A technique for creating critical-size defects in the metatarsus of sheep for use in investigation of healing of long-bone defects

OBJECTIVE: To develop a technique for use in investigation of healing of long-bone defects by creation of a critical-size defect in the left metarsal III and IV bone (metatarsus) of sheep. ANIMALS: 18 healthy adult sheep. PROCEDURE: Sheep were allocated to 4 groups (3, 3, 5, and 7 sheep in groups 1 to 4, respectively). An ostectomy with various segmental length-to-diaphyseal diameter ratios (0.5, 1.0, 2.0, and 2.0 for groups 1 to 4, respectively) was performed on the left metatarsus of each sheep. The defect was left empty in sheep of groups 1, 2, and 3, whereas the defect was filled with a massive corticocancellous bone autograft in sheep of group 4. RESULTS: All sheep tolerated the surgical procedure well and were able to use the affected limb the day after surgery. Radiographic and histologic examinations conducted 16 weeks after surgery revealed nonunion in all sheep of groups 1, 2, and 3, whereas consistent bone healing with abundant bone formation was observed in all sheep of group 4. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these findings suggests that the sheep metatarsal model is a critical-size defect model with low morbidity. It should allow the assessment of new technologies for bone regeneration in conditions closely mimicking the clinical setting. IMPACT FOR HUMAN MEDICINE: Use of this technique in sheep should be of benefit for the preclinical study of osteoconductive, osteoinductive, or osteogenic biomaterials for use in humans.     

Isolation of Babesia divergens from carrier cattle blood using in vitro culture

Babesia divergens, the main causative agent of bovine babesiosis in Western Europe, was isolated from naturally infected cattle. Ninety-six blood samples were examined by means of an in vitro culture technique in sheep erythrocytes: 19 of them were collected from animals in the acute phase of the disease with visible parasitemia on blood smears, while the 77 remaining animals showed no microscopically detectable parasites. B. divergens was cultured from the 19 first blood samples as well as from 31 samples collected from asymptomatic animals. The time period before parasites could be detected in the culture varied in the latter samples from 6 to 20 days. The effects of sampling condition (anticoagulant used) and storage length were tested. A good correlation was obtained between immunofluorescent antibody test and culture, with identical results (positive or negative) for 89.6% of the samples collected from asymptomatic animals. The sensitivity of the in vitro culture method was determined and was about 10 parasites/mL of whole blood from three independent experiments performed with three different isolates, confirming its suitability to detect and culture diverse B. divergens isolates from carrier cattle. The parasites could indeed be isolated 9 months after the acute babesiosis phase in the blood of naturally infected animals. The 50 isolates collected in this study were successfully subcultured, cryopreserved and resuscitated using the same culture medium. The in vitro isolation of B. divergens from asymptomatic carrier cattle was achieved and will allow the analysis of parasite diversity within cattle herds.     

Bovine herpesvirus 1 glycoprotein D expression in bovine upper respiratory tract mediated by a human adenovirus type 5

Bovine herpesvirus 1 glycoprotein D (gD) gene expression by recombinant replication defective human adenovirus type 5 (HAdV-5) was investigated in calves using indirect immunofluorescence microscopy (IIFM), confocal laser scanning microscopy (CLSM) and RT-PCR. One fold intranasal instillation of HAdV-5-expressing gD in the cattle upper respiratory tract showed a short term expression of at least 5 days, but not 10 days, limited only to epithelial cells localised in the epithelium of the nasal mucosa in one out of six calves. Observed limited gene transfer into well differentiated cattle airway epithelial cells must be taken into consideration in order to enhance transfection efficiency, and consequently the vaccine potential of this vector.     

Global mineralogical and aqueous mars history derived from OMEGA/Mars Express data

Global mineralogical mapping of Mars by the Observatoire pour la Mineralogie, l'Eau, les Glaces et l'Activité (OMEGA) instrument on the European Space Agency's Mars Express spacecraft provides new information on Mars' geological and climatic history. Phyllosilicates formed by aqueous alteration very early in the planet's history (the "phyllocian" era) are found in the oldest terrains; sulfates were formed in a second era (the "theiikian" era) in an acidic environment. Beginning about 3.5 billion years ago, the last era (the "siderikian") is dominated by the formation of anhydrous ferric oxides in a slow superficial weathering, without liquid water playing a major role across the planet.     

Shedding routes of Coxiella burnetii in dairy cows: implications for detection and control

Reliable detection of Coxiella burnetii shedders is a critical point for the control of the spread of this bacterium among animals and from animals to humans. Coxiella burnetii is shed by ruminants mainly by birth products (placenta, birth fluids), but may also be shed by vaginal mucus, milk, and faeces, urine and semen. However, the informative value of these types of samples to identify shedders under field conditions is unknown. Our aim was then to describe the responses obtained using a real-time PCR technique applied to milk, vaginal mucus and faeces samples taken from 242 dairy cows in commercial dairy herds known to be naturally infected with Coxiella burnetii, and to assess their putative associations. Positive results were found in all types of tested samples even in faeces. No predominant shedding route was identified. Among the shedder cows, 65.4% were detected as shedders by only one route. By contrast, cows with positive results for all three samples were scarce (less than 7%). Testing a cow based on only one type of biological sample may lead to misclassify it with regards to its shedding of Coxiella burnetii and thereby underestimate the risk of bacterial spread within a herd.     

Mapping of the <Emphasis Type="Italic">ms8</Emphasis> male sterility gene in sweet pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) on the chromosome P4 using PCR-based markers useful for breeding programmes

The nuclear male sterility gene ms8 is expected to facilitate the production of sweet pepper (Capsicum annuum L.) hybrids as it provides means for hybridization without the labor-intensive hand emasculation of female inbred lines. The development of molecular markers linked to ms8 locus will help the breeding practice for the selection of hybrid parental lines. In this study, F₂ population resulting from a cross between the sweet pepper male sterile line 320 and the male fertile variety Elf was used to identify DNA markers linked to the ms8 locus. With the use of RAPD–BSA technique, seven markers linked to the ms8 locus were found. Four of them were converted into SCAR markers. In addition, two COSII/CAPS markers linked to the ms8 locus were identified. Comparative mapping with reference pepper maps indicated that the ms8 locus is located on the lower arm of the pepper chromosome P4. Identified markers are useful for molecular breeding, however, at present markers tightly linked to ms8 locus are still lacking. Identification of molecular markers linked to the ms8 locus and determination of its chromosomal localization are useful for fine mapping and also provide the perspective for ms8 gene cloning.     

Thinning effect on light regime and growth of underplanted red oak and black cherry in post-agricultural forests of south-eastern Canada

New Forests - Over the last century, north-eastern North America has seen the gradual abandonment of much agricultural land that had become unsuitable for modern practices. This shift in land-use...     

Effectiveness of vaccination and antibiotics to control Coxiella burnetii shedding around calving in dairy cows

Effectiveness of phase 1 vaccine, combined or not with tetracycline, to control Coxiella burnetii vaginal shedding at calving in cows was assessed through a 13 months study in 22 Q fever clinically affected commercial dairy herds. Four medical strategies implemented at herd level but randomly assigned to cows (vaccination, vaccination and tetracycline, tetracycline, nothing) were compared. There was no significant interaction effect between vaccination and antibiotherapy. Tetracycline used once at drying off was associated with a lower risk of being detected shedder at calving (OR=0.40, CI 95% [0.21-0.75]), but had no significant effect on the bacterial load shed. Vaccination did not significantly prevent shedding but was significantly (OR=0.15, CI 95% [0.03-0.85]) associated with lower bacterial load shed. Thus, vaccination using a phase 1 vaccine and antibiotherapy using tetracycline is associated with a decrease in shedding in dairy cows and could contribute to reduce the bacterial load generated in the environment. To the best of our knowledge, this is the first study providing useful information for evidenced-based and rational use of medical strategy combining antibiotic and vaccination in infected dairy cattle herds.     

Experimental and theoretical studies of the thermal degradation of a phenolic dibenzodioxocin lignin model

A large part of biphenyl structures in lignin are etherified by α- and β-carbons of another phenylpropane unit to give an eight-member ring called dibenzodioxocin. The behavior of a phenolic dibenzodioxocin lignin model, 4-(4,9-dimethoxy-2,11-n-dipropyl-6,7-dihydro-5,8-dioxa-dibenzo[a,c]cycloocten-6-yl)-2-methoxyphenol (DBDOH, 1), was studied by different mass spectrometry and thermal methods, leading to the conclusion that dibenzodioxocins are thermally unstable products. Both semi-empirical and density functional theory quantum calculations show that both C–O bonds, which connect the biphenyl part of the dibenzodioxocin molecule to the phenolic group, can be broken under increasing temperature. However, they do not play the same role since their dissociation occurs through different barrier heights. The C–O bond directly connected to the phenolic group (α–O-4) will dissociate first since its barrier energy for scission is lower than the other one (β–O-4), by roughly 12 kcal mol^–1 (≈50 kJ mol^?1). This conclusion is likely applicable to thermal degradation of DBDO units in lignin polymer.