Chicken anemia virus induced apoptosis: underlying molecular mechanisms

In 1990, the chicken anemia virus (CAV) genome was cloned by us and proven to be representative for CAV isolates worldwide. This genome contains unique promoter/enhancer replication elements and genes. Upon infection of its target cells, CAV replicates via a double-stranded (ds) DNA intermediate. From this ds CAV molecule, a single mRNA is transcribed, which encodes for three distinct proteins VP1, VP2, and VP3 or apoptin. Its capsid contains only the VP1 protein. However, for the production of the neutralizing epitope, co-synthesis of VP1 and VP2 are needed. CAV genomes with mutations in the 12 bp insert of the promoter/enhancer region were shown to produce immunogenic functional CAV particles. Mutations in these and other regulatory elements of CAV might also decrease its virus load resulting in a reduced pathogenic effect. CAV causes fatal cytopathogenic effects in e.g. chicken thymocytes via apoptosis. Under in vitro conditions, CAV replicates only in transformed chicken cell lines, which indicates that at least a part of the CAV life-cycle requires transformed-like cellular events. In these transformed cell lines, the synthesis of the apoptin protein alone mimics the CAV-induced apoptosis, whereas the VP2 protein also harbors some apoptotic activity. Extensive studies on apoptin resulted in the characterization of domains essential for its apoptotic activity and nuclear localization, which seems to be related with its ability to induce apoptosis. Therefore, both VP2 and apoptin are of interest in reducing the pathogenicity of CAV infections. A series of biomedical studies on apoptin have been carried out in human cell systems, which are informative about the mechanism of CAV-induced apoptosis in chicken (transformed) cells. Synthesis of apoptin alone induces apoptosis in various human transformed and/or tumorigenic cell lines, but not in normal human diploid cells. A striking difference in the cellular localization of apoptin was observed in human normal diploid cells versus tumor cells. In all tumor cells, apoptin is located mainly in the heterochromatic regions of the nucleus, whereas in normal cells it is present in peri-nuclear structures. Apoptin contains a bipartite nuclear localization signal, and one domain that resemble a nuclear export signal. Elucidation of parts of the apoptin-induced apoptotic pathway revealed unique characteristics: apoptin-induced apoptosis is independent of the tumor suppressor p53. The anti-apoptotic protein Bcl-2 does not inhibit but even accelerates apoptin-induced apoptosis in tumor cells, whereas over expression of Bcl-2 in normal cells has no effect on the apoptin activity. Upstream caspases are not involved, whereas downstream caspase 3 is, but seems not to be essential. A number of novel proteins were shown to interact with apoptin in transformed cells. Future studies of apoptin, VP2 and related cellular proteins in chicken cells will unravel the regulatory aspects of CAV-induced apoptosis.     

Treatment of experimental ureteral strictures by endourological ureterotomy and implantation of stents in the porcine animal model

The objective of this study is to evaluate the dilation of the ureter using endoureterotomy and an expanding-sheath double pigtail ureteral stent in the treatment of experimentally induced ureteral strictures in the porcine animal model. This is a new treatment in the ureteral strictures resolution in Veterinary Urology, although it is not a common affection, it usually appears as a consequence of ureteritis and in the iatrogenic female genital surgery. The experimental study is design in three phases: induction of experimental stricture, diagnosis and treatment of the stricture and follow-up. We have used 10 healthy Large White female pigs. The internal ureteral diameter was measured prior to laparoscopic ligature stricture induction using retrograde ureteropyelography (RUPG). Experimental stricture was diagnosed 4 weeks after intervention, using RUPG and ultrasound, and treated by endoureterotomy and subsequent placement of a double pigtail ureteral stent, which was removed 6 weeks later. The study finished 4 weeks later with measurement of ureteral diameters using RPUG and ultrasound evaluation. Except in one case, all ureters displayed permanent dilation of the strictured area for 10 weeks after treatment (6 weeks with ureteral stent and 4 more weeks without stent). Finally, this technique proved to be effective in cases of short-length and short-living ureteral strictures, and represents a viable alternative to conventional surgery in animals.     

Chemoattraction and chemorepulsion of Strongyloides stercoralis infective larvae on a sodium chloride gradient is mediated by amphidial neuron pairs ASE and ASH, respectively

Depending on its concentration, sodium chloride acts as either an attractant or a repellant to the infective larvae (L3i) of Strongyloides stercoralis. On a concentration gradient, L3i are attracted to 0.05 M NaCl, but repelled by 2.8M. To test the hypothesis that amphidial neurons ASE and ASH might mediate attraction and repulsion, respectively, these neurons, and control neurons as well, were ablated in hatchling larvae with a laser microbeam. After the larvae attained infectivity (L3i), they were tested on a NaCl gradient. When placed at low salinity, 73.5% of normal controls migrated "up" the gradient, while 26.4% crawled randomly. In contrast, only 20.6% of ASE-ablated L3i migrated "up" the gradient, while 79.4% migrated randomly. Ablation-control ASK-ablated L3i (58.8%) migrated "up" the gradient while 41.1% crawled randomly. When placed at a region of high salinity, 100% of normal control L3i migrated "down" the gradient, whereas 62.5% of ASH-ablated L3i migrated randomly, the remaining 37.5% migrating "down" the gradient. In sharp contrast with ASH-ablated L3i, 94.1% of ablation-control larvae, i.e. ASK-ablated L3i, migrated "down" the gradient. Migration behavior of ASE- and ASH-ablated L3i was significantly different (P < 0.001) from that of ASK-ablated L3i and normal controls. It is noteworthy that 87.5% of ASE-ablated L3i that failed to exhibit chemoattractive behavior were actively chemorepelled from high salinity. Also, 70.0% of ASH-ablated L3i that failed to be chemorepelled from high salinity were capable of chemoattractive behavior, indicating that the worms had retained their behavioral responses except for those associated with the targeted neurons.     

Identification of a novel hemotropic mycoplasma in a splenectomized dog with hemic neoplasia

A 3-year-old sexually intact male Bull Mastiff underwent splenectomy for splenic thrombosis; prior to and after splenectomy, multiple blood transfusions were administered. Two weeks after the procedure, T-cell lymphoproliferative disease was diagnosed. Treatment with prednisone and chlorambucil was initiated, and 2 weeks later, cytologic examination of a blood smear revealed small (0.3 microm), coccoid basophilic bodies on the surface of approximately 70% of the RBCs. Morphologically, these resembled "Candidatus Mycoplasma haemominutum." A polymerase chain reaction assay was used to amplify a partial 16S rRNA sequence in blood obtained from the dog; the product was sequenced and compared with 16S rRNA gene sequences of other hemotropic mycoplasmas. The sequence was 98% homologous to that of "Candidatus M haemominutum", but only 77% homologous to that of M haemocanis and M haemofelis.     

Veterinary school admission interviews, part 3: strategies for increasing interview validity

The veterinary school admission interview is a widely used selection tool, yet concerns persist about its reliability, validity, and cost. Relative to medicine, optometry, and dentistry schools, veterinary schools have been more likely to conduct panel interviews and to fix the interview's weight in selection decisions, strategies that increase interview validity. This article provides strategies for further increasing the veterinary school interview's validity. Interview reliability and validity studies point to key strategies the veterinary school admissions committee can implement before the interview: (1) establishing the interview's purpose(s); (2) conducting a "job" analysis to identify desirable candidate skills, knowledge, and attributes; (3) developing a structured and panel interview where interviewers, if possible, are blind to other admission data; (4) training interviewers; (5) setting a reasonable interview schedule; and (6) determining methods for analyzing applicant data. During the interview, interviewers should proceed through a structured series of steps: (1) open the interview with a specified agenda; (2) probe for information using structured questions and anchored rating scales; (3) close the interview to allow for candidate questions; and (4) evaluate the interview data. After the interview, the admissions committee should (1) analyze the interview data within and across interviewers and (2) analyze the data across all selection tools in order to assign relative weights to the selection tools.     

Effects of contact dermatitis on hepatic gene expression in broilers

1.?Severe foot and hock dermatitis in broiler chickens can reduce growth rate and increase susceptibility to bacterial infection, affecting both profitability and welfare. However, little is known about the underlying physiological changes associated with foot and hock lesions.

2.?This study compared global hepatic gene expression in control birds and those with ammonia-induced foot and hock lesions using Agilent 44?K chicken oligonucleotide microarrays (8 birds per group).

3.?In total, 417 genes were differentially expressed of which 174 could be mapped onto the genome. Genes associated with energy metabolism, thyroid hormone activity and cellular control were affected, while there was also evidence of an up-regulation of genes linked to a pro-inflammatory response.

4.?It is conceivable that pain is the underlying cause for the observed changes in energy metabolism genes.

5.?Changes in hepatic gene expression provide new information on how a chicken's physiological mechanisms alter to cope with foot and hock lesions. The findings support other data indicating that birds with increased severity of lesions are likely to be in pain and that growth will be compromised. Reduction of the incidence of dermatitis by improved husbandry should therefore benefit both welfare and commercial performance.     

Investigation of the Concurrent Emergence of Salmonella enteritidis in Humans and Poultry in British Columbia,Canada, 2008–2010

An increase in the rate of human infections with Salmonella enteritidis (SE) occurred between 2007 and 2010 in British Columbia (BC). During the same time period, an increase in SE from poultry‐sourced isolates and increased clinical severity in poultry were also observed in BC. This article describes a multi‐sectoral collaboration during a 3‐year investigation, and the actions taken by public health and animal health professionals. Human cases were interviewed, clusters were investigated, and a case–control study was conducted. Environmental investigations were conducted in food service establishments (FSE). Suspect foods were tested. Laboratory data from poultry‐sourced isolates were analysed. Five hundred and eighty‐four human cases of SE with the same pulsed‐field gel electrophoresis pattern were identified between May 2008 and August 2010. Seventy‐three percentage of cases reported consumption of eggs. The odds of egg consumption were 2.4 times higher for cases than controls. Implicated FSE were found to use ungraded eggs, which had been distributed illegally. Investigation suggested that there were multiple suppliers of these eggs. Collaboration between public health and animal health professionals led to data sharing, improved understanding of SE, engagement with the poultry industry and public communication. Multi‐disciplinary, multi‐sectoral and multi‐pronged investigations are recommended to identify the likely source of illness in large, protracted foodborne outbreaks caused by commonly consumed foods.     

Proximal hindlimb flexion in the horse: Effect on movement symmetry and implications for defining soundness

Reasons for performing the study: Flexion tests are a common tool during the prepurchase and clinical lameness examination, yet studies quantifying the effect of flexion, apart from distal forelimb flexion in sound horses, are sparse. Objectives: To investigate the effect of proximal hindlimb flexion on perceivable and measurable changes in movement symmetry in horses with objective movement symmetry falling within the margins of ‘sound’. Methods: Thirteen horses, selected based on objective movement symmetry, were instrumented with inertial sensors on left and right tuber coxae and over the os sacrum. Vertical movement symmetry was quantified at trot before and after proximal hindlimb flexion, with a repeat of flexion after 5 min. Video recordings of the horses were assessed visually. Results: Proximal hindlimb flexion introduced additive changes in movement symmetry to the individual baseline movement, with a tendency towards smaller effects with increasing stride number. The main systematic effect was a decrease in upward movement of the os sacrum following mid‐stance of the flexed limb and an increase in upward movement following mid‐stance of the nonflexed limb, also manifesting in a ‘hip hike’ of the flexed limb; these findings reflected increased movement asymmetry following flexion. Depending on individual baseline asymmetry, flexion can also increase movement symmetry. Conclusions: Proximal hindlimb flexion can exacerbate subtle asymmetry when performed carefully. Variation in measured symmetry following flexion within and between horses showed that the individual response to flexion is highly variable. Potential relevance: Proximal hindlimb flexion may elevate the asymmetry of a slightly lame limb above the threshold for visibility, thus assisting in the clinical gait examination. Further work is needed to examine the causes for a positive response to flexion and possible differences between sound and lame horses as well as horses of different athletic disciplines.