Immunoaffinity column coupled with solution fluorometry or liquid chromatography postcolumn derivatization for determination of aflatoxins in corn, peanuts, and peanut butter: collaborative study

An AOAC/IUPAC (International Union of Pure and Applied Chemistry) collaborative study was conducted to evaluate the effectiveness of an immunoaffinity column for the determination of aflatoxin. The test portion is extracted with methanol-water (7 + 3), filtered, diluted to less than 30% methanol with water, and applied to the affinity column. The column is washed with water and the concentrated aflatoxins are eluted with methanol. Total aflatoxins are determined by solution fluorometry with bromine (SFB), and individual toxins are determined by reverse-phase liquid chromatography with postcolumn derivatization with iodine (PCD). Corn naturally contaminated with aflatoxins, and peanuts, peanut butter, and corn containing added aflatoxins (B1:B2:G1:G2 = 7:1:3:1) were sent to 24 collaborators in the United States, France, Canada, and the Republic of South Africa. Twelve collaborators used the SFB method, 9 used the PCD method, and 3 used both SFB and PCD methods. Twenty collaborators completed the study (10 used the SFB method, 7 used the PCD method, and 3 used both SFB and PCD methods). Test portions were spiked at 10, 20, and 30 ng/g. For SFB analyses, recoveries of total aflatoxins were 123, 105, and 107%, respectively; the relative standard deviation for repeatability (RSDr) ranged from 11.75 to 16.57%, and the relative standard deviation for reproducibility (RSDR) ranged from 10.97 to 33.09%. For PCD analyses, recoveries were 81, 81, and 83%, respectively; the RSDr ranged from 5.20 to 17.22%, and the RSDR ranged from 4.68 to 50.77%. The RSDr for aflatoxins B1 and G1 for spiked test portions ranged from 5.45 to 23.55%, and the RSDR ranged from 4.21 to 57.28%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of organic and conventional crop rotation, fertilization, and crop protection practices on metal contents in wheat (Triticum aestivum)

The effects of organic versus conventional crop management practices (crop rotation, crop protection, and fertility management strategies) on wheat yields and grain metal (Al, Cd, Cu, Ni, Pb, and Zn) concentrations were investigated in a long-term field trial. The interactions between crop management practices and the season that the crop was grown were investigated using univariate and redundancy analysis approaches. Grain yields were highest where conventional fertility management and crop protection practices were used, but growing wheat after a previous crop of grass/clover was shown to partially compensate for yield reductions due to the use of organic fertility management. All metals except for Pb were significantly affected by crop management practices and the year that the wheat was grown. Grain Cd and Cu levels were higher on average when conventional fertility management practices were used. Al and Cu were higher on average when conventional crop protection practices were used. The results demonstrate that there is potential to manage metal concentrations in the diet by adopting specific crop management practices shown to affect crop uptake of metals.     

Suspect novel adverse drug reactions to trimethoprim-sulphonamide combinations in horses: a case series

Adverse drug reactions to trimethoprim-sulphonamide combinations are common in many species, manifesting as gastrointestinal tract disorders, dermatopathies and blood dyscrasias. In this case series, neurological abnormalities in 4 horses being treated with trimethoprim-sulphonamide combinations at normal dosages and in one foal that received an overdose are described. The horses developed hypermetric gait, agitation and erratic behaviour. All signs resolved once medication was withdrawn, and no horse had residual deficits. No other cause for observed neurological deficits could be determined. These clinical signs appear to represent a novel adverse drug reaction to some commonly used antimicrobial combinations.     

Preliminary findings on the experimental transmission of chronic wasting disease agent of mule deer to cattle.

To determine the transmissibility of chronic wasting disease (CWD) to cattle and to provide information about clinical course, lesions, and suitability of currently used diagnostic procedures for detection of CWD in cattle, 13 calves were inoculated intracerebrally with brain suspension from mule deer naturally affected with CWD. Between 24 and 27 months postinoculation, 3 animals became recumbent and were euthanized. Gross necropsies revealed emaciation in 2 animals and a large pulmonary abscess in the third. Brains were examined for protease-resistant prion protein (PrP(res)) by immunohistochemistry and Western blotting and for scrapie-associated fibrils (SAFs) by negative-stain electron microscopy. Microscopic lesions in the brain were subtle in 2 animals and absent in the third case. However, all 3 animals were positive for PrP(res) by immunohistochemistry and Western blot, and SAFs were detected in 2 of the animals. An uninoculated control animal euthanized during the same period did not have PrP(res) in its brain. These are preliminary observations from a currently in-progress experiment. Three years after the CWD challenge, the 10 remaining inoculated cattle are alive and apparently healthy. These preliminary findings demonstrate that diagnostic techniques currently used for bovine spongiform encephalopathy (BSE) surveillance would also detect CWD in cattle should it occur naturally.     

Idiopathic disseminated intracytoplasmic neuronal vacuolation in a neonatal Holstein calf born in the USA.

Histopathologic, immunohistochemical, and ultrastructural evaluations were made of a 6-day-old Holstein calf with severe vacuolation of the neuronal perikarya that was widely distributed throughout the central nervous system. No evidence of storage material within the vacuoles was revealed by histopathologic and ultrastructural examinations. Immunohistochemical and electron microscopic examinations were negative for protease-resistant prion protein and scrapie-associated fibrils, respectively. These results indicate that the clinical signs in this calf were not associated with transmissible spongiform encephalopathy. Neuronal vacuolation has not previously been documented in calves.     

Resistance of domestic cats to a US sheep scrapie agent by intracerebral route.

Feline spongiform encephalopathy (FSE) is thought to have resulted from consumption of food contaminated with bovine spongiform encephalopathy and the latter is believed to result from the consumption of food contaminated with scrapie. However, no direct experimental documentation exists to indicate that the scrapie agent is capable of amplifying in cats, and, therefore, crossing the species barrier. During 1979, 6 cats ranging in age from 3.5 to 18 months were intracerebrally inoculated with sheep scrapie (inoculum G-639-PP) and were observed for an extended period. Inoculated cats did not develop neurologic disease, and microscopic lesions of spongiform encephalopathy were not evident. Immunohistochemistry and Western blot techniques failed to detect the abnormal form of prion protein (PrP(res)). These results indicate that the sheep scrapie agent (G-639-PP) used in this study was not capable of amplifying in cats and therefore was unable to cross the species barrier to produce FSE.     

Survey of the seroprevalence of brucellosis in ruminants in Tajikistan

A cross-sectional serological survey of the prevalence of brucellosis in ruminants in the Region of Republican Subordination and Khatlon oblasts (provinces) in Tajikistan was conducted in May 2003. Sera from 13,625 ruminants involving 3513 households in 172 kishlaks (villages) were collected and screened by the rose bengal test. Doubtful and positive results were further tested with competitive and indirect elisas. The overall serological prevalences (95 per cent confidence intervals [cis]) were 5.8 per cent (5.2 to 6.4 per cent) for sheep, 5.5 per cent (5.0 to 6.0 per cent) for goats and 2.1 per cent (1.0 to 3.2 per cent) for cattle. The results show that brucellosis was a common disease of ruminants that was widely but unevenly distributed throughout the two oblasts. Seropositive animals were found in 119 of the 172 kishlaks (69.2 per cent [95 per cent ci 61.9 to 75.6 per cent]) and 14.4 per cent (95 per cent ci 13.3 to 15.6 per cent) of the 3513 households. Evidence of infection was also found in cattle kept for milk production in urban kishlaks in two major cities and in state-owned dairy farms.     

Reactions of hard red spring wheat to common root rot under field conditions of Northern United States of America

Common root rot (CRR), primarily caused by Cochliobolus sativus, is one of the important soil borne diseases of wheat (Triticum aestivum L). To gain information about resistance of hard red spring wheat (HRSW) to the disease, forty HRSW cultivars and advanced breeding lines were grown and evaluated in a completely randomized block design for reaction to CRR from 2000 to 2002 in a field plot with high natural inoculum of C. sativus at Williston, ND. Plant root samples during the soft to mid-dough stage were collected and CRR incidences and severities were scored using the subcrown internode index method. Disease severities (DS) varied from 24–61% in 2000 and 2002, but were relatively low in 2001 (19–40%). None of the wheat cultivars was immune to CRR. The genotypes Canon, Forge, Dandy, Ember, Butte 86, Oxen, AC Barrie, Parshall, and 2371 had higher DS (>45%) and exhibited susceptible disease reactions. However, the genotypes ND 722, AC Cadillac, HJ 98, Argent and Scholar had lower levels of DS throughout the study period, and these were better than, or similar to Amidon, the resistant check. The HRSW cultivars with consistent resistance to CRR during the three year evaluation could be valuable resistance sources for use in breeding programs.