Microbiological assays for antibiotics and vitamins: considerations for assuring accuracy.

Factors that may influence the accuracy and precision of microbiological manual and semiautomated turbidimetric methods as well as diffusion assays are discussed. Influence of kind of equipment, media, test bacteria, sample preparation, form of dose response lines, operations, and personnel on quality of assays is examined with the objective of reducing to insignificance those factors under control of the analyst that are responsible for low quality assays.     

Rapid method for extracting aldrin, dieldrin, and endrin from visceral material.

A simple method is described for purifying aldrin, dieldrin, and endrin from visceral material by using a Celite column. Celite retains all interfering materials including Lipids whereas the chlorinated Insecticides are eluted from the column with n-hexane. This extract can be used directly for gas chromatographic analysis.     

Chitinase production by an Arthrobacter sp. lysing cells of Fusarium roseum

An Arthrobacter sp. which actively lysed Fusarium roseum was found to liberate chitinase (E.C. 3.2-1.14, chitin glycanohydrolase), an enzyme essential for the hydrolysis of chitin, a major component of fusarial hyphal walls. Factors involved in the production of chitinase were investigated by modifying culture conditions and assaying for enzyme activity. Production occurred on colloidal chitin as well as on native chitin supplemented with yeast extract or peptone. Enzyme production paralleled growth; liberation of enzyme took place during the log phase with the maximum yield being obtained at the stationary phase. Addition of the non-ionic surfactant, polyoxyethylene sorbitan monooleate (Tween 80) increased enzyme yield. An inverse relationship was found between the amount of enzyme produced and the quantity of n-acetyl-glucosamine liberated. The enzyme was generally not produced when grown on various other carbohydrates. These findings suggest that chitinase is inducible and that chitin breakdown is regulated by a repressor-inducer mechanism.Initial hydrolysis rates of colloidal chitin were proportional to the concentration of chitinase used. Optimal pH and temperature for enzyme activity were 4.9 and 50°C, respectively. Purification of the chitinase was obtained by (NH₄)₂SO₄ precipitation followed by DEAE-cellulose and Sephadex chromatography, achieving a 12-fold increase in specific activity.     

Horizontal transmission of Mycobacterium avium subsp. paratuberculosis in cattle in an experimental setting: calves can transmit the infection to other calves

In September 2001, two subsequent transmission experiments both lasting 3 months were carried out to study cow-calf transmission of Mycobacterium avium subsp. paratuberculosis (Map) (Period 1), followed by calf-calf transmission of the infection (Period 2). Every 2 weeks, serum, heparinised blood and faecal samples were collected from all animals. After these experiments, the 20 calves were housed individually for more than 3 years to be able to detect the infection status and excretion pattern of each animal. In autumn 2004, the animals were inseminated, to observe a possible increase in faecal excretion of Map shortly before expected calving. One month before the expected calving date in 2005, animals were slaughtered and several tissues per cow and unborn calf were sampled for culture. The results indicate that horizontal cow-calf transmission is readily achieved (Period 1). At the highest infection pressure (six shedding cows of which three high shedders in Period 1) all five calves excreted Map in their faeces during Period 1 (shortly after infection), and four of these calves during Period 2 (when the shedding cows were absent). After that, excretion became less frequently. Horizontal calf-calf transmission did take place (Period 2), as the four donor-calves infected two receiver-calves. Transmission rates during the 3 months periods were quantified as a reproduction ratio R. The R [95% CI] of cow-calf and calf-calf transmission were estimated as 2.7 [1.1, 6.6] and 0.9 [0.1, 3.2] new infections per infectious animal during 3 months.     

Remediation of Soils Contaminated with Molybdenum using Soil Amendments and Phytoremediation

For ruminants, there is a narrow span betweennutritional deficiency of Molybdenum and its potential toxicity. Molybdenosis occurs among cattlefeeding on forage with Mo concentrations above10 μg g^-1 or a Cu:Mo ratio <2. In the areaunder investigation forage Mo contents in the valleyare as high as 180 μg g^-1 due to industrialpollution, while the alpine pastures, where cattlegraze during summer, are nutrient (e.g. Cu) deficient.When driven to the valley pastures in fall, animalsoften fell ill with molybdenosis, and several died.The aim of the study was to remediate this 300 ha area. Two approaches were attempted: (1) to reduce theMo contents of the forage by immobilizing soil Mo, and(2) to increase plant Mo contents by mobilizing soilMo to increase plant Mo which may then be removed fromthe system (phytoremediation).In a soil extraction experiment we demonstrated thatphosphate fertilization, ammonium sulfate amendment,vermiculite, humic acid and sewage sludge increased Moleaching by 30 to 110%. Fe-humate and Mn-humateapplication decreased Mo in the leachate from96 μg L^-1 to 55 and 7 μg L^-1,respectively. Plant Mo contents were increased up to170% by P-fertilizer and up to 150% by vermiculite.Conversely, sewage sludge, Fe- and Mn-humate decreasedplant Mo contents by 60, 40 and 75% in thegreenhouse. In the field study, the effects weresmaller, but Mo export through plant harvest increasedby 88% after P-fertilization and 84% aftervermiculite amendment. Mn-humate and sewage sludgedecreased plant Mo content by 25 and 40%,respectively, rendering the forage suitable forfeeding of ruminants. P-fertilization and vermiculitemay thus be recommended for the severely contaminatedsites to enhance phytoremediation through Mo export,and Mn-humate and sewage sludge application appearsuited to remediate the less severely contaminated sites.