Crotalaria species containing hepatotoxic pyrrolizidine alkaloids grow widely in pastures in northern Australia and have sporadically poisoned grazing livestock. The diverse Crotalaria taxa present in these pastures include varieties, subspecies, and chemotypes not previously chemically examined. This paper reports the pyrrolizidine alkaloid composition and content of 24 Crotalaria taxa from this region and assesses the risk of poisoning in livestock consuming them. Alkaloids present in C. goreensis , C. aridicola subsp. densifolia, and C. medicaginea var. neglecta lack the esterified 1,2-unsaturated functionality required for pyrrole adduct formation, and these taxa are not hepatotoxic. Taxa with high levels of hepatotoxic alkaloids, abundance, and biomass pose the greatest risk to livestock health, particularly C. novae-hollandiae subsp. novae-hollandiae, C. ramosissima , C. retusa var. retusa, and C. crispata . Other species containing moderate alkaloid levels, C. spectabilis and C. mitchellii , also pose significant risk when locally abundant.
Sorghum ergot should not exceed 0.3% (1 mg alkaloid/kg) in diets of multiparous sows fed before farrowing, and should be limited to 0.1% for primiparous sows, or avoided completely.
Austral bracken Pteridium esculentum contains three unstable norsesquiterpene glycosides: ptaquiloside, ptesculentoside, and caudatoside, in variable proportions. The concentration of each of the glycosides was determined in this study as their respective degradation products, pterosin B, pterosin G and pterosin A, by HPLC-UV analysis. Samples of P. esculentum collected from six sites in eastern Australia contained up to 17 mg of total glycoside/g DW, with both ptaquiloside and ptesculentoside present as major components accompanied by smaller amounts of caudatoside. Ratios of ptaquiloside to ptesculentoside varied from 1:3 to 4:3, but in all Australian samples ptesculentoside was a significant component. This profile differed substantially from that of P. esculentum from New Zealand, which contained only small amounts of both ptesculentoside and caudatoside, with ptaquiloside as the dominant component. A similar profile with ptaquiloside as the dominant glycoside was obtained for Pteridium aquilinum subsp. wightianum (previously P. revolutum ) from northern Queensland and also P. aquilinum from European sources. Ptesculentoside has chemical reactivity similar to that of ptaquiloside and presumably biological activity similar to that of this potent carcinogen. The presence of this additional reactive glycoside in Australian P. esculentum implies greater toxicity for consuming animals than previously estimated from ptaquiloside content alone.
Agalactia and feed refusal are classical signs of poisoning by rye ergot (C purpurea), but this is the first time that sorghum ergot has been associated with a similar syndrome.
Australian isolates of Fusarium pseudograminearum (Fp = F. graminearum Group 1) and F. graminearum (Fg = F. graminearum Group 2) can produce mycotoxins including zearalenone (ZEA), 4-deoxynivalenol (DON), and nivalenol (NIV). Fp isolates from wheat and barley tillers in southern Queensland all produced ZEA and DON in culture, and one typical isolate also produced 3-acetyldeoxynivalenol. Most Fg isolates from wheat and sorghum grains in southern Queensland produced ZEA and DON and one typical isolate also produced 15-acetyldeoxynivalenol. Fg isolates from maize plants in northern Queensland were all ZEA and NIV producers, which was consistent with previous reports, and they also produced high concentrations of acetyl-nivalenols. ZEA and either DON or NIV production by cultures derived from different conidia (and ascospores in Fg isolates) varied by 4-18-fold for ZEA and 2-4-fold for DON/NIV production, and there were significant negative correlations between ZEA and either DON or NIV, indicating a common controlling process. The pattern of ZEA production was quite different between Fp and Fg, with ZEA production being relatively delayed in Fg. After 7 days incubation at 28°C on maize meal, one Fp isolate produced 49 mg ZEA/kg, but in both DON-producing and NIV-producing isolates of Fg, ZEA concentrations after 7 days were <1 mg/kg. ZEA and DON were produced on sorghum and combined wheat-barley grains as well as maize meal, although there were trends for maize meal to be more productive, probably due to greater surface area or different gaseous exchange. Low temperature incubation of a Fg DON-type isolate increased ZEA production, but did not affect either a Fg NIV-type isolate or a Fp isolate. Relationships between these patterns of mycotoxin production, pathogenicity, and implications for crop contamination are discussed.
Austral bracken, Pteridium esculentum , occurs widely in Australian grazing lands and contains both the known carcinogen ptaquiloside and its hydroxy analogue, ptesculentoside, with untested carcinogenic potential. Calves were fed a diet containing 19% P. esculentum that delivered 1.8 mg of ptaquiloside and 4.0 mg of ptesculentoside per kilogram of body weight (bw) per day to explore the carcass residue potential of these compounds. Concentrations of ptaquiloside and ptesculentoside in the liver, kidney, skeletal muscle, heart, and blood of these calves were determined as their respective elimination products, pterosin B and pterosin G, by HPLC-UV analysis. Plasma concentrations of up to 0.97 μg/mL ptaquiloside and 1.30 μg/mL ptesculentoside were found, but were shown to deplete to <10% of these values within 24 h of bracken consumption. Both glycosides were also detected in all tissues assayed, with ptesculentoside appearing to be more residual than ptaquiloside. Up to 0.42 and 0.32 μg/g ptesculentoside was present in skeletal muscle and liver, respectively, 15 days after bracken consumption ended. This detection of residual glycosides in tissues of cattle feeding on Austral bracken raises health concerns for consumers and warrants further investigation.
Abstract. Assay methods for the alkaloids of sorghum ergot (Claviceps africana) are described and compared. Sorghum ergot bodies (sclerotia/sphacelia) from various regions of Queensland and New South Wales were collected in 1997 and 2001 and assayed by spectrophotometry, thin layer chromatography, or high performance liquid chromatography (HPLC). All contained dihydroergosine (DHES) as the main alkaloid component (about 80%), with smaller amounts of dihydroelymoclavine and festuclavine. The preferred method of assay for infected sorghum and mixed feeds involved extraction into dichloromethane:methanol:ethyl acetate:ammonium hydroxide (50:5:25:1) using an ultrasonic bath. After solvent removal, the extract was dissolved in diethyl ether and partitioned into 0.5 M hydrochloric acid. After adjusting the pH to 8-10 with ammonium hydroxide, the alkaloids were extracted into dichloromethane, the solvent evaporated, and the residue dissolved in methanol. HPLC separation was on a C18 column, 150 by 3.9 mm, run isocratically at 40°C, with acetonitrile : 0.1% ammonium acetate:methanol (31:50:20) as the mobile phase. Detection was either by UV at 280 nm or by fluorescence with excitation at 235 nm and absorbance at 340 nm. Levels of quantitation for DHES in sorghum approached 0.1 mg/kg (UV) and 0.01 mg/kg (fluorescence). Method recoveries for DHES in the range of 0.025-7 mg/kg averaged 75%. The total alkaloid content of ergot bodies (sclerotia/sphacelia) from different batches of grain varied from 100 to 7900 mg/kg (0.79%). Within batches, there was much less variation in the alkaloid content of ergot bodies, but larger ergots tended to contain more alkaloid than smaller ergots, and those infected with Cerebella species contained even less; this probably related to the ratio of sclerotial/sphacelial tissue present. Honeydew also contained DHES (1-10 mg/kg) and might contaminate clean grain at significant levels. Tests on 4 farms showed that substantial amounts of ergot bodies and alkaloids were removed during grain harvesting. . B l a n e y e t a l .
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