The effect of select seminal plasma proteins on endometrial mRNA cytokine expression in mares susceptible to persistent mating‐induced endometritis

In the horse, breeding induces a transient endometrial inflammation. A subset of mares are unable to resolve this inflammation, and they are considered susceptible to persistent mating‐induced endometritis PMIE Select seminal plasma proteins cysteine‐rich secretory protein‐3 (CRISP‐3) and lactoferrin have been shown to affect the innate immune response to sperm in vitro. The objective of this study was to determine whether the addition of CRISP‐3 and lactoferrin at the time of insemination had an effect on the mRNA expression of endometrial cytokines in susceptible mares after breeding. Six mares classified as susceptible to PMIE were inseminated during four consecutive oestrous cycles with treatments in randomized order of: 1 mg/ml CRISP‐3, 150 μg/ml lactoferrin, seminal plasma (positive control) or lactated Ringer's solution (LRS; negative control) to a total volume of 10 ml combined with 1 × 10⁹ spermatozoa pooled from two stallions. Six hours after treatment, an endometrial biopsy was obtained for qPCR analysis of selected genes associated with inflammation (pro‐inflammatory cytokines interleukin (IL)‐1β, IL‐8, tumour necrosis factor (TNF)‐α, interferon (INF)‐γ, anti‐inflammatory cytokines IL‐1RN and IL‐10, and inflammatory‐modulating cytokine IL‐6). Seminal plasma treatment increased the mRNA expression of IL‐1β (p = .019) and IL‐8 (p = .0068), while suppressing the mRNA expression of TNF (p = .0013). Lactoferrin also suppressed the mRNA expression of TNF (p = .0013). In conclusion, exogenous lactoferrin may be considered as one modulator of the complex series of events resulting in the poorly regulated pro‐inflammatory response seen in susceptible mares.     

Spatial sap flow and xylem anatomical characteristics in olive trees under different irrigation regimes

The compensation heat pulse (CHP) method is widely used to estimate sap flow and transpiration in conducting organs of woody plants. Previous studies have reported a natural azimuthal variability in sap flow, which could have practical implications in locating the CHP probes and integrating their output. Sap flow of several olive trees (Olea europaea L. cv. 'Arbequina') previously grown under different irrigation treatments were monitored by the CHP method, and their xylem anatomical characteristics were analyzed from wood samples taken at the same location in which the probes were installed. A significant azimuthal variability in the sap flow was found in a well-irrigated olive tree monitored by eight CHP probes. The azimuthal variability was well related to crown architecture, but poorly to azimuthal differences in the xylem anatomical characteristics. Well-irrigated and deficit-irrigated olive trees showed similar xylem anatomical characteristics, but they differed in xylem growth and in the ratio of nocturnal-to-diurnal sap flow (N/D index). The results of this work indicate that transpiration cannot be accurately estimated by the CHP method in olive trees if a small number of sensors are employed and that the N/D index could be used as a sensitive water status indicator.     

Modulation of diversity by grazing and mowing in native tallgrass prairie

Species diversity has declined in ecosystems worldwide as a result of habitat fragmentation, eutrophication, and land-use change. If such decline is to be halted ecological mechanisms that restore or maintain biodiversity are needed. Two long-term field experiments were performed in native grassland to assess the effects of fire, nitrogen addition, and grazing or mowing on plant species diversity. In one experiment, richness declined on burned and fertilized treatments, whereas mowing maintained diversity under these conditions. In the second experiment, loss of species diversity due to frequent burning was reversed by bison, a keystone herbivore in North American grasslands. Thus, mowing or the reestablishment of grazing in anthropogenically stressed grasslands enhanced biodiversity.     

Mortality supposedly due to intoxication by pyrrolizidine alkaloids from Heliotropium indicum in a horse population in Costa Rica: a case report.

This article describes a case of massive mortality among horses which was probably due to intoxication by pyrrolizidine alkaloids from Heliotropium indicum. Over 4 years more than 75% of a population of about 110 horses on a farm in Costa Rica died after showing nervous neurological symptoms. Two clinical manifestations were encountered, an acute and a chronic one, both with a fatal outcome. Pathological findings in 2 horses coincided with those reported in the literature for intoxication by pyrrolizidine alkaloids and were not specific for VEE. However Venezuelan equine encephalitis (VEE) was the main differential diagnosis and could not completely be excluded because this disease was endemic in the region and VEE titres were found to be high. Taxonomic and toxicological investigations implicated Heliotropium indicum as the most probable principal cause of the intoxication.     

Kinetics of Plasma Cell‐Free DNA and Creatine Kinase in a Canine Model of Tissue Injury

Background

Cell‐free DNA (cfDNA) comprises short, double‐stranded circulating DNA sequences released from damaged cells. In people, cfDNA concentrations correlate well with disease severity and tissue damage. No reports are available regarding cfDNA kinetics in dogs.

Objectives/Hypothesis

Cell‐free DNA will have a short biological half‐life and would be able to stratify mild, moderate, and severe tissue injury. Our study aims were to determine the kinetics and biological half‐life of cfDNA and to contrast them with those of creatine kinase (CK).

Animals

Three groups of 10 dogs undergoing open ovariohysterectomy, surgery for cranial cruciate ligament rupture (CCLR), or hemilaminectomy.

Methods

Plasma for cfDNA and CK analysis was collected at admission, at induction of anesthesia, postsurgery (time 0) and at 6, 12, 24, 36, 48, 60, and 72 hours after surgery.

Results

The biological half‐life of plasma cfDNA and CK were 5.64 hours (95% confidence interval [CI 95], 4.36–7.98 hours) and 28.7 hours (CI95, 25.3–33.3 hours), respectively. In the hemilaminectomy group, cfDNA concentrations differed significantly from admission at 6–12 hours after surgery. Creatine kinase activity differed among the surgical groups and reached a peak 6 hours after surgery. In the ovariohysterectomy and CCLR groups, plasma CK activity 72 hours after surgery did not differ from admission activity of the ovariohysterectomy group. In contrast, in the hemilaminectomy group, plasma CK activity after 72 hours did not return to the ovariohysterectomy group admission activity.

Conclusions and Clinical Importance

Plasma CK activity has a longer biological half‐life than previously thought. In contrast to plasma CK activity, cfDNA has a short half‐life and could be a useful marker for peracute severe tissue injury.     

Genetic engineering of plant volatile terpenoids: effects on a herbivore,a predator and a parasitoid

BACKGROUND: Most insect‐resistant transgenic crops employ toxins to control pests. A novel approach is to enhance the effectiveness of natural enemies by genetic engineering of the biosynthesis of volatile organic compounds (VOCs). Before the commercialisation of such transgenic plants can be pursued, detailed fundamental studies of their effects on herbivores and their natural enemies are necessary. The linalool/nerolidol synthase gene FaNES1 was constitutively expressed from strawberry in three Arabidopsis thaliana accessions, and the behaviour of the aphid Brevicoryne brassicae L., the parasitoid Diaeretiella rapae McIntosh and the predator Episyrphus balteatus de Geer was studied. RESULTS: Transgenic FaNES1‐expressing plants emitted (E)‐nerolidol and larger amounts of (E)‐DMNT and linalool. Brevicoryne brassicae was repelled by the transgenic lines of two of the accessions, whereas its performance was not affected. Diaeretiella rapae preferred aphid‐infested transgenic plants over aphid‐infested wild‐type plants for two of the accessions. In contrast, female E. balteatus predators did not differentiate between aphid‐infested transgenic or wild‐type plants. CONCLUSION: The results indicate that the genetic engineering of plants to modify their emission of VOCs holds considerable promise for facilitating biological control of herbivores. Validation for crop plants is a necessary next step to assess the usefulness of modified volatile emission in integrated pest management. Copyright © 2012 Society of Chemical Industry     

Spring and autumn animal treading effects on pre‐grazing herbage mass and tiller density on two contrasting pasture types in Ireland

A perennial ryegrass (Lolium perenne L.)‐dominated sward on a well‐drained soil (Experiment 1) and a creeping bent (Agrostis stolonifera L.)‐dominated sward on a poorly drained soil (Experiment 2) were subjected to four treading treatments: control (C, no damage), light damage (L), moderate damage (M) or severe damage (S) to quantify the effects on herbage dry‐matter (DM) production and tiller density. In Experiment 1, treading damage was imposed in spring. In Experiment 2, one‐third of the site was damaged in autumn, one‐third in spring and one‐third in both spring and autumn. Both sites were rotationally grazed after treading treatments. Pre‐grazing herbage mass was measured eight times in Experiment 1 and seven times in Experiment 2 on each plot, and tiller density was assessed four times in each experiment. In Experiment 1, pre‐grazing herbage mass was reduced by 30% in S plots at the first harvest after damage, but cumulative pre‐grazing herbage DM production was not different between treatments (12·7 t DM ha^?1). In Experiment 2, annual cumulative pre‐grazing herbage mass was reduced by between 14 and 49%, depending on intensity of treading damage event and season when damage occurred. Tiller density was not affected by treatment in either experiment. A perennial ryegrass‐dominated sward on a well‐drained soil was resilient to heavy treading damage. A creeping bent‐dominated sward on poorly drained soil requires a more careful grazing management approach to avoid major losses in cumulative pre‐grazing herbage mass production during wet weather grazing events.     

The Use of Liveweight Change as an Indicator of Oestrus in a Seasonally Calving,Pasture‐Fed Dairy Herd

This was an observational study of 828 lactations in 542 mixed‐age dairy cows that calved seasonally in a single, pasture‐fed herd in New Zealand in 2008 and 2009. The study objectives were to: (i) document daily liveweight change (?LW) before and after observed oestrus for cows subsequently diagnosed pregnant or non‐pregnant and (ii) quantify the sensitivity and specificity of ?LW as a test for oestrus. The sensitivity and specificity of ?LW when combined with other commonly used oestrous detection methods was also evaluated. In cows that conceived as a result of service at detected oestrus, liveweight loss began 1 day before the day of detection and was greatest on the day of detection (?9.6 kg, 95% CI ?11.3 kg to ?7.8 kg; p < 0.01) compared with LW recorded 2 days before the day of detection. In cows that did not conceive to a service at a detected oestrus, the lowest liveweights were recorded 1 day before the day oestrus was detected (?4.3 kg, 95% CI ?7.7 to ?0.8 kg; p = 0.02) compared with LW recorded 4 days before the day of detection. The sensitivity and specificity of ?LW as a means of oestrous detection were 0.42 (95% CI 0.40–0.45) and 0.96 (95% CI 0.95–0.97), respectively. When ?LW was combined with tail paint and visual observation, the oestrous detection sensitivity and specificity were 0.86 and 0.94, respectively. Monitoring LW change holds promise to enhance the sensitivity and specificity of oestrous detection in combination with other oestrous detection methods.     

Distinct Effects of Boar Seminal Plasma Fractions Exhibiting Different Protein Profiles on the Functionality of Highly Diluted Boar Spermatozoa

The aim of this study was to evaluate how different protein profiles of seminal plasma (SP) fractions affect sperm functionality in vitro. Ejaculates from three boars were separated into six fractions. The fractions differed from each other in their sperm content, in their total SP protein content, and their spermadhesin PSP-I/PSP-II and heparin-binding protein (HBP) concentrations. Spermatozoa were mainly recovered in fraction 2 (sperm-rich fraction, >1800 × 10⁶ spermatozoa/ml), whereas the pre-sperm fraction 1 and the post-sperm fractions 4–6 contained low numbers of spermatozoa (<500 × 10⁶/ml). Except in fraction 2, the total SP protein concentration and the concentration of both, spermadhesin PSP-I/PSP-II and the HBPs increased with fraction order. Distinct time-dependent effects were observed on motility characteristics and membrane integrity of highly diluted boar spermatozoa upon incubation with a 10% dilution of the SP from each fraction. The highest sperm viability was recorded after exposure for 5 h to fraction 2, followed by fractions 1 and 3. The percentages of motile spermatozoa also differed significantly among fractions after 5 h of incubation. Spermatozoa incubated with SP of fractions 1–3 showed the highest percentage motility. We conclude that different SP fractions exert distinct effects on the functionality of highly diluted boar spermatozoa. Fractions 1–3 appear to promote sperm survival, whereas fractions 4–6 seem to be harmful for preserving the physiological functions of highly diluted boar spermatozoa.