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In recent years, livestock producers have been supplementing animal diets with fish meal (FM) to produce value-added products for health conscious consumers. As components of FM have unique neuroendocrine-immunomodulatory properties, we hypothesize that livestock producers may be influencing the overall health of their animals by supplementing diets with FM. In this study, 40 pregnant ewes were supplemented with rumen protected (RP) soybean meal (SBM: control diet) or RP FM, commencing gestation day 100 (gd100), in order to evaluate the impact of FM supplementation on the innate and acquired immune response and neuroendocrine response of sheep during pregnancy and lactation. On gd135, half the ewes from each diet (n = 10 FM, n = 10 SBM) were challenged iv with lipopolysaccharide (LPS) to simulate a systemic bacterial infection and the febrile, respiratory and neuroendocrine responses were monitored over time; the other half (n = 10 FM, n = 10 SBM) of the ewes received a saline injection as control. On lactation day 20 (ld20), all ewes (n = 20 FM, n = 20 SBM) were sensitized with hen egg white lysozyme (HEWL) and the serum haptoglobin (Hp) response was measured over time. The cutaneous hypersensitivity response (CHR) to HEWL challenge was measured on ld30 (n = 20 FM, n = 20 SBM), and blood samples were collected over time to measure the primary and secondary immunoglobulin G (IgG) response to HEWL. There was an attenuated trend in the LPS-induced febrile response by the FM treatment when compared with the SBM treatment (P = 0.06), as was also true for the respiratory response (P = 0.07), but significant differences in neuroendocrine function (serum cortisol and plasma ACTH) were not observed between treatments. Basal Hp levels were significantly lower in the FM supplemented ewes when compared with the SBM supplemented ewes (P < 0.01), and the Hp response to HEWL sensitization differed significantly over time between treatments (P < 0.01). The CHR to HEWL was also significantly attenuated in the FM treatment compared with the SBM (P < 0.01); however, treatment differences in the primary and secondary IgG responses to HEWL were not observed. These results indicate that FM supplementation differentially affects the innate and acquired immune responses in pregnant and lactating sheep compared with a typical SBM diet of commercial flocks. The long-term implications of this immunomodulation warrant further investigation.

Despite its growing prevalence in westernised countries, our current understanding of the molecular mechanism(s) of food allergy remains incomplete. In this study, transcriptome profile of spleen from BALB/c mice was analysed in response to egg ovomucoid (OVM) sensitisation and challenge. Microarray analysis revealed 87 genes (±1.5 fold, p≤0.05); 28 up-regulated and 59 down-regulated. Gene ontology analysis revealed several biological processes related to hypersensitivity, inflammation or immune response. Biological network analysis illustrated connections among differentially expressed genes and several immune or hypersensitivity-related processes. Expression of five genes from microarray experiment was validated by real-time RT-PCR. Microarray analysis identified several differentially expressed genes that were not previously characterised in food allergic conditions. Results from this study may help to understand the underlying molecular events that take place in the spleen after OVM-stimulati...

An experiment was conducted to explore the effects of Fusarium mycotoxins, common animal feed contaminants, on intestinal immune responses to coccidia (Eimeria) in chickens. Effects of feed-borne Fusarium mycotoxins and a polymeric glucomannan mycotoxin adsorbent (GMA) on immune cell populations were studied in the jejunum of broiler breeder pullets using an Eimeria maxima infection model. Birds were fed a control diet, a diet naturally contaminated with Fusarium mycotoxins, contaminated diet plus 0.2% GMA, or control diet plus 0.2% GMA. Contaminated diets contained up to 6.5μg/g deoxynivalenol (DON), 0.47μg/g 15-acetyl-DON and 0.73μg/g zearalenone. Birds received a primary oral inoculation (1000 oocysts/bird) with E. maxima USDA strain 68 at 2 weeks of age and a secondary oral inoculation (30,000 oocysts/bird) with the same strain at 4 weeks of age. Diet-related differences in CD4(+) cell, CD8(+) cell and macrophage recruitment pattern into the jejunum were observed following both the primary and secondary infections. It was concluded that feed-borne Fusarium mycotoxins and GMA have the potential to modulate immune response to coccidial infections.

An experiment was conducted to investigate the effects of dietary Fusarium mycotoxins on gut immunity, cell proliferation, and histology of avian lymphoid organs. The efficacy of a polymeric glucomannan mycotoxin adsorbent (GMA) was also determined. Seventy-two one-day-old male turkey poults were fed corn, wheat, and soybean meal-based diets for 21 days. Diets included control grains, contaminated grains and contaminated grains +0.2% GMA. The major contaminant was deoxynivalenol (3.9 μg/g) with lesser amounts of zearalenone (0.67-0.75 μg/g), 15-acetyl-deoxynivalenol (0.34 μg/g) and HT-2 toxin (0.078-0.085 μg/g). T- and B-lymphocyte populations and crypt cellular proliferation in duodenum, jejunum, ileum and cecal tonsil were measured immunohistochemically on day 14 and 21. Histological changes were recorded after 14 and 21 days of feeding. Feeding contaminated grains significantly increased the percentage of B-lymphocytes in ileum on day 14, and reduced (P0.05) the percentages of CD8(+)-lymphocytes in cecal tonsil on day 21. GMA supplementation prevented these effects. The feeding of contaminated diets also caused a reduction (P0.05) in ileal crypt proliferating cells and a significant increase in spleen secondary follicle on day 21. It was concluded that the feeding of grains naturally contaminated with Fusarium mycotoxins results in adverse effects on gut immunity and mucosal cell proliferation.

Interleukin-12 (IL-12) and interleukin-23 (IL-23) are proinflammatory cytokines produced by macrophages and dendritic cells in response to infection with intracellular pathogens. The IL-12 receptor (IL-12R) is a heterodimer composed of 2 subunits, β1 and β2. The IL-23 receptor (IL-23R) is a heterodimer composed of the IL-12Rβ1 subunit and a unique IL-23R subunit. Given the importance of IL-12 and IL-23 for modulating inflammation and the host immune response, the IL-12 and IL-23 receptor genes may be suitable candidate genes for studying disease resistance in dairy cattle. We hypothesize that single nucleotide polymorphisms (SNP) exist within these genes and that they contribute to variation in health and production traits in dairy cattle. To investigate this, a selective DNA pool was constructed using bull semen based on the estimated breeding values for somatic cell score (SCS), an indicator trait used to achieve genetic improvement for resistance to mastitis. Gene segments were amplified from this pool by PCR and the amplicons were sequenced to reveal SNP. A total of 10 SNP, including 2 in IL-12Rβ1, 5 in IL-12Rβ2, and 3 in IL-23R were identified. The SNP (n=5) were found in the 5' untranslated region (UTR) putative promoter regions of the genes, and SNP IL-23R c.1714A>C was a nonsynonymous SNP. Canadian Holstein bulls (n=492) were genotyped using Sequenom MassARRAY (Sequenom Inc., San Diego, CA). No association was found with SCS based on bull deregressed estimated breeding values for SCS; however, associations of SNP in the IL-12Rβ2 gene (c.-511A>G, c.87A>G, c.2957A>C) were found with milk and protein yield. Further investigation will be required to elucidate the biological and practical relevance of these SNP.

A study was conducted to determine the cytochrome (CYP) P450 enzymes responsible for the bioactivation of aflatoxin B1 (AFB1) into its epoxide form (AFBO) in duck liver microsomes. Six male and six female 6-week-old Pekin ducks were used. The biochemical toxicology strategies applied included the use of selective inhibitors, prototype substrate activity for specific human P450s, correlation between aflatoxin bioactivation and enzymatic activity of prototype substrates, and the expression of specific CYP450 enzymes using antibodies against human CYP450s. Enzymatic activity was detected for the duck orthologues CYP1A1/2, CYP2A6 and CYP3A4 but not for the CYP2D6 orthologue. Immunoreactive proteins for CYP1A1, CYP2A6 and CYP3A4 were also detected. Inhibition studies suggested that the duck turkey CYP2A6 orthologue and, to a lesser extent, the CYP1A1 orthologue are involved in the bioactivation of AFB1. Correlation studies, however, suggest that CYP3A4, CYP2A6 and CYP1A1/2 are all involved in AFBO formation. The finding that four CYP enzymes may be involved in AFB1 bioactivation in ducks could explain the high sensitivity of this species to AFB1. Further studies are needed to fully elucidate the phase I hepatic metabolism of AFB1 in ducks, the only poultry species that develops hepatic cancer from AFB1 exposure.

Bi-directional communication between the neuroendocrine and immune systems is designed, in part, to maintain or restore homeostasis during physiological stress. Exposure to endotoxin during Gram-negative bacterial infection for example, elicits the release of pro-inflammatory cytokines that activate the hypothalamic-pituitary-adrenal axis (HPAA). The secretion of adrenal glucocorticoids subsequently down regulates the host inflammatory response, minimizing potential tissue damage. Sequence and epigenetic variants in genes involved in regulating the neuroendocrine and immune systems are likely to contribute to individual differences in the HPAA response, and this may influence the host anti-inflammatory response to toxin exposure and susceptibility to inflammatory disease. In this study, high (HCR) and low (LCR) cortisol responders were selected from a normal population of 110 female sheep challenged iv with Escherichia coli endotoxin (400 ng/kg) to identify potential determinants that contribute to variation in the cortisol response phenotype. This phenotype was stable over several years in the HCR and LCR animals, and did not appear to be attributed to differences in expression of hepatic immune-related genes or systemic pro-inflammatory cytokine concentrations. Mechanistic studies using corticotrophin-releasing factor (0.5 microg/kg body weight), arginine vasopressin (0.5 microg/kg), and adrenocorticotropic hormone (0.5 microg/kg) administered iv demonstrated that variation in this phenotype is largely determined by signalling within the HPAA. Future studies will use this ovine HCR/LCR model to investigate potential genetic and epigenetic variants that may contribute to variation in cortisol responsiveness to bacterial endotoxin.

Inbred rodent studies have demonstrated that cutaneous hypersensitivity reactions are exacerbated in stress-susceptible, and attenuated in stress-resistant strains of mice. This physiological response was, in part, mediated by activation of the hypothalamic-pituitary-adrenal axis during the acute restraint stress. A study was conducted to examine whether or not cutaneous hypersensitivity reactions are also associated with variable cortisol responsiveness to inflammatory stress in an outbred ovine population. High (H), medium (M), and low (L) cortisol responsive sheep were identified from a population of 110 females based on their estimated breeding values for cortisol concentration measured 4 h post-systemic challenge with Escherichia coli endotoxin (400 ng kg(-1)). Cutaneous hypersensitivity reactions to phytohaemagglutinin (PHA), 1-chloro-2, 4-dinitrobenzene (DNCB), and Candida albicans cellular antigen (CAA) were measured in these variable cortisol-responding sheep, in addition to serum interleukin (IL)-6, interferon (IFN)-gamma, and ovalbumin (OVA)-specific IgG concentrations. When compared to the M cortisol responders, both H and L cortisol responders had significantly greater cutaneous swelling during the elicitation phase in response to DNCB (P0.05) and CAA (P0.05); a similar but not significant trend was observed during the PHA challenge. The primary, but not the secondary, IgG response to OVA was significantly lower in the H and L cortisol responders when compared to the M cortisol responders. Differences in serum IL-6 or IFN-gamma concentration were not observed across variable cortisol-responsive groups. Together, these results demonstrate that cutaneous hypersensitivity reactions are enhanced in outbred H and L cortisol-responding sheep, independent of systemic modulation by IL-6 and IFN-gamma.

An experiment was conducted to investigate the effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on performance, hematology, metabolism, and immunological parameters of turkeys. The efficacy of polymeric glucomannan mycotoxin adsorbent (GMA) in preventing these adverse effects was also evaluated. Three hundred 1-d-old male turkey poults were fed wheat-, corn-, and soybean meal-based starter (0 to 3 wk), grower (4 to 6 wk), developer (7 to 9 wk), and finisher (10 to 12 wk) diets formulated with uncontaminated grains, contaminated grains, and contaminated grains + 0.2% GMA. Feeding contaminated grains significantly decreased BW gains during the grower and developer phases, and GMA supplementation prevented these effects. There was no effect of diet, however, on feed intake or feed efficiency. The feeding of contaminated grains reduced total lymphocyte counts at wk 3 (P < 0.05). Dietary supplementation with GMA increased plasma total protein concentrations compared with controls and birds fed the contaminated diet. Plasma uric acid concentrations in birds fed contaminated grains were increased at the end of the experiment compared with controls, and the feeding of GMA prevented this effect. Feeding contaminated grains significantly increased the percentage of CD4(+) lymphocyte populations during wk 6; however, there was no change in the percentage of CD8(+) and B-lymphocyte populations. Contact hypersensitivity to dinitrochlorobenzene, which is a CD8(+) T cell-mediated delayed-type hypersensitivity response, was significantly decreased after 24 and 72 h by feedborne mycotoxins compared with controls. Supplementation of the contaminated diet with GMA prevented the decrease in response after 24 h. Secondary antibody (IgG titer) response against SRBC antigens (CD4(+) T cell-dependent) was significantly decreased after feeding contaminated grains compared with controls. It was concluded that turkey performance and some blood and immunological parameters were adversely affected by feedborne Fusarium mycotoxins, and GMA prevented many of these effects.

Mycotoxin contamination in pet food poses a serious health threat to pets, causing an emotional and economical concern to the pet owners. Aflatoxins, ochratoxins, trichothecenes, zearalenone, fumonisins and fusaric acid have been found in the ingredients and final products of pet food, resulting in both acute toxicity and chronic health problems in pets. Toxicological interaction among mycotoxins as a natural mixture further complicates the issue. The concepts of "risk assessment", using hazard identification, dose-response assessment, no observable adverse effect level (NOAEL), and lowest observed adverse effect level (LOAEL), should be applied to assess the risk and safety of mycotoxins in pet food, thereby instilling public confidence in the pet food industry.

Objective—To investigate the effects of feeding cereal-based diets that are naturally contaminated with Fusarium mycotoxins to dogs and assess the efficacy of a polymeric glucomannan mycotoxin adsorbent (GMA) in prevention of Fusarium mycotoxicosis. Animals—12 mature female Beagles. Procedures—Dogs received each of 3 cereal-based diets for 14 days. One diet was uncontaminated (control diet), and the other 2 contained contaminated grains; one of the contaminated diets also contained 0.2% GMA. Contaminants included deoxynivalenol, 15-acetyl deoxynivalenol, zearalenone, and fusaric acid. Food intake and nutrient digestibility, body weight, blood pressure, heart rate, and clinicopathologic variables of the dogs were assessed at intervals during the feeding periods. Results—Food intake and body weight of dogs fed the contaminated diet without GMA were significantly decreased, compared with effects of the control diet. Reductions in blood pressure; heart rate; serum concentrations of total protein, globulin, and fibrinogen; and serum activities of alkaline phosphatase and amylase as well as increases in blood monocyte count and mean corpuscular volume were detected. Consumption of GMA did not ameliorate the effects of the Fusarium mycotoxins. For the GMA-contaminated diet, digestibility of carbohydrate, protein, and lipid was significantly higher than that associated with the control diet, possibly because of physiologic adaptation of the recipient dogs to reduced food intake. Conclusions and Clinical Relevance—Results indicated that consumption of grains naturally contaminated with Fusarium mycotoxins can adversely affect dogs' feeding behaviors and metabolism. As a food additive, GMA was not effective in prevention of Fusarium mycotoxicosis in dogs.

Bovine macrophages (BoMacs) were exposed to the following Penicillium mycotoxins (PM): citrinin (CIT), ochratoxin A (OTA), patulin (PAT), mycophenolic acid (MPA) and penicillic acid (PA). PM exposure at the concentration that inhibits proliferation by 25% (IC25) differentially for 24h altered the gene expression of various cytokines. OTA significantly induced IL-1α expression (p

Feeding grains naturally contaminated with Fusarium mycotoxins has been shown to alter metabolism and performance of laying hens. The objectives of the current experiment were to examine the effects of feeding grains naturally contaminated with Fusarium mycotoxins on hematology and immunological indices and functions of laying hens and the possible protective effect of feeding a polymeric glucomannan mycotoxin adsorbent (GMA). One hundred forty-four laying hens were fed for 12 wk with diets formulated with (1) uncontaminated grains, (2) contaminated grains, or (3) contaminated grains + 0.2% GMA. Fusarium mycotoxins such as deoxynivalenol (DON, 12 mg/kg), 15-acetyl-DON (0.5 mg/kg), and zearalenone (0.6 mg/kg) were identified in the contaminated diets arising from contaminated grains grown in Ontario, Canada. The concentrations of DON arising from naturally contaminated grains in this study were similar to purified mycotoxin fed to experimental mice. The chronic feeding of Fusarium mycotoxins induced small decreases in hematocrit values, total numbers of white blood cells, lymphocytes including both CD4+ and CD8+ T lymphocytes and B lymphocytes, and biliary IgA concentration. Supplementation of diets containing feedborne mycotoxins with GMA prevented the reduction in total number of B lymphocytes in the peripheral blood and the reduction in biliary IgA concentration. In addition, the delayed-type hypersensitivity response to dinitrochlorobenzene was increased by feed-borne mycotoxins, whereas IgG and IgM antibody titers to sheep red blood cells were not affected by diet. We concluded that chronic consumption of grains naturally contaminated with Fusarium mycotoxins at levels likely to be encountered in practice were not systemically immunosuppressive or hematotoxic; however, mucosal immunocompetence needs to be explored further.

Feeding grains naturally-contaminated with Fusarium mycotoxins has been shown to alter the metabolism and performance of turkeys. The objectives of the current experiment were to examine the effects of feeding turkeys with grains naturally contaminated with Fusarium mycotoxins on their hematology and immunological indices (including functions), and the possible protective effect of feeding a polymeric glucomannan mycotoxin adsorbent (GMA). Two hundred twenty-five 1-d-old male turkey poults were fed corn, wheat, and soybean meal-based starter (0 to 3 wk), grower (4 to 6 wk), developer (7 to 9 wk), and finisher (10 to 12 wk) diets formulated with uncontaminated grains, contaminated grains, or contaminated grains with 0.2% GMA. The chronic consumption of Fusarium mycotoxins caused minor and transient changes in hematocrit (0.33 L/L) and hemoglobin (10(6) g/L) concentrations as well as in blood basophil (0.13 x 10(9)/L) and monocyte counts (3.42 x 10(9)/L) compared with controls. Supplementation of the contaminated diet with GMA prevented these effects on blood cell counts. Biliary IgA concentrations were significantly increased (4.45-fold) when birds were fed contaminated grains compared with controls, but serum IgA concentrations were not affected. Contact hypersensitivity to dinitrochlorobenzene, which is a CD8+ T-cell-mediated delayed-type hypersensitivity response, was decreased (48%) by feed-borne mycotoxins compared with the control. By contrast, the primary and secondary antibody response to sheep red blood cells, a CD4+ T-cell-mediated response, was not affected. It was concluded that chronic consumption of grains naturally contaminated with Fusarium mycotoxins exerts only minor adverse effects on the hematology and some immunological indices of turkeys. Consumption of grains naturally contaminated with Fusarium mycotoxins may, however, increase the susceptibility of turkeys to infectious agents against which CD8+ T cells play a major role in defense.

Experiments were conducted to determine the effects of feeding grains naturally contaminated with Fusarium mycotoxins on performance, metabolism, hematology, and immune competence of ducklings. Four hundred sixty-four 1-d-old White Pekin male ducklings were fed starter (0 to 2 wk), grower (3 to 4 wk), and finisher (5 to 6 wk) diets formulated with uncontaminated grains, a low level of contaminated grains, a high level of contaminated grains, or the higher level of contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent. Body weight gains, feed consumption, and feed efficiency were not affected by diet. However, consumption of contaminated grains decreased plasma calcium concentrations after 2 wk and plasma uric acid concentrations at the 4-wk assessment point. Mean corpuscular hemoglobin concentrations and hematocrit decreased when ducks were fed contaminated grains for 4 or 6 wk, respectively. In contrast, total numbers of white blood cells and lymphocytes increased transiently in birds fed contaminated grains for 4 wk. The antibody response to sheep red blood cells (CD4+ T cell dependent) and the cell-mediated response to phytohemagglutinin-P (also CD4+ T cell dependent) were not affected by diet, but consumption of contaminated grains for 6 wk decreased the duration of peak cell-mediated response to dinitrochlorobenzene (CD8+ T cell dependent) assessed in a skin test. Feeding grains naturally contaminated with Fusarium mycotoxins, even at levels widely regarded as high, exerted only minor adverse effects on plasma chemistry and hematology of ducklings, and production parameters were unaffected in this avian species. Mycotoxin-contaminated feeds may, however, render these animals susceptible to infectious agents such as viruses against which the CD8+ T cell provides necessary defence. Glucomannan mycotoxin adsorbent was not effective in preventing alterations caused by Fusarium mycotoxins.

Objective—To characterize hematologic and clinical consequences of chronic dietary consumption of freeze-dried garlic at maximum voluntary intake in horses.Animals—4 healthy sex- and age-matched horses.Procedure—An initial garlic dose (0.05 g/kg, twice daily) was fed to 2 horses in a molasses carrier as part of their normal ration and was gradually increased to maximum voluntary intake (0.25 g/kg, twice daily) over 41 days. Dietary supplementation then continued for a total of 71 days. Two control horses were fed molasses with no garlic with their ration. Blood samples were collected weekly and analyzed for hematologic and biochemical changes, including the presence of Heinz bodies. Recovery of affected blood values was followed for 5 weeks after termination of dietary supplementation with garlic.Results—At a daily dose of > 0.2 g/kg, horses fed garlic developed hematologic and biochemical indications of Heinz body anemia, as characterized by increases in Heinz body score (HBS), mean corpuscular volume (MCV), mean corpuscular hemoglobin, platelet count, and serum unconjugated and total bilirubin concentrations and decreases in RBC count, blood hemoglobin concentration, mean corpuscular hemoglobin concentration, and serum haptoglobin concentration. Recovery from anemia was largely complete within 5 weeks after termination of dietary supplementation with garlic. Heinz body score and MCV remained high at the end of the 5-week recovery period.Conclusions and Clinical Relevance—Horses will voluntarily consume sufficient quantities of garlic to cause Heinz body anemia. The potential for garlic toxicosis exists when horses are chronically fed garlic. Further study is required to determine the safe dietary dose of garlic in horses. (Am J Vet Res2005;66:457–465)

An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on growth and immunological parameters of broiler chickens. Three hundred sixty, 1-d-old male broiler chicks were fed 1 of 4 diets containing grains naturally contaminated with Fusarium mycotoxins for 56 d. The diets included (1) control; (2) low level of contaminated grains (5.9 mg/kg deoxynivalenol (DON), 19.1 mg/kg fusaric acid (FA), 0.4 mg/kg zearalenone, and 0.3 mg/kg 15-acetyldeoxynivalenol; (3) high level of contaminated grains (9.5 mg/kg DON, 21.4 mg/kg FA, 0.7 mg/kg zearalenone, and 0.5 mg/kg 15-acetyldeoxynivalenol); and (4) high level of contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent (GM polymer). Body weight gains and feed consumption of chickens fed contaminated grains decreased linearly with the inclusion of contaminated grains during the grower phase (d 21 to 42). Efficiency of feed utilization, however, was not affected by diet. Production parameters were not significantly affected by the supplementation of GM polymer to the contaminated grains. Peripheral blood monocytes decreased linearly in birds fed contaminated grains. The feeding of contaminated diets linearly reduced the B-cell count at the end of the experiment, whereas the T-cell count on d 28 responded quadratically to the contaminated diets. The feeding of contaminated diets did not significantly alter serum or bile immunoglobulin concentrations, contact hypersensitivity to dinitrochlorobenzene, or antibody response to SRBC. Supplementation with GM polymer in the contaminated diet nonspecifically increased white blood cell count and lymphocyte count, while preventing mycotoxin-induced decreases in B-cell counts. It was concluded that broiler chickens are susceptible during extended feeding of grains naturally contaminated with Fusarium mycotoxins.

Semipermeable membrane devices (SPMDs), RTL-W1 and H4IIE cell line-derived 2,3,7,8-TCDD equivalents (TEQs) and hepatic 7-ethoxyresorufin O-deethylase (EROD) activity were used to show that following 28-days in creosote-dosed microcosms, rainbow trout (Oncorhynchus mykiss) were exposed to biologically-active polycyclic aromatic hydrocarbons (PAHs), despite the fact that PAHs were not detected at high concentrations in the trout tissue. Rainbow trout and SPMDs were exposed to creosote at doses of 0, 5, 9, 17, 31 and 56 μL/L in microcosms. Total PAH concentration in SPMDs increased with creosote dose (r 2 = 0.910, p < 0.05). In contrast, concentrations of PAHs in rainbow trout liver extracts did not increase with creosote dose, likely due to metabolism of these compounds. SPMD TEQs derived using both cell line bioassays (BD-TEQs) or toxic equivalent factors (TEFs; CD-TEQs) increased with creosote dose, revealing that more Ah receptor-active compounds were accumulated at higher creosote doses. Trout ...

The effects of glyphosate ((N-phosphonomethyl)glycine) and triclopyr (((3,5,6-trichloro-2-pyridinyl)oxy)acetic acid) on the embryonic development of Xenopus laeviswere evaluated using Frog Embryo Teratogenesis Assay— Xenopus(FETAX). Rodeot, the isopropylamine (ipa) salt of glyphosate formulated without a surfactant was found to be the least toxic, with a LC5 and LC50 of 3,779 and 5,407 mg acid equivalent (AE)/L, respectively. The LC5 and LC50 of Roundupt, the ipa salt of glyphosate formulated with a surfactant, was 6.4 and 9.4 mg AE/L, respectively. The surfactant component of Roundup, polyoxyethyleneamine (POEA), had a LC5 and LC50 of 2.2 and 2.7 mg/L, respectively. Garlont 3A, the triethylamine salt of triclopyr, had a LC5 and LC50 of 119 and 162.5 mg AE/L, respectively. The LC5 and LC50 of Garlon 4t, the butoxyethyl ester of triclopyr, was 6.7 and 9.3 mg AE/L, respectively. Considering a theoretical worst case scenario when the highest rates recommended for glyphosate (12 L of Roundup/ha) or triclopyr (8 L of Garlon/ha) are applied to water 15 cm in depth, the expected environmental concentrations calculated on the basis of AE would be 2.8 and 2.6 mg AE/L, respectively. The margins of safety (LC5/expected environmental concentrations) for frog embryos exposed to these concentrations would be approximately 2, 2, 47, and 1,312 for Roundup, Garlon 4, Garlon 3A, and Rodeo, respectively.

1. The individual and combined effects of T‐2 toxin and 4,15‐diacetoxyscirpenol (DAS) on laying hens were investigated in an experiment consisting of a 2 X 2 completely randomised factorial design with dietary concentrations of 0 and 2 mg/kg T‐2 toxin and 0 and 2 mg/kg DAS. 2. Individually, T‐2 toxin and DAS induced oral lesions in half of the hens and decreased significantly egg production and food intake. 3. The effects of T‐2 toxin and DAS were additive for reduced food consumption and incidence of oral lesions. However, a synergism for reduced egg production was observed during the last experimental period. 4. No effects on body weight were observed during this study. Mild changes in selected plasma enzymes activities and no change in liver malondialde‐hyde content were detected. 5. The combination of T‐2 toxin and DAS was more toxic than the single mycotoxins, for some parameters, and therefore, may pose a greater economic threat to the poultry industry than either of the toxins individually.

Mallard (Anas platyrhynchos) ducklings were led for distances of 50, 150 and 300 m through plots of upland vegetation that had been sprayed with a flowable formulation of carbofuran at rates of 0, 132 and 264 g active ingredient/ha. Duckling behavior was observed and quantified immediately after exposure by using an approach-response apparatus. Brain and plasma cholinesterase (ChE) activities were determined for a subsample of ducklings from each treatment group; the remaining ducklings were monitored for growth effects. No mortality occurred during field exposure, but toxic signs were observed in ducklings moving 150 m, 17 and 100% of the ducklings had difficulty keeping pace due to anti-ChE toxicosis in the low and high spray plots, respectively. Duckling approach-response behavior became significantly slower with increasing spray rate and exposure distance (p = 0.0001). Inhibition of brain ChE activity was directly related to both spray rate and exposure distance; ducklings receiving maximum carbofuran exposure had 29% of the mean ChE activities of controls. Plasma ChE activity was inversely related to spray rate but was not affected by exposure distance and did not correlate well with brain ChE activity (R2 = 0.36). Subsequent growth rates were not affected in a dose-related manner.

A rapid, two-stain (fluorescein diacetate and ethidium homodimer-1) flow cytometric assay to evaluat viability and cytotoxicity of the alga Selenastrum capricornutum in preserved samples is described. For storage, stained cells were fixed in glutaraldehyde, flash frozen in liquid nitrogen, and stored frozen (−20°C) for assessment at a later date. Weekly analysis of frozen samples showed that fluorescence was stable for 7 weeks. A mixture of 50% healthy and 50% heat-killed cells of S. capricornutum showed 36.1% healthy cells, 13.2% compromised cells, 12% nonstained cells, and 38.7% dead cells. This technique was tested using sodium dodecyl sulfate (SDS) and phenol as toxicants. Threshold concentrations for toxicant impact were in the range of 1 to 10 mg/L for SDS and 10 to 100 mg/L for phenol. Estimates of mortality showed 96-h median lethal concentration (LC50) values of 2,340 mg/L and 6,970 mg/L for SDS and phenol, respectively. This two-stain flow cytometric procedure has proven to be a reliable, sensitive assay for determining viability of S. capricornutum in preserved samples. The sensitivity of this dual-color assay and the ability to store samples for later analysis are significant improvements over current techniques.

We administered oral doses of carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl methyl carbamate) twice daily to mallard (Anas platyrhynchos) ducklings during a 10-day exposure period. Daily dosage levels included 0.00 (control), 0.25, 0.45, and 0.85 mg of carbofuran per kg of body mass. At the highest dosage 68.8% of the ducklings died, with 40.6% mortality occurring after the initial dose. Clinical signs of cholinesterase (ChE) inhibition were observed in ducklings from the high and medium dosage groups. Growth rates of these 2 groups were lower than the control rate. After an initial decline, mean body mass recovered to the level of controls 1 week after exposure in the 0.45 mg/kg/day group, but remained lower in the 0.85 mg/kg/day group. Although ducklings in the highest dosage group fledged at a mass similar to controls, they fledged 6-18 days later. Brain ChE activity of ducklings that died following dosing was 6-38% of that of controls but was not different among dosage groups. Cholinesterase activity of ducklings that survived dosing and were euthanized 6 hours later was not different from controls