A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

Improved method for determination of pectin degree of esterification by diffuse reflectance Fourier transform infrared spectroscopy

An improved method for the determination of pectin degree of esterification (DE) by diffuse reflectance Fourier transform infrared spectroscopy (DRIFTS) was developed. Pectin samples with a range of DE as determined by gas chromatography were used for developing a calibration curve by DRIFTS. A linear relationship between the DE of pectin standards and FTIR peak ratio for ester carboxyl peak area to total carboxyl peak area was found (R(2) = 0.97). Pectin DE of various samples was calculated from the linear fit equation developed by DRIFTS. Accuracy of the DRIFTS method was determined by comparing the DE values of four commercial pectins obtained by DRIFTS methods to the values obtained by the gas chromatography method. Greater precision was obtained for the FTIR measurement of test pectin samples when the ester peak ratio was used relative to the ester peak area.     

Determination of potassium guaiacolsulfonate by ion-pairing partition chromatography.

Potassium guaiacolsulfonate, a highly polar, acidic substance, is readily eluted by chloroform from a pH 4.5 Celite column in the form of its ion-pair with trihexylamine, thereby effecting facile separation from other pharmaceuticals. The sulfonic acid is then back-extracted into aqueous alkali and determined spectrophotometrically. Assay of standard solutions by this procedre averaged 100.44 plus or minus 0.81%. The method was applied successfully to 4 commercial cough preparations containing a variety of other drugs.     

Complementary host–pathogen genetic analyses of the role of fumonisins in the Zea mays–Gibberella moniliformis interaction

Zea mays often is colonized with the fungus Gibberella moniliformis, which produces fumonisin toxins. The role of fumonisins in seedling colonization and blight was studied using complementary genetic analyses of host and pathogen. Only one of two fumonisin B1 (FB1)-insensitive maize backcross lines was more resistant than the FB1-sensitive parent to seedling blight, indicating that the increase in FB1-insensitivity was not associated with an increase in resistance. FB1-producing and nonproducing isogenic fungal strains did not differ in ability to cause seedling blight, but the FB1-producing strain was more effective in systemic colonization of seedlings in reciprocal strain challenge tests. Together, these and previous results indicate that the role of fumonisins depends on complex environmental and genetic contexts in this host–pathogen interaction.     

Acute cortisol responses of calves to scoop dehorning using local anaesthesia and/or cautery of the wound

Objective To measure plasma cortisol responses in calves dehorned using a scoop after administration of local anaesthesia and/or cautery of the wounds.
Design A physiological study with controls.
Procedure There were six treatments: control handling with and without local anaesthesia, dehorning, dehorning after local anaesthesia, dehorning followed by wound cautery, and dehorning after local anaesthesia followed by wound cautery. Blood samples were taken before and after dehorning.
Results Dehorning caused an increase in plasma cortisol concentrations, which decreased a little to plateau values and then declined to pretreatment values 3 to 4 h after dehorning. The peak was smaller after local anaesthesia was administered but when its effects wore off, cortisol concentrations increased and thereafter were similar to those in the dehorned animals. The combination of local anaesthesia and cautery resulted in a plasma cortisol response similar to those in control calves with or without local anaesthesia.
Conclusions If plasma cortisol concentrations reflect the distress being experienced by the calves, then local anaesthesia reduces the acute distress for about 3 h after dehorning but not during the subsequent 3 to 4 h. Combining local anaesthetic and cautery prevented the significant increase in plasma cortisol following dehorning and may eliminate the acute distress caused by scoop dehorning.     

Timing of shedding seeds and cones, and production in different stands of Scots pines at Abernethy Forest, Scotland

A study was carried out over 11 seed years on the timing ofshedding of seeds and cones, and annual seed fall and cone productionin three stands of native Scots pinewood and a Scots pine plantationin Abernethy Forest, Scotland. Peaks in seed fall took placemainly in May, and cones were shed mainly between June and August.There were few residual seeds remaining in shed cones. Synchronizedpeaks in seed fall and cone production (mast years) took placeat 3-year intervals across the different stands. The differencebetween cohorts of high and low cone production ranged fromfactors of 5 to 20 among sites. Coefficients of variation forcone production ranged from 62 to 84 per cent among sites. Therewere no significant differences in cone production among sites,but there were site-related differences in seed fall. The largercanopy cover in the plantation probably accounted for the higherseed fall per square metre there, though variations in the amountof seed eaten by birds and mammals may also have been important.Canopy cover needs to be considered when converting cone densitiesunder crowns to cone density per unit of woodland area. A similarcalculation is difficult for seeds because they are lighterthan cones and many fall outside the area under the crowns.The results are discussed in relation to the potential for treeregeneration and the availability of food for birds and mammalsprior to seed dispersal.     

Kinetics of photoirradiation-induced synthesis of soy oil-conjugated linoleic acid isomers

Photoirradiation of soy oil with UV/visible light has been shown to produce significant amounts of trans,trans conjugated linoleic acid (CLA) isomers through conversion of various synthesized intermediate cis,trans isomers. The objective of this study was to determine the kinetics of CLA isomers synthesis to better understand the production of various isomers. Soy oil was irradiated with UV/visible light for 144 h in the presence of an iodine catalyst and CLA isomers analyzed by gas chromatography (GC). Arrhenius plots were developed for the conversion of soy oil linoleic acid (A) to form cis-, trans/trans-, cis-CLA (B), conversion of cis-, trans/trans-, cis-CLA to form trans,trans-CLA (C) with respect to B, and formation of trans,trans-CLA isomers with respect to C. The kinetics of consumption of linoleic acid (LA) to form cis-, trans/trans-, cis-CLA was found to be of second-order with a rate constant of 9.01 x 10-7 L/mol s. The rate of formation of cis-, trans/trans-, cis-CLA isomers depends on the rate of formation from LA and its rate of consumption to form trans,trans-CLA isomers. The conversion of cis-, trans/trans-, cis-CLA isomers to trans,trans-CLA isomers was found to be of first-order with a rate constant of 2.75 x 10-6 s-1. However, the formation of thermodynamically stable trans,trans-CLA isomers (C) with respect to C was found to be a zero-order reaction with a rate constant of 10.66 x 10-7 mol/L s. The consumption of LA was found to be the rate-determining step in the CLA isomers formation reaction mechanism. The findings provide a better understanding of the mechanism of CLA isomers synthesis by photoirradiation and the factors controlling the ratio of various isomers.     

Expression of Perilipin 2 (PLIN2) in Porcine Oocytes During Maturation

Perilipins have been reported to limit the interaction of lipases with neutral lipids within the droplets, thereby regulating neutral lipid accumulation and utilization. This study aimed to identify the location and expression of PLIN1 and PLIN2 in porcine oocytes during maturation. Quantitative real‐time polymerase chain reaction (qRT‐PCR), immunostaining and Western blot methods were used to characterize the expression and distribution patterns of PLIN1 and PLIN2 in porcine oocytes. The results showed that PLIN1 was not detectable in porcine oocytes. PLIN2 and BODIPY 493/503‐detected neutral lipid droplets appeared identical distribution patterns and extensive colocalization in both GV and MII porcine oocytes. PLIN2 protein expression was higher in GV oocytes than that in MII oocytes (p < 0.05), although PLIN2 mRNA expression was similar in both groups. These findings suggested that PLIN2 was a major lipid droplet‐associated protein in porcine oocytes.