of iophenoxic acid derivatives and evaluation as markers of oral baits to deliver pharmaceuticals to wildlife

Iophenoxic acid and its derivatives (methyl, ethyl, and propyl) are organic chemicals used as markers in baiting campaigns to deliver vaccines, pharmaceuticals, contraceptives or poisons to wildlife. In this study we develop a method of detection of IPA derivatives by LC/ESI-MS (using butyl-IPA as internal standard) obtaining a limit of detection and quantification in wild boar ( Sus scrofa) serum of 0.037 μg/ml and 0.123 μg/ml, respectively. The average recovery of IPA derivatives was 88% at levels >0.2 μg/ml, with coefficients of variation <15%. Wild boars in captivity were orally treated with 5 mg/kg b.w. (three adults) or 15 mg/kg b.w (two piglets and three adults) of methyl-, ethyl- and propyl-IPA and the serum levels of these were monitored during 18 months after dosing. Ethyl- and propyl-IPA were detected up to 18 months after a single oral dose in wild boar, especially at 15 mg/kg. Methyl-IPA was detected until 9 months after dosing. Half-lives of methyl-, ethyl- and propyl-IPA were (mean ± SD) 41 ± 5, 183 ± 85 and 165 ± 45 days, respectively. One control piglet not exposed to IPA, but housed in the same facility than treated animals showed detectable IPA levels in serum. Piglets born from mothers exposed to marked baits also showed detectable IPA levels in serum. The high persistence of Et- and Pr-IPA must be considered in the field trials, because the presence of the product at low levels in one animal may not reflect a real ingestion of the marked bait.

Iophenoxic Acid as a Long‐Term Marker for Wild Boar

ABSTRACT Chemical markers are increasingly used to investigate consumption of baits used to deliver vaccines, toxicants, and contraceptives. We evaluated whether ethyl‐iophenoxic acid (Et‐IPA) and propyl‐iophenoxic acid (Pr‐IPA) can be used as long‐lasting systemic bait markers for wild boar (Sus scrofa). We presented captive wild boar with baits treated with either Et‐IPA or Pr‐IPA at 5 mg/kg (low dose), 10 mg/kg (medium dose), and 20 mg/kg (high dose) of body weight. We collected serum from each boar at 5 time points: 5 days, 3 weeks, 6 weeks, 11 weeks, and 39 weeks following ingestion of iophenoxic acid‐treated baits. We detected both Et‐IPA and Pr‐IPA for ≥39 weeks after ingestion. Throughout the trial, the Et‐IPA we found in serum was proportional to the amount eaten. At each time point, animals in the high‐dose group had significantly more Et‐IPA than animals in the low‐dose group. We concluded that both compounds can be used as long‐lasting markers in wild boar and that Et‐IPA can also be employed as quantitative marker to indicate multiple bait uptake. Both compounds have potential applications in the context of vaccination, fertility, and population control campaigns, where baits are used to deliver pharmaceuticals, and in behavioral studies to establish spatial and temporal patterns of bait uptake.

Vaccination of feral pigs (Sus scrofa) using iophenoxic acid as a simulated vaccine

To develop an encapsulation method for delivery of vaccines to feral pigs, and quantify the effect of iophenoxic acid on captive feral pig blood iodine concentrations to assist in investigation of factors affecting vaccine uptake. Feral pigs were administered iophenoxic acid by oral gavage, and consumption was assessed for different encapsulation methods in baits. Blood iodine concentrations were monitored for eight days after consumption. The relationship between dose rate, time since dosing and blood iodine concentration was assessed for gavaged and baited captive feral pigs. Wild feral pigs were baited with PIGOUT baits containing 20 mg of encapsulated iophenoxic acid to simulate a vaccination program. Using knowledge from the pen studies, bait uptake and factors affecting bait uptake were investigated. Bait-delivered iophenoxic acid led to variable and inconsistent changes in blood iodine concentrations, in contrast to pigs receiving iophenoxic acid by gavage. This precluded accurate assessment of the quantity consumed, but still allowed a conservative determination of bait uptake. Iophenoxic acid in smaller capsules was consumed readily. Increasing baiting intensity appeared to increase bait uptake by wild feral pigs, and pigs of varying sexes, ages and weights appeared equally likely to consume baits. Encapsulated liquids can be delivered to feral pigs within baits, should the need to vaccinate feral pigs for fertility or disease management arise. High baiting intensities may be required.

19F NMR spectroscopic characterization of the interaction of niflumic acid with human serum albumin

The interaction of a non-steroidal anti-inflammatory drug, niflumic acid (NFA), with human serum albumin (HSA) has been investigated by (19)F nuclear magnetic resonance (NMR) spectroscopy. A (19)F NMR spectrum of NFA in a buffered (pH 7.4) solution of NaCl (0.1 mol L(-1)) contained a single sharp signal of its CF(3) group 14.33 ppm from the internal reference 2,2,2-trifluoroethanol. Addition of 0.6 mmol L(-1) HSA to the NFA buffer solution caused splitting of the CF(3) signal into two broadened signals, shifted to the lower fields of 14.56 and 15.06 ppm, with an approximate intensity ratio of 1:3. Denaturation of HSA by addition of 3.0 mol L(-1) guanidine hydrochloride (GU) restored a single sharp signal of CF(3) at 14.38 ppm, indicating complete liberation of NFA from HSA as a result of its denaturation. These results suggest that the binding is reversible and occurs in at least two HSA regions. Competitive (19)F NMR experiments using warfarin, dansyl-L: -asparagine, and benzocaine (site I ligands), and L: -tryptophan and ibuprofen (site II ligands) revealed that NFA binds to site I at two different regions, Ia and Ib, in the ratio 1:3. By use of (19)F NMR with NFA as an (19)F NMR probe the nonfluorinated site I-binding drugs sulfobromophthalein and iophenoxic acid were also found to bind sites Ia and Ib, respectively. These results illustrate the usefulness and convenience of (19)F NMR for investigation of the HSA binding of both fluorinated and nonfluorinated drugs.

Effects of bait type and deployment strategy on uptake by free-living badgers

Baits are increasingly used in wildlife management to deliver orally administered vaccines and contraceptives. The efficacy and cost-effectiveness of vaccination or fertility-control campaigns can be substantially affected by bait uptake rates. This study assessed whether bait type and deployment strategy affected bait uptake by free-living badgers (Meles meles L.). Six social groups of badgers were presented with three bait types (meat, fruit, cereals) and two deployment strategies (dispersed single baits versus aggregated multiple baits at fixed baiting stations) for six weeks. In each social group, the type of bait and deployment strategy were rotated every week so that by the end of the test every group had experienced all combinations. On three days, biomarkers (ethyl iophenoxic acid, propyl iophenoxic acid and rhodamine B) were added to the baits to determine the proportion of badgers ingesting these baits. The results indicated that both bait type and deployment strategy affected the proportion of baits eaten by badgers and the number of badgers gaining access to baits. Meat and fruit baits were taken significantly more frequently than cereals, and dispersed meat baits had the highest rates of disappearance. Biomarker levels suggested that the proportion of badgers that gained access to all baits was substantially lower when baits were aggregated, although small sample sizes prevented statistical assessment of this effect. The results suggest that dispersed single baits are likely to be consumed in greater proportions by a higher number of individual badgers than multiple baits at fixed stations.

Using Baits to Deliver Pharmaceuticals to Feral Swine in Southern Texas

Few studies have evaluated oral delivery systems of pharmaceuticals (e.g., vaccines, fertility control agents, and toxicants) to feral swine (Sus scrofa) in the United States. Our objective was to assess, through a field trial, the percentage of feral swine and nontarget animals that remove and consume baits intended to transport pharmaceuticals to feral swine in southern Texas, USA. We hand-placed 1,178 iophenoxic acid (IA)—marked baits distributed over 1,721 ha (68 baits/km2) in April 2005 and monitored species-specific bait removal and consumption using track stations, automated camera systems, and serum IA values from captured animals. Ninety percent of baits were removed after 72 hours. For baits for which we determined the species that “definitely” or “likely” removed bait using track stations and cameras, 51% were taken by raccoons (Procyon lotor), 22% were taken by feral swine, and 20% were taken by collared peccaries (Tayassu tajacu). We found elevated serum IA values in 74% of trapped feral swine, 89% of raccoons, and 43% of opossums (Didelphis virginiana). Our oral delivery system was successful in marking a substantial proportion of feral swine. However, our observed removal rates suggest that the majority of the baits were taken by nontarget species and, therefore, unsuitable for most pharmaceutical applications in their current form.

Liquid chromatography–electrospray ionization mass spectrometry for direct identification and quantification of iophenoxic acid in serum

A liquid chromatographic-electrospray ionization mass spectrometric technique was developed for direct quantitation of iophenoxic acid (IA) in serum. IA was spiked into canine, feline, bovine, equine, and porcine sera, extracted, and quantified using negative ion monitoring following chromatographic separation on a Luna C18(2) 3 microm (100 mm x 2.1mm) reversed-phase column. The limit of detection was 25 ng/mL and the limit of quantification was 50 ng/mL. Inter- and intra-assay accuracy (86-113% and 87-115%, respectively) and precision (1.8-7.7%) were calculated. Analysis of serum collected from feral pigs, raccoons, and opossums following ingestion of IA-marked baits confirmed the appropriateness of this method for bait acceptance studies.