Serodiagnosis of Burkholderia mallei Infections in Horses: State‐of‐the‐art and Perspectives

Burkholderia mallei causes glanders or farcy in solipeds, a disease that must be reported to the OIE (Office International des Epizooties, Paris, France). The number of reported outbreaks has increased steadily during the last decade. Serodiagnosis is hampered by the considerable number of false‐positives and ‐negatives of the internationally prescribed tests. The major problem leading to low sensitivity and specificity of complement fixation test (CFT) and enzyme‐linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e. crude preparations of whole cells. Future perspectives for the development and evaluation of serological test kits using well‐characterized single antigens are discussed in the light of recent molecular research on B. mallei and the closely related saprozoonotic agent B. pseudomallei.     

Trade-offs among establishment success,stem morphology and productivity of underplanted Toona ciliata: Effects of nurse-species and thinning density

Toona ciliata (Australian red cedar), a valuable hardwood species, requires a nurse-tree overstorey to prevent damage from frost and drought in some regions of north-eastern Argentina. Different nurse-species offer different degrees of protection, resource competition, and thinning revenues. In addition, choice of nurse-species will influence trade-offs between T. ciliata establishment success, stem quality and productivity as well as revenue from intermediate and final harvests. We tested the effects of overstorey species and planting density on underplanted T. ciliata survival, stem morphology and growth. Initial stand densities of nurse-species Pinus taeda (625 stems/ha), P. elliottii × P. caribaea (625 stems/ha), and Grevillea robusta (833 stems/ha) were thinned by 0%, 25%, 50%, and 75% in three experiments, each using a randomized block design. Initial mortality, light availability and growth of T. ciliata, as well as growth of its nurse-trees, were measured before and two years after thinning. In the third year, we characterized T. ciliata stem morphology (straightness and height-to-diameter-ratio) and destructively harvested a subsample of T. ciliata in order to investigate T. ciliata biomass production and allocation patterns in response to overstorey species, thinning density and light availability. T. ciliata mortality was lower when planted under G. robusta than the pines, and increased in plots with greater exposure to solar radiation. T. ciliata productivity was also greater under G. robusta than under the pines, and was generally greatest where any of the nurse-species were thinned by 75%. Stem form was best when growing under P. taeda and in stands thinned by 50%, regardless of nurse-species. With the exception of early mortality, light levels appeared to have limited effects on T. ciliata growth dynamics. Neither growth rate nor stem morphology was strongly correlated with light availability. Total overstorey basal-area was highest in P. taeda stands and individual overstorey tree growth generally increased with decreasing stand densities, except for slower height growth of G. robusta in more open stands. Thinning operations resulted in sawn-wood revenue for the pine species only. Because stand productivity and stem quality were optimized under different nurse-species and thinning treatments, we conclude that the choice of nurse-species and thinning intensity will depend on management objectives and will change with market preferences and conditions.     

Influence of long-time transportation stress on re-activation of Salmonella typhimurium DT104 in experimentally infected pigs

In this study a Salmonella Typhimurium infection model in swine was used in order to investigate the influence of pre-mortal stress induced by long time period transportation on the re-activation of Salmonella in experimentally infected pigs. Salmonella free pigs were exposed to a highly virulent strain of Salmonella Typhimurium DT104 by direct intragastrical administration. Clinical parameters were monitored and the shedding rate in faeces was qualitatively and quantitatively determined by standard bacteriological procedures for 21 days. The distribution of the challenge organism in 14 different internal organs of transported and nontransported animals was determined. All infected animals developed clinical signs of salmonellosis 12 to 24 hours post infection. About 88 to 100% of the fecal samples were culture-positive up to post exposure day 6, and then varied from 71 to 92% until slaughter, respectively. At necropsy S. Typhimurium was recovered most frequently from caecum and ileocolic lymph nodes (83%), colon (79%), palatine tonsils (71%) and mandibular lymph nodes (62.5%). A negative impact of transportation stress on the shedding rate and the general condition of the animals was observed.     

Evaluation of porcine ileum models of enterocyte infection by Lawsonia intracellularis

The early interaction of Lawsonia intracellularis with host cells was examined with the use of porcine ileum models. Two conventional swine were anesthetized, and ligated ileum loops were prepared during abdominal surgery. The loops were inoculated with 108 L. intracellularis or saline. After 60 min, samples of each loop were processed for routine histologic and electron microscopic study. Histologic and ultrathin sections of all the loops appeared normal, with no apposition of bacteria and host cells or bacterial entry events in any loop. Portions of ileum from a single gnotobiotic piglet were introduced as xenografts into the subcutis of each flank of 5 weaned mice with severe combined immunodeficiency disease. After 4 wk, 108 L. intracellularis were inoculated into each of 4 viable xenografts with a sterile needle; the other 3 viable xenografts received saline. Histologic and ultrathin sections of all the xenografts 3 wk after inoculation showed relatively normal porcine intestinal architecture, with normal crypts, crypt cell differentiation, and low villous structures; the xenografts treated with the bacteria also showed intracytoplasmic L. intracellularis within crypt and villous epithelial cells. Thus, entry of L. intracellularis into target epithelial cells and multiplication may not be sufficient alone to directly cause cell proliferation. A proliferative response may require active division of crypt cells and differentiation in conjunction with L. intracellularis growth.     

Performance of a foot-and-mouth disease virus reverse transcription-polymerase chain reaction with amplification controls between three real-time instruments.

The foot-and-mouth disease virus (FMDV) is a member of the picornavirus family, possessing an 8-kb single-stranded RNA genome of positive polarity. It is highly contagious among several livestock species and can lead to severe economic consequences, as evidenced by the UK outbreak in 2001. The usage of real-time polymerase chain reaction has facilitated rapid detection of FMDV. Several real-time PCR instruments are available with various capabilities, such as portability and high sample volume analysis. Primers and a dual-labeled TaqMan probe were optimized to detect a single, highly conserved 88-bp segment of the FMDV 3D (RNA polymerase) gene. To increase the confidence of the RT-PCR result, a positive amplification control was synthesized to detect potential false-positive results due to contamination if a wild-type virus is used as positive control. In addition, a preventative measure against false-negative results was developed in which endogenous beta actin mRNA is coamplified by RT-PCR. Assay performance was compared on the LightCycler1.2 (Roche), the SmartCyclerII (Cepheid), and the SDS 7900HT (ABI). These assays successfully identified the FMDV genome and beta actin mRNA from several sources of infected nasal and oral swabs, as well as probang samples.     

Induction dose and recovery quality of propofol and alfaxalone with or without midazolam coinduction followed by total intravenous anesthesia in dogs

Objectives

To compare propofol and alfaxalone, with or without midazolam, for induction of anesthesia in fentanyl-sedated dogs, and to assess recovery from total intravenous anesthesia (TIVA).