Successful treatment of bifenthrin toxicosis using therapeutic plasma exchange.

OBJECTIVE: To describe a case of bifenthrin toxicosis in a dog with a successful outcome following the use of therapeutic plasma exchange (TPE) and intralipid therapy. CASE SUMMARY: An 8-month-old female neutered poodle mix dog ingested an unknown amount of powered bifenthrin, which resulted in acutely altered mentation, cranial nerve deficits, and intractable tremors that persisted in severity despite aggressive medical management to include intravenous fluids, intravenous lipid emulsion, anticonvulsant medications, and methocarbamol. TPE was initiated after lack of significant clinical improvement 12 hours after initial presentation. The dog underwent cardiopulmonary arrest (CPA) following approximately 1 plasma volume equivalent exchange. The dog was successfully resuscitated and showed marked improvement 12 hours postarrest and post-TPE treatment. Serum bifenthrin concentrations were analyzed prior to TPE (445.38 ng/mL) and ∼10 hours after TPE (51.18 ng/mL), which resulted in an 89% reduction in serum bifenthrin concentration. NEW INFORMATION: TPE may be a promising adjunctive therapeutic modality for bifenthrin toxicosis in dogs.

Diagnostic survey of analytical methods used to determine bone mineralization in pigs.

Pigs from 64 commercial sites across 14 production systems in the Midwest United States were evaluated for baseline biological measurements used to determine bone mineralization. There were three pigs selected from each commercial site representing: 1) a clinically normal pig (healthy), 2) a pig with evidence of clinical lameness (lame), and 3) a pig from a hospital pen that was assumed to have recent low feed intake (unhealthy). Pigs ranged in age from nursery to market weight, with the three pigs sampled from each site representing the same age or phase of production. Blood, urine, metacarpal, fibula, 2nd rib, and 10th rib were collected and analyzed. Each bone was measured for density and ash (defatted and non-defatted technique). A bone × pig type interaction (P < 0.001) was observed for defatted and non-defatted bone ash and density. For defatted bone ash, there were no differences among pig types for the fibulas, 2nd rib, and 10th rib (P > 0.10), but metacarpals from healthy pigs had greater (P < 0.05) percentage bone ash compared to unhealthy pigs, with the lame pigs intermediate. For non-defatted bone ash, there were no differences among pig types for metacarpals and fibulas (P > 0.10), but unhealthy pigs had greater (P < 0.05) non-defatted percentage bone ash for 2nd and 10th ribs compared to healthy pigs, with lame pigs intermediate. Healthy and lame pigs had greater (P < 0.05) bone density than unhealthy pigs for metacarpals and fibulas, with no difference observed for ribs (P > 0.10). Healthy pigs had greater (P < 0.05) serum Ca and 25(OH)D3 compared to unhealthy pigs, with lame pigs intermediate. Healthy pigs had greater (P < 0.05) serum P compared to unhealthy and lame pigs, with no differences between the unhealthy and lame pigs. Unhealthy pigs excreted significantly more (P < 0.05) P and creatinine in the urine compared to healthy pigs with lame pigs intermediate. In summary, there are differences in serum Ca, P, and vitamin D among healthy, lame, and unhealthy pigs. Differences in bone mineralization among pig types varied depending on the analytical procedure and bone, with a considerable range in values within pig type across the 14 production systems sampled.

There is little literature or data comparing bone diagnostic results for healthy, lame, and unhealthy pigs. Typically, diagnosticians assessing clinical lameness cases in pigs will measure bone mineralization along with histopathological evaluation to diagnose and assess the severity of metabolic bone disease. Bone ash is the primary method to determine bone mineralization, with the removal of the lipid in the bone (defatting) before the bone is ashed, compared to not removing the lipid before the ashing (non-defatted). Defatting the bone reduces the amount of variation across the bones compared to non-defatting. In this diagnostic survey, there was no difference among the healthy, lame, or unhealthy pigs when comparing defatted bone ash, however, unhealthy pigs had an increased bone ash percentage compared to the healthy and lame pigs when the bones were assessed using the non-defatted procedure. There was variation across production systems and pig types for serum vitamin D. When comparing the pig types, healthy pigs had increased serum Ca, P, and vitamin D [25(OH)D3] compared to the unhealthy pigs, with the lame pigs intermediate.

Blue Green Algae.

Blue green algae cyanotoxins have become increasingly more prevalent due to environmental, industrial, and agricultural changes that promote their growth into harmful algal blooms. Animals are usually exposed via water used for drinking or bathing, though specific cases related to equines are very limited. The toxic dose for horses has not been determined, and currently only experimental data in other animals can be relied upon to aid in case interpretation and treatment. Treatment is mostly limited to supportive care, and preventative control methods to limit exposures are more likely to aid in animal health until more research has been performed.

Equine Mycotoxins.

The main mycotoxins involved in adverse equine health issues are aflatoxins, fumonisins, trichothecenes, and probably ergovaline (fescue grass endophyte toxicosis). Most exposures are through contaminated grains and grain byproducts, although grasses and hays can contain mycotoxins. Clinical signs are often nonspecific and include feed refusal, colic, diarrhea, and liver damage but can be dramatic with neurologic signs associated with equine leukoencephalomalacia and tremorgens. Specific antidotes for mycotoxicosis are rare, and treatment involves stopping the use of contaminated feed, switching to a "clean" feed source, and providing supportive care.

The effect of bone and analytical methods on the assessment of bone mineralization response to dietary phosphorus, phytase, and vitamin D in nursery pigs.

A total of 360 pigs (DNA 600 × 241, DNA; initially 11.9 ±â€…0.56 kg) were used in a 28-d trial to evaluate the effects of different bones and analytical methods on the assessment of bone mineralization response to dietary P, vitamin D, and phytase in nursery pigs. Pens of pigs (six pigs per pen) were randomized to six dietary treatments in a randomized complete block design with 10 pens per treatment. Dietary treatments were designed to create differences in bone mineralization and included: (1) 0.19% standardized total tract digestibility (STTD) P (deficient), (2) 0.33% STTD P (NRC [2012] requirement) using monocalcium phosphate, (3) 0.33% STTD P including 0.14% release from phytase (Ronozyme HiPhos 2700, DSM Nutritional Products, Parsippany, NJ), (4) 0.44% STTD P using monocalcium phosphate, phytase, and no vitamin D, (5) diet 4 with vitamin D (1,653 IU/kg), and (6) diet 5 with an additional 50 µg/kg of 25(OH)D3 (HyD, DSM Nutritional Products, Parsippany, NJ) estimated to provide an additional 2,000 IU/kg of vitamin D3. After 28 d on feed, eight pigs per treatment were euthanized for bone (metacarpal, 2nd rib, 10th rib, and fibula), blood, and urine analysis. The response to treatment for bone density and ash was dependent upon the bone analyzed (treatment × bone interaction for bone density, P = 0.044; non-defatted bone ash, P = 0.060; defatted bone ash, P = 0.068). Thus, the response related to dietary treatment differed depending on which bone (metacarpal, fibula, 2nd rib, or 10th rib) was measured. Pigs fed 0.19% STTD P had decreased (P < 0.05) bone density and ash (non-defatted and defatted) for all bones compared to 0.44% STTD P, with 0.33% STTD P generally intermediate or similar to 0.44% STTD P. Pigs fed 0.44% STTD P with no vitamin D had greater (P < 0.05) non-defatted fibula ash compared to all treatments other than 0.44% STTD P with added 25(OH)D3. Pigs fed diets with 0.44% STTD P had greater (P < 0.05) defatted second rib ash compared to pigs fed 0.19% STTD P or 0.33% STTD P with no phytase. In summary, bone density and ash responses varied depending on bone analyzed. Differences in bone density and ash in response to P and vitamin D were most apparent with fibulas and second ribs. There were apparent differences in the bone ash percentage between defatted and non-defatted bone. However, differences between the treatments remain consistent regardless of the analytic procedure. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for the detection of lesions.

Lameness is defined as impaired movement or deviation from normal gait. There are many factors that can contribute to lameness, including but not limited to: infectious disease, genetic and conformational anomaly, and toxicity that affects the bone, muscle, and nervous systems. Metabolic bone disease is another cause of lameness in swine production and can be caused by inappropriate levels of essential vitamins or minerals. To understand and evaluate bone mineralization, it is important to understand the differences in diagnostic results between different bones and analytical techniques. Historically, percentage bone ash has been used as one of the procedures to assess metabolic bone disease as it measures the level of bone mineralization; however, procedures and results vary depending on the methodology and type of bone measured. Differences in bone density and ash in response to dietary P and vitamin D were most apparent in the fibulas and second ribs. There were apparent differences in the percentage of bone ash between defatted and non-defatted bone; however, the differences between the treatments remain consistent regardless of the analytic procedure. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for detection of lesions associated with metabolic bone disease.

Pharmacokinetics, Milk Residues, and Toxicological Evaluation of a Single High Dose of Meloxicam Administered at 30 mg/kg per os to Lactating Dairy Cattle.

Adverse effects associated with overdose of NSAIDs are rarely reported in cattle, and the risk level is unknown. If high doses of NSAIDs can be safely administered to cattle, this may provide a longer duration of analgesia than using current doses where repeated administration is not practical. Meloxicam was administered to 5 mid-lactation Holstein dairy cows orally at 30 mg/kg, which is 30 times higher than the recommended 1 mg/kg oral dose. Plasma and milk meloxicam concentrations were determined using high-pressure liquid chromatography with mass spectroscopy (HPLC-MS). Pharmacokinetic analysis was performed by using noncompartmental analysis. The geometric mean maximum plasma concentration (Cmax) was 91.06 µg/mL at 19.71 h (Tmax), and the terminal elimination half-life (T1/2) was 13.79 h. The geometric mean maximum milk concentration was 33.43 µg/mL at 23.74 h, with a terminal elimination half-life of 12.23 h. A thorough investigation into the potential adverse effects of a meloxicam overdose was performed, with no significant abnormalities reported. The cows were humanely euthanized at 10 d after the treatment, and no gross or histologic lesions were identified. As expected, significantly higher plasma and milk concentrations were attained after the administration of 30 mg/kg meloxicam with similar half-lives to previously published reports. However, no identifiable adverse effects were observed with a drug dose 30 times greater than the industry uses within 10 days of treatment. More research is needed to determine the tissue withdrawal period, safety, and efficacy of meloxicam after a dose of this magnitude in dairy cattle.

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.

Investigation of suspected gadolinium neurotoxicity in a dog.

Gadolinium-based contrast agents are used across species to better visualize abnormalities during MRI and are considered generally safe in clinical practice. The aim of this study was to investigate central nervous system (CNS) gadolinium deposition in 11 dogs that had an MRI performed, received 0.22 mL/kg (0.1 mmol/kg) of gadopentetate dimeglumine, and were necropsied on the same hospital admission. The index case was a 5-year-old castrated male Australian Shepherd that presented for ataxia and following MRI developed seizure-like activity that became refractory to anticonvulsant therapy. Gadolinium concentration was quantified in CNS tissues by inductively-coupled plasma mass spectrometry and was 43-fold higher in the index case. These findings suggest the possibility of gadolinium toxicity in select patients.

Detecting and quantifying marijuana metabolites in serum and urine of 19 dogs affected by marijuana toxicity.

Veterinarians diagnose marijuana toxicity based on clinical signs and history, or in conjunction with an over-the-counter (OTC) human urine drug screen. With the legalization of recreational marijuana use becoming more prevalent in the United States, a more accurate test to aid in the diagnosis of canine marijuana toxicity is needed. We collected urine and serum samples from 19 dogs with confirmed or suspected marijuana toxicosis from multiple veterinary hospitals and analyzed them with a novel UPLC-MS/MS method. Calibrations from 0.1 to 100 ng/mL and QC materials were prepared. Samples were extracted, purified, and eluted with solid-phase extraction. Urine samples were tested with an OTC human urine drug screen. The limit of detection (LOD) and lower limit of quantification (LLOQ) ranges for marijuana metabolites in serum were 0.05-0.25 ng/mL and 0.1-0.5 ng/mL, respectively. In urine, the LOD and LLOQ ranges for the metabolites were 0.05-0.1 ng/mL and 0.1-0.5 ng/mL, respectively. In serum, median and range of metabolite concentrations (ng/mL) detected included: THC, 65.0 (0.14-160); 11-OH-Δ9-THC, 4.78 (1.15-17.8); 11-nor-9-carboxy-Δ9-THC, 2.18 (0.71-7.79); CBD, 0.28 (0.11-82.5); and THC-glucuronide, 2.05 (0.72-18.3). In the 19 urine samples, metabolite: creatinine (ng: mg) values detected included: THC, 0.22 (0.05-0.74); 11-OH-Δ9-THC, 0; 11-nor-9-carboxy-Δ9-THC, 1.32 (0.16-11.2); CBD, 0.19 (0.12-0.26); THC-COOH-glucuronide, 0.08 (0.04-0.11); and THC-glucuronide, 0.98 (0.25-10.7). Twenty of 21 urine samples tested negative for THC on the urine drug screen. All 19 serum samples contained quantifiable concentrations of THC using our novel UPLC-MS/MS method. Utilizing a UPLC-MS/MS method can be a useful aid in the diagnosis of marijuana toxicosis in dogs, whereas using an OTC human urine drug test is not a useful test for confirming marijuana exposure in dogs because of the low concentration of THC-COOH in urine.

Zn-based physiometacomposite nanoparticles: distribution, tolerance, imaging, and antiviral and anticancer activity.

The aim of this study was to investigate the distribution, tolerance, and anticancer and antiviral activity of Zn-based physiometacomposites (PMCs). Manganese, iron, nickel and cobalt-doped ZnO, ZnS or ZnSe were synthesized. Cell uptake, distribution into 3D culture and mice, and biochemical and chemotherapeutic activity were studied by fluorescence/bioluminescence, confocal microscopy, flow cytometry, viability, antitumor and virus titer assays. Luminescence and inductively coupled plasma mass spectrometry analysis showed that nanoparticle distribution was liver >spleen >kidney >lung >brain, without tissue or blood pathology. Photophysical characterization as ex vivo tissue probes and LL37 peptide, antisense oligomer or aptamer delivery targeting RAS/Ras binding domain (RBD) was investigated. Treatment at 25 µg/ml for 48 h showed ≥98-99% cell viability, 3D organoid uptake, 3-log inhibition of ß-Galactosidase and porcine reproductive respiratory virus infection. Data support the preclinical development of PMCs for imaging and delivery targeting cancer and infectious disease.

Impact of increasing levels of fumonisin on performance, liver toxicity, and tissue histopathology of finishing beef steers.

To address the gaps in current scientific knowledge, the objective of the present study was to investigate the impact of fumonisin exposure on feedlot cattle intake and performance. Fifty steers were received (day 0; 361 ± 6.4 kg), housed individually and fed once daily at 0800 hours. All steers were transitioned to a dry-rolled corn-based finishing diet from days 0 to 21 and then were fed the control finishing diet until day 50. Treatment diets were formulated to achieve ≤5 (CON), 15 (15PPM), 30 (30PPM), 60 (60PPM), or 90 ppm (90PPM) of total dietary fumonisin. Steers were fed the fumonisin treatment diets from day 50 until harvest on day 160; individual animal body weights (BW) were measured on days 0, 50, 100, 150, 159, and 160. Liver, kidney, and skeletal muscle tissue samples were collected at harvest for histopathological analyses, and liver samples were further analyzed for sphinganine (SA) and sphingosine (SO) concentration. Animal performance, carcass data, and liver enzyme concentration were analyzed using a mixed model; categorical data were analyzed via nonparametric models. Contrasts were used to test for linear and quadratic responses. Throughout the study, there was no effect of treatment (P > 0.60), or a linear response (P > 0.16) from increasing fumonisin levels, on BW or dry matter intake (DMI). However, CON tended to have a lower average daily gain (ADG) than the fumonisin treatments during the fumonisin treatment period (P = 0.10), and there was a positive linear response (P = 0.02) of ADG to fumonisin during the treatment period. There were no treatment differences in hot carcass weight, dressing percentage, marbling score, ribeye area, or yield grade. There were no effects of treatment on either liver abscesses (P = 0.95) or telangiectasis (P = 0.13). We observed a treatment difference for SA and SA:SO (P < 0.01), as well as a quadratic response (P < 0.02); both SA and SA:SO increased as dietary fumonisin increased. There were no observed differences between treatments for histopathology scores of kidney (P = 0.16), liver (P = 0.25), or skeletal muscle (P = 0.59) tissue. No adverse effects were observed in steers fed increasing dietary levels of fumonisin for 110 d prior to harvest. While elevated liver amino alcohol concentration did occur, negative effects on growth and carcass characters were not observed.

Evaluating Mineral Status in Ruminant Livestock.

Determining mineral status of production animals is important when developing an optimum health program. Nutrition is the largest expense in food animal production and has the greatest impact on health and productivity of the animals. Knowing the bioavailability of minerals in the diet is difficult. Evaluating fluid or tissues from animals is the optimum method to determine bioavailability. Evaluating the diet provides some information. Serum/blood or liver from the animal needs to be analyzed to determine bioavailability of vitamin and minerals in the diet. This article reviews how to sample and the function of these minerals in cattle.

Ionophore Use and Toxicosis in Cattle.

Ionophores are a commonly used feed additive for animals and when used properly are safe. When feed mis-mixing occurs and an elevated dose of ionophore is given, a toxicosis can develop. Myocardial and skeletal muscles are the targets of a toxicosis. In many species there is a delay from the time of ingestion of a toxic dose in feed to when clinical signs occur. This makes it difficult to collect the feed in question that was at an elevated concentration. Cardiac troponins in serum can be used to make a diagnosis of an ionophore toxicosis.

Biofuels Co-Products Tolerance and Toxicology for Ruminants: An Update.

Corn co-products are a co-product of the dry and wet corn-milling ethanol manufacturing industry. The dry mill corn co-product is distiller's grains. Distillers grain can be further categorized into dry distillers grains (DDG), DDG with solubles, wet distillers grains with solubles (WDGS), modified WDGS, and corn syrup (solubles). Wet mill ethanol production produces 2 main feed stuffs: corn gluten (wet and dry) and heavy steep water.

Quantitative Analysis of Ethyl Carbamate in Distillers Grains Co-products and Bovine Plasma by Gas Chromatography-Mass Spectrometry.

Ethyl carbamate (EC) is a fermentation byproduct in foods and beverages and classified as a Group 2A probable human carcinogen. Each year, greater than 40 million metric tons of fermentation co-products from the U.S. ethanol industry are fed to food animals. A gas chromatography-mass spectrometry assay was developed to quantify EC extracted from various distillers grains co-products with a limit of detection at 0.7 ng/g (on an as-fed basis). EC was detected in all the distillers grains co-products surveyed in this study. Corn condensed distillers solubles contained the highest concentration of EC, ranging from 1618 to 2956 ng/g. Concentrations of EC in other types of distillers grains co-products varied from 17 to 917 ng/g. Cattle fed distillers grains co-products that constituted 19-38% of the total feed (as-fed) were found to contain 2-3 ng/mL of EC in blood plasma. No EC was detected in blood plasma from grass-fed control cattle.

Utility of a PCR-based method for rapid and specific detection of toxigenic Microcystis spp. in farm ponds.

Microcystis is a widespread freshwater cyanobacterium that can produce microcystin, a potent hepatotoxin harmful to animals and humans. Therefore, it is crucial to monitor for the presence of toxigenic Microcystis spp. to provide early warning of potential microcystin contamination. Microscopy, which has been used traditionally to identify Microcystis spp., cannot differentiate toxigenic from non-toxigenic Microcystis. We developed a PCR-based method to detect toxigenic Microcystis spp. based on detection of the microcystin synthetase C (mcyC) gene and 16S rRNA gene. Specificity was validated against toxic and nontoxic M. aeruginosa strains, as well as 4 intergeneric freshwater cyanobacterial strains. Analytical sensitivity was as low as 747 fg/µL genomic DNA (or 3 cells/µL) for toxic M. aeruginosa. Furthermore, we tested 60 water samples from 4 farm ponds providing drinking water to swine facilities in the midwestern United States using this method. Although all water samples were positive for Microcystis spp. (i.e., 16S rRNA gene), toxigenic Microcystis spp. were detected in only 34 samples (57%). Seventeen water samples contained microcystin (0.1-9.1 µg/L) determined with liquid chromatography-mass spectrometry, of which 14 samples (82%) were positive for mcyC. A significant correlation was found between the presence of toxigenic Microcystis spp. and microcystin in water samples (p = 0.0004). Our PCR method can be a low-cost molecular tool for rapid and specific identification of toxigenic Microcystis spp. in farm ponds, improving detection of microcystin contamination, and ensuring water safety for farm animals.

Inductively coupled plasma mass spectrometry determination of hepatic copper, manganese, selenium, and zinc concentrations in relation to sample amount and storage duration.

Trace mineral status is a critical component of bovine health. Impairment of physiological processes, caused by trace mineral toxicities or deficiencies, can be potential underlying factors of disease. Historically, the status of critical trace minerals, such as copper, manganese, selenium, and zinc, has been evaluated through the analysis of hepatic tissue. We assessed variation of these 4 elements between homogenized liver and samples of 0.02 g, 0.1 g, 0.5 g, and 1.0 g. We also evaluated concentration differences in copper, manganese, selenium, and zinc among samples stored under different durations. No differences in concentrations of copper, manganese, selenium, or zinc were observed among samples stored frozen for 3, 7, and 14 d post-collection. Statistical differences in concentrations of selenium and zinc were observed between 0.02-g biopsy samples and larger samples. Moisture content differed between 0.02-g biopsies and larger samples and over time. Results indicate that as little as 0.02 g of hepatic tissue dried to ~0.006 g is reliable for interpretation of trace mineral status and determination of toxicities and deficiencies in cattle pertaining to copper, manganese, selenium, and zinc, despite the small differences observed.

High-performance liquid chromatography and Enzyme-Linked Immunosorbent Assay techniques for detection and quantification of aflatoxin B1 in feed samples: a comparative study.

OBJECTIVE: Comparison was done between high-performance liquid chromatography (HPLC) and a competitive enzyme-linked immunosorbent assay (ELISA) for detection and quantification of aflatoxin B1 (AFB1) in feed samples. The two procedures were standardized and validated before the actual experiment. Five concentrations (0, 5, 10, 20 and 30 ppb) of feed samples were used for both methods. For the HPLC technique, the samples were extracted in acetonitrile/water (90/10) solution, cleaned-up using solid phase extraction (SPE) column, and derivatized by water/trifluoroacetic acid/glacial acetic acid (35/10/5) solution before instrument analysis. The samples were extracted in 70% methanol for the ELISA technique. RESULTS: The two tests showed very strong linearity with correlation coefficient value of > 0.99 using standard solutions. The mean recovery rate was 92.42% (with relative standard deviation (RSD) of 5.97) and 75.64% (RSD = 34.88) for HPLC and ELISA, respectively. There was no statistically significant difference in recovery rate between the two methods. There was a positive correlation (r = 0.84) between them which indicated that the two techniques can be used to detect and quantify aflatoxin B1 in feed samples. However, there were variations among replicates for the ELISA method, which shows that this method is more applicable for screening purposes.

Intra-laboratory Development and Evaluation of a Quantitative Method for Measurement of Aflatoxins B1, M1 and Q1 in Animal Urine by High Performance Liquid Chromatography with Fluorescence Detection.

Mycotoxins negatively impact animal health. Aflatoxins (AFs) are the most common mycotoxins affecting both large and small animals and are a common cause of toxin-related pet food recalls. Definitive diagnosis of aflatoxicosis is constrained by a lack of validated ante-mortem analytical methods for detection and quantitation of AFs and their metabolites in biological specimens. Herein, we developed and evaluated a urine-based quantitative method for measurement of aflatoxin B1 (AFB1) and its metabolites aflatoxin M1 (AFM1) and aflatoxin Q1 (AFQ1) in animal urine. (Some of the results have been presented at 59th AAVLD conference, Greensboro, North Carolina, October 13-19th, 2016.) This method uses an immuno-affinity column for clean-up and pre-column derivatization followed by high performance liquid chromatography analysis with fluorescence detection. The method has high selectivity, recovery (>81%) and sensitivity with an instrument limit of detection of 0.20-1.02 pg; instrument limit of quantitation of 0.77-4.46 pg; and a method lower limit of quantitation of 0.30-2.5 ng/mL. The method has high accuracy, repeatability, and is rugged against minor changes. However, because of poor sensitivity of AFQ1 at low concentrations we recommend this method for quantitative determination of AFB1 and AFM1, and for qualitative measurement of AFQ1 in animal urine for diagnosis of aflatoxicosis.