Mutation and virulence assessment of chromosomal genes of Rhodococcus equi 103

Rhodococcus equi can cause severe or fatal pneumonia in foals as well as in immunocompromised animals and humans. Its ability to persist in macrophages is fundamental to how it causes disease, but the basis of this is poorly understood. To examine further the general application of a recently developed system of targeted gene mutation and to assess the importance of different genes in resistance to innate immune defenses, we disrupted the genes encoding high-temperature requirement A (htrA), nitrate reductase (narG), peptidase D (pepD), phosphoribosylaminoimidazole-succinocarboxamide synthase (purC), and superoxide dismutase (sodC) in strain 103 of R. equi using a double-crossover homologous recombination approach. Virulence testing by clearance after intravenous injection in mice showed that the htrA and narG mutants were fully attenuated, the purC and sodC mutants were unchanged, and the pepD mutant was slightly attenuated. Complementation with the pREM shuttle plasmid restored the virulence of the htrA and pepD mutants but not that of the narG mutant. A single-crossover mutation approach was simpler and faster than the double-crossover homologous recombination technique and was used to obtain mutations in 6 other genes potentially involved in virulence (clpB, fadD8, fbpB, glnA1, regX3, and sigF). These mutants were not attenuated in the mouse clearance assay. We were not able to obtain mutants for genesfurA, galE, and sigE using the single-crossover mutation approach. In summary, the targeted-mutation system had general applicability but was not always completely successful, perhaps because some genes are essential under the growth conditions used or because the success of mutation depends on the target genes.     

Susceptibility to and transmission of H5N1 and H7N1 highly pathogenic avian influenza viruses in bank voles (Myodes glareolus)

The study of influenza type A (IA) infections in wild mammals populations is a critical gap in our knowledge of how IA viruses evolve in novel hosts that could be in close contact with avian reservoir species and other wild animals. The aim of this study was to evaluate the susceptibility to infection, the nasal shedding and the transmissibility of the H7N1 and H5N1 highly pathogenic avian influenza (HPAI) viruses in the bank vole (Myodes glareolus), a wild rodent common throughout Europe and Asia. Two out of 24 H5N1-infected voles displayed evident respiratory distress, while H7N1-infected voles remained asymptomatic. Viable virus was isolated from nasal washes collected from animals infected with both HPAI viruses, and extra-pulmonary infection was confirmed in both experimental groups. Histopathological lesions were evident in the respiratory tract of infected animals, although immunohistochemistry positivity was only detected in lungs and trachea of two H7N1-infected voles. Both HPAI viruses were transmitted by direct contact, and seroconversion was confirmed in 50% and 12.5% of the asymptomatic sentinels in the H7N1 and H5N1 groups, respectively. Interestingly, viable virus was isolated from lungs and nasal washes collected from contact sentinels of both groups. The present study demonstrated that two non-rodent adapted HPAI viruses caused asymptomatic infection in bank voles, which shed high amounts of the viruses and were able to infect contact voles. Further investigations are needed to determine whether bank voles could be involved as silent hosts in the transmission of HPAI viruses to other mammals and domestic poultry.     

Synthesis and resistance to in vitro proteolysis of transglutaminase cross-linked phaseolin, the major storage protein from Phaseolus vulgaris

The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin.     

First report of Enterocytozoon bieneusi from dairy cattle in Argentina

Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.     

Assessment of attenuated Salmonella vaccine strains in controlling experimental Salmonella Typhimurium infection in chickens

Salmonella hold considerable promise as vaccine delivery vectors for heterologous antigens in chickens. Such vaccines have the potential additional benefit of also controlling Salmonella infection in immunized birds. As a way of selecting attenuated strains with optimal immunogenic potential as antigen delivery vectors, this study screened 20 novel Salmonella Typhimurium vaccine strains, differing in mutations associated with delayed antigen synthesis and delayed attenuation, for their efficacy in controlling colonization by virulent Salmonella Typhimurium, as well as for their persistence in the intestine and the spleen. Marked differences were observed between strains in these characteristics, which provide the basis for selection for further study as vaccine vectors.     

Grazing behaviour,physical activity and metabolic profile of two Holstein strains in an organic grazing system

The challenge for sustainable organic dairy farming is identification of cows that are well adapted to forage‐based production systems. Therefore, the aim of this study was to compare the grazing behaviour, physical activity and metabolic profile of two different Holstein strains kept in an organic grazing system without concentrate supplementation. Twelve Swiss (H_CH; 566 kg body weight (BW) and 12 New Zealand Holstein‐Friesian (H_NZ; 530 kg BW) cows in mid‐lactation were kept in a rotational grazing system. After an adaptation period, the milk yield, nutrient intake, physical activity and grazing behaviour were recorded for each cow for 7 days. On three consecutive days, blood was sampled at 07:00, 12:00 and 17:00 h from each cow by jugular vein puncture. Data were analysed using linear mixed models. No differences were found in milk yield, but milk fat (3.69 vs. 4.05%, P = 0.05) and milk protein percentage (2.92 vs. 3.20%, P < 0.01) were lower in H_CH than in H_NZ cows. Herbage intake did not differ between strains, but organic matter digestibility was greater (P = 0.01) in H_CH compared to H_NZ cows. The H_CH cows spent less (P = 0.04) time ruminating (439 vs. 469 min/day) and had a lower (P = 0.02) number of ruminating boli when compared to the H_NZ cows. The time spent eating and physical activity did not differ between strains. Concentrations of IGF‐1 and T3 were lower (P ≤ 0.05) in H_CH than H_NZ cows. In conclusion, H_CH cows were not able to increase dry matter intake in order to express their full genetic potential for milk production when kept in an organic grazing system without concentrate supplementation. On the other hand, H_NZ cows seem to compensate for the reduced nutrient availability better than H_CH cows but could not use that advantage for increased production efficiency.     

肠道微生物区系中与肉鸡生产性能相关菌属的鉴定

肠道菌群对家禽健康与营养具有潜在的影响,其对宿主能量代谢的调节作用可能与饲料转化率的变化有关。本文在简述肠道微生物功能,家禽肠道微生物的形成与分布,影响肠道微生物区系平衡因素的基础上,鉴定肠道微生物区系中与肉鸡生产性能相关的特定菌属。     

肠道微生物区系中与肉鸡生产性能相关菌属的鉴定

正肉鸡的胃肠系统在提高饲料转化率方面发挥着极其重要的作用,它参与分解日粮组分、吸收营养物质及将机体代谢物排出体外。肉鸡肠道具有独特的微生物生态系统,可与肉鸡形成共生关系。肠道微生物可使宿主完全消化日粮成分,是细菌密度最高的生态系统,含有有益菌、有害菌和良性菌等,因而可对动物健康与营养造成潜在的影响。日粮营养物质摄入的速度、宿主的免疫应答状态及肠道结构等均可影响肠道微生物;同时日粮组分、宿主免疫或肠道结构     

Expert knowledge for translating land cover/use maps to General Habitat Categories (GHC)

Monitoring biodiversity at the level of habitats and landscape is becoming widespread in Europe and elsewhere as countries establish international and national habitat conservation policies and monitoring systems. Earth Observation (EO) data offers a potential solution to long-term biodiversity monitoring through direct mapping of habitats or by integrating Land Cover/Use (LC/LU) maps with contextual spatial information and in situ data. Therefore, it appears necessary to develop an automatic/semi-automatic translation framework of LC/LU classes to habitat classes, but also challenging due to discrepancies in domain definitions. In the context of the FP7 BIO_SOS (www.biosos.eu) project, the authors demonstrated the feasibility of the Food and Agricultural Organization Land Cover Classification System (LCCS) taxonomy to habitat class translation. They also developed a framework to automatically translate LCCS classes into the recently proposed General Habitat Categories classification system, able to provide an exhaustive typology of habitat types, ranging from natural ecosystems to urban areas around the globe. However discrepancies in terminology, plant height criteria and basic principles between the two mapping domains inducing a number of one-to-many and many-to-many relations were identified, revealing the need of additional ecological expert knowledge to resolve the ambiguities. This paper illustrates how class phenology, class topological arrangement in the landscape, class spectral signature from multi-temporal Very High spatial Resolution (VHR) satellite imagery and plant height measurements can be used to resolve such ambiguities. Concerning plant height, this paper also compares the mapping results obtained by using accurate values extracted from LIght Detection And Ranging (LIDAR) data and by exploiting EO data texture features (i.e. entropy) as a proxy of plant height information, when LIDAR data are not available. An application for two Natura 2000 coastal sites in Southern Italy is discussed.