Salmonella enterica serovar Choleraesuis derivatives harbouring deletions in rpoS and phoP regulatory genes are attenuated in pigs, and survive and multiply in porcine intestinal macrophages and fibroblasts, respectively

Live attenuated Salmonella enterica strains have been extensively studied as potential vectors for the oral delivery of heterologous antigens. Due to its ability to target immune cells, its specific mechanism for crossing the intestinal barrier, and its swine-restricted tropism, S. enterica subspecies enterica serovar Choleraesuis (S. Choleraesuis) has attracted a great deal of interest for the production of bacterial-based oral carriers specifically adapted to swine. In this study, two mutants of S. Choleraesuis were constructed and their attenuation and intracellular fate analysed with the purpose of engineering new attenuated live strains with improved properties as oral vaccine carriers. Those strains harboured a specific deletion either within the phoP or rpoS genes, which encode virulence-related regulators in S. Typhimurium. In comparison to the wild-type parental S. Choleraesuis, the mutant strains, especially DeltaphoP, were extremely low in virulence in the murine model and in the natural host, the pig. Moreover, when compared with a commercial live vaccine strain, SC-54, the two mutants showed a higher level of attenuation in mice and DeltaphoP also in pigs. In addition, DeltarpoS and DeltaphoP presented a proliferation and survival phenotype within swine intestinal primary fibroblast and macrophage cell cultures, respectively. Collectively, the present results indicate that the DeltarpoS and DeltaphoP strains of S. Choleraesuis gather adequate features to be potential candidates for vaccine vectors for the specific delivery of heterologous antigens adapted to pigs.     

A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.     

On the potential of documenting decadal-scale avifaunal change from before-and-after comparisons of museum and observational data across North America

Studies of biodiversity dynamics have been cast on either long(systematics)or short(ecology)time scales,leaving a gap in coverage for moderate time scales of de...     

Anatomy,Morphometry and Radiography in the thoracic limb bones of the Patagonian Huemul Deer (Hippocamelus bisulcus)

The aim of this study was to provide morphometric, anatomic and radiographic data of the thoracic limb bones of the Patagonian Huemul (Hippocamelus bisulcus) including a functional interpretation of this, as a reference for clinical use, biomedical research and teaching purposes. Currently, the Patagonian huemul deer is in danger of becoming extinct due to multiple causes. Research carried out for its conservation has focused mainly on its ecology and pathology, leaving gaps in biological knowledge, which is basic and important for its comprehension. This study was conducted to reveal the gross osteology and radiology features of the thoracic limb bones of the Patagonian huemul deer. The osteological findings suggest the presence of powerful flexor muscles in the scapulohumeral and elbow joints, useful to cushion the jumps. Also, the principal nutrient foramen of Patagonian huemul differs in position with respect to domestic ungulates, which may be important to consider during surgical procedures. Finally, the radiographic data can provide new information about the tissue loading conditions in Patagonian huemul, so that this new knowledge can be of great importance for a better understanding of mechanically induced or adaptive changes in bone produced by habitat or other ecological phenomena.     

Salmonella serotypes in wild boars (Sus scrofa) hunted in northern Italy

Background

Salmonella species (spp.) are zoonotic enteric bacteria able to infect humans, livestock and wildlife.However, little is known about the prevalence and the presence of the different serovars in wildlife. Considering the wide distribution of wild boars and the feeding behaviour (omnivorous scavengers), wild boars may be a good indicator for environmental presence of Salmonella spp. The aims of this study were to determine the presence of Salmonella spp. in hunted wild boars and to determine the serotype the isolated strains.

Findings

Over three hunting seasons, the intestinal contents of 1,313 boars hunted in northern Italy were sampled and cultured. Salmonella spp. were isolated from 326 boars (24.82%). Thirty different serovars belonging to three different S. enterica spp. were found. Twenty-one serovars of S. enterica subsp. Enterica were found including the human pathogens S. Typhimurium and S. Enteritidis. In addition, nine serovars belonging to S.enterica subsp. diarizonae and S. enterica subsp. houtenae were detected.

Conclusions

Considering the widespread occurrence of wild boars in Europe, the epidemiological role of this species in relation to salmonellosis might be relevant and should be further investigated. Wild boars may act as healthy carriers of a wide range of Salmonella serotypes.     

Diet composition, herbage intake and digestibility in Inner Mongolian Cashmere goats grazing on native Leymus chinensis plant communities

The botanical composition, intake and digestibility of the diet selected by Inner Mongolian Cashmere goats grazing on a native Leymus chinensis plant community were measured in two grazing periods (June and August) using the n-alkane markers. In each grazing period, 48 Inner Mongolian Cashmere goats (24 wethers and 24 does with an average live weight of 22.1 ± 1.2 kg and 19.6 ± 0.8 kg, respectively) were divided in two groups (G1: grazing group; G2: cage-fed group) of 24 animals each. Based on the observation of G1 grazing behaviour, G2 goats were fed on herbage obtained by a simulative method and were housed in metabolism cages to determine the alkane faecal recoveries. Diet composition was estimated by comparing the odd-chain n-alkane pattern (C₂₅–C₃₅) of the selected plant species with the n-alkane faecal concentrations, corrected for their incomplete faecal recovery. The alkane pair C₃₂:C₃₃ and C₃₆ alkane were used to estimate intake and diet digestibility, respectively. The results showed that the increase of herbage availability from June to August, resulted in higher (P < 0.05) DM, CP, ME, ADF and NDF intake of both wethers and does, with no evident changes in ADF and NDF intake in does., The intake of nutrients differed significantly between wethers and does in both experimental periods, except for NDF in EP1 and ME in EP2. Differences between grazing seasons and goats' sex in nutrients digestibility were not observed. Diet composition indicated seasonal differences, being Sonchus brachyotus, Artemisia frigida and Phragmites communis the main components of the diet in June. However, Suaeda corniculata, Chloris virgata, Xanthium sibiricum and Echinochloa crusgalli became the main diet components in August. Moreover, the sex affected significantly the diet choices in both grazing periods.     

Dairy goat production systems

Goat production concentrated in developing countries (tropics, dry areas), contributes largely to the livelihoods of low and medium income farmers. Farming systems in these areas have evolved to cope with the formidable constraints imposed by harsh natural and economic conditions by adapting integrated crop/livestock production strategies. In Asia, Africa and Latin America, due to its almost exclusive extensive nature, goat production relies mainly on grazing on communal lands that hardly provide the minimum nutrient requirements due to overstocking and degradation. While some of these production systems are becoming semi-intensive, appropriate breeding strategies should be designed to promote conservation and improvement of their unique attributes, such as adaptability, water use efficiency and suitability under harsh climatic conditions. In Europe, dairy goat production is more common around the Mediterranean basin, where it is important from an economic, environmental and sociological perspective to the Mediterranean countries: Spain, France, Italy and Greece. Europe owns only 5.1 % of the world’s dairy goat herds, but produces 15.6 % of the world’s goat milk; this is the only continent where goat milk has such an economic importance and organization. In developing countries the dairy goat sector requires a systemic approach, whereby nutrition, animal health, breeding, know-how, inputs and technologies must be assembled. This would allow the optimization of natural and local resources and would promote the transition from a risk reduction strategy towards an increased productivity strategy. Such an increase would privilege production efficiency based on clean, green and ethical practices for responsible innovation.     

Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.     

Early detection of Neophysopella tropicalis in grapevine leaves and on spore traps by qPCR

Neophysopella tropicalis, one of the causal agents of Asian grapevine leaf rust (AGLR), can cause severe epidemics in Brazil that lead to yield losses in commercial vineyards. An early detection of the pathogen by air sampling of urediniospores on spore traps or in symptomless leaves would be valuable to multiple studies, such as epidemics modelling, risk forecasting, monitoring of pathogen introductions in rust-free areas, and predicting the beginning of epidemics. This study developed a quantitative PCR (qPCR) protocol to quantify N. tropicalis urediniospores attached to adhesive tapes and in grapevine leaves before symptom appearance. A specific primer pair was designed based on the internal transcribed spacer (ITS) sequence region of the AGLR pathogen. Standard amplification curves using genomic DNA from urediniospores of N. tropicalis and from urediniospores attached to adhesive tapes were established. Grapevine leaves inoculated with N. tropicalis were collected at 2, 5, and 7 days postinoculation (dpi). One primer pair (580F/720R) amplified a 140 bp product in all AGLR isolates but did not amplify products of other rust genera, such as Phakopsora, Puccinia, Hemileia, Tranzschelia, Cerotelium, and Coleosporium. As little as 0.1 pg DNA and 10 urediniospores of N. tropicalis attached to adhesive tapes could be detected. qPCR enabled the detection of the pathogen as early as 2 dpi, before symptom appearance. This method can be used to monitor N. tropicalis inoculum in grapevine-growing areas and to quantify symptomless infections of the AGLR pathogen.