Evaluation of elutriation and magnetic microbead purification of canine monocytes

An elutriation technique was developed to obtain large quantities of pure canine monocytes. Firstly, peripheral blood mononuclear cells (PBMC) were isolated from whole blood by Ficoll gradient. Then, the PBMC were separated by an elutriation procedure. We demonstrated that these techniques allow the isolation of canine peripheral blood monocytes with a purity of 64% +/- 7.9 when labelled with anti-CD14 antibody. This purity increased to 83% +/- 2.2 after separation by magnetic anti-CD14 microbeads. The cell viability was more than 95% and apoptotic cells were less than 10%. The monocytes purified by these methods were functionally active in a mixed leukocyte reaction (MLR). A lymphocyte fraction was obtained directly only by elutriation with an average of 79.9% +/- 10.7 of CD5+, 7.9% +/- 3.5 of CD21+ and 1.78% +/- 2.53 of CD14+. Our results indicate that this elutriation procedure is a safe method to purify monocytes as well as lymphocytes, useful in MLR.     

Radio-telemetry as a tool to define protected areas for seabirds in the marine environment

Although most Manx shearwaters (Puffinus puffinus) in the UK are protected within Special Protection Areas (SPAs) at their breeding colonies, such site-based protection does not currently extend into the marine environment. Manx shearwaters form rafts (dense flocks of birds) on the sea adjacent to their breeding colony from late afternoon onwards, before coming ashore after nightfall. The ecological and protection requirements of Manx shearwaters in the UK would be more completely addressed if existing SPA boundaries were extended into adjacent marine areas to include those used for rafting behaviour. However the spatial extent of rafts has never been assessed accurately. Because rafting behaviour continues after nightfall, radio-telemetry was used to investigate rafting distributions of Manx shearwaters around the three largest UK colonies: Skomer (south-west Wales), Bardsey (north-west Wales) and Rum (north-west Scotland). Totals of 30, 28 and 30 birds were radio-tagged, with 174, 264 and 385 rafting locations estimated for Skomer, Rum and Bardsey, respectively. Kernel contour analysis was used to describe the ‘rafting range’ around each colony. The maximum extent of the 95% kernel contours was used to determine the area which would encompass most rafting birds. The maximum extents of rafting ranges were 4, 6 and 9 km, respectively for Skomer, Rum and Bardsey. We believe this is the first successful illustration of how radio-telemetry data can potentially be used to define protected areas for birds in the marine environment.     

IMAGING DIAGNOSIS—NUCLEAR SCINTIGRAPHIC,RADIOLOGICAL, AND PATHOLOGIC CHARACTERISTICS OF METASTATIC PILOMATRICOMA IN A DOG

A 6‐year‐old castrated Goldendoodle dog was presented for left‐sided lameness of 3 weeks’ duration. Focal, moderate to marked increased ^99mTc‐methylene diphosphonate (^99mTc‐MDP) uptake was detected in the right caudal lung lobe, caudal angle of the left scapula, and the distal aspect of the left femur with whole body bone phase scintigraphy. Radiographs identified a well‐circumscribed, oval‐shaped soft tissue opaque mass in the right caudal lung lobe; a suspect oval‐shaped osteolytic lesion in the proximal third of the left scapula; and an osteolytic lesion in the distal aspect of the left femur. Metastatic pilomatricoma was confirmed histologically at all three sites.     

High prevalence of Cryptosporidium bovis and the deer-like genotype in calves compared to mature cows in beef cow-calf operations

Recent studies have identified the novel, host adapted Cryptosporidium bovis and the deer-like genotype in dairy cattle from farms in the United States, China, India and Europe. This novel species and genotype appear to be more prevalent in older, post-weaned dairy cattle than previously thought. However, little information is available on their prevalence in beef cow-calf operations. In the present study, we determined the prevalence of Cryptosporidium species in 98 calves (6-8 months old) and 114 cows (>2 years old) in seven beef cow-calf herds in western North Dakota. DNA was extracted from fecal samples and Cryptosporidium spp. were identified by amplification of the 18S rRNA gene followed by sequencing or RFLP analysis. All seven herds tested positive for Cryptosporidium. Overall, 43/212 (20.3%) animals were positive. Only five of these positives were from cows. C. bovis, the deer-like genotype and C. andersoni were identified in 9.4, 6.6 and 1.4% of animals sampled, respectively. C. parvum was not identified in any of the positive samples. C. bovis, the deer-like genotype and C. andersoni were detected in 6/7, 5/7 and 2/7 herds, respectively. C. bovis and the deer-like genotype were primarily detected in calves, while C. andersoni was only detected in cows. Six isolates could not be typed. These results show a relatively high prevalence of C. bovis and the deer-like genotype in 6-8-month-old beef calves compared to cows older than 2 years in the seven herds studied.     

Dermatosparaxis in two Limousin calves

Background

An unusual presentation of skin disease was identified in two related neonatal Pedigree Limousin calves presented to University Veterinary Hospital, University College Dublin, following detailed post mortem examination a diagnosis of dermatosparaxis was made. Dermatosparaxis in animals or Ehlers Danlos Syndrome, which is the analogous condition seen in humans, is a connective tissue disorder characterised by extreme skin fragility. To the authors’ knowledge this is the first report of such a diagnosis in the Limousin breed and the features of this lethal phenotype were severe in comparison to previous reports of the condition.

Case presentation

Two calves, which were full siblings, a pedigree Limousin bull (Calf A) and pedigree Limousin heifer (Calf B) were examined clinically after presenting collapsed since birth, both had grossly abnormal skin with multiple skin fissures visible and both calves were subsequently euthanised. Both calves underwent gross post mortem examination, after which histological samples were reviewed and electron microscopical examination of selected skin samples was carried out. Histological features of dysplastic dermal collagen were identified. The diagnosis of dermatosparaxis in the Limousin breed was confirmed. Genetic testing was conducted to determine if the current cases had the same mutation as has previously been described in Belgian Blue cattle. Some common parentage was traced but genetic testing did not show a similar mutation to that previously described in cattle. The specific genetic cause in this case is unknown.

Conclusions

This is the first report of dermatosparaxis in the Limousin and the presentation of the dermatosparaxis phenotype has some noteworthy features thus further genetic testing is required to pinpoint the causative mutation or other genetic defect. Given the popularity of the breed and the lethal nature of the phenotype in this case it is important to raise awareness of the condition.

     

Above- and belowground responses to tree thinning depend on the treatment of tree debris

Mechanical mastication is increasingly prescribed for wildfire mitigation, yet little is known about the ecological impacts of this fuels treatment. Mastication shreds trees into woodchips as an alternative to tree thinning and burning the resulting slash, which can create soil disturbances that favor exotic plants. Previous research on mastication has not simultaneously considered both the responses of soil organisms and understory plant communities. We compared mastication to slash pile burning (both 6-months and 2.5-years post-treatment) and untreated controls in pinyon–juniper (Pinus edulis–Juniperus osteosperma) woodland and measured soil properties, arbuscular mycorrhizal fungi (AMF) and understory plant composition. Our results showed that slash pile burns had severely degraded soil properties and low AMF abundance and richness compared to untreated or mastication plots. Pile burns were dominated by exotic plant species and had approximately 6× less understory plant abundance and richness than untreated plots. Only two variables differed between mastication and untreated plots 6-months post-treatment: mastication had lower soil temperature and higher soil moisture. Mastication plots 2.5-years post-treatment had more plant cover and richness than untreated plots or pile burns, although non-native Bromus tectorum cover was also greater and AMF spore richness was lower than untreated plots. The structural equation model (SEM) we developed showed that plant cover strongly influenced AMF abundance (0.50) and both plant cover (0.36) and AMF (0.31) positively influenced soil stability. In the short-term, mastication is a preferable method as it creates fewer disturbances than pile burning; however long-term impacts of mastication need further study as this practice could affect native plant communities. Our results suggest that the manner in which woody debris is treated following tree thinning has an important influence on soil stability and native plant biodiversity.     

Diversification of Pinus halepensis forests by sowing Quercus ilex and Quercus pubescens acorns: testing the effects of different vegetation and soil treatments

The goal of this study was to develop management strategies favouring establishment and survival of holm oak (Quercus ilex L.) and downy oak (Quercus pubescens Willd.)??two species co-occurring in Southern France??in mature Allepo pine forests (Pinus halepensis Mill subsp. halepensis). An experimental design was assessed in a partially cut mature stand of Aleppo pine in which five soil and vegetation treatments??chopping, chopping followed by scarification in one or two directions, prescribed burning, control??and two slash treatments (presence/absence) were applied. A total of 1,600 sowing points, each composed of 3 Q. ilex or Q. pubescens acorns, were installed in the different treatments in November of two consecutive years at 6 and 18 months, after the end of treatments. Survival was monitored 3 and 2 years after sowing, soil surface at the sowing points was characterized at different dates, and predawn leaf water potentials were measured during the dry season. High mortality occurred after the first summer, but survival after 1 year was 2.3?C5.2 higher in Q. ilex than in Q. pubescens, confirming that Q. ilex was better adapted to the drier parts of the Mediterranean area. Survival was significantly influenced by the treatments, but there was a variable response between the two sowing years under most of the treatments. Only intense fire proved the most beneficial treatment for seedling survival in both years. The micro-local soil cover conditions induced by the treatments played a major role in explaining oak survival. In particular, grass cover (mainly Brachypodium retusum) proved to be largely unfavourable to seedling survival and growth, and this detrimental effect was also confirmed by lower predawn leaf water potential values with increasing grass cover. Acorn introduction designed to diversify mature Aleppo pine forest after soil and vegetation treatments therefore has to be considered for treatments that most efficiently impair the pre-existing competing grass cover such as prescribed high-intensity fire treatment.     

Genetic and genomic approaches for improving biofuel production from maize

Grasses, which are currently at the basis of cattle feeding, will, in the near future, be a major source of cell wall carbohydrates for sustainable biofuel production. The association of lignins with other matrix components, together with linkages between cell wall carbohydrates, greatly influences cell wall properties, including the degradability of structural polysaccharides by micro-organisms in animal rumen or industrial fermenters. The improvement in biofuel production from plants is based on the understanding of the cell wall composition and assembly, and on the discovery of genetic and genomic mechanisms involved in each component biosynthesis and their depositions in each lignified tissue. While nearly 40 QTL have been shown for lignin content, only seven locations appeared of greater importance in investigated genetic resources. Expression studies highlighted that several genes in the lignin pathway are less expressed in lines with higher cell wall degradability. However, only a few lignin pathway genes mapped in QTL positions, and the fully relevant candidates might be genes involved in regulation of lignin pathway genes, or in regulation of lignified tissue assembly.