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.     

Inhalation toxicology.

Two kinds of problems associated with developing standardized procedures for the safety evaluation of new compounds are identified. The first of these is the question of the desirability of using such standard methods. It is concluded that a basic set of procedures is to be recommended, but this should be supplemented with special tests as may be indicated. The second problem is connected with the technical difficulties of any given type of assay and is normally dealt with in terms of the state of the art at the time. Assays by the inhalation route tend to be custom designed and do not follow standard protocols. One of the causes of this situation is the propensity of individual investigators to design de novo the equipment used to effect exposure of animals to airborne substances. Second, some confusion exists because investigators do not always appreciate that the concentration-time product is not the same as the true dose received by the exposed subjects and this may lead to anomalies when dose-response relationships are being characterized. It is suggested that interlaboratory studies be undertaken to ascertain the variability that might be expected in independent assays of inhalation toxicity.     

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.     

Semiautomated method for riboflavin in food products: collaborative study

A collaborative study was conducted comparing a semiautomated riboflavin method with a manual riboflavin method for 10 food products. Six laboratories provided results from the semiautomated method and 16 laboratories used the manual technique. The semiautomated method was more repeatable within a laboratory than was the manual method. The semiautomated method results compared favorably with the manual method for all 10 products.     

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.     

Spatial scale effects on conservation network design: trade-offs and omissions in regional versus local scale planning

Ecological patterns and processes operate at a variety of spatial scales. Those which are regional in nature may not be effectively captured through the combination of conservation plans derived at the local level, where land use planning frequently takes place. Conversely, regional conservation plans may not identify resources important for conservation of intraregional ecological variation. We compare modeled conservation networks derived at regional and local scales from the same area in order to analyze the impact of scale effects on conservation planning. Using the MARXAN reserve selection algorithm and least cost corridor analysis we identified a potential regional conservation network for the Central Valley ecoregion of California, USA, from which we extracted those portions found within five individual counties. We then conducted the same analysis for each of the five counties. An overlay of the results from the two scales shows a general pattern of large differences in the identified networks. Especially noteworthy are the trade-offs and omissions evident at both scales of analysis and the disparateness of the identified corridors that connect core reserves. The results suggest that planning efforts limited to one scale will neglect biodiversity patterns and ecological processes that are important at other scales. An intersection of results from the two scales can potentially be used to prioritize areas for conservation found to be important at several spatial scales.     

Bacterial culture of septic synovial structures of horses: Does a positive bacterial culture influence prognosis?

Reasons for performing study: The influence of synovial fluid culture on short‐ and long‐term prognosis of cases with septic synovitis requires study. Hypotheses: Horses with a positive bacterial culture from septic synovial fluid are less likely to survive or return to successful athletic function than those with a negative bacterial culture from septic synovial fluid. Methods: Records of mature horses presented to 2 equine referral hospitals for investigation of suspected septic synovitis were examined. Horses (n = 206) were included in the study if synovial fluid was submitted for full laboratory examination, including bacterial culture. A diagnosis of septic synovitis was based on a nucleated cell count >30 × 10⁹ cells/l or >90% neutrophils and other clinical, cytological and bacteriological parameters. Long‐term follow‐up was obtained by telephone questionnaire. Univariate analysis, using the Fisher's exact test, was used for all outcomes. Results: Fourteen (20.9%) of 67 horses with a positive bacterial culture from synovial fluid were subjected to euthanasia because of persistent synovial sepsis compared to 2 (1.44%) of 139 with negative bacterial cultures (P<0.001). Overall survival and successful long‐term return to function in horses with a positive bacterial culture was 50% (24/48 horses) compared to 70.5% (74/105) in culture negative horses (P = 0.01). In horses that survived to be discharged, successful long‐term return to function was not significantly different between culture positive and culture negative groups. Growth of Staphylococcus aureus from synovial fluid did not affect short‐term survival to discharge from the hospital compared to other positive bacterial culture; however, successful long‐term return to function was only 30.4% (4/13) in horses from which S. aureus was cultured compared to 73.9% (17/23) of horses in which other bacteria were cultured (P = 0.015). Conclusions and potential clinical relevance: Horses with a positive bacterial culture from a septic synovitis have a poorer prognosis for survival to discharge from hospital and overall long‐term return to function than horses that yielded no bacterial growth. When S. aureus was cultured, the long‐term prognosis was poorer.