Substrate utilization by Aspergillus flavus in inoculated whole corn kernels and isolated tissues

Utilization of the major corn (Zea mays) reserve materials (free saccharides, starch, triglycerides, and zein) was monitored during infection of detached kernels by Aspergillus flavus (A. flavus) over a 12-day period. Inoculated whole kernels were compared to noninoculated kernels. Concentrations of sucrose and raffinose in inoculated seed decreased to nearly zero at 6 days, whereas concentrations of these saccharides in noninoculated seed dropped at a considerably slower rate, and significant levels remained at the end of the incubation period. Triglyceride concentrations remained unchanged in the noninoculated seed but dropped continuously after 2 days in the inoculated seed. Starch and zein concentrations did not change during the 12-day incubation period. Aflatoxin B1 was first detected after 2 days and increased to about 20 microg/g (20,000 ppb) after 12 days. Very low aflatoxin concentrations were detected in the noninoculated seed. Significant concentrations of erythritol, arabitol, and mannitol were produced during infection, with peak concentrations occurring at 8 days. Whole seed and germ tissue appeared to support good fungal growth and aflatoxin production, whereas ground tissues and endosperm did not. A. flavus preferentially utilized saccharides as initial carbon substrates followed by triglycerides. When invading nonwounded corn kernels, the fungus selectively targets the germ tissue where these materials are localized in the highest concentrations.     

Liver biopsy techniques for adult horses and neonatal foals to assess copper status

Objective To evaluate standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals, as techniques for obtaining tissue for assessment of copper status. The techniques were evaluated with respect to ease of use and effect on the animal.
Procedure Twenty of 24 Thoroughbred mares and 21 of their foals were biopsied. The animals were part of a larger study of the effect of copper supplementation on copper status and the prevalence of developmental orthopaedic disease. Livers were also collected from unrelated horses and sampled to investigate the variability in the distribution of copper in liver tissue.
Result The biopsy technique caused no lasting effect on the mares, but there was an increased risk of viscus penetration associated with taking multiple biopsy cores. The use of ultrasonography to scan the target area for the liver identified four cases that were not appropriate candidates for liver biopsy, because of large intestine being located in the biopsy area. In the foals there were no serious postoperative adverse effects, nor was there any evidence of problems caused by the procedure when the abdomen was examined post-mortem at 5 months of age. In livers collected to investigate the variability of copper concentration, copper appeared to be relatively evenly distributed through the liver.
Conclusion Standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals are convenient procedures for obtaining liver tissue for assessing copper status in horses. The use of ultrasound to identify liver tissue is recommended, especially in older mares.     

Naphthalophos combinations with benzimidazoles or levamisole as effective anthelmintics for sheep

Objective: To investigate the relative efficacy and safety of the anthelmintic naphthalophos in sheep, either given alone or in combination with benzimidazole (fenbendazole and albendazole) or levamisole anthelmintics.
Design: A parasitological study using faecal egg count reduction tests, a validating slaughter trial and field safety trials.
Procedure: Faecal egg count reduction tests were carried out on 13 farms. Naphthalophos and combinations of naphthalophos with levamisole and fenbendazole were included in the drench tests. On one property a controlled efficacy study was carried out to validate faecal egg count reduction test findings. In this trial, sheep were slaughtered 10 days after treatment and the remaining parasites recovered from the gastro-intestinal tract. Safety trials were carried out on eight farms where approximately 50 000 sheep were treated with naphthalophos and albendazole that were tank mixed in the backpack.
Results: The efficacy of naphthalophos alone in faecal egg count reduction tests ranged from 59 to 98% with one test showing 95% reduction. The efficacy of naphthalophos and levamisole ranged from 74 to 100%, with 5 farms showing 95% reduction. The efficacy of naphthalophos and fenbendazole ranged between 88 and 100% with 95% reduction achieved on 10 farms. The controlled efficacy study showed a good correlation between the faecal egg count reduction tests and numbers of parasites recovered, except for Nematodirus where the faecal egg count reduction tests overestimated efficacy. The mortality rate in the safety trials was 0.05%, with most fatalities occurring on one farm.
Conclusion: The combination of naphthalophos and fenbendazole was more effective than a combination of naphthalophos and levamisole, and will provide a sufficiently safe drench rotation option.     

Toxicity of (+)- and (-)-gossypol to the plant pathogen,Rhizoctonia solani

The dimeric sesquiterpene gossypol occurs naturally in cottonseed and other parts of the cotton plant. Gossypol exists as enantiomers because of the restricted rotation around the central binaphthyl bond. The (-)-enantiomer is toxic to nonruminant animals while the (+)-enantiomer exhibits little, if any, toxicity to these animals. Developing cotton plants with low levels of the (-)-gossypol could expand the use of cottonseed as a feed source. Gossypol also may play a role in protecting the plant from pathogens. The relative toxicity of (+)- and (-)-gossypol to plant pathogens has not been reported. We measured the concentration of (+)- and (-)-gossypol in roots from cotton seedlings that were treated with the biocontrol agent Trichoderma virens that induces biosynthesis of gossypol and related terpenoids in cotton roots. (-)-Gossypol was the minor enantiomer in control and treated roots, but levels were slightly higher in roots from T. virens-treated seed. We also determined the toxicity of the gossypol enantiomers and the racemate to the seedling disease pathogen Rhizoctonia solani. The (+)- and (-)-enantiomers of gossypol and the racemate are equally effective in inhibiting growth of this pathogen. The lethal doses of the gossypols required to kill the pathogen appeared to be similar, but their toxicities are significantly less than those of related cotton and kenaf sesquiterpenes. The results indicate that altering the enantiomeric ratio in cotton roots will not adversely affect the resistance of seedlings to the seedling pathogen R. solani.     

Developmental changes in trans-permethrin and α-naphthyl acetate ester hydrolysis during the last larval instar of Pseudoplusia includens

The hydrolysis of trans-permethrin and α-naphthyl acatate by midgut, fat body, and cuticle homogenates from Pseudoplusia includens (Walker) was monitored during the development of the last instar. The midgut homogenates appeared to have two pH optima (7.6 and 8.6) for the hydrolysis of trans-permethrin, the fat body homogenates had one optimum (7.4–7.8), and the cuticle homogenates had a major optimum at 6.6. Hydrolysis of both substrates peaked during the late feeding stages for midgut and cuticle homogenates, although relative changes were not the same. Hydrolysis of trans-permethrin peaked during the late feeding stage in fat body homogenates, while hydrolysis of α-naphthyl acetate continually increased through the prepupal stage. Thus, the hydrolysis of α-naphthyl acetate is not necessarily associated with the hydrolysis of trans-permethrin. The LD₅₀ values for trans-permethrin on the different stages appeared to reflect the influence of hydrolysis.