Persistence and molecular evolution of Mycobacterium bovis population from cattle and wildlife in Doñana National Park revealed by genotype variation

The role of wildlife in tuberculosis epidemiology is being widely studied since it can affect the effectiveness of eradication campaigns in cattle. The health problem is enhanced when it concerns also wildlife welfare and biodiversity conservation. This study was performed to understand the epidemiology of Mycobacterium bovis population affecting livestock and wild animals in the Do?ana National Park using bacteriology and molecular characterisation techniques. Tuberculosis research was performed on 1209 cattle and wild animals (artiodactyla and carnivore) collected over 6 years in the Park. One hundred and sixty-three animals were found to be infected with M. bovis, comprising 7.96% of the cattle and 20.53% of the wild animals tested. Spoligotyping revealed nine patterns, being SB1232 and SB1230 the most prevalent (77.30% and 15.34% of infected animals, respectively). MIRU-VNTR analysis of a selected panel of 92 isolates showed eight different profiles, including several spoligotypes within the same MIRU-VNTR profile. The discriminatory capacity of both techniques in this panel was similar. The results obtained by combination of both techniques corroborate that wildlife species are infected with the M. bovis strains which are more prevalent in cattle and reveal their persistence. Genotype variation between isolates strongly suggests micro-evolutionary events in the M. bovis population in the same area. This study in the Do?ana National Park exposes the risk of introduction of domestic animals into wildlife areas when there is not a warranty of disease freedom, appropriate diagnostic techniques and control measures.     

Genotyping of Mycoplasma mycoides subsp. mycoides SC by multilocus sequence analysis allows molecular epidemiology of contagious bovine pleuropneumonia

Mycoplasma mycoides subsp. mycoides SC (MmmSC) is the etiological agent of contagious bovine pleuropneumonia (CBPP). Although eradicated in most developed countries, the disease reappeared in Europe in the 1990s. This reappearance may have been caused either by importation from sub-Saharan Africa, where CBPP is still endemic, or by the reemergence of virulent strains in Europe, as suggested by earlier studies. A multilocus sequence analysis scheme has been developed to address this issue and, most importantly, to be able to monitor new epidemics. The alignment of the full genome sequence of the reference strain PG1 and the partial genome sequence of a pathogenic strain allowed the identification of polymorphic sites. Nineteen initial loci were selected within housekeeping genes, genes of unknown function and non coding sequences. The suitability of these loci for genotyping MmmSC strains was first tested on six strains of diverse geographic origin. The analyses showed that the published PG1 sequence contained a number of specific polymorphisms that were therefore of no use for molecular typing. Among the eight informative polymorphic loci finally selected, only one (ftsY) was positioned within a housekeeping gene. Three main groups and 31 different allelic profiles were identified among 51 strains and strain variants examined. Cluster analysis confirmed that European strains from the 1990s did not originate from Africa. It also showed a genetic link between a European strain isolated in 1967 and those found in southern Africa and Australia. This was in agreement with historical data showing that CBPP was introduced in these regions during colonisation in the 19th century.     

Phage display selection and characterization of single-chain recombinant antibodies against Eimeria tenella sporozoites

A single-chain antibody library against Eimeria tenella sporozoites was constructed by phage display. Antibody-displaying phage was selected in five panning rounds against cryopreserved E. tenella sporozoites. A 1000-fold increase in phage output and a 3000-fold enrichment were obtained after three rounds of panning, as the binding clones became the dominant population in the library. Ten clones were randomly selected from the last selection round, and their nucleotide sequences were aligned and compared to chicken germ-line sequences. Analysis of the light chain variable regions revealed possible donor pseudogenes which act as donors in gene conversion events, and contribute to the diversification of the V(L) immune repertoire. Possible somatic hypermutation events, a consequence of affinity maturation, were also identified. Soluble antibody was produced in a non-suppressor E. coli strain, purified by nickel affinity chromatography, and characterized by immunoblotting. In an immunofluorescence assay, this recombinant antibody showed specific binding to E. tenella sporozoites.     

Growth performance,carcass quality characteristics and colonic microbiota profiles in finishing pigs fed diets with different inclusion levels of rice distillers’ by‐product

The aim of this study was to evaluate the effects of diets containing rice distillers’ by‐product (RDP) on growth performance, carcass characteristics, meat quality, and gut microbiota of fattening pigs. Twenty‐four crossbred finishing pigs (Duroc × Landrace × Yorkshire), 56.9 ± 3.1 kg initial body weight, were randomly allocated to three groups. For 56 days, pigs were fed one of three diets including RDP0 (control), RDP15 (15% RDP in DM), and RDP30 (30% RDP in DM). With RDP level in diet, average daily gain and backfat thickness linearly increased (p < 0.05), and drip loss tended to increase (p ≤ 0.08). In addition, 16S ribosomal RNA gene amplicon profiling showed that RDP was associated with modulation of colonic microbiota composition, especially at family and genus levels. Relative abundance of Porphyromonadaceae and Erysipelotrichaceae families in colonic digesta increased with inclusion of RDP, while that of Enterobacteriaceae decreased. The proportion of genera unclassified Erysipelotrichaceae, and Butyrivibrio increased as inclusion of RDP. These results indicate that up to 30% inclusion in diet of finishing pigs, RDP can modulate colonic microbiota composition, and induces an improvement of animal growth and fat deposition.     

Sward characteristics, grass dry matter intake and milk production performance is affected by timing of spring grazing and subsequent stocking rate

The objective of this study is to investigate the effect of contrasting spring grazing dates (GD) and stocking rate (SR) on sward characteristics, grass dry matter intake and milk production performance of autumn calving dairy cows during the spring period. Two swards were created by grazing in March (early grazing; E) or by delaying first grazing until mid-April (late grazing; L). Two stocking rates, high (H; 5.5) and medium (M; 4.5) were applied across each sward. Forty eight autumn calving Holstein cows (160 ± 35 days in milk) were assigned to one of four (n = 12) different grazing treatments. The experiment began on April 17th and finished after 2 grazing rotations on June 20th. Later spring grazing significantly increased herbage mass (kg DM/ha) above ground level (+ 933, P < 0.05) and > 50 mm (+ 738, P < 0.05). Compressed sward height (+ 22.1 mm, P < 0.05), extended tiller height (+ 73 mm, P < 0.001) and pseudostem height (+ 35 mm, P < 0.001) were also significantly higher for later grazed swards. In the grazing horizon (> 80 mm— extended tiller height), later grazed swards had significantly lower leaf proportion (− 0.09, P < 0.05) and higher dead material (+ 0.05, P < 0.001). Daily herbage allowance (> 50 mm) was on average 12.7, 15.9, 18.2 and 21.9 kg DM/cow for EH, EM, LH and LM, respectively. Daily leaf allowance (> 80 mm) was 10.1, 12.3, 13.3 and 14.5 kg DM/cow for EH, EM, LH and LM, respectively. The EM (16.2 kg DM/cow), LH (+ 0.1 kg) and LM (0.8 kg) treatments all had similar grass DM intake, however there was evidence of an interaction (P < 0.10) between GD and SR, this was due to the low grass DM intake of the EH (13.9 kg DM/cow) treatment. When expressed as UFL (Fill unit) intake the EM treatment recorded the highest value. There was a significant interaction between GD and SR (P < 0.01) for milk, protein yield, 4% fat corrected milk yield (P < 0.05) and protein concentration (P < 0.001). Cows grazing the EM treatment produced 23.9 kg of milk, 876 and 685 g of fat and protein yield. The difference in milk production (cow/day) between EM and EH treatments was + 3.6 kg milk, + 98 g fat and + 107 g protein. The production yield difference between LM and LH treatments was + 1.1 kg milk, + 27 g fat and + 29 g protein in favour of the LM treatment (23.9 kg of milk, 877 and 687 g fat and protein yield). Herbage quality and morphological characteristics are clearly improved with early spring grazing as herbage mass is reduced on subsequent rotations. Swards grazed in early spring allow higher grass utilisation and high milk production performance when grazed at a medium stocking rate. Improved milk production from herbage can be achieved provided herbage mass and allowance are maintained at levels where herbage quality decreases are minimised.     

Cardiorespiratory effects of a 7° reverse Trendelenburg position in anaesthetized horses: a randomized clinical trial

Objective

To evaluate the cardiorespiratory effects of a 7° reverse Trendelenburg position (RTP) in anaesthetized horses.

Granulocyte and granulocyte macrophage colony-stimulating factors as therapy in human and veterinary medicine

Granulocyte colony-stimulating factors (G-CSFs) and granulocyte macrophage colony-stimulating factors (GM-CSFs) are endogenous cytokines that regulate granulocyte colonies and play a major role in the stimulation of granulopoiesis (neutrophils, basophils and eosinophils) and in the regulation of microbicidal functions. There are numerous pathological conditions in which neutrophils are decreased, the most common being neutropenia associated with cancer chemotherapy, which increases the risk of serious microbial infections developing with the potential for high morbidity and mortality. New methods in molecular biology have led to the identification and cloning of CSF genes and biopharmaceutical production. Since then, CSFs have been widely used for the prevention and treatment of neutropenia associated with cancer chemotherapy, for mobilising haematopoietic cell precursors, and for other neutropenia-related pathologies. This review focuses on the use of CSFs within both human and veterinary medicine. Clinical applications, pharmacology, tolerability and the potential role of these factors in veterinary medicine are considered.     

Prediction of β‐Glucan Concentration Based on Viscosity Evaluations of Raw Oat Flours from High β‐Glucan and Traditional Oat Lines

Rheological properties of raw oat flour slurries were determined in experimental high β‐glucan (≤7.8%) and traditional oat lines (4–5% β‐glucan) grown in two consecutive years. Three different media were used to disperse oat flours: deionized water, silver nitrate solution (to inactivate endogenous enzymes), and alkali solution (to solubilize both water‐soluble and water‐insoluble β‐glucans). Significant correlations (P < 0.05) between viscosity of slurries and β‐glucan concentration obtained in either deionized water (r = 0.833), silver nitrate (r = 0.940), or alkali (r = 0.896) solutions showed that β‐glucans were the main contributor to oat extract viscosity. The highest correlation was obtained in silver nitrate solution, suggesting that inactivating endogenous enzymes is important to obtain high correlations. Predictive models of oat β‐glucan concentration based on the viscosity profile were developed using partial least squares (PLS) regression. Prediction of β‐glucan concentration based on viscosity was most effective in the silver nitrate solution (r = 0.949, correlation coefficient of predicted vs. analyzed β‐glucans) and least effective in the alkali solution (r = 0.870). These findings demonstrate that the β‐glucan in oat could be predicted by measuring the viscosity of raw flours in silver nitrate solution, and this method could be used as a screening tool for selective breeding.     

Pasting and Thermal Properties of Flours from Oat Lines with High and Typical Amounts of β‐Glucan

Experimental oat lines high in β‐glucan (6–7.8%) and traditional lines (3.9–5.7% β‐glucan) were used to evaluate the effect of β‐glucan on pasting (by rapid viscoanalysis) and thermal properties (by differential scanning calorimetry) of oat flours. Significant correlations established between β‐glucan concentration and the pasting parameters after amylolysis demonstrated the role of β‐glucans in oat pasting. The relative decrease of peak viscosity (PV) observed after enzymatic removal of β‐glucans was correlated with β‐glucan concentration (r = 0.880, P < 0.010) and reconfirmed their contribution to pasting. A significant increase of PV with β‐glucan concentration obtained under conditions of either autolysis (deionized water used for dispersion) (r = 0.89, P < 0.010) or inhibition (silver nitrate solution used for dispersion) (r = 0.91, P < 0.001) might be explained by an increase in water retention capacity caused by the β‐glucans. Predictive models of β‐glucan concentration based on the whole pasting profile or selected profile regions were developed using partial least squares (PLS) regression.Prediction of β‐glucan based on the whole profile obtained in the silver nitrate solution was the most effective (r = 0.93, correlation coefficient of predicted vs. analyzed β‐glucans, P < 0.050). No correlations were observed between the thermal properties of oat flours and the β‐glucan concentration.