Cantharidin.

Cantharidin is the toxic component of blister beetles of the genus Epicauta. Cantharidin is a potent vesicant which causes blisters, erosions, and ulcerations in the gastrointestinal and urinary tracts, and can cause myocardial necrosis. Blister beetles are found over most of North America and specifically contaminate alfalfa at harvest. History of alfalfa feeding, with colic, dysuria, hypocalcemia, and hypomagnesemia are suggestive of blister beetle toxicosis. Myocardial damage causes increased serum cardiac troponin 1. Tentative diagnosis can be made by finding the beetles in feed or ingesta. Definitive diagnosis requires detection of cantharidin in urine or gastric contents. Treatment involves ending exposure, decreasing absorption, controlling pain, using gastroprotectants, and fluids and electrolyte replacement. Prognosis is guarded to poor.

Pyrogallol Toxicosis in Horses.

Plants in the maple genus, Acer, and pistachio genus, Pistacia, have been reported to cause acute hemolysis in horses. The cause of hemolysis seems to be metabolism of gallic acids to the potent oxidant pyrogallol by enteric bacteria of the horse. Diagnosis is often tentative and circumstantial. Treatment is symptomatic and supportive and can include detoxification, fluid and electrolyte therapy, supplemental oxygen, and pain control. Corticosteroid and antioxidant therapies do not improve prognosis. Prognosis is guarded to poor but horses that survive 6 days postexposure are expected to recover.

The role of the veterinary diagnostic toxicologist in apiary health.

Susceptibility of individuals and groups to toxicants depends on complex interactions involving the host, environment, and other exposures. Apiary diagnostic investigation and honey bee health are truly population medicine: the colony is the patient. Here we provide basic information on the application of toxicology to the testing of domestic honey bees, and, in light of recent research, expand on some of the challenges of interpreting analytical chemistry findings as they pertain to hive health. The hive is an efficiently organized system of wax cells used to store brood, honey, and bee bread, and is protected by the bee-procured antimicrobial compound propolis. Toxicants can affect individual workers outside or inside the hive, with disease processes that range from acute to chronic and subclinical to lethal. Toxicants can impact brood and contaminate honey, bee bread, and structural wax. We provide an overview of important natural and synthetic toxicants to which honey bees are exposed; behavioral, husbandry, and external environmental factors influencing exposure; short- and long-term impacts of toxicant exposure on individual bee and colony health; and the convergent impacts of stress, nutrition, infectious disease, and toxicant exposures on colony health. Current and potential future toxicology testing options are included. Common contaminants in apiary products consumed or used by humans (honey, wax, pollen), their sources, and the potential need for product testing are also noted.

Pesticide contamination of beeswax from managed honey bee colonies in New York State.

The New York State (NYS) beekeeping industry generated >$11M worth of honey in 2020 and >$300M in pollination services to agriculture annually. Bees are frequently exposed to pesticides through foraging and husbandry practices. Lipophilic pesticides can remain in beeswax for extended periods. We analyzed for pesticides in wax comb samples collected from NYS apiaries at the end of the growing season, comparing residue numbers and concentrations among beekeepers of different operation scales: commercial beekeepers (>300 colonies), sideliners (50-299 colonies), and hobbyists (<50 colonies). We analyzed samples collected from 72 managed honey bee colonies for 92 insecticides, herbicides, and fungicides by liquid chromatography-tandem mass spectrometry. Pesticides were detected in all samples and included 34 fungicides, 33 insecticides, and 22 herbicides. Each wax sample contained 7-35 different residues (x¯ = 17.8 residues). Wax from colonies managed by commercial beekeepers contained the most residues (x¯ = 21.9 residues), hobbyists were second (x¯ = 16.3 residues), and sideliners had the fewest (x¯ = 11.7 residues). Nearly all wax samples (98.6%) contained the pesticide synergist piperonyl butoxide, most samples (86%) contained common varroacides used to control honey bee parasites, including coumaphos and amitraz breakdown products, and 93.1% contained the fungicide difenoconazole. We detected 34 fungicides, 7 of which were found in 50% or more of the samples. We detected 22 herbicides. We found pesticide contamination of beeswax to be common, with commercial beekeepers experiencing the greatest contamination.

Marijuana toxicosis in 2 donkeys.

Marijuana toxicosis is typically seen by companion animal veterinarians. However, with increased marijuana availability, there is a greater potential for toxicosis in other species. Herein we describe a case of suspected marijuana toxicosis in a female and a male American Mammoth donkey, aged 8 y and 20 y, respectively, fed cannabis buds. Both cases were presented because of depression and lethargy. However, the jenny had ataxia, mild colic, tachycardia, tachypnea, and decreased tongue tone. Plasma samples from the jenny on presentation and 3 d following hospitalization were submitted to the Kansas State Veterinary Diagnostic Laboratory to be screened for cannabinoids using high-pressure liquid chromatography coupled with tandem mass spectroscopy (HPLC-MS/MS). A single serum sample from the jack was taken on presentation and submitted to the Animal Health Diagnostic Center at Cornell University for Δ9-tetrahydrocannabinol (THC) and cannabidiol analysis using HPLC-MS/MS. THC was detected in all samples. Clinical signs were noted 24-36 h after ingestion, which included mild-to-moderate neurologic deficits, mild colic, tachycardia, tachypnea, and decreased tongue tone. Both donkeys recovered uneventfully within 24 h of peak effects. Utilizing a cannabinoid screening assay in collaboration with a veterinary diagnostic laboratory may be useful when an equine practitioner suspects marijuana toxicosis in a patient.

The urban lead (Pb) burden in humans, animals and the natural environment.

Centuries of human activities, particularly housing and transportation practices from the late 19th century through the 1980's, dispersed hundreds of millions of tons of lead into our urban areas. The urban lead burden is evident among humans, wild and domesticated animals, and plants. Animal lead exposures closely mirror and often exceed the lead exposure patterns of their human partners. Some examples: Pigeons in New York City neighborhoods mimicked the lead exposures of neighborhood children, with more contaminated areas associated with higher exposures in both species. Also, immediately following the lead in drinking water crisis in Flint MI in 2015, blood lead levels in pet dogs in Flint were 4 times higher than in surrounding towns. And combining lead's neurotoxicity with urban stress results in well-characterized aggressive behaviors across multiple species. Lead pollution is not distributed evenly across urban areas. Although average US pediatric lead exposures have declined by 90% since the 1970s, there remain well defined neighborhoods where children continue to have toxic lead exposures; animals are poisoned there, too. Those neighborhoods tend to have disproportionate commercial and industrial lead activity; a history of dense traffic; older and deteriorating housing; past and operating landfills, dumps and hazardous waste sites; and often lead contaminated drinking water. The population there tends to be low income and minority. Urban wild and domesticated animals bear that same lead burden. Soil, buildings, dust and even trees constitute huge lead repositories throughout urban areas. Until and unless we begin to address the lead repositories in our cities, the urban lead burden will continue to impose enormous costs distributed disproportionately across the domains of the natural environment. Evidence-based research has shown the efficacy and cost-effectiveness of some US public policies to prevent or reduce these exposures. We end with a series of recommendations to manage lead-safe urban environments.

Analysis of neurofilament concentration in healthy adult horses and utility in the diagnosis of equine protozoal myeloencephalitis and equine motor neuron disease.

Neurofilaments (NFs) are structural proteins of neurons that are released in significant quantities in the cerebrospinal fluid and blood as a result of neuronal degeneration or axonal damage. Therefore, NFs have potential as biomarkers for neurologic disorders. Neural degeneration increases with age and has the potential to confound the utility of NFs as biomarkers in the diagnosis of neurologic disorders. We investigated this relationship in horses with and without neurological diagnosis. While controlling for horse type (draft, pleasure, and racing), we evaluated the relationship between serum heavy-chain phosphorylated neurofilaments (pNF-H) and age, sex, and serum vitamin E concentrations. Serum pNF-H concentrations increased by 0.002 ng/ml for each year increase in age. There were significant differences in the serum pNF-H concentration among the type of activity performed by the horse. The highest serum pNF-H concentration was found in horses performing heavy work activity (racehorse) and with lower serum pNF-H concentration found among light (pleasure riding) and moderate (draft) activity. There was no significant association between the pNF-H concentration and sex or vitamin E concentration. Serum pNF-H concentration was elevated among horses afflicted with EMND and EPM when compared with control horses without evidence of neurologic disorders. Accordingly, serum pNF-H concentration can serve as a useful biomarker to complement the existing diagnostic work-up of horses suspected of having EPM or EMND.

Pet Food Recalls and Pet Food Contaminants in Small Animals: An Update.

Commercial pet foods are usually safe, but incidents of contamination can have a devastating impact on companion animals and their owners. There are numerous possible contaminants ranging from natural contaminants to nutrient imbalances to chemical adulteration, making it impossible to predict what will cause the next pet food recall. Veterinarians involvement with pet food recalls includes examining and treating affected animals, documentation and sample collection, and communicating with pet food manufacturers and regulatory agencies.

Equine feed contamination and toxicology.

Feed as a cause of poisoning in horses can occur on small or large scales. It is challenging to work up cases of suspected feed contamination, but there are resources available to veterinarians and owners. Feed contamination can be chemical or biological. This article focuses on and provides examples of chemical feed contamination including misformulation, adulteration, and natural contaminants. Additionally, recommendations for feed sampling and diagnostic submission, including legal documentation, are included.

Zinc Toxicosis in a Boxer Dog Secondary to Ingestion of Holiday Garland.

INTRODUCTION: Increased admissions occur in small animal veterinary emergency clinics during some holidays, and some of the increased caseload is due to ingestion of toxic substances. This report documents zinc toxicosis contributing to the death of a dog after ingestion of holiday tinsel garland. CASE STUDY: A mature boxer dog presented with a 4-day history of vomiting and diarrhea. Radiodense foreign material was detected in the stomach and removed via gastrotomy. The patient clinically worsened over the next several days with evidence of hemolytic anemia, severe hypernatremia, and an elevated WBC count with a suspected dehiscence of the surgical site and acute renal failure. The serum zinc concentration was moderately elevated. Postmortem findings included surgical dehiscence from the gastrotomy and enterotomy sites, hepatic extramedullary hematopoiesis, hemoglobinuric nephrosis, and pancreatic fibrosis. The foreign material removed from the stomach also contained zinc. DISCUSSION: Ingestion of holiday tinsel garland made from metal-coated plastic film has not previously been implicated in zinc toxicosis. Zinc toxicosis has a good prognosis in veterinary medicine when diagnosed and treated promptly, but the unique source of zinc in this dog contributed to the delay in diagnosis and grave outcome in this case.

Severe Lead Toxicosis in a Lionhead Rabbit.

INTRODUCTION: Lead toxicosis occurs in veterinary patients, with few reports involving rabbits, and no previous reports using oral calcium disodium EDTA. CASE REPORT: A 7-year-old male castrated Lionhead rabbit presented to the Cornell University Hospital for Animals (CUHA) for evaluation after a 2-day history of lethargy and a 2-week history of hyporexia. The patient had been observed pulling paint from the walls of the home, a house built circa 1900, in the months prior to presentation. The patient was moderately anemic with a hematocrit of 21% with red blood cell morphological changes consistent with lead toxicosis, including basophilic stippling, nucleated red blood cells, and polychromasia. Radiographic images of the abdomen revealed excessive accumulation of gas in the gastrointestinal tract in a pattern consistent with gastric stasis and numerous small mineral to metallic opacities in the cecum. The blood lead concentration was 792 µg/dL, confirming the diagnosis of lead toxicosis with secondary gastrointestinal stasis. The rabbit was hospitalized for treatment with oral and subcutaneous calcium disodium EDTA for 4 days and then discharged home to the care of the owners. DISCUSSION: Severe lead toxicosis in a rabbit can be treated successfully with oral and subcutaneous calcium disodium EDTA and aggressive supportive treatment.

Comparison of blood and tissue lead concentrations from cattle with known lead exposure.

Blood lead (Pb) is used to diagnose Pb poisoning and exposure in cattle, but there are limited data comparing circulating Pb with concentrations in beef from the same cattle. This study determines whether there is a correlation between blood Pb and tissue Pb concentrations in accidentally exposed cattle. Pb analyses were carried out on ante-mortem blood and post-mortem tissues (including, if available, brain, liver, skeletal muscle, bone, gastrointestinal contents and kidney, and also foetal tissues from one cow) collected from 13 cattle known to have accidental Pb exposure and from three control cows with no known exposure. Variables from cattle were analysed statistically using a Shapiro-Wilk normality test and non-parametric descriptive and association statistics. Blood Pb from exposed cattle rank-correlated with liver, bone and kidney Pb concentrations, but not with the Pb concentrations of brain, skeletal muscle or gastrointestinal contents. The lowest blood Pb concentration associated with a detectable skeletal muscle Pb concentration (> 0.1 mg kg-1 dry matter) was 4.57 µg dl-1. Based on these findings, we recommend that cattle with blood Pb > 2.5 µg dl-1 be withheld from slaughter and that liver, bone and kidney from all cattle with known Pb exposure be withheld from the human food chain.

Treatment of pieris ingestion in goats with intravenous lipid emulsion.

Seven goats and one ram presented with clinical signs including regurgitation, obtundation, anorexia, apparent pain, and bloat. The animals had escaped from their barn, and it was discovered that they had ingested leaves of Pieris japonica, Japanese pieris, a grayanotoxin-containing plant. Animals were treated with antibiotics, calcium borogluconate, B vitamins, and activated charcoal within the first 24-h postexposure, which was followed by the recovery of the ram and two goats and the death of two goats. Approximately 36 h after Japanese pieris ingestion, one of the three remaining anorectic goats was dosed with intravenous lipid emulsion (ILE). This goat recovered within a few hours. The remaining two goats were given ILE the next day and appeared to recover, but one died a week later of aspiration pneumonia.

A rapid screen for four corticosteroids in equine synovial fluid.

Most antidoping method development in the equine industry has been for plasma and urine, though there has been recent interest in the analysis of synovial fluid for evidence of doping by intra-articular corticosteroid injection. Published methods for corticosteroid analysis in synovial fluid are primarily singleplex methods, do not screen for all corticosteroids of interest and are not adequately sensitive. The purpose of this study is to develop a rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) screening method for the detection of four of the most common intra-articularly administered corticosteroids--betamethasone, methylprednisolone, methylprednisolone acetate and triamcinolone acetonide. Sample preparation consisted of protein precipitation followed by a basified liquid-liquid extraction. LC-MS-MS experiments consisted of a six-min isocratic separation using a Phenomenex Polar-RP stationary phase and a mobile phase consisting of 35% acetonitrile, 5 mM ammonium acetate and 0.1% formic acid in nanopure water. The detection system used was a triple quadrupole mass analyzer with thermospray ionization, and compounds were identified using selective reaction monitoring. The method was validated to the ISO/IEC 17025 standard, and real synovial fluid samples were analyzed to demonstrate the application of the method in an antidoping context. The method was highly selective for the four corticosteroids with limits of detection of 1-3 ng/mL. The extraction efficiency was 50-101%, and the matrix effects were 14-31%. These results indicate that the method is a rapid and sensitive screen for the four corticosteroids in equine synovial fluid, fit for purpose for equine antidoping assays.

Lead excretion in milk of accidentally exposed dairy cattle.

Lead (Pb) exposure in dairy cattle is associated with economic losses due to mortality and treatment costs, but with production animals there is also risk to the human food chain. The first objective of this study was to quantify the Pb concentration in milk from Pb-exposed cattle. The second objective was to correlate blood and milk Pb concentrations from individual cows. The third objective was long-term monitoring to determine the duration of milk contamination after exposure ceased. A dairy herd of more than 100 cows was accidentally exposed to Pb-contaminated feed. Milk and blood were collected for Pb analysis. Serial collection of milk samples continued for 2.5 years. The initial concentration of Pb in bulk tank milk was 0.0999 mg l⁻¹. The highest milk Pb concentration from an individual cow was 0.4657 mg l⁻¹ and the highest blood Pb concentration was 1.216 mg l⁻¹. One milk sample collected at the end of the study (day 922) contained 0.0117 mg Pb l⁻¹ of Pb. The calculated relationship between milk (y) and blood (x) Pb concentration was ln(y) = 3.4(x) - 2.21 (R² = 0.98).

Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay.

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 mg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 mg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Identification of protoxins and a microbial basis for red maple (Acer rubrum) toxicosis in equines.

The leaves of Acer rubrum (red maple), especially when wilted in the fall, cause severe oxidative damage to equine erythrocytes, leading to potentially fatal methemoglobinemia and hemolytic anemia. Gallic acid and tannins from A. rubrum leaves have been implicated as the toxic compounds responsible for red maple toxicosis, but the mechanism of action and toxic principle(s) have not been elucidated to date. In order to investigate further how red maple toxicosis occurs, aqueous solutions of gallic acid, tannic acid, and ground dried A. rubrum leaves were incubated with contents of equine ileum, jejunum, cecum, colon, and liver, and then analyzed for the metabolite pyrogallol, as pyrogallol is a more potent oxidizing agent. Gallic acid was observed to be metabolized to pyrogallol maximally in equine ileum contents in the first 24 hr. Incubation of tannic acid and A. rubrum leaves, individually with ileum contents, produced gallic acid and, subsequently, pyrogallol. Ileum suspensions, when passed through a filter to exclude microbes but not enzymes, formed no pyrogallol, suggesting a microbial basis to the pathway. Bacteria isolated from ileum capable of pyrogallol formation were identified as Klebsiella pneumoniae and Enterobacter cloacae. Therefore, gallotannins and free gallic acid are present in A. rubrum leaves and can be metabolized by K. pneumoniae and E. cloacae found in the equine ileum to form pyrogallol either directly or through a gallic acid intermediate (gallotannins). Identification of these compounds and their physiological effects is necessary for the development of effective treatments for red maple toxicosis in equines.

Production of a highly group-specific monoclonal antibody against zearalenone and its application in an enzyme-linked immunosorbent assay.

A monoclonal antibody (mAb) against zearalenone (ZEN) was produced using ZEN-carboxymethoxylamine and -BSA conjugates. Antibody produced by one clone showing a very high binding ability was selected and found to have a higher affinity for ZEN compared to a commerciall ZEN antibody. We developed two direct competitive ELISA systems using the selected antibody (ZEN-coated and anti-ZEN antibody-coated ELISA). Quantitative ranges for the anti-ZEN antibody coated ELISA and ZEN-coated ELISA were from 25 to 750 ppb and from 12.5 to 100 ppb, respectively. The detection limit of both methods as measured with standard solutions was 10 ppb. The intra-plate and inter-well variation of both ELISAs were less than 10%. The IC(50) values for α-zearalenol, ß-zearalenol, α-zearalanol, and ß-zearalanol compared to ZEN were 108.1, 119.3, 114.1, and 130.3% for the ZEN-coated ELISA. These values were 100.7, 120.7, 121.6, and 151.6% for the anti-ZEN antibody-coated ELISA. According to the anti-ZEN antibody-coated ELISA, the average recovery rates of ZEN from spiked animal feed containing 150 to 600 ng/mL of ZEN ranged from 106.07 to 123.00% with 0.93 to 2.28% coefficients of variation. Our results demonstrate that the mAb developed in this study could be used to simultaneously screen for ZEN and its metabolites in feed.

Pet food recalls and pet food contaminants in small animals.

Most pet foods are safe, but incidents of chemical contamination occur and lead to illness and recalls. There were 11 major pet food recalls in the United States between 1996 and 2010 that were due to chemical contaminants or misformulations: 3 aflatoxin, 3 excess vitamin D3, 1 excess methionine, 3 inadequate thiamine, and 1 adulteration with melamine and related compounds and an additional 2 warnings concerning a Fanconilike renal syndrome in dogs after ingesting large amounts of chicken jerky treat products. This article describes clinical findings and treatment of animals exposed to the most common pet food contaminants.

Declines in blood lead concentrations in clinically affected and unaffected cattle accidentally exposed to lead.

Lead (Pb) poisoning remains a common cause of morbidity in dairy and beef cattle. Although Pb toxicosis is typically diagnosed in cattle with clinical signs of acute or subacute Pb poisoning, it has been hypothesized that subclinical chronic exposure of cattle to Pb, which often goes undiagnosed, poses more of a risk to the human consumer. There is not adequate information on Pb kinetics to determine when or if Pb-exposed cattle can safely enter the food chain. The objectives of the current study were to determine whether subclinical elevations in blood Pb (bPb) were present in cattle from herds where 1 or more individuals had clinical Pb poisoning and to determine the half-life (t(1/2)) of bPb in Pb-exposed cattle. Samples of blood were collected and analyzed for Pb from 126 cattle from 9 farms. Blood lead concentrations ranged from below the detection limit (2.50 µg/dl) to 423.0 µg/dl. Only 11 of the 94 cattle with detectable bPb had clinical signs such as diarrhea, blindness, bruxism, or seizures. When possible, cattle with detectable bPb had serial samples taken. The mean t(1/2) calculated from 44 serially sampled cattle was 135 days (standard deviation: 125 days, range: 3-577 days). A source of Pb on the farm was determined for all but one herd.