Antibody responses of calves to Histophilus somni recombinant IbpA subunits

Histophilus somni causes bovine pneumonia and septicemia, but protective immune responses are not well understood and immunodiagnostic methods are not well defined. We previously showed that antibody to a new virulence factor, IbpA, neutralizes cytotoxicity and immunization with a recombinant IbpA domain protects calves against experimental H. somni pneumonia. To further define immune responses to IbpA, we determined isotypic serum antibody responses to three IbpA domains (IbpA3, an N-terminal coiled coil region; IbpA5, a central region of 200 bp repeats and IbpA DR2, a C-terminal cytotoxic domain). ELISA was used to quantitate IgG1 or IgG2 antibodies to each of the IbpA subunits as well as H. somni whole cells (WCs) or culture supernatant (SUP). Calves experimentally infected with H. somni and monitored for up to 10 weeks had the least "0 time" (background) antibody levels to IbpA5, as well as the earliest and highest responses of greatest duration to the IbpA5 subunit. Responses of these calves were high to WC or SUP antigens but with higher "0 time" (background) antibody levels. We concluded that IbpA5 may be a useful immunodiagnostic antigen. Calves immunized with H. somni WC vaccine had antibody responses to WC antigens, but not to IbpA subunits before challenge. After challenge with H. somni, vaccinated calves had slight anamnestic responses to IbpA3 and IbpA5, but not to IbpA DR2. Since IbpA DR2 is a protective antigen, the data suggest the IbpA DR2 would be a useful addition to H. somni vaccines.     

Microwave-assisted headspace solid-phase microextraction for the analysis of bioemissions from Eucalyptus citriodora leaves

Microwave-assisted headspace solid-phase microextraction (MA-HS-SPME) was developed as a simple and effective method for fast sampling of volatile organic compounds (VOCs) from Eucalyptus citriodora Hook (E. citriodora) leaves. During microwave heating, a simple shielding device made of aluminum foil was used to protect the SPME fiber from microwave irradiation while allowing the sample to be heated. A room temperature water bath was also used to allow microwave heating to be conducted in a more controlled manner. The inner heating caused by microwave irradiation dramatically accelerated the emission of VOCs from the sample, but no marked change in headspace temperature in the sample vial was found. Under optimum conditions, the extraction efficiencies obtained with microwave heating were much higher than those obtained without microwave heating for all fibers used, namely, 7-microm polydimethylsiloxane (PDMS), 100-microm polydimethylsiloxane (PDMS), 65-microm polydimethylsiloxane/divinylbenzene (PDMS/DVB), and 75-microm carboxen/polydimethylsiloxane (CAR/PDMS). The improvement of extraction efficiency using MA-HS-SPME allowed more VOC events to be detected, with more balanced extraction of VOCs of lower and higher molecular masses. Moreover, a good linear relationship was found between sample size and GC-FID response (total peak area of VOCs), indicating the usefulness of MA-HS-SPME for quantitative analysis of individual volatile compounds in E. citriodora leaves.     

Precursors to damascenone: synthesis and hydrolysis of isomeric 3,9-dihydroxymegastigma-4,6,7-trienes

A series of four isomeric 3,9-dihydroxymegastigma-4,6,7-trienes, 8, has been prepared. The (3S,6R,9S) isomer of 8 proved to be identical to an isomer of this compound tentatively identified as an intermediate in the formation of damascenone from an allene triol. Each of the four isomers, when hydrolyzed independently of each other at pH 3.0 and 25 degrees C, produced product mixtures in which the major product was damascenone (1). Contrary to expectation, 3-hydroxydamascone (5) was not observed in any of the hydrolyses. Consequently, the mechanism of formation of damascenone proposed earlier requires modification. In each hydrolysis, the product mixtures showed the presence of a second isomer of 8, produced by epimerization during hydrolysis. Chiral analysis on a Cyclosil B column revealed that this epimerization was occurring at C(3) in each of the hydrolyses.     

Determination of capsaicinoids in habanero peppers by chemometric analysis of UV spectral data

A novel spectrophotometric method for the determination of capsaicinoids in habanero pepper extracts is described that does not require prior analyte separation. The method uses partial-least-squares (PLS-1) multivariate regression modeling techniques in conjunction with ordinary UV absorption spectral data obtained on alcoholic extracts of habanero peppers (Capsicum chinese). The PLS-1 regression models were developed by correlating the known total concentration of the two major capsaicinoids (capsaicin and dihydrocapsaicin) in the extracts as determined by high-performance liquid chromatography with the spectral data. The regression models were subsequently validated with laboratory-prepared test sets. The validation studies revealed that the root-mean-square error of prediction varied from 4 to 8 ppm, based on the results obtained from models prepared from nine test sets. Once a regression model has been developed and validated, analyses of the extracts can be accomplished rapidly by ordinary spectrophotometric procedures without any prior separation steps.     

USE OF 99mTc-MERCAPTOACETYLTRIGLYCINE TO EVALUATE RENAL FUNCTION IN HORSES

Ten healthy horses were injected intravenously with 99mTc-MAG3 and the disappearance of radioactivity from the blood was measured. The total body clearance (Cl(B)) and elimination half-life (t1/2(beta)) were 7.9 +/- 1.5 ml/kg/minute and 32.8 +/- 4.1 minutes, respectively. The disappearance of 99mTc-MAG3 from the blood of 2 horses with compromised renal function was also measured. The data suggest that 99mTc-MAG3 is a useful and clinically applicable radiopharmaceutical for measurement of effective renal blood flow in the horse.     

A decision-analytic approach to the optimal allocation of resources for endangered species consultation

The resources available to support conservation work, whether time or money, are limited. Decision makers need methods to help them identify the optimal allocation of limited resources to meet conservation goals, and decision analysis is uniquely suited to assist with the development of such methods. In recent years, a number of case studies have been described that examine optimal conservation decisions under fiscal constraints; here we develop methods to look at other types of constraints, including limited staff and regulatory deadlines. In the US, Section Seven consultation, an important component of protection under the federal Endangered Species Act, requires that federal agencies overseeing projects consult with federal biologists to avoid jeopardizing species. A benefit of consultation is negotiation of project modifications that lessen impacts on species, so staff time allocated to consultation supports conservation. However, some offices have experienced declining staff, potentially reducing the efficacy of consultation. This is true of the US Fish and Wildlife Service’s Washington Fish and Wildlife Office (WFWO) and its consultation work on federally-threatened bull trout (Salvelinus confluentus). To improve effectiveness, WFWO managers needed a tool to help allocate this work to maximize conservation benefits. We used a decision-analytic approach to score projects based on the value of staff time investment, and then identified an optimal decision rule for how scored projects would be allocated across bins, where projects in different bins received different time investments. We found that, given current staff, the optimal decision rule placed 80% of informal consultations (those where expected effects are beneficial, insignificant, or discountable) in a short bin where they would be completed without negotiating changes. The remaining 20% would be placed in a long bin, warranting an investment of seven days, including time for negotiation. For formal consultations (those where expected effects are significant), 82% of projects would be placed in a long bin, with an average time investment of 15 days. The WFWO is using this decision-support tool to help allocate staff time. Because workload allocation decisions are iterative, we describe a monitoring plan designed to increase the tool’s efficacy over time. This work has general application beyond Section Seven consultation, in that it provides a framework for efficient investment of staff time in conservation when such time is limited and when regulatory deadlines prevent an unconstrained approach.     

Comparison of Oil,Shortening, and a Structured Shortening on Wheat Dough Rheology and Starch Pasting Properties

Monoacylglycerol‐stabilized oil in water emulsion (MAG gel) is an alternate shortening that is free of trans fatty acids, and low in saturated fatty acids. However, the behavior of MAG gels in comparison to other lipids has not been studied. This study investigated effects of structured MAG gel, a mixture of MAG gel unstructured components (Mixture), canola oil (Oil), or interesterified soy shortening (IE Soy) at different levels (6–24%) on hard or soft wheat dough properties. Doughs were prepared with different lipid types at equivalent lipid contents. Dough mixing and water absorption parameters were evaluated using a farinograph; gluten behavior was measured using a gluten peak tester (GPT); and pasting characteristics were measured using a micro‐viscoamylograph (MVAG). Water absorption values decreased with increasing lipid content. Dough development times were similar between the MAG gel and IE Soy, but farinogram curve characteristics during mixing were similar between MAG gel, Mixture, and Oil. The trend for peak max time in GPT was similar between MAG gel and IE Soy exhibiting delayed gluten aggregation; whereas Mixture and Oil exhibited earlier gluten aggregation. In MVAG, starch interaction with monoglyceride component of MAG gel and Mixture appeared to be the dominating factor resulting in increased pasting temperature and a second viscosity peak during cooling at higher levels of lipid addition.