Lessons learned from three avian influenza simulation exercises in the southwest of France.

The Southwest of France raises different species of poultry. These production activities present structural vulnerabilities to severe infectious diseases such as highly pathogenic avian influenza. Indeed, many farms have free-range flocks, multi-age and multi-species productions, while being located near wild bird migratory corridors. These factors may partly explain the H5 virus epidemics that occurred between 2015 and 2021. Their serious economic and technical consequences and psychological impact have generated solidarity, collective learning and operational cohesiveness among all poultry professionals. Consequently, a decision was made to conduct annual simulation exercises for a major health event in order to maintain a high level of vigilance and responsiveness within different poultry sectors. Three exercises took place, in 2017, 2018 and 2019, in semi-real conditions (real dates and compressed time) and according to different scenarios. They took place outside an epidemic context and have in common to focus on the initial phase of the crisis (suspicions, results of preliminary analyzes), which is critical to assess the reactivity of industry personnel in order to mitigate infectious disease spread. The preparation of the simulation exercises was based on a common methodology. They were created by an organizing team and each included up to 60 people (industry personnel, observers and auditors). These simulations highlighted several critical points: poultry professionals have detailed knowledge of the field, but this information can only be effectively obtained and used if there is already a poultry industry decision-making structure in place (with good networking); there is a need (1) for better information sharing within the industry; (2) to develop an assistance structure for producers directly involved in a crisis; and (3) to increase collaboration with State services in peacetime. Finally, several technical issues were raised regarding control zones; blocking poultry movements; production site quarantine; depopulation strategies; self-financing capacity of the poultry industry in the absence of governmental involvement; and enhanced mapping tools with real-time traceability of animal transportation.

Assessing most practical and effective protocols to sanitize hands of poultry catching crew members.

Catching crew members can heavily contaminate their hands with organic material. They can act as mechanical vector and spread diseases between farms. Hand hygiene is an important issue for the industry as a whole and for human health by reducing contamination risks. Many studies, in human medicine, tend to make hand rub a standard for hand hygiene. However, few studies have tested the effectiveness of hand hygiene products on visibly contaminated hands. The objective of this study was to evaluate the effectiveness of practical hand sanitization protocols: water and soap, degreasing cream and hand wipes, all combined with alcohol-based hand gel. The use of alcohol-based gel alone was also evaluated. For the reduction of coliforms after washing, there was no statistically significant difference between protocols when the initial level of bacterial contamination was low to moderate. When hands were highly contaminated, the alcohol-based gel alone was less effective than the degreasing cream combined with the alcohol-based gel (p=0.002). As for the reduction in total aerobic bacteria counts, there was no difference between protocols when the initial level of bacterial contamination was low. The water, soap and alcohol-based gel protocol was more effective than the scrubbing wipes and alcohol-based gel protocol when hands were moderately (p=0.002) and highly contaminated (p=0.001). All protocols were effective in neutralizing Salmonella on hands. Reducing the level of bacterial contamination on hands before using an alcohol-based gel seems important to ensure effective hand sanitation for highly and moderately contaminated hands. This can be done by using a degreasing cream or water and soap. Based on the survey, catching crew members preferred using warm water and soap compared to a degreasing cream.

Canadian experiences with avian influenza: a look at regional disease control--past, present, and future.

Over the past 5 yr, the poultry industry in Canada has had a few H5 or H7 avian influenza (AI) epidemics. An analysis of these outbreaks by government officials highlighted the need to establish a better partnership between those responsible for controlling the disease and public health officials responsible for protecting the public and those participating in eradication efforts. These officials also agreed that compensations had to be reviewed, that national biosecurity standards needed to be established to better prevent AI, that a national mortality disposal plan was needed, and finally that the current emergency disease management protocols had to be reviewed. Industry representatives stressed the need for early detection and reporting; for more effective tools for decision making, including using local expertise for trace-back activities and quick interventions; for better communications within industry, but mainly between industry and governmental authorities at the federal, provincial, and municipal levels; and finally, for better planning to minimize the impact of eradication efforts on poultry production and for the recovery following the epidemic. These observations triggered a series of initiatives. A National Office of Animal Biosecurity was created by federal authorities, with the mandate to establish national biosecurity standards. A Canadian Animal Health Surveillance Network was also put in place to improve the capacity of early detection of the disease and to increase the surge capacity of the Canadian laboratory system. Wildlife and commercial poultry AI surveillance programs have also been put in place. Provincial poultry grower organizations have established AI control and eradication plans that are increasing their ability to intervene early and to assist government authorities once AI is confirmed in the field. This includes the creation of industry incident command centers with emphasis on confidentiality agreements between government and industry organizations and effective grower assistance before, during, and after an epidemic.

Experimental infection of chickens and turkeys with Mycoplasma gallisepticum reference strain S6 and North Carolina field isolate RAPD type B.

During an epidemic of mycoplasmosis in chicken and turkey flocks in North Carolina between 1999 and 2001, isolates of Mycoplasma gallisepticum (MG) from affected flocks were characterized by random amplification of polymorphic DNA (RAPD), and eight distinct RAPD types were identified. MG RAPD type B accounted for more than 90% of the isolates and was associated with moderate-to-severe clinical signs and mortality. The virulence of MG RAPD type B for chickens and turkeys was compared with sham-inoculated negative controls and MG S6 (a virulent strain)-inoculated positive controls. Clinical signs occurred in chickens and turkeys inoculated with either MG RAPD type B or MG S6. However, they were not as frequent or severe as those seen in naturally affected flocks, and there was no mortality in the experimental groups. Based on gross and microscopic findings, MG RAPD type B was equal to or more virulent than MG S6. All MG-inoculated birds were culture and PCR positive at 7 and 14 days postinoculation (PI). Among serological tests, the serum plate agglutination test was positive for the majority of chickens and turkeys (58%-100%) infected with either strain of MG at both 7 and 14 days PI. The hemagglutination inhibition test was negative for all birds at 7 days PI and positive for a few chickens (8%-17%) and several turkey sera (40%-60%) at 14 days PI. Only a single serum was positive by enzyme-linked immunosorbent assay (an MG S6-infected turkey) at 14 days PI.

Enhancement of enteropathogenic Escherichia coli pathogenicity in young turkeys by concurrent turkey coronavirus infection.

In a previous study, turkey coronavirus (TCV) and enteropathogenic Escherichia coli (EPEC) were shown to synergistically interact in young turkeys coinfected with these agents. In that study, inapparent or mild disease was observed in turkeys inoculated with only TCV or EPEC, whereas severe growth depression and high mortality were observed in dually inoculated turkeys. The purpose of the present study was to further evaluate the pathogenesis of combined TCV/EPEC infection in young turkeys and determine the role of these agents in the observed synergistic interaction. Experiments were conducted to determine 1) effect of EPEC dose, with and without concurrent TCV infection, and 2) effect of TCV exposure, before and after EPEC exposure, on development of clinical disease. Additionally, the effect of combined infection on TCV and EPEC shedding was determined. No clinical sign of disease and no attaching and effacing (AE) lesions characteristic of EPEC were observed in turkeys inoculated with only EPEC isolate R98/5, even when turkeys were inoculated with 10(10) colony forming units (CFU) EPEC (high dose exposure). Only mild growth depression was observed in turkeys inoculated with only TCV; however, turkeys inoculated with both TCV and 10(4) CFU EPEC (low dose exposure) developed severe disease characterized by high mortality, marked growth depression, and AE lesions. Inoculation of turkeys with TCV 7 days prior to EPEC inoculation produced more severe disease (numerically greater mortality, significantly lower survival probability [P < 0.05], increased frequency of AE lesions) than that observed in turkeys inoculated with EPEC prior to TCV or simultaneously inoculated with these agents. Coinfection of turkeys with TCV and EPEC resulted in significantly increased (P < 0.05) shedding of EPEC, but not TCV, in intestinal contents of turkeys. These findings indicate that TCV infection predisposes young turkeys to secondary EPEC infection and potentiates the expression of EPEC pathogenicity in young turkeys.

Prevalence of enteropathogenic Escherichia coli in naturally occurring cases of poult enteritis-mortality syndrome.

Enteropathogenic Escherichia coli (EPEC) previously were identified in poult enteritis-mortality syndrome (PEMS)-affected turkeys and associated as a cause of this disease. In the present study, the prevalence of EPEC in PEMS-affected turkeys was examined retrospectively with archived tissues and intestinal contents collected from 12 PEMS-affected turkey flocks in 1998. Formalin-fixed intestinal tissues were examined by light and electron microscopy for attaching and effacing (AE) lesions characteristic of EPEC, and frozen (-75 C) intestinal contents were examined for presence of EPEC. Escherichia coli isolates were characterized on the basis of epithelial cell attachment, fluorescent actin staining (FAS) test, and presence of E. coli attaching/effacing (EAE), shigalike toxin (SLT) type I, SLT II, and bundle-forming pilus (BFP) genes by polymerase chain reaction procedures. EPEC isolates were examined for pathogenicity and ability to induce AE lesions in experimentally inoculated young turkeys. AE lesions were identified by light microscopy in Giemsa-stained intestines from 7 of 12 PEMS-affected turkey flocks. Lesions consisted of bacterial microcolonies attached to epithelial surfaces with epithelial degeneration at sites of attachment and inflammatory infiltration of the lamina propria. Electron microscopy confirmed the identity of AE lesions in six of seven flocks determined to have AE lesions by light microscopy. EPEC were identified in 4 of 12 flocks on the basis of the presence of EAE genes a nd absence of SLT I and SLT II genes; all isolates lacked BFP genes. EPEC isolates produced AE lesions and variable mortality in turkeys coinfected with turkey coronavirus. In total, EPEC were associated with 10 of 12 (83%) naturally occurring PEMS cases on the basis of identification of AE lesions and/or EPEC isolates. These findings provide additional evidence suggesting a possible role for EPEC in the pathogenesis of PEMS.

Mortality patterns associated with poult enteritis mortality syndrome (PEMS) and coronaviral enteritis in turkey flocks raised in PEMS-affected regions.

Poult enteritis mortality syndrome (PEMS) is an economically devastating disease. To date, many questions about the syndrome remain unanswered, including its cause, transmission of causative agent(s), and control methods. Turkey coronavirus (TCV) infection has been associated with some outbreaks of PEMS, with areas having a higher prevalence of TCV infection also experiencing an increased incidence of PEMS. This study was designed to establish mortality patterns for flocks experiencing excess mortality and TCV infection in PEMS-affected regions and to delineate the possible role of TCV in PEMS-affected flocks. Fifty-four commercial turkey flocks on farms in areas with and without a history of TCV infection were monitored for weekly mortality and for antibodies to TCV. Flocks were chosen on the basis of placement dates and were monitored from day of placement until processing. All flocks were tested for TCV by an indirect fluorescent antibody assay. PEMS status was determined with the use of the clinical definition of mortality greater than 2% during any 3-wk period from 2 wk of age through the end of brooding due to unknown cause. Of the 54 flocks, 24 remained healthy, 23 experienced PEMS, and 7 tested positive for TCV but did not experience PEMS. Ten flocks experienced PEMS and tested positive for TCV, whereas 13 flocks experienced PEMS and did not test positive for TCV. Four health status groups were evident: healthy, PEMS positive, TCV positive, and PEMS + TCV positive. Distinct mortality patterns were seen for each of the four health status groups. Whereas TCV was associated with PEMS in 43% of PEMS cases, 13 cases (57%) of PEMS did not involve TCV. Additionally, 7 out of 17 cases of TCV (41%) did not experience excess mortality (PEMS) at any time during brooding of the flock. The results of this study indicate that TCV can be associated with PEMS but is neither necessary nor sufficient to cause PEMS.

Bacteriological and histological profile of turkeys condemned for cyanosis.

Canadian Food Inspection Agency (CFIA) has adopted the term cyanosis to describe a category of condemnation for poultry that is dark but has no other condemnable lesions. Two case-control studies (n = 30 pairs; n = 65 pairs) of 18-wk-old tom turkeys were conducted. A case was defined as a carcass condemned by the veterinary inspector for cyanosis, and a control carcass was one that passed inspection. Microbiological tests were conducted on samples of Pectoralis major and Gastrocnemius lacteralis. A modified Rappaport Vassiliadis medium was used for Salmonella, and a Petrifilm method was used to assess aerobic counts, coliform counts, and Escherichia coli. The Salmonella (qualitative) test was negative for all cases and controls, and there were no significant differences between the aerobic counts, coliform counts, and E. coli counts of case and control carcasses. Two pathologists conducted a blind histopathological study: there were no lesions compatible with those of septicemia-toxemia, as defined by CFIA and the USDA, nor any significant histopathological differences between the skin, P. major, G. lateralis, kidney, liver, spleen, small intestine, pancreas, lung, and heart of cases and controls. The inter-rater agreement between pathologists ranged from good to excellent (Kappa = 0.7 to 1.0). In the absence of important lesions and microbial contamination, carcasses with this color change alone should be suitable for human consumption.

Poult enteritis complex.

Poult enteritis complex (PEC) is a general term that encompasses the infectious intestinal diseases of young turkeys. Some diseases, such as coronaviral enteritis and stunting syndrome, are relatively well characterised, while others, such as transmissible viral enteritis, poult growth depression and poult enteritis mortality syndrome, remain ill-defined. All forms of PEC are multifactorial, transmissible and infectious. Salient clinical features include stunting and poor feed utilisation that result from enteritis. In the more severe forms, runting, immune dysfunction and mortality are reported. Gross and microscopic lesions of enteritis are present in all forms but tend to be non-specific. Other lesions may be present, depending on the agents involved. The basic pathogenesis involves the following: a) alteration of the intestinal mucosa, generally by one or more viruses infecting enterocytes; b) inflammation; c) proliferation of secondary agents, usually bacteria. Non-infectious factors interplay with infectious agents to modulate the course and severity of disease. Diarrhoea is believed to be primarily osmotic because of maldigestion and malabsorption, but may also have a secretory component. Transmission is primarily faecal-oral. No public health significance is recognised or suspected. Prevention is based on eliminating the infectious agents from contaminated premises and preventing introduction into flocks. This is accomplished by an effective cleaning, disinfection and biosecurity programme. All-in/all-out production or separate brooding and finishing units are helpful. Control may require regional co-ordination among all companies producing turkeys, especially if the production is highly concentrated, and a quarantine programme for more severe forms of PEC. No vaccines or specific measures for controlling the organisms involved in PEC are available. Treatment is supportive for the viral component, while antibiotics, especially those with efficacy against Gram positive bacteria, may help to reduce the impact to bacterial infections. Evidence suggests that PEC occurs wherever turkeys are raised commercially, but this is not well documented and distribution of the various organisms that have been associated with PEC is largely unknown. The disease causes enormous economic loss, mostly from failure of the turkey to reach its genetic potential.

Risk factors for abattoir condemnation of turkey carcasses due to cyanosis in southern Ontario.

Cyanosis is a category of condemnation for poultry, as defined by Agriculture and Agrifood Canada. A retrospective study to examine the seasonal patterns and trends for turkey cyanosis condemnation was conducted for the years 1987 to 1995 with the use of condemnation records made available by a single abattoir in Ontario. Condemnation for cyanosis had a significant seasonal pattern, with major losses occurring in the colder months (October to March). A prospective longitudinal study was conducted during January to March and October to December 1996 in the same abattoir and the 75 turkey farms that contracted to process their birds through it and all the 913 truckloads of turkeys processed in these two 3-mo periods. The data from this prospective study were used for multivariate modeling of the effects of potential risk factors on the incidence of cyanosis. Risk factors (at P < or = 0.05) included bird type (broiler age turkeys, mature toms, and mature hens compared to toms and hens), ambient temperature (cold: -9.9 to 0.0 C and very cold: < or =-10.0 C compared to mild: >0.0 C), clean-out lot (the last shipped from a poultry house), shipping time > or =8 h, and emaciation (proportion of turkeys in the lot condemned for insufficient muscle mass). The crate density was a sparing factor, and an increase in turkey density during shipping resulted in a lower incidence of cyanosis. This study suggested that the number of turkeys condemned for cyanosis was associated with cold, shipping stress, and subclinical syndromes.

Roaster breast meat condemned for cyanosis: a dark firm dry-like condition?

A case-control study (n = 68) of roaster chickens condemned for cyanosis was conducted. Color (CIE L*a*b*) and pH were measured at slaughter, and after 24 h aging on ice, at four predetermined sites of the Pectoralis major. Cyanotic carcasses (dark) had a higher pH than controls at the time of slaughter and at 24 h postmortem (P < 0.01). Perimortem pH was significantly correlated with pH at 24 h postmortem (r = 0.64) and also was correlated with lightness (L*) perimortem and postmortem (24 h; r = -0.36 and -0.50, respectively). Perimortem pH was not correlated with meat redness (a*) at slaughter time and after 24 h. Ultimate pH and lightness at 24 h postmortem were also correlated. Tests based on pH, L*, and a* of the P. major were assessed: the sensitivity and specificity at various cut-off points were, respectively, pH(6.3) = 76.47 and 88.24%, L*(41) = 91.18 and 79.41%, and a*(3) = 76.47 and 97.06%. The repeatability (p) of pH and color measurements was excellent and ranged from 0.87 to 0.98. Breast meat from roasters condemned for cyanosis had dark, firm, and dry (DFD)-like traits, and accurate tests based on color and pH could be described as a means of identifying chickens condemned for cyanosis.

Spatial distribution of cannibalism mortalities in commercial laying hens.

The distribution of cannibalism cases in a flock of 19,776 Babcock White Leghorns was monitored from 21 to 54 wk of age. The hens were kept in a single-floor house consisting of four banks of two-deck stair-step cages. Each of the 4,944 cages held four hens at a density of 152 cm(2) (60 inches(2)) per hen. Each cage was assigned a number from 1 to 4,944, and each dead bird was tagged according to its cage of origin. Dead birds were collected daily, kept in a freezer, and necropsied weekly. Farm personnel routinely transferred a live hen from an end cage to a cage where a mortality had occurred. The cause of death, age, cage number, and cage location were recorded for each dead hen. Of the 1,173 hens that died during the study period, 253 (21.6%) died from egg peritonitis, 184 (15.7%) from hypocalcemia, 167 (14.1%) from cannibalism, 164 (14%) from neoplastic disease, and the rest from various other causes. Cannibalism cases were analyzed statistically for clustering. Cannibalism was defined as death from tissue trauma and hemorrhage inflicted by cage mates. A spatial analysis showed that cannibalism is not a random event but one that occurs in clusters. The incidence of cannibalism was also found to be significantly higher on the top rows of cages as compared with the bottom rows.

A dark, firm dry-like condition in turkeys condemned for cyanosis.

A case-control study (n = 130) was conducted on toms condemned for cyanosis. Color (CIE L*a*b*), pH, and physical characteristics were measured on the Pectoralis major at slaughter and after 24 h. Meat from carcasses condemned for cyanosis had dark, firm, dry-like traits. It was darker and redder and showed higher water-holding capacity, lower cooking loss, and higher gel strength than did controls. Perimortem pH was negatively correlated with the lightness (L*) of meat at the time of slaughter (r = -0.58) and at 24 h postmortem (r = -0.64), positively correlated with water-holding capacity (r = 0.73) and gel strength (r = 0.43) and negatively correlated with cooking loss (r = -0.50). Ultimate pH was negatively correlated with lightness (L*) of meat at slaughter time (r = -0.62) and at 24 h postmortem (r = 0.79) was positively correlated with water-holding capacity (r = 0.87) and gel strength (r = 0.61) and negatively correlated with cooking loss (r = -0.52). Tests based on pH and L* of the P. major were also assessed; tests based on pH had a sensitivity in the range of 0.79 to 0.89 and specificity (Sp) of 0.60 to 0.94. Tests based on L* showed sensitivity of 0.75 to 0.92, and specificity of 0.79 to 0.97. The repeatability of measurements varied from good (L*: rho = 0.6) to excellent (pH: rho = 0.92). Overall, turkey breast condemned for cyanosis showed dark, firm, dry-like traits. Tests based on color and pH are described as a means of identifying turkeys condemned for cyanosis.

High mortality and growth depression experimentally produced in young turkeys by dual infection with enteropathogenic Escherichia coli and turkey coronavirus.

Six-day-old turkeys were inoculated with turkey coronavirus (TCV) and an enteropathogenic Escherichia coli (EPEC) (isolate R98/5) that were isolated from poult enteritis and mortality syndrome (PEMS)-affected turkeys. Turkeys inoculated with only R98/5 did not develop clinically apparent disease, and only mild disease and moderate growth depression were observed in turkeys inoculated with only TCV. Turkeys dually inoculated with TCV and R98/5 developed severe enteritis with high mortality (38/48, 79%) and marked growth depression. R98/5 infection resulted in attaching/effacing (AE) intestinal lesions characteristic of EPEC: adherence of bacterial microcolonies to intestinal epithelium with degeneration and necrosis of epithelium at sites of bacterial attachment. AE lesions were more extensive and were detected for a prolonged duration in dually inoculated turkeys compared with turkeys inoculated with only R98/5. An apparent synergistic effect in dually inoculated turkeys was indicated by increased mortality, enhanced growth depression, and enhanced AE lesion development. The results suggest that TCV promoted intestinal colonization by R98/5; however, R98/5 did not appear to alter TCV infection. The present study provides a possible etiologic explanation for PEMS.

Purification and catalytic properties of human caspase family members.

Members of the caspase family of cysteine proteases are known to be key mediators of mammalian inflammation and apoptosis. To better understand the catalytic properties of these enzymes, and to facilitate the identification of selective inhibitors, we have systematically purified and biochemically characterized ten homologues of human origin (caspases 1 - 10). The method used for production of most of these enzymes involves folding of active enzymes from their constituent subunits which are expressed separately in E. coli, followed by ion exchange chromatography. In cases where it was not possible to use this method (caspase-6 and -10), the enzymes were instead expressed as soluble proteins in E. coli, and partially purified by ion exchange chromatography. Based on the optimal tetrapeptide recognition motif for each enzyme, substrates with the general structure Ac-XEXD-AMC were used to develop continuous fluorometric assays. In some cases, enzymes with virtually identical tetrapeptide specificities have kcat/Km values for fluorogenic substrates that differ by more than 1000-fold. Using these assays, we have investigated the effects of a variety of environmental factors (e.g. pH, NaCl, Ca2+) on the activities of these enzymes. Some of these variables have a profound effect on the rate of catalysis, a finding that may have important biological implications.

Involvement of caspases in proteolytic cleavage of Alzheimer's amyloid-beta precursor protein and amyloidogenic A beta peptide formation.

The amyloid-beta precursor protein (APP) is directly and efficiently cleaved by caspases during apoptosis, resulting in elevated amyloid-beta (A beta) peptide formation. The predominant site of caspase-mediated proteolysis is within the cytoplasmic tail of APP, and cleavage at this site occurs in hippocampal neurons in vivo following acute excitotoxic or ischemic brain injury. Caspase-3 is the predominant caspase involved in APP cleavage, consistent with its marked elevation in dying neurons of Alzheimer's disease brains and colocalization of its APP cleavage product with A beta in senile plaques. Caspases thus appear to play a dual role in proteolytic processing of APP and the resulting propensity for A beta peptide formation, as well as in the ultimate apoptotic death of neurons in Alzheimer's disease.

Cell death attenuation by 'Usurpin', a mammalian DED-caspase homologue that precludes caspase-8 recruitment and activation by the CD-95 (Fas, APO-1) receptor complex.

Apoptotic cell suicide initiated by ligation of CD95 (Fas/APO-1) occurs through recruitment, oligomerization and autocatalytic activation of the cysteine protease, caspase-8 (MACH, FLICE, Mch5). An endogenous mammalian regulator of this process, named Usurpin, has been identified (aliases for Usurpin include CASH, Casper, CLARP, FLAME-1, FLIP, I-FLICE and MRIT). This protein is ubiquitously expressed and exists as at least three isoforms arising by alternative mRNA splicing. The Usurpin gene is comprised of 13 exons and is clustered within approximately 200 Kb with the caspase-8 and -10 genes on human chromosome 2q33-34. The Usurpin polypeptide has features in common with pro-caspase-8 and -10, including tandem 'death effector domains' on the N-terminus of a large subunit/small subunit caspase-like domain, but it lacks key residues that are necessary for caspase proteolytic activity, including the His and Cys which form the catalytic substrates diad, and residues that stabilize the P1 aspartic acid in substrates. Retro-mutation of these residues to functional caspase counterparts failed to restore proteolytic activity, indicating that other determinants also ensure the absence of catalytic potential. Usurpin heterodimerized with pro-caspase-8 in vitro and precluded pro-caspase-8 recruitment by the FADD/MORT1 adapter protein. Cell death induced by CD95 (Fas/APO-1) ligation was attenuated in cells transfected with Usurpin. In vivo, a Usurpin deficit was found in cardiac infarcts where TUNEL-positive myocytes and active caspase-3 expression were prominent following ischemia/reperfusion injury. In contrast, abundant Usurpin expression (and a caspase-3 deficit) occurred in surrounding unaffected cardiac tissue, suggesting reciprocal regulation of these pro- and anti-apoptotic molecules in vivo. Usurpin thus appears to be an endogenous modulator of apoptosis sensitivity in mammalian cells, including the susceptibility of cardiac myocytes to apoptotic death following ischemia/ reperfusion injury.

Validation of a poultry biosecurity survey.

A questionnaire for farm managers was designed, to obtain information regarding biosecurity on Ontario commercial broiler chicken and turkey operations, and then pre-tested. The questions that could be validated were verifiable by seeing the facility, by using farm records or by interviewing technical personnel other than the survey respondent. The survey was validated using a convenience sample of 24 farms from two companies. For 15 questions with dichotomous responses, the sensitivity ranged from 16.7 to 100%; the specificity ranged from 0 to 100%. For example, fences and gates seen during the farm visit were not accurately reported on the survey (poor sensitivity). Chance-corrected agreement was low (kappa < 0.4) for 34 questions, fair to good (0.4 < kappa < 0.8) for 25 questions, and excellent (kappa > 0.8) for seven questions. The percent agreement for questions where only one of the possible options was observed on validation ranged from 60.9 to 100%. Five questions with continuous numeric variables were analysed. A difference was observed (P < 0.1) between the survey and validation data for three questions regarding the number of birds, the bird sources and the downtime between flocks. In spite of pre-testing, the lack of clear wording and the absence of definitions for technical terms appeared to reduce validity. Response bias seems to be an issue with biosecurity surveys. The value of validating questionnaires before their use in epidemiologic research is confirmed.