Development of RT‐PCR assays using fluorogenic‐3′ minor groove binder DNA probes for detection of fruit tree viruses*

In phytosanitary certification, there is currently a need for the development of reliable, sensitive and rapid tests for the routine detection of Ilarviruses and latent viruses from fruit trees during the dormant season. We have developed real‐time RT‐PCR assays that allow the reliable detection of Prunus necrotic ringspot ilarvirus, Apple chlorotic leafspot trichovirus and Apple stem pitting virus in bark tissues of dormant wood. These assays are well adapted for the routine detection of these three viruses because they eliminate one risk of contamination by performing the whole test in a single closed tube.     

Calf and colostrum management practices on New Zealand dairy farms and their associations with concentrations of total protein in calf serum

AIMS: To gather information on management practices and farmer attitudes to management of cows and calves during the immediate post-partum period on dairy farms in New Zealand, and to assess these practices for associations with concentrations of total protein (TP) in serum of calves 1–8 days-old.

METHOD: Between July and September 2015 blood samples were collected from calves aged between 24 hours and 7 days, from dairy farms (n=105) in nine areas in New Zealand, on three occasions throughout the calving period. Concentrations of TP were determined in all serum samples. At each visit technicians collected 1?L of the pooled colostrum that was intended for feeding to newborn calves that day. These samples were assessed for Brix, coliform and total bacterial counts. After the last sampling visit, the calf rearer or farm manager were asked to complete a questionnaire describing calf and colostrum management practices on the farm. Potential farm-level variables associated with concentrations of TP in serum of sampled calves were identified using univariable and multivariable linear mixed models.

RESULTS: Mean concentration of TP in serum of calves across all farms was 59.8 (95% CI=59.4–60.2) g/L, and was associated with region and herd size in the final multivariable model. Concentrations of TP were lower in calves from farms in Otago (56.2 (95% CI=53.4–58.9) g/L) and Southland (56.9 (95% CI=54.1–59.7) g/L) compared to calves on farms in the Far North (62.6 (95% CI=59.8–65.3) g/L), and were lower in calves from farms with a herd size >600 (58.3 (95% CI=56.7–59.8) g/L) than ≤600 (61.3 (95% CI=60.1–62.5) g/L) cows. After accounting for fixed effects, farm accounted for only 8.4% of the unexplained variation. There was no association between any of the measures of colostrum quality and concentrations of TP in serum (p>0.2).

CONCLUSION AND CLINICAL RELEVANCE: Very few herd-level variables were associated with concentrations of TP in serum. Risk factors that have been shown to be of importance in previous studies outside New Zealand were not identified as important in the current study. It is possible that, in the situation where calves are kept at pasture with their dams for prolonged periods, variables which influence how well a cow can feed its calf in the first 12–24 hours have a larger influence on concentrations of TP in serum than the collection and management of calves once they reach the rearing shed.     

Formation of micella containing solubilized sterols during rehydration of active dry yeasts improves their fermenting capacity

During their rehydration in aqueous media, active dry yeasts (ADY) may be supplemented with inactive yeasts, yeast derivatives, or other optional complementary nutrients to improve their fermentation capacity. We found that yeast sterols solubilized in situ during ADY rehydration were particularly efficient for stimulating the fermenting capacity of ADY. Spontaneous solubilization of sterols during rehydration occurred by the formation of micelles by membrane phospholipids and specific cell wall polysaccharides and sterols, both compounds being provided by inactive dry yeasts (IDY). These micelles contained a specific distribution of the initial sterols from the inactive yeasts. Above a concentration of 100 mg L(-1) in the rehydration medium, these micelles acted as emulsifiers. Their critical micellar concentration (cmc) was found to be about 4 g L(-1). During rehydration, purified micelles, at a concentration near the cmc, were able to interact quickly with yeast cell membranes by modifying the yeast plasma membrane order [monitored by steady-state fluorescence anisotropy of 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene-p-toluenesulfonate (TMA-DPH) probe] and by increasing the sterol contents of ADY. Such an enrichment of ADY by very low concentrations of solubilized sterols was very efficient for the completion of fermentations. This is useful when musts are limited in available phytosterols or when micro-oxygenation is not desirable during fermentation.     

Use of an avoidance test for the assessment of microbial degradation of PAHs

An avoidance test using the soil springtail Folsomia candida was used to assess changes in contamination levels at low doses of PAHs following incubation with indigenous microflora. A soil from a former coke site was diluted to 1% in an unpolluted soil from the same site, which was used as a control, then both substrates were remoistened to 80% field capacity. The diluted soil was previously shown to be strongly repellent to F. candida, although not toxic. After 2-month incubation at 20 °C, the mixture lost its repellence capacity and became attractive to the test animal, while the global 16 PAHs content had decreased to a great extent (50%). PAH disappearance was linked to the occurrence of indigenous microbiota able to degrade hydrocarbons.     

Earthworm excreta (mucus and urine) affect the distribution of springtails in forest soils

The collembolan species Heteromurus nitidus, exclusively located in soils at pH>5, can be cultured in acidic humus. As this species is attracted to the excreta of earthworms from calcic mull, its distribution is supposed to be determined only indirectly by soil pH through the distribution of earthworms. Higher densities and biomasses of Lumbricidae were observed in a calcic mull (pH 7.8) than in an acidic mull (pH 4.8) and a moder humus (pH 4.2). Choice experiments were performed to compare the attraction of H. nitidus to the mucus-urine mix of five earthworm species from the calcic mull and the acidic mull. H. nitidus was attracted to the excreta of the five species, whatever their ecological category and the humus form from which they originated. The collembolan Heteromurus major, which was indifferent to soil pH, was not attracted to earthworm excreta, which emphasizes the significance of this phenomenon for the distribution of H. nitidus over a pH range. The attraction of H. nitidus to earthworm excreta tended to be weaker and more variable when earthworms originated from acidic mull compared to calcic mull, particularly in the case of Lumbricus terrestris. Increased earthworm density reinforced by better mucus quality and quantity could determine the distribution of H. nitidus according to soil acidity. The only urine compound capable of attracting H. nitidus was NH₃ at a low concentration (0.03 g l^-1). The NH₃ content of the mucus-urine mix which attracted H. nitidus was 0.037 g l^-1, and was therefore responsible, at least partly, for the attraction.     

Lymphocyte stimulation: selective destruction of cells during blastogenic response to transplantation antigens

The blastogenic response of human lymphoid cells toward any individual's transplantation antigens can be deleted by the addition of tritiated thymidine of high specific activity during the incubation of the lymphoid cells in mixed leukocyte culture. After the immunocompetent clones which responded to histocompatibility antigens had been destroyed, the remaining population still retained its capacity to respond to unrelated antigens, including other transplantation antigens.