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Brain functional networks identified from resting functional magnetic resonance imaging (fMRI) data have the potential to reveal biomarkers for brain disorders, but studies of complex mental illnesses such as schizophrenia (SZ) often yield mixed results across replication studies. This is likely due in part to the complexity of the disorder, the short data acquisition time, and the limited ability of the approaches for brain imaging data mining. Therefore, the use of analytic approaches which can both capture individual variability while offering comparability across analyses is highly preferred. Fully blind data-driven approaches such as independent component analysis (ICA) are hard to compare across studies, and approaches that use fixed atlas-based regions can have limited sensitivity to individual sensitivity. By contrast, spatially constrained ICA (scICA) provides a hybrid, fully automated solution that can incorporate spatial network priors while also adapting to new subjects. However, scICA has thus far only been used with a single spatial scale (ICA dimensionality, i.e., ICA model order). In this work, we present an approach using multi-objective optimization scICA with reference algorithm (MOO-ICAR) to extract subject-specific intrinsic connectivity networks (ICNs) from fMRI data at multiple spatial scales, which also enables us to study interactions across spatial scales. We evaluate this approach using a large N (N > 1,600) study of schizophrenia divided into separate validation and replication sets. A multi-scale ICN template was estimated and labeled, then used as input into scICA which was computed on an individual subject level. We then performed a subsequent analysis of multiscale functional network connectivity (msFNC) to evaluate the patient data, including group differences and classification. Results showed highly consistent group differences in msFNC in regions including cerebellum, thalamus, and motor/auditory networks. Importantly, multiple msFNC pairs linking different spatial scales were implicated. The classification model built on the msFNC features obtained up to 85% F1 score, 83% precision, and 88% recall, indicating the strength of the proposed framework in detecting group differences between schizophrenia and the control group. Finally, we evaluated the relationship of the identified patterns to positive symptoms and found consistent results across datasets. The results verified the robustness of our framework in evaluating brain functional connectivity of schizophrenia at multiple spatial scales, implicated consistent and replicable brain networks, and highlighted a promising approach for leveraging resting fMRI data for brain biomarker development., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Plasmodium falciparum resistance to artemisinin-based combination therapy (ACT) is a global threat to malaria control and elimination efforts. Mutations in the P. falciparum kelch13 gene (Pfk13) that are associated with delayed parasite clearance have emerged on the Thai-Cambodian border since 2008. There is growing evidence of widespread Pfk13 mutations throughout South-East Asia and they have independently emerged in other endemic regions. In Papua New Guinea (PNG), Pfk13 "C580Y" mutant parasites with reduced in vitro sensitivity to artemisinin have been isolated in Wewak, a port town in East Sepik Province. However, the extent of any local spread of these mutant parasites in other parts of PNG is unknown. We investigated the prevalence of Pfk13 mutations in multiple malaria-endemic regions of PNG. P. falciparum isolates (n = 1152) collected between 2016 and 2018 and assessed for Pfk13 variation by sequencing. Of 663 high quality Pfk13 sequences a total of five variants were identified. They included C580Y, a mutation at a previously documented polymorphic locus: N499K, and three previously undescribed mutations: R471C, K586E and Y635C. All variants were found in single isolates, indicating that these Pfk13 mutations were rare in the areas surveyed. Notably, C580Y was absent from Maprik district, which neighbours Wewak where C580Y mutant parasites were previously identified. The single C580Y isolate was found in the port town of Lae, Morobe Province, a potential entry site for the importation of drug resistant parasites into PNG. Although sample size in this location was small (n = 5), our identification of a C580Y mutant in this second location is concerning, highlighting the urgent need for further surveillance in Lae. Other Pfk13 mutants were rare in PNG between 2016 and 2018. Continued surveillance for molecular markers of drug resistance is critically important to inform malaria control in PNG., (Copyright © 2021. Published by Elsevier Ltd.)

In a recent machine learning study classifying "brain age" based on cross-sectional neuroanatomical data, clinical high-risk (CHR) individuals were observed to show deviation from the normal neuromaturational pattern, which in turn was predictive of greater risk of conversion to psychosis and a pattern of stably poor functional outcome. These effects were unique to cases who were between 12 and 17 years of age when their prodromal and psychotic symptoms began, suggesting that neuroanatomical deviance observable at the point of ascertainment of a CHR syndrome marks risk for an early onset form of psychosis. In the present study, we sought to clarify the pattern of neuroanatomical deviance linked to this "early onset" form of psychosis and whether this deviance is associated with poorer premorbid functioning. T 1 MRI scans from 378 CHR individuals and 190 healthy controls (HC) from the North American Prodrome Longitudinal Study (NAPLS2) were analyzed. Widespread smaller cortical volume was observed among CHR individuals compared with HC at baseline evaluation, particularly among the younger group (i.e., those who were 12 to 17 years of age). Moreover, the younger CHR individuals who converted or presented worsened clinical symptoms at follow-up (within 2 years) exhibited smaller surface area in rostral anterior cingulate, lateral and medial prefrontal regions, and parahippocampal gyrus relative to the younger CHR individuals who remitted or presented a stable pattern of prodromal symptoms at follow-up. In turn, poorer premorbid functioning in childhood was associated with smaller surface area in medial orbitofrontal, lateral frontal, rostral anterior cingulate, precuneus, and temporal regions. Together with our prior report, these results are consistent with the view that neuroanatomical deviance manifesting in early adolescence marks vulnerability to a form of psychosis presenting with poor premorbid adjustment, an earlier age of onset (generally prior to the age of 18 years), and poor long-term outcome., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: The profile of cortical neuroanatomical abnormalities in schizophrenia is not fully understood, despite hundreds of published structural brain imaging studies. This study presents the first meta-analysis of cortical thickness and surface area abnormalities in schizophrenia conducted by the ENIGMA (Enhancing Neuro Imaging Genetics through Meta Analysis) Schizophrenia Working Group., Methods: The study included data from 4474 individuals with schizophrenia (mean age, 32.3 years; range, 11-78 years; 66% male) and 5098 healthy volunteers (mean age, 32.8 years; range, 10-87 years; 53% male) assessed with standardized methods at 39 centers worldwide., Results: Compared with healthy volunteers, individuals with schizophrenia have widespread thinner cortex (left/right hemisphere: Cohen's d = -0.530/-0.516) and smaller surface area (left/right hemisphere: Cohen's d = -0.251/-0.254), with the largest effect sizes for both in frontal and temporal lobe regions. Regional group differences in cortical thickness remained significant when statistically controlling for global cortical thickness, suggesting regional specificity. In contrast, effects for cortical surface area appear global. Case-control, negative, cortical thickness effect sizes were two to three times larger in individuals receiving antipsychotic medication relative to unmedicated individuals. Negative correlations between age and bilateral temporal pole thickness were stronger in individuals with schizophrenia than in healthy volunteers. Regional cortical thickness showed significant negative correlations with normalized medication dose, symptom severity, and duration of illness and positive correlations with age at onset., Conclusions: The findings indicate that the ENIGMA meta-analysis approach can achieve robust findings in clinical neuroscience studies; also, medication effects should be taken into account in future genetic association studies of cortical thickness in schizophrenia., (Copyright © 2018 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

Background: Individuals at clinical high risk (CHR) who progress to fully psychotic symptoms have been observed to show a steeper rate of cortical gray matter reduction compared with individuals without symptomatic progression and with healthy control subjects. Whether such changes reflect processes associated with the pathophysiology of schizophrenia or exposure to antipsychotic drugs is unknown., Methods: In this multisite study, 274 CHR cases, including 35 individuals who converted to psychosis, and 135 healthy comparison subjects were scanned with magnetic resonance imaging at baseline, 12-month follow-up, or the point of conversion for the subjects who developed fully psychotic symptoms., Results: In a traveling subjects substudy, excellent reliability was observed for measures of cortical thickness and subcortical volumes. Controlling for multiple comparisons throughout the brain, CHR subjects who converted to psychosis showed a steeper rate of gray matter loss in the right superior frontal, middle frontal, and medial orbitofrontal cortical regions as well as a greater rate of expansion of the third ventricle compared with CHR subjects who did not convert to psychosis and healthy control subjects. Differential tissue loss was present in subjects who had not received antipsychotic medications during the interscan interval and was predicted by baseline levels of an aggregate measure of proinflammatory cytokines in plasma., Conclusions: These findings demonstrate that the brain changes are not explained by exposure to antipsychotic drugs but likely play a role in psychosis pathophysiology. Given that the cortical changes were more pronounced in subjects with briefer durations of prodromal symptoms, contributing factors may predominantly play a role in acute-onset forms of psychosis., (Copyright © 2015 Society of Biological Psychiatry. All rights reserved.)

Schizophrenia is a psychotic disorder characterized by functional dysconnectivity or abnormal integration between distant brain regions. Recent functional imaging studies have implicated large-scale thalamo-cortical connectivity as being disrupted in patients. However, observed connectivity differences in schizophrenia have been inconsistent between studies, with reports of hyperconnectivity and hypoconnectivity between the same brain regions. Using resting state eyes-closed functional imaging and independent component analysis on a multi-site data that included 151 schizophrenia patients and 163 age- and gender matched healthy controls, we decomposed the functional brain data into 100 components and identified 47 as functionally relevant intrinsic connectivity networks. We subsequently evaluated group differences in functional network connectivity, both in a static sense, computed as the pairwise Pearson correlations between the full network time courses (5.4 minutes in length), and a dynamic sense, computed using sliding windows (44 s in length) and k-means clustering to characterize five discrete functional connectivity states. Static connectivity analysis revealed that compared to healthy controls, patients show significantly stronger connectivity, i.e., hyperconnectivity, between the thalamus and sensory networks (auditory, motor and visual), as well as reduced connectivity (hypoconnectivity) between sensory networks from all modalities. Dynamic analysis suggests that (1), on average, schizophrenia patients spend much less time than healthy controls in states typified by strong, large-scale connectivity, and (2), that abnormal connectivity patterns are more pronounced during these connectivity states. In particular, states exhibiting cortical-subcortical antagonism (anti-correlations) and strong positive connectivity between sensory networks are those that show the group differences of thalamic hyperconnectivity and sensory hypoconnectivity. Group differences are weak or absent during other connectivity states. Dynamic analysis also revealed hypoconnectivity between the putamen and sensory networks during the same states of thalamic hyperconnectivity; notably, this finding cannot be observed in the static connectivity analysis. Finally, in post-hoc analyses we observed that the relationships between sub-cortical low frequency power and connectivity with sensory networks is altered in patients, suggesting different functional interactions between sub-cortical nuclei and sensorimotor cortex during specific connectivity states. While important differences between patients with schizophrenia and healthy controls have been identified, one should interpret the results with caution given the history of medication in patients. Taken together, our results support and expand current knowledge regarding dysconnectivity in schizophrenia, and strongly advocate the use of dynamic analyses to better account for and understand functional connectivity differences.

In the food safety arena, the decision-making process can be especially difficult. Decision makers are often faced with social and fiscal pressures when attempting to identify an appropriate balance among several choices. Concurrently, policy and decision makers in microbial food safety are under increasing pressure to demonstrate that their policies and decisions are made using transparent and accountable processes. In this article, we present a multi-criteria decision analysis approach that can be used to address the problem of trying to select a food safety intervention while balancing various criteria. Criteria that are important when selecting an intervention were determined, as a result of an expert consultation, to include effectiveness, cost, weight of evidence, and practicality associated with the interventions. The multi-criteria decision analysis approach we present is able to consider these criteria and arrive at a ranking of interventions. It can also provide a clear justification for the ranking as well as demonstrate to stakeholders, through a scenario analysis approach, how to potentially converge toward common ground. While this article focuses on the problem of selecting food safety interventions, the range of applications in the food safety arena is truly diverse and can be a significant tool in assisting decisions that need to be coherent, transparent, and justifiable. Most importantly, it is a significant contributor when there is a need to strike a fine balance between various potentially competing alternatives and/or stakeholder groups.

This study estimated the health burden and costs associated with gastroenteritis in the City of Hamilton (Ontario, Canada). The number of cases, number of different resource units used, and cost per resource unit were represented by probability distributions and point estimates. These were subsequently integrated in a stochastic model to estimate the overall burden and cost in the population and to depict the uncertainty of the estimates. The estimated mean annual cost per capita was Can dollar 115. The estimated mean annual cost per case was Can dollar 1,089 and was similar to other published figures. Gastroenteritis represented a significant burden in the study population, with costs high enough to justify prevention efforts. These results, currently the most accurate available estimates for a Canadian population, can inform future economic evaluations to determine the most cost effective measures for reducing the burden and cost of gastroenteritis in the community.

The adaptation of a standard Escherichia coli O157 isolation method involving immunomagnetic separation and a period of frozen storage was investigated. A series of experiments was designed to test the recovery of a bovine strain of E. coli O157 from buffered peptone water after a period of frozen storage at -80 degrees C. The effects of the addition of glycerol at 5 and 10%, freezing time, the number of freeze-thaw cycles, the method of freezing and the method of thawing, the inclusion of a resuscitation-and-incubation step, and the sensitivity of the isolation method were investigated. The most effective method of storing frozen samples for 6 months and recovering strains of E. coli O157 after storage was found to involve 6 h of incubation of sample material in buffered peptone water at 37 degrees C before frozen storage at -80 degrees C with 10% glycerol, a rapid thaw after frozen storage, and resuscitation at 27 degrees C for 1 h and incubation at 37 degrees C for 1 h to allow freeze-injured and stressed bacteria to recover with a period of growth prior to immunomagnetic separation isolation. There was no significant decrease in log counts of a bovine strain E. coli O157 over 6 months of frozen storage in buffered peptone water with 10% glycerol. With this method, it was possible to isolate E. coli O157 from naturally infected bovine carcasses after a period of frozen storage.

In this paper we describe dissociations of implicit versus explicit access to semantic information in a patient with deep dyslexia. This acquired reading disorder is characterized by the production of morphological (e.g., SLEEP read as SLEEPING) and semantic errors (e.g., HEART read as BLOOD) and consequently provides a potential window into the operation of both aspects of the language system. The deep dyslexic patient in this study (JO) demonstrated implicit semantic access to items in a number of tasks despite the fact that she was unable to correctly read these items aloud. The findings from this study are consistent with a model of lexical deficits that distinguishes between explicit and implicit access to lexical representations on the basis of inhibitory processes.

The purpose of this study was to evaluate an enzyme-linked immunosorbent assay (ELISA) and an immunoblot procedure for detection and isolation of Shiga toxin-producing Escherichia coli (STEC) from beef, and to correlate the presence of STEC in beef with E. coli and total coliform counts. A total of 120 samples of boneless beef supplied to a meat processor in southern Ontario were tested for the presence of STEC, E. coli, and total coliforms. Following enrichment in modified tryptic soy broth, samples were screened for Shiga toxin (Stx) by a Stx-ELISA and a Vero cell assay (VCA). Samples that were positive in the Stx-ELISA were subjected to the Stx-immunoblot for STEC isolation. Overall, 33.3% of samples were positive in the VCA, and 34.2% were positive in the Stx-ELISA. There was almost complete agreement between the Stx-ELISA and the VCA results (kappa = 0.98). The sensitivity and specificity of the Stx-ELISA with respect to the VCA were 100% and 98.75%, respectively. STEC were isolated by the Stx-immunoblot from 87.8% of the samples that were positive in the Stx-ELISA. The STEC isolates belonged to 19 serotypes, with serotype O113:H21 accounting for 10 of 41 isolates. No STEC of serotype O157:H7 were isolated. There was a significant correlation between E. coli counts and total coliform counts (Spearman correlation coefficient = 0.68, P < 0.01). The E. coli count was positively correlated with detection of STEC by both the Stx-ELISA and the VCA (P < 0.01).

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.

The Reveal (Neogen Corp., Lansing, Mich.) and SafePath (SafePath Laboratories LLC, St. Paul, Minn.) tests were evaluated for their performance as beef fecal and beef carcass Escherichia coli O157:H7 monitoring tests. Agreement between these tests and a reference test system was determined using naturally contaminated bovine feces and beef carcasses. The reference system utilized immunomagnetic separation with plating onto cefixime, tellurite, sorbitol MacConkey agar, followed by colony testing using a serum agglutination test for the O157 antigen. Relative to this reference method, the Reveal test showed a sensitivity of 46% and a specificity of 82% on bovine feces and a specificity of 99% on carcass samples. The SafePath test, demonstrated a significantly higher sensitivity at 79% and a similar specificity of 79%. On carcass samples the SafePath test performed similarly to the Reveal test, demonstrating a specificity of 100% relative to the reference system. There was an insufficient number of E. coli O157-positive carcass samples to estimate precisely the sensitivity of these two methods. Both methods show promise as rapid carcass monitoring tests, but further field testing to estimate sensitivity is needed to complete their evaluation. The proportion of positive fecal samples for E. coli O157:H7 by the reference method ranged from 10.2% to 36% in 10 lots of cattle with an overall mean of 17.3% (39/225). Quarter carcass sponging of 125 carcasses revealed 1.6% positive for the pathogen (2/125).

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.

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.

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.

A study was conducted to provide a quantitative description of the amount of tag (mud, soil, and bedding) adhered to the hides of feedlot beef cattle and to appraise the statistical reliability of a subjective rating system for assessing this trait. Initially, a single rater obtained baseline data by assessing 2,417 cattle for 1 month at an Ontario beef processing plant. Analysis revealed that there was a strong tendency for animals within sale-lots to have a similar total tag score (intralot correlation = 0.42). Baseline data were summarized by fitting a linear model describing an individual's total tag score as the sum of their lot mean tag score (LMTS) plus an amount representing normal variation within the lot. LMTSs predicted by the linear model were adequately described by a beta distribution with parameters nu = 3.12 and omega = 5.82 scaled to fit on the 0-to-9 interval. Five raters, trained in use of the tag scoring system, made 1,334 tag score observations in a commercial abattoir, allowing reliability to be assessed at the individual level and at the lot level. High values for reliability were obtained for individual total tag score (0.84) and lot total tag score (0.83); these values suggest that the tag scoring system could be used in the marketing and slaughter of Ontario beef cattle to improve the cleanliness of animals presented for slaughter in an effort to control the entry of microbial contamination into abattoirs. Implications for the use of the tag scoring system in research are discussed.

The Lactek test, marketed for antimicrobial residue detection in milk, was validated for the detection of antimicrobial residues in tissues. A previous study found that the LacTek test could confidently identify tissue samples spiked with antimicrobial residues. However, the test could not reliably distinguish violative from nonviolative spiked samples relative to Canadian maximum residue limits (MRLs). The objectives of this study were to assess and compare the performance of the LacTek tests for beta-lactams, tetracyclines, gentamicin, and sulfamethazine on samples containing naturally incurred residues by running the test in parallel with the standard microbial inhibition test (MIT) presently used for the routine testing of tissues at our facility and to assess the agreement with high pressure liquid chromatographic (HPLC) determinative methods. Parallel testing with the official MIT found that the Lactek tests could be confidently used for testing tissue samples containing incurred residues. Among 1,008 MIT-positive samples, the LacTek test found that 90% contained beta-lactams and/or tetracyclines. A further 7.3% of violative residues could not be identified to an antimicrobial class. In addition, 9% of samples testing negative on the MIT were found to contain an antimicrobial residue by the LacTek tests. Comparative testing with HPLC methods found that there was very good agreement between the two tests and that most violations were due to penicillin G and oxytetracycline. Although the LacTek test cannot be used to distinguish violative from nonviolative residue levels, it does offer several advantages over the present MIT. These include speed, ease of use, the ability to identify residues to a specific class, and an improved sensitivity at the MRL level for the most commonly found antimicrobials in tissue.

This paper presents a historical review of antimicrobial use in food animals, the causes of residues in meat and milk, the types of residues found, their regulation in Canada, tests used for their detection, and test performance parameters, with an emphasis on immunoassay techniques. The development of residue detection methods began shortly after the introduction of antimicrobials to food animal production in the late 1940s. From initial technical concerns expressed by the dairy industry to the present public health and international trade implications, there has been an ongoing need for reliable, sensitive, and economical methods for the detection of antimicrobial residues in food animal products such as milk and meat. Initially there were microbial growth inhibition tests, followed by more sensitive and specific methods based on receptor binding, immunochemical, and chromatographic principle. An understanding of basic test performance parameters and their implications is essential when choosing an analytical strategy for residue testing. While each test format has its own attributes, none test will meet all the required analytical needs. Therefore the use of a tiered or integrated system employing assays designated for screening and confirmation is necessary to ensure that foods containing violative residues are not introduced into the food chain.

A field trial using cattle from a commercial feedlot was conducted to quantify the effect of duration of fasting and a temporary change in ration on the concentration of Escherichia coli biotype 1 in feces. A nested hierarchical design with repeated measures through time was used. Two groups of 20 British x European breed beef steers having reached slaughter weight (mean live weight 685 kg; SD 50 kg) were fed entirely on a high-energy ration typical of that used in the Ontario beef finishing industry or were switched for 4 days onto a high-roughage ration. This was followed by a period of fasting and water deprivation to mimic that which occurs prior to slaughter. Fecal samples were collected at 0, 24, and 48 h of fasting, and for each sample the total presumptive E. coli (biotype 1) CFU/g of feces was enumerated by spiral plating. Estimates of effect for the design factors were obtained by restricted maximum likelihood, and these were compared to robust counterparts obtained from generalized estimating equations. Results indicated that the ration, the duration of fasting, and their interaction had significant effects on total log E. coli concentration in feces. Cattle on the high-roughage ration for four days had a significantly lower initial log E. coli CFU/g of feces compared to cattle on the normal ration, but after 48 h of fasting they had a significantly higher concentration. It is concluded that while a temporary change in ration and duration of fasting does affect E. coli concentration in feces, these changes do not seem large enough to deliver a drastic improvement in beef carcass hygiene should they be incorporated in hazard analysis and critical control point (HACCP) plans for the preslaughter period of beef production.

The Petrifilm HEC test (3M Canada Inc., London, Ontario), a quantitative microbiological test for Escherichia coli O157:H7, was evaluated for its performance as a beef-carcass monitoring test. Test repeatability and agreement and agreement with an E. coli O157:H7 detection method using a hydrophobic grid membrane filter (HGMF) overlaid onto cefixime-tellurite-sorbitol MacConkey agar (CT-SMAC) followed by a latex agglutination test for the O157 antigen were determined by using pure cultures of E. coli O157:H7, beef samples experimentally contaminated with bovine feces containing E. coli O157:H7, and naturally contaminated beef carcasses of unknown E. coli O157:H7 status from a local abattoir. The Petrifilm HEC test showed excellent repeatability and excellent agreement with the HGMF-CT-SMAC method when test samples were obtained from pure cultures and experimentally contaminated meat. All 125 naturally contaminated beef carcasses surveyed were negative for E. coli O157:H7 with both microbial methods. The Petrifilm HEC test, however, demonstrated a significantly lower proportion of cross-reactive organisms (false-positive reactions) than the HGMF-CT-SMAC method. Given the performance of this test coupled with its ease of use and compact size, it shows considerable promise for carcass testing where abattoir laboratory facilities are limited and as a substitute for more complex laboratory testing methods used in established laboratories.

Sulfamethazine-spiked chicken liver and kidney were tested for residues using the Delvotest SP. The results were compared to a standard plate assay using Bacillus subtilis as the test organism. The Delvotest SP gave positive responses to all homogenized liver replicates spiked with sulfamethazine at or above 1.0 μg/g of liver. Mixed responses were obtained at 0.5 μg/g and negative responses at ≤0.25 μg/g. The plate assay had a minimum sulfamethazine detection limit of 1 μg/g and 0.5 μg/g of liver and kidney, respectively. Chickens were dosed with sulfamethazine (100 mg/kg of body weight) daily for 5 days, and tissues were tested for residues after treatment stopped. The Delvotest SP was positive for all serum and kidney samples from the end of treatment to 24 h, and for muscle and liver samples up to 8 h. The plate assay detected the drug up to 24 h in serum and kidney samples and up to 8 h in liver and muscle samples. Kidney and serum samples both appeared to be good tissues for testing sulfamethazine residues in chickens. Serum could be used for antemortem screening, whereas kidney samples would appear to be the best for postmortem screening of residues in chickens.

Oxytetracycline spiked chicken liver and kidney were tested for residues using Delvotest P and the results compared to standard plate assay using Bacillus cereus as test organism. Delvotest P detected all replicates at concentrations of ≥0.62 μg/g in liver and kidney. Between 0.41-0.2 μg/g, the responses were mixed (positive, doubtful, and negative); at 0.1 ≥ concentration, the responses were negative. The limit of detection of the plate assay was 0.2 and 0.31 μg/g for liver and kidney, respectively. Residue analysis on tissues from chickens dosed twice daily (for 4 d) with 25 mg/kg oxytetracycline was carried out using the Delvotest P and the plate assay. Delvotest P responses were all positive for 1-4 h after treatment in kidney and 1 and 2 h in liver and serum. Muscle never achieved concentrations that were consistently positive. The plate assay detected drug from 0.5-6 h in serum, from 0.5-8 h in liver and kidney, and at 4-6 h in muscle. Kidney tissue appeared to be the best for detecting oxytetracycline residues in chicken. The Delvotest P method was simpler to use and required less time; 3-4 h compared to about 18 h for the plate assay.