融合表达淋巴细胞脉络丛脑膜炎病毒和卵清白蛋白CD8+T细胞表位的减毒鼠伤寒沙门氏菌的构建与鉴定

依据淋巴细胞脉络丛脑膜炎病毒(LCMV)主要保护性抗原CD8 T细胞表位VRRPQASGVYMGNLTAQ和卵清白蛋白(OVA)CD8 T细胞表位SIINFEKL,设计、合成两条编码LCMV和OVA CD8 T细胞表位的寡核苷酸片段,经退火后,克隆入绿色荧光蛋白表达质粒pYAGFP,经PCR扩增和序列测定分析,证实成功构建T细胞表位与绿色荧光蛋白融合表达的重组质粒pYAGFPL-O,将此重组质粒pYAGFPL-O转化减毒鼠伤寒沙门氏菌X4550,获得重组沙门氏菌X4550(pYAGFPL-O).用SDS-PAGE电泳测得重组菌表达的融合蛋白约为30kD.同时,荧光显微镜下观察到X4550(pYAGFPL-O)发出黄绿色荧光.这些结果表明LCMV和OVA T细胞表位已成功表达.重组菌X4550(pYAGFPL-O)的获得为研究沙门氏菌载体携带外源抗原的T细胞应答规律及调控机理打下了重要基础.     

Antimicrobial resistance and recovery of Salmonella,Campylobacter, and Escherichia coli from chicken egg layer flocks in Canadian sentinel surveillance sites using 2 types of sample matrices

Eggs are important to the diet of Canadians. This product is one of the supply-managed commodities in Canada, but unlike other commodities, where food safety risks are extensively explored and reported, information on the prevalence of enteric organisms (e.g., Salmonella, Campylobacter) and antimicrobial resistance (AMR) in layers in Canada are limited. This study was conducted to determine the prevalence of select bacteria and the associated AMR patterns in layer flocks using 2 sample matrices. Farms were located within FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance sentinel sites (SS). Fecal samples (Ontario: ON_SS1a, ON_SS1b) and environmental sponge swabs (British Columbia: BC_SS2a) were collected. Salmonella prevalence was 29% and 8% in ON_SS1a and ON_SS1b, respectively, and 7% in BC_SS2a. S. Kentucky and S. Livingstone were the most frequently isolated serovars and no S. Enteritidis was detected. Campylobacter was not detected in the BC sponge swabs but was isolated from 89% and 53% of Ontario fecal samples (ON_SS1a and ON_SS1b, respectively). Seven C. jejuni from Ontario were ciprofloxacin-resistant. Escherichia coli prevalence was high in both sample types (98%). Overall, tetracycline resistance among E. coli ranged from 26% to 69%. Resistance to ceftiofur (n = 2 isolates) and gentamicin (n = 2) was relatively low. There were diverse resistance patterns (excludes susceptible isolates) observed among E. coli in Ontario (10 patterns) and British Columbia (14 patterns). This study revealed that fecal samples are more informative for farm-level monitoring of pathogen and AMR prevalence. Without further validation, sponge swabs are limited in their utility for Campylobacter detection and thus, for public health surveillance.     

TRUNCATION ARTIFACT IN MAGNETIC RESONANCE IMAGES OF THE CANINE SPINAL CORD

The truncation artifact in magnetic resonance (MR) images is a line of abnormal signal intensity that occurs parallel to an interface between tissues of markedly different signal intensity. In order to demonstrate the truncation artifact in sagittal images of the canine spinal cord and the effect of changing spatial resolution, we conducted an experimental in vitro study. A section of fixed canine spinal cord was imaged using a 1.5T magnet. Spatial resolution was increased by increasing the acquisition matrix and reconstruction matrix, producing series of T2‐weighted (T2w) images with the following pixel sizes: A, 1.6 (vertical) × 2.2 mm² (horizontal); B, 1.2 × 1.7 mm²; C, 0.8 × 1.1 mm²; D, 0.4 × 0. 6 mm². Plots of mean pixel value across the cord showed variations in signal intensity compatible with truncation artifact, which appeared as a single, wide central hyperintense zone in low‐resolution images and as multiple narrower zones in high spatial resolution images. Even in images obtained using the highest spatial resolution available for the MR system, the edge of the spinal cord was not accurately defined and the central canal was not visible. The experiment was repeated using an unfixed spinal cord specimen with focal compression applied to mimic a pathologic lesion. Slight hyperintensity was observed within the spinal cord at the site of compression although the cord was normal histologically. Results of this study suggest that caution should be applied when interpreting hyperintensity affecting the spinal cord in T2w sagittal images of clinical patients because of the possibility that the abnormal signal could represent a truncation artifact.     

Physiological response of one of South Africa’s premier freshwater sport angling species,the Orange-Vaal smallmouth yellowfish Labeobarbus aeneus,to catch-and-release angling

The practice of catch-and-release fishing has been widely promoted by angling fraternities as a fisheries manage- ment tool. The aim of this investigation was to determine the physiological response of Orange-Vaal smallmouth yellowfish, Labeobarbus aeneus, to catch-and-release angling in the Vaal River, South Africa. Fish were collected using standard fly-fishing techniques, anaesthetised in clove oil and blood was drawn from the caudal vein; thereafter the fish were weighed, measured, revived and released. Blood plasma was analysed for concentra- tions of glucose, cortisol and lactate to determine the effects of angling duration, fish size and water tempera- ture. Larger fish were angled for a longer duration compared with smaller fish. Levels of glucose were affected by water temperature (influenced by time of year). Plasma glucose concentrations decreased with greater angling duration. Few individuals (n = 12) showed increased plasma cortisol concentrations. In extended-capture fish (angled for >1 min), lactate concentrations increased significantly above values for rapid-capture fish (angled for >30 s). These data suggest that catch-and-release causes physiological stress to fish, but nonetheless this practice can be a valuable fisheries management tool to ensure the sustainability of fish populations. Other factors beyond the ‘angling’ time are likely to contribute to physiological disruptions in homeostasis and therefore handling and air exposure of angled fish should be included in future catch-and–release angling studies. In addition, the longer-term impact of angling on fish health should also be determined.     

Conjunctival habronemiasis in a square‐lipped rhinoceros (Ceratotherium simum)

A captive female square‐lipped rhinoceros born in 1993 had been showing intermittent signs of bilateral conjunctivitis and conjunctival proliferation since 1998. Periodic improvement was noted, especially in winter, but overall the condition had deteriorated over the years. Treatment with various topical, intralesional, and systemic antibiotics and glucocorticosteroids was largely ineffective, as were repeated dewormings. No primary cause for these lesions was found in biopsies taken in 2000 and 2006, although a severe infiltrate of numerous eosinophils was observed in the latter. As the condition worsened, secondary corneal changes were noted, and eventually vision was lost due to proliferative conjunctival tissue. Aggressive resection of the proliferating tissue in 2013 restored vision and submitted biopsies yielded a diagnosis of severe allergic conjunctivitis, eosinophilic granuloma, and habronematid (Habronema or Draschia) larval infection. As no other rhinoceros in the herd was affected, including two calves born to the patient who were in close contact with their mother, it was concluded the presentation was most likely due to a hypersensitivity reaction to the dead or dying larvae. Fly repellent is now regularly applied around the eye of this rhinoceros, and a protective face mask has been fitted. Ongoing periodic relapses are treated with oral ivermectin, topical antibiotics, and steroids.