Effects of tumour necrosis factor‐alpha and interleukin‐1 beta on in vitro development of bovine secondary follicles

The objective of this study was to determine the effects of TNF‐α and IL‐1β on development and survival of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (~0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5°C, with 5% CO₂ in air, for 18 days, in TCM‐199⁺ alone (cultured control) or supplemented with 10 ng/ml IL‐1β, 10 ng/ml TNF‐α or both TNF‐α and IL‐1β. The effects of these treatments on growth, follicular survival, antrum formation, viability, ultrastructure and mRNA levels for GDF‐9, c‐MOS, H1foo and Cyclin B1 were evaluated. The results showed that addition of TNF‐α to culture medium increased follicular diameter and rate of antrum formation, whereas that of IL‐1β and a mixture of IL‐1β and TNF‐α did not do so. Ultrastructural analysis showed that, among the tested cytokine treatments, follicles cultured in the presence of TNF‐α had the best‐preserved oocytes and granulosa cells. The presence of TNF‐α, IL‐1β or both did not influence the expression of mRNAs analysed. In conclusion, in contrast to IL‐1β, TNF‐α promotes growth of and antrum formation in in vitro cultured bovine secondary follicles, while their ultrastructure and viability were maintained.     

Spittlebugs as vectors of <Emphasis Type="Italic">Xylella fastidiosa</Emphasis> in olive orchards in Italy

The recent introduction of Xylella fastidiosa in Europe and its involvement in the Olive Quick Decline Syndrome (OQDS) in Apulia (Salento, Lecce district, South Italy) led us to investigate the biology and transmission ability of the meadow spittlebug, Philaenus spumarius, which was recently demonstrated to transmit X. fastidiosa to periwinkle plants. Four xylem-sap-feeding insect species were found within and bordering olive orchards across Salento during a survey carried out from October 2013 to December 2014: P. spumarius was the most abundant species on non-olive vegetation in olive orchards as well as on olive foliage and was the only species that consistently tested positive for the presence of X. fastidiosa using real-time PCR. P. spumarius, whose nymphs develop within spittle on weeds during the spring, are likely to move from weeds beneath olive trees to olive canopy during the dry period (May to October 2014). The first X. fastidiosa-infective P. spumarius were collected in May from olive canopy: all the individuals previously collected on weeds tested negative for the bacterium. Experiments demonstrated that P. spumarius transmitted X. fastidiosa from infected to uninfected olive plants. Moreover, P. spumarius acquired X. fastidiosa from several host plant species in the field, with the highest acquisition rate from olive, polygala and acacia. Scanning electron microscopy (SEM) revealed bacterial cells resembling X. fastidiosa in the foreguts of adult P. spumarius. The data presented here are essential to plan an effective IPM strategy and limit further spread of the fastidious bacterium.     

Predatory ability of the ant <Emphasis Type="Italic">Crematogaster scutellaris</Emphasis> on the brown marmorated stink bug <Emphasis Type="Italic">Halyomorpha halys</Emphasis>

The pest Halyomorpha halys is a recent example of severe worldwide biological invasion. In Italy, after the first detection in 2012, it has rapidly spread and economic damage has already been reported in commercial fruit orchards, eliciting enormous concern for Italian agricultural production. As a long-term solution, biological control investigations are focusing on the potential of natural antagonists, such as generalist predators, in the invaded areas. Due to their biology, ecology and behavior, ants represent one such generalist predator potentially suitable for inflicting mortality on H. halys. In this research, Crematogaster scutellaris was used as a study model. Specifically, the effect of ant predatory behavior on the survival of all H. halys pre-imaginal stages was tested in the laboratory environment. The results suggest that C. scutellaris has a significant predatory impact on all H. halys pre-imaginal stages except for eggs. The reduction of survivorship is between 95 and 85% for instars 1 and 2 and between 38 and 52% for instars 3, 4 and 5. The different levels of predation success suggest that these ants could act as useful generalist predators when associated with other antagonists and/or other control strategies. Although further field investigations are needed, the potential of ants as biocontrol agents of invasive species in agroecosystems deserves consideration in integrated pest management programs.     

An integrated method for high-resolution definition of new diameter-based fine root sub-classes of <Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.

Key message

Compared to the traditional approach, applying micrometric image analysis to fine root samples of Fagus sylvatica with subsequent data treatment through principal component and cluster analysis yielded specific diameter sizes for fine root sub-classes having better resolution of the corresponding branching orders, and a more coherent relationship with the values of annual production and turnover rate.

Context

Fine root traits are poorly understood, impeding an accurate representation of terrestrial biogeochemical models. Traditionally used, arbitrary diameter thresholds lead to a misestimation of fine root traits such as branching order, environmental relationship, annual production, and turnover rate.

Aims

Here, we present, as modification of the traditional method, an integrated approach to segregate, at high-resolution, fine root populations of Fagus sylvatica into new diameter sub-classes that better correspond with the traits mentioned above.

Methods

Samples, collected with a sequential soil coring method, were subjected to a micrometric image analysis, and resultant data were treated with principal component and cluster analysis.

Results

Results showed that fine roots were distributed into diameter-size sub-classes (0–0.3 mm, 0.3–1 mm, and 1–2 mm) different from those determined by traditional methods (0–0.5 mm, 0.5–1 mm, and 1–2 mm). New sub-classes provided a better resolution of the corresponding branching-orders, and the values of annual production and turnover rate were more coherent with diameter class and soil depth. Moreover, new sub-classes provided a more precise match with soil temperature than traditional methods.

Conclusion

Our method may help to unveil fine root dynamics and development, reduce data analysis time, and make the diameter-based classification more precise and trustworthy even in the case of non-intact samples.

     

Autosomal trisomy 20 (61,XX,+20) in a malformed bovine fetus.

A 240-day-gestation female bovine fetus with severe anasarca, palatoschisis, cheiloschisis, mild cranioschisis, and a flattened facies was collected at a slaughterhouse, and a fibroblast line was established from the fetal skin. Chromosome preparations were Q-banded, and chromosome counts were taken that indicated the presence of 61 chromosomes in cells of the fetus (the normal diploid number for domestic cattle is 60). Q-band karyotypes were constructed, and Q-band analysis revealed the presence of three copies of chromosome 20. Trisomy 20 (61,XX,+20) was confirmed through the use of two-color fluorescence in situ hybridization of bovine bacterial artificial chromosome clones that were specific to chromosome 20 and the X chromosome.     

Comparative efficacy of several disinfectants in suspension and carrier tests against Haemophilus parasuis serovars 1 and 5

The comparative efficacy of 16 active compounds (including the most commonly used chemical groups) and 10 commercial formulations against Haemophilus parasuis serovars 1 and 5 was studied. These organisms were tested in suspension and carrier tests in the presence and absence of serum as representative of organic matter. Chloramine-T and half of the formulations from commercial sources (most of them including quaternary ammonium compounds) were effective in both in vitro tests, regardless of the presence or absence of organic load. All 26 disinfectants except for an iodophor (0.1% available iodine) resulted in at least 3-log₁₀ reduction in colony-forming units in suspension test, and most of them resulted in the maximal level of detection (>6-log₁₀ reduction). On the other hand, disinfectants were not as effective in carrier test as in suspension test, and the presence of serum considerably reduced the activities of most of the compounds tested, especially in carrier test. These results suggest the importance of selecting suitable disinfection for routine use on surfaces contaminated with H. parasuis, particularly when organic matter is present. Chloramine-T and formulations 2 and 7–10 are recommended for a complete inactivation of H. parasuis in swine herds.     

Acute-phase protein response in pigs experimentally infected with Haemophilus parasuis

The acute-phase protein (APP) response to an infection caused by Haemophilus parasuis, the etiological agent of Glässer's disease in pigs, was characterized measuring serum concentrations of pig major acute-phase protein (pig MAP), haptoglobin (HPT), C-reactive protein (CRP) and apolipoprotein A-I (ApoA-I) in colostrum-deprived pigs. They were divided into six experimental groups: non-immunized control group (I); immunized with a non-commercial bacterin (II); with an OMP-vaccine (III); with a sublethal dose (IV); and with two commercial bacterins (V and VI). All groups were challenged intratracheally with 5 × 10⁹ CFU of H. parasuis 37 days after immunisation. The highest levels of the positive APPs (pig MAP, HPT and CRP) and the lowest levels of the negative APPs (ApoA-I) were observed in the animals that died as a consequence of the infection, both those in the non-inmunized and in the immunized groups. However, the surviving animals (all of them in groups II, V and VI, two pigs in group III, and three in group IV) showed a minor variation in APP response, mainly on day 1 post-challenge (p.c.), and then tended to recover the initial values. APP response was still less pronounced in the groups of pigs previously immunized with bacterins. In conclusion, APP response can reflect Glässer-disease ongoing, showing a correlation between the severity and duration of the clinical signs and lesions and the magnitude of changes in the APP levels.     

Accounting for extensive topographic and pedologic secondary information to improve soil mapping

Soils generally vary in a quite complex way. To assess and model spatial variability, several variables can be sampled in addition to the target used to quantify and describe the main phenomenon under study. One of the major advantages of geostatistics over simpler spatialization methods is that sparsely observations of the primary attribute can be complemented by secondary attributes that are more densely sampled. Two methods were applied to incorporate dense secondary information: multicollocated cokriging, which restricts the neighbourhood to the only secondary data collocated with the available data of the primary variable and simple cokriging with varying local means related to crisp classes. The objective of this paper was to find the method that best improves the estimation of primary attributes through dense secondary information for the study area, which was the province of Siena in central Italy, an area of about 3820 km². Soil samples were taken at 742 locations and depth, sand and clay contents and available water capacity were determined.     

Effects of added sodium caseinate on the formation of particles in heated milk

The effects of heat at temperatures in the range of 80-90 degrees C on mixtures of reconstituted skim milk powder (RSMP) and sodium caseinate have been determined. In the absence of caseinate, the action of heat on RSMP produces soluble complexes of whey proteins and kappa-casein, as well as complexes of whey protein with the casein micelles. When sodium caseinate was added to RSMP at levels of 0.5 and 1.0%, the denaturation of the whey protein and the production of the soluble complexes in the serum were hardly affected, either in rate or in amount. However, during the heating, the caseinate disappeared from the serum. Further studies on model mixtures of the different components showed that it was probable that the bulk of the caseinate associated with the casein micelles during heating, probably by binding inside the surface layer of kappa-casein, because no increase in the diameters of the casein micelles could be observed.