Proteomic approach to identify candidate effector molecules during the in vitro immune exclusion of infective Teladorsagia circumcincta in the abomasum of sheep

In the present study we have employed an in vitro organ challenge model to study the post-challenge responses in parasite na?ve and immune gastric tissue of sheep, in an attempt to identify the host derived factors involved in immune exclusion of Teladorsagia circumcincta larvae. Proteins present in the epithelial cells and mucus from ovine abomasa following parasite challenge in previously na?ve and immune animals were analysed through Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-Tof)-MS and shotgun proteomics. MALDI-ToF analysis of epithelial cell lysates revealed that a number of proteins identified were differentially expressed in na?ve and immune cells. These included intelectin and lysozymes, which were present at higher levels in epithelial cell lysates derived from immune samples. A large number of proteins were identified in the mucosal wash from immune tissue which were not present in the mucosal wash of the na?ve tissue. Some of these proteins were present in washes of immune tissue prior to the parasite challenge including immunoglobulin A, galectin 14 and 15 and sheep mast cell protease 1. However, other proteins, such as calcium activated chloride channel and intelectin were only detected in the washings from the challenged tissue. The latter may be related to an enhanced mucus release, which may result in entrapment of infective larvae and thus reduced establishment in tissue that has been previously challenged with the parasite. In conclusion, several proteins have been identified which may be involved, either directly or indirectly, in the exclusion and immune elimination of incoming infective larvae. In the present study, the usefulness of the in vitro model has been confirmed, and the global proteomic approach has identified proteins that had not previously been associated with parasite exclusion from abomasal mucosa, such as the calcium activated chloride channel.     

Assessment of blood pressure in cats presented with urethral obstruction

Objective: To determine the arterial blood pressure at presentation in male cats with acute urethral obstruction, and to determine whether there was any correlation between these measurements and concurrent metabolic abnormalities. Design: Prospective, single cohort, observational study. Setting: Private, small animal, after‐hours emergency clinic. Animals: Twenty‐eight client‐owned male cats with acute urethral obstruction and no other known coexisting disease. Interventions: Indirect oscillometric blood pressure measurements obtained before blood sampling and treatment. Measurements and main results: Mean arterial blood pressure (MAP) measurements, physical examination parameters, serum blood urea nitrogen (BUN), creatinine, potassium, phosphorus, total calcium and magnesium concentration, venous pH, lead II electrocardiogram, and urine volume in bladder were evaluated. No cats were hypotensive at presentation; 71% (20/28) were normotensive (median MAP=100 mmHg, range 93–140 mmHg); and 29% (8/28) were hypertensive (median MAP=153 mmHg, range 145–176 mmHg). Compared with hypertensive cats, normotensive cats had significantly lower heart rates (P=0.0201) and lower calcium (P=0.0152). For all 28 cats, MAP correlated with serum potassium and total calcium (P=0.0033). Conclusions: Though potassium and total calcium were inversely and directly correlated respectively with blood pressure in cats with urethral obstruction, none of the cats were hypotensive on presentation. Normotension on admission does not support the absence of biochemical and physical abnormalities in obstructed cats.     

Strong differentiation within Diplocarpon rosae strains based on microsatellite markers and greenhouse-based inoculation protocol on Rosa

The haploid ascomycete Diplocarpon rosae is the causal agent of black spot disease on roses, a widespread and devastating disease in the outdoor landscape. In this study, we established a Eurasian collection of 77 monoconidial strains of D. rosae: 50 strains collected on cultivated roses in Europe and Asia, and 27 strains on wild roses in Kazakhstan. To provide tools to describe its biology and to study its genetic diversity, we sequenced two strains of D. rosae using Illumina paired-end technology. The genome sizes of these two strains were estimated at 31.1 and 35.2 Mb, which are two times smaller than the genome size of the unique strain previously published. A BUSCO analysis confirmed a genome duplication of the strain previously sequenced and partial gene duplication of strains analysed in this study. Using the two genome sequences, 27 polymorphic microsatellite markers were identified. Polymorphism analysis of the 77 strains revealed a strong genetic differentiation between strains from cultivated and wild roses, and a lower diversity within the fungal population from cultivated roses compared to the population from wild roses. Pathogenicity of 10 strains was evaluated on 9 rose cultivars inoculated in the greenhouse. Disease scoring allowed the classification of strains into three groups and the characterization of resistance of rose cultivars. Good correlation observed between resistance scoring in greenhouse conditions and in the field indicates that pathogenicity assays in controlled conditions could be very useful in the near future to rapidly characterize the resistance of new rose varieties to black spot disease.     

Monoethanolamine extraction of copper-preservative-treated wood and reuse of the extract for wood preservation

Remediation of copper-based alkaline copper quat (ACQ) and micronized copper quat (MCQ) wood-preservative-treated weathered southern pine mulch samples by monoethanolamine (Mea) extraction was studied. The potential reuse of Mea extract in ACQ wood preservative application was also evaluated. Continuous column extraction of copper-preservative-treated samples for 32 h, using 12 % Mea removed 66–86 % (between top and bottom of the column) and 65–80 % of Cu, respectively, from ACQ- and MCQ-treated wood. Mea extracts of treated wood could be reused in ACQ-treating solutions that met the American Wood Protection Association standard specifications for ACQ wood preservatives. This was achieved either by mixing the extract with industrial Cu–Mea solution at 1:4 ratio or by adding cupric acetate salt directly into the Mea extract. ACQ-treating solutions reformulated using Mea extracts of ACQ- and MCQ-treated wood performed similarly to wood treated with industrial ACQ solution and freshly prepared solution using cupric acetate. All treated wood had similar penetrations of Cu in wood during preservative treatment, Cu leaching from the treated wood in contact with water, corrosion of nails in contact with treated wood, and ability to protect wood from mold growth.     

Evaluation of a technique to measure heart rate variability in anaesthetised cats

Analysis of heart rate (HR) and heart rate variability (HRV) are powerful tools to investigate cardiac diseases, but current methods, including 24-h Holter monitoring, can be cumbersome and may be compromised by movement artefact. A commercially available data capture and analysis system was used in anaesthetised healthy cats to measure HR and HRV during pharmacological manipulation of HR. Seven healthy cats were subjected to a randomised crossover study design with a 7 day washout period between two treatment groups, placebo and atenolol (1 mg/kg, IV), with the efficacy of atenolol to inhibit β₁ adrenoreceptors challenged by epinephrine. Statistical significance for the epinephrine challenge was set at P < 0.0027 (Holm–Bonferroni correction), whereas a level of significance of P < 0.05 was set for other variables.Analysis of the continuous electrocardiography (ECG) recordings showed that epinephrine challenge increased HR in the placebo group (P = 0.0003) but not in the atenolol group. The change in HR was greater in the placebo group than in the atenolol group (P = 0.0004). Therefore, compared to cats pre-treated with placebo, pre-treatment with atenolol significantly antagonised the tachycardia while not significantly affecting HRV. The increased HR in the placebo group following epinephrine challenge was consistent with a shift of the sympathovagal balance towards a predominantly sympathetic tone. However, the small (but not significant at the critical value) decrease in the normalised high-frequency component (HF_norm) in both groups of cats suggested that epinephrine induced a parasympathetic withdrawal in addition to sympathetic enhancement (increased normalised low frequency component or LF_norm). In conclusion, this model is a highly sensitive and repeatable model to investigate HRV in anaesthetised cats that would be useful in the laboratory setting for short-term investigation of cardiovascular disease and subtle responses to pharmacological agents in this species.     

EVALUATION OF STANDARD MAGNETIC RESONANCE CHARACTERISTICS USED TO DIFFERENTIATE NEOPLASTIC,INFLAMMATORY, AND VASCULAR BRAIN LESIONS IN DOGS

Magnetic resonance (MR) imaging characteristics are commonly used to help predict intracranial disease categories in dogs, however, few large studies have objectively evaluated these characteristics. The purpose of this retrospective study was to evaluate MR characteristics that have been used to differentiate neoplastic, inflammatory, and vascular intracranial diseases in a large, multi‐institutional population of dogs. Medical records from three veterinary teaching hospitals were searched over a 6‐year period for dogs that had diagnostic quality brain MR scans and histologically confirmed intracranial disease. Three examiners who were unaware of histologic diagnosis independently evaluated 19 MR lesion characteristics totaling 57 possible responses. A total of 75 dogs with histologically confirmed intracranial disease were included in analyses: 51 with neoplasia, 18 with inflammatory disease, and six with cerebrovascular disease. Only strong contrast enhancement was more common in neoplasia than other disease categories. A multivariable statistical model suggested that extra‐axial origin, T2‐FLAIR mixed intensity, and defined lesion margins were also predictive of neoplasia. Meningeal enhancement, irregular lesion shape, and multifocal location distinguished inflammatory diseases from the other disease categories. No MR characteristics distinguished vascular lesions and these appeared most similar to neoplasia. These results differed from a previous report describing seven MR characteristics that were predictive of neoplasia in dogs and cats. Findings from the current study indicated that the high performance of MR for diagnosing canine intracranial diseases might be due to evaluator recognition of combinations of MR characteristics vs. relying on any one MR characteristic alone.     

Genetic dissection of complex traits with chromosome substitution strains of mice

Chromosome substitution strains (CSSs) have been proposed as a simple and powerful way to identify quantitative trait loci (QTLs) affecting developmental, physiological, and behavioral processes. Here, we report the construction of a complete CSS panel for a vertebrate species. The CSS panel consists of 22 mouse strains, each of which carries a single chromosome substituted from a donor strain (A/J) onto a common host background (C57BL/6J). A survey of 53 traits revealed evidence for 150 QTLs affecting serum levels of sterols and amino acids, diet-induced obesity, and anxiety. These results demonstrate that CSSs greatly facilitate the detection and identification of genes that control the wide diversity of naturally occurring phenotypic variation in the A/J and C57BL/6J inbred strains.     

How does drought tolerance compare between two improved hybrids of balsam poplar and an unimproved native species?

Poplars are one of the woody plants that are very sensitive to water stress, which may reduce the productivity of fast-growing plantations. Poplars can exhibit several drought tolerance strategies that may impact productivity differently. Trees from two improved hybrids, Populus balsamifera?×?Populus trichocarpa Torr. & Gray (clone B?×?T) and P. balsamifera?×?Populus maximowiczii A. Henry (clone B?×?M), having P. balsamifera L. as a parent and trees from native and unimproved P. balsamifera were subjected to a 1-month drying cycle in a growth chamber and then rewatered. The unimproved and native B clone maintained higher stomatal conductance (g(s)) than the hybrids, and high photosynthetic activity and transpiration, even when soil water content was nearly zero. As a result, both instantaneous water use efficiency (WUE(i)) and leaf carbon isotope composition (δ(13)C) indicated that this clone was less affected by drought than both hybrids at maximal drought stress. However, this clone shed its leaves when the drought threshold was exceeded, which implied a greater loss of productivity. The B?×?M hybrid showed a relatively conservative response to water stress, with the greatest decrease in transpiring versus absorbing surface (total leaf area to root biomass ratio). This clone was also the only one to develop new leaves after rewatering, and its total biomass production was not significantly decreased by drought. Among the two hybrids, clone B?×?T was the most vigorous, with the greatest transpiration (E(i)) and net CO(2) assimilation (A) rates, allowing for high biomass production. However, it had a more risky strategy under drought conditions by keeping its stomata open and high E(i) rates under moderate drought, resulting in a lower recovery rate after rewatering. The opposite drought response strategies of the two hybrids were reflected by clone B?×?T having lower WUE(i) values than clone B?×?M at maximal drought, with a very low Ψ(min) value of -3.2 MPa, despite closed stomata and stopped photosynthetic activity. Positive linear relationships between A and g(s) for the three hybrids indicated strong stomatal control of photosynthesis. Moreover, the three poplar clones showed anisohydric behaviour for stomatal control and their use under long-term drought should be of interest, especially the B?×?M clone.     

Assessment of the biological control capability of <Emphasis Type="Italic">Hippodamia variegata</Emphasis> (Col.: Coccinellidae) using functional response experiments

Lady beetles are among the most successful predators of aphids in different environments. The functional responses of different life stages of Hippodamia variegata (Goeze) towards cotton aphids were examined in two different set-ups, a two-dimensional Petri dish set-up with detached leaves and a three-dimensional set-up with whole plants. In addition, the functional responses in two-dimensional set-ups towards cotton aphids and the pea aphids were compared. H. variegata exhibited a functional type II response to both cotton aphids and pea aphids irrespective of life stage and spatial scale of the set-up. Females and fourth instars generally had higher search rates than third instars whereas handling times were consistently lower for the fourth instar stage compared with the preceding juvenile stage and with females. The spatial dimensions did not have any significant influence on the functional responses towards cotton aphids, except for third instars which in the three-dimensional set-up reduced their search rate and increased their handling time. Fourth instars reacted in the same way to both Aphis gossypii and Acyrthosiphon pisum whereas the functional response parameters for third instars and females were significantly different on the two prey species. Our study, a part of this first-step evaluation of H. variegata as a biocontrol agent against A. gossypii and A. pisum under field condition, suggest that the voracity of all tested stages of H. variegata towards both aphid species hold good promises for a use of especially fourth instars and females in inundative biocontrol.