Steve Ensley

Lead Contamination of Chicken Eggs and Tissues from a Small Farm Flock

Twenty mixed-breed adult laying hens from a small farm flock in Iowa were clinically normal but had been exposed to chips of lead-based paint in their environment. These chickens were brought to the Iowa State University Veterinary Diagnostic Laboratory, Ames, Iowa, where the concentration of lead in blood, eggs (yolk, albumen, and shell), and tissues (liver, kidney, muscle, and ovary) from 5 selected chickens was determined over a period of 9 days. Blood lead levels ranged from less than 50 to 760 ppb. Lead contamination of the yolks varied from less than 20 to 400 ppb, and shells were found to contain up to 450 ppb lead. Albumen contained no detectable amount. Lead content of the egg yolks strongly correlated with blood lead levels. Deposition of lead in the shells did not correlate well with blood lead levels. Mean tissue lead accumulation was highest in kidneys (1,360 ppb), with livers ranking second (500 ppb) and ovarian tissue third (320 ppb). Muscle contained the lowest level of lead (280 ppb). Lead contamination of egg yolks and edible chicken tissues represents a potential public health hazard, especially to children repeatedly consuming eggs from contaminated family-owned flocks.

Assessment of ruminal hydrogen sulfide or urine thiosulfate as diagnostic tools for sulfur induced polioencephalomalacia in cattle

To determine if ruminal hydrogen sulfide, urine thiosulfate, or blood sulfhemoglobin could be used as diagnostic indicators for sulfur-induced polioencephalomalacia, 16 steers (8 cannulated, 368 ± 12 kg; 8 unmodified, 388 ± 10 kg; mean ± standard error) were fed 1 of 2 dietary treatments. Diets consisted of a low sulfate (0.24% S; control) wheat midd–based pellet or the control pellet with sodium sulfate added to achieve a high-sulfate (0.68% S) pellet. As designed, intake did not differ (P = 0.80) between treatments. At 8 hr postfeeding, ruminal hydrogen sulfide was not affected by cannulation (P = 0.35) but was greater (P < 0.01) in high S (6,005 ± 475 mg/l) than control (1,639 ± 472 mg/l) steers. Time of day of sampling affected (P = 0.01) ruminal hydrogen sulfide, with peak concentrations occurring 4–12 hr after feeding. Urine was collected prefeeding (AM) and 7–9 hr postfeeding (PM). Urine thiosulfate concentrations of high S steers sampled in the PM were greater (P > 0.01) than in the AM. However, there was no difference due to time of sampling for control. In both the AM and PM, urine thiosulfate concentrations of high S were greater (P > 0.01) than control. Although hydrogen sulfide and thiosulfate were elevated by increased dietary S intake, a concentration at which polioencephalomalacia is likely to occur could not be determined. Sampling urine for thiosulfate or rumen gas for hydrogen sulfide of nonsymptomatic pen mates 4–8 hr after feeding may be useful to assess sulfur exposure and differentiate between causes of polioencephalomalacia.

Evaluation of two mycotoxin mitigation strategies in grow-finish swine diets containing corn dried distillers grains with solubles naturally contaminated with deoxynivalenol

A total of 1,040 growing pigs (initially, 22.9 ± 4.3 kg) were used in a 115-d study to evaluate the effects of 2 mycotoxin mitigation strategies, a preservative blend (PB) and a yeast product (YP), on the growth performance of swine fed diets containing corn dried distillers grains with solubles naturally contaminated with deoxynivalenol (DON). The PB consists of preservatives, antioxidants, AA, and direct-fed microbials and is included in diets to help pigs cope with the toxic effects of ingested mycotoxins. The YP works as an adsorbent to bind and prevent the absorption of mycotoxins in the gastrointestinal tract. Pigs were allotted to pens by initial BW and sex; pens were then assigned to treatments in a randomized block design with initial BW and sex serving as the blocking factors. Pens were randomly allotted to 1 of 4 dietary treatments consisting of a positive control (PC) containing <1 mg kg(-1) DON, a negative control (NC) formulated to contain 4 mg kg(-1) DON, NC with PB, and NC with YP. From d 0 to 42 and 42 to 84, no effect of diets containing PB or YP were observed for any of the growth criteria evaluated. From d 84 to 115, pigs fed PC or diets containing PB had improved (P < 0.05) ADG compared to pigs fed NC or diets containing YP, whereas pigs fed YP had improved (P < 0.05) ADG compared to those fed NC. Pigs fed diets containing PB or YP had improved (P < 0.05) ADFI and G:F compared to pigs fed NC. Overall (d 0 to 115), pigs fed diets containing PB had improved (P < 0.05) ADG, ADFI, and G:F compared to pigs fed NC. These results indicate that PB may be a suitable mycotoxin mitigation strategy in growing swine fed diets naturally contaminated with DON.

Evaluation of XPC and prototypes on aflatoxin-challenged broilers

Various products and prototypes were added to poultry diets during an aflatoxin challenge on growth and histological parameters. Male broiler chicks were randomly assigned to 8 treatment groups with 8 replicates/treatment and 3 birds/replicate. Treatments were as follows: 1) negative control containing no aflatoxin (NC); 2) positive control containing aflatoxin (PC); 3) 0.1% glucomannan mycotoxin standard industry ameliorator (STD); 4) 0.1% prototype A, a proprietary mixture of a Saccharomyces cerevisiae product and diatomaceous clay; 5) 0.2% prototype A; 6) 0.15% prototype B, a proprietary mixture of a S. cerevisiae product and diatomaceous clay (PB); 7) 0.0625% XPC (S. cerevisiae fermentation product); and 8) 0.125% XPC (XPC2). All treatments except NC contained 2,280 +/- 102 ng/g of aflatoxin and were fed for 28 d. Body weight and feed intake were measured weekly. Livers were collected on d 28, weighed, and used for histopathological evaluation. Beginning weights were similar across treatments, but BW were lower (P </= 0.05) than NC for all treatments except PB and XPC2. Total feed intake was lower (P </= 0.05) only for PC and STD compared with all other groups. Feed efficiency was not significantly (P >/= 0.05) different among the treatment groups. Liver weights relative to BW were higher (P </= 0.05) for all treatments compared with NC. Liver vacuolar lesions were significantly higher (P </= 0.05) than NC for the PC, STD, 0.1% prototype A, and 0.0625% XPC groups. However, vacuolar lesions in the 0.15% prototype B, PB, and XPC2 treatments were not significantly different (P >/= 0.05) compared with NC. Overall, BW gain in treatment groups PB and XPC2 was not different from NC and that corresponded to protective effects against liver lesions. Benefits observed during an aflatoxin challenge when broilers were supplemented with XPC, a fermentation product that does not contain any adsorbents, may be attributed to something other than adsorption as a primary mechanism.

Lasalocid Toxicosis in Neonatal Calves

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Rickets case series and diagnostic review of hypovitaminosis D in swine

Rickets can be attributed to nutritional, genetic, hormonal, or toxic disturbances and is classified as a metabolic bone disease. Rickets is most often associated with inappropriate dietary levels of calcium, phosphorus, and/or vitamin D. During a 27-month period (January 2010 through March 2012), the Iowa State University Veterinary Diagnostic Laboratory investigated causes of sudden, unexpected death and lameness in growing pigs throughout the Midwestern United States. Clinical observations from 17 growing pig cases included weakness, lameness, reluctance to move, muscle fasciculations and/or tremors, tetany, and death. Ribs were weak, soft, and bent prior to breaking; rachitic lesions were apparent at costochondral junctions in multiple cases. Acute and/or chronic bone fractures were also noted in multiple bones. Failure of endochondral ossification, expanded physes, infractions, thin trabeculae, and increased osteoclasts were noted microscopically. Decreased bone ash and serum 25(OH)D(3), combined with clinical and microscopic evaluation, confirmed a diagnosis of vitamin D-dependent rickets in all cases. In 3 cases, disease was linked to a specific nutrient supplier that ultimately resulted in a voluntary feed recall; however, most cases in the current investigation were not associated with a particular feed company. The present report describes vitamin D-associated rickets and its importance as a potential cause of weakness, lameness, muscle fasciculations, recumbency or sudden unexpected death in swine, and describes appropriate samples and tests for disease diagnosis.

Ergot alkaloid intoxication in perennial ryegrass (Lolium perenne): an emerging animal health concern in Ireland?

Four primary mycotoxicosis have been reported in livestock caused by fungal infections of grasses or cereals by members of the Clavicipitaceae family. Ergotism (generally associated with grasses, rye, triticale and other grains) and fescue toxicosis (associated with tall fescue grass, Festuca arundinacea) are both caused by ergot alkaloids, and referred to as ‘ergot alkaloid intoxication’. Ryegrass staggers (associated with perennial ryegrass Lolium perenne) is due to intoxication with an indole-diperpene, Lolitrem B, and metabolites. Fescue-associated oedema, recently described in Australia, may be associated with a pyrrolizidine alkaloid, N-acetyl norloline.Ergotism, caused by the fungus Claviceps purpurea, is visible and infects the outside of the plant seed. Fescue toxicosis and ryegrass staggers are caused by Neotyphodium coenophalium and N. lolii, respectively. Fescue-associated oedema has been associated with tall fescue varieties infected with a specific strain of N. coenophialum (AR542, Max P or Max Q). The name Neotyphodium refers to asexual derivatives of Epichloë spp., which have collectively been termed the epichloë fungi. These fungi exist symbiotically within the grass and are invisible to the naked eye.The primary toxicological effect of ergot alkaloid involves vasoconstriction and/or hypoprolactinaemia. Ingestion of ergot alkaloid by livestock can cause a range of effects, including poor weight gain, reduced fertility, hyperthermia, convulsions, gangrene of the extremities, and death. To date there are no published reports, either internationally or nationally, reporting ergot alkaloid intoxication specifically associated with perennial ryegrass endophytes. However, unpublished reports from the Irish Equine Centre have identified a potential emerging problem of ergot alkaloid intoxication with respect to equines and bovines, on primarily perennial ryegrass-based diets. Ergovaline has been isolated in varying concentrations in the herbage of a small number of equine and bovine farms where poor animal health and performance had been reported. Additionally, in some circumstances changes to the diet, where animals were fed primarily herbage, were sufficient to reverse adverse effects. Pending additional information, these results suggest that Irish farm advisors and veterinarians should be aware of the potential adverse role on animal health and performance of ergot alkaloids from perennial ryegrass infected with endophytic fungi.

Multilaboratory Survey To Evaluate Salmonella Prevalence in Diarrheic and Nondiarrheic Dogs and Cats in the United States between 2012 and 2014

ABSTRACT Eleven laboratories collaborated to determine the periodic prevalence of Salmonella in a population of dogs and cats in the United States visiting veterinary clinics. Fecal samples (2,965) solicited from 11 geographically dispersed veterinary testing laboratories were collected in 36 states between January 2012 and April 2014 and tested using a harmonized method. The overall study prevalence of Salmonella in cats (3 of 542) was <1%. The prevalence in dogs (60 of 2,422) was 2.5%. Diarrhea was present in only 55% of positive dogs; however, 3.8% of the all diarrheic dogs were positive, compared with 1.8% of the nondiarrheic dogs. Salmonella-positive dogs were significantly more likely to have consumed raw food (P = 0.01), to have consumed probiotics (P = 0.002), or to have been given antibiotics (P = 0.01). Rural dogs were also more likely to be Salmonella positive than urban (P = 0.002) or suburban (P = 0.001) dogs. In the 67 isolates, 27 unique serovars were identified, with three dogs having two serovars present. Antimicrobial susceptibility testing of 66 isolates revealed that only four of the isolates were resistant to one or more antibiotics. Additional characterization of the 66 isolates was done using pulsed-field gel electrophoresis and whole-genome sequencing (WGS). Sequence data compared well to resistance phenotypic data and were submitted to the National Center for Biotechnology Information (NCBI). This study suggests an overall decline in prevalence of Salmonella-positive dogs and cats over the last decades and identifies consumption of raw food as a major risk factor for Salmonella infection. Of note is that almost half of the Salmonella-positive animals were clinically nondiarrheic.

Influence of Soil Properties and Test Conditions on Sorption and Desorption of Testosterone

In this study, batch sorption and desorption experiments were conducted for testosterone using four agricultural soils and five clay minerals. Significant differences in sorption behavior were observed between abiotic and biotic systems. The Freundlich sorption coefficient Kf (µg per g)/(µg per mL) ranged from 8.53 to 74.46 for soils and from 35.28 to 1243 for clays. The maximum sorption capacity (µg per g) of soils ranged from 25.25 to 440.61 for soils and 168.46 to 499.84 for clays. Correlation of sorption model parameters with soil properties indicated that both clay content and soil organic matter are important variables in predicting testosterone sorption behavior. Observed testosterone desorption from agricultural soils ranged from approximately 14 to 100 percent after 3 desorption cycles, and the desorption percentage decreased as the initial testosterone concentration decreased. Temperature, ionic strength, the water/soil ratio and soil depth were determined to influence sorption and desorption of testosterone. Desorption significantly increase with the soil depth and with the increase in the water to soil ratio. Temperature had an inverse effect on the sorption capacity of the soils tested. Thermodynamic calculations showed that the enthalpy change of the soils tested were the range of 12.9-20.7 kJ per mol, indicating weak interaction between testosterone and soil. Our results suggest that additional studies on how soil particles with different size fractions affect hormones fate and transport are needed in order to determine the potential risk of testosterone leaching or runoff.

Intralaboratory development and evaluation of a high-performance liquid chromatography–fluorescence method for detection and quantitation of aflatoxins M1, B1, B2, G1, and G2 in animal liver

Aflatoxins are potent mycotoxins with effects that include hepatotoxicity, immunosuppression, and suppression of animal growth and production. The etiologic diagnosis of aflatoxicosis, which is largely based on analysis of contaminated feed matrices, has significant disadvantages given the fact that representative feed samples may not be available and feed-based test methods are not confirmatory of an etiologic diagnosis. A tissue-based analytical method for biomarkers of exposure would be valuable for confirmation of aflatoxicosis. We describe in-house development and evaluation of a high-performance liquid chromatographic method with fluorescence detection and precolumn derivatization for determination of aflatoxins M1, B1, B2, G1, and G2 in animal liver. The method demonstrates good selectivity for the tested aflatoxins in the liver matrix. The overall range was 0.03–0.10 ng/g for limit of detection and 0.09–0.18 ng/g for limit of quantitation. The correlation coefficient (R2) of calibration curves was >0.9978 for AFM1, 0.9995 for AFB1, 0.9986 for AFB2, 0.9983 for AFG1, and 0.9980 for AFG2. For fortification levels of 0.2–10 ng/g, repeatability was 10–18% for AFM1, 7–14% for AFB1, 5–14% for AFB2, 6–16% for AFG1, and 10–15% for AFG2. Recovery was 52–57% for AFM1, 54–62% for AFB1, 55–61% for AFB2, 57–67% for AFG1, and 61–65% for AFG2. There was no liver matrix effect found. The method is rugged against minor changes based on the selected factors. The results indicate that the proposed method is suitable for quantitative determination of aflatoxins M1, B1, B2, G1, and G2 in liver.