Genome-Wide Identification of Tomato Xylem Sap Fitness Factors for Three Plant-Pathogenic Ralstonia Species.

Plant-pathogenic Ralstonia spp. colonize plant xylem and cause wilt diseases on a broad range of host plants. To identify genes that promote growth of diverse Ralstonia strains in xylem sap from tomato plants, we performed genome-scale genetic screens (random barcoded transposon mutant sequencing screens [RB-TnSeq]) in three strains spanning the genetic, geographical, and physiological range of plant-pathogenic Ralstonia: Ralstonia solanacearum IBSBF1503, Ralstonia pseudosolanacearum GMI1000, and Ralstonia syzygii PSI07. Contrasting mutant fitness phenotypes in culture media versus in xylem sap suggest that Ralstonia strains are adapted to ex vivo xylem sap and that culture media impose foreign selective pressures. Although wild-type Ralstonia grew in sap and in rich medium with similar doubling times and to a similar carrying capacity, more genes were essential for growth in sap than in rich medium. Each strain required many genes associated with envelope remodeling and repair processes for full fitness in xylem sap. These genes were associated with peptidoglycan peptide formation (murI), secretion of periplasmic proteins (tatC), periplasmic protein folding (dsbA), synthesis of osmoregulated periplasmic glucans (mdoGH), and lipopolysaccharide (LPS) biosynthesis. Mutant strains with mutations in four genes had strong, sap-specific fitness defects in all strain backgrounds: murI, thiC, purU, and a lipoprotein (RSc2007). Many amino acid biosynthesis genes were required for fitness in both minimal medium and xylem sap. Multiple mutants with insertions in virulence regulators had gains of fitness in culture media and neutral fitness in sap. Our genome-scale genetic screen identified Ralstonia fitness factors that promote growth in xylem sap, an ecologically relevant condition. IMPORTANCE Traditional transposon mutagenesis genetic screens pioneered molecular plant pathology and identified core virulence traits like the type III secretion system. TnSeq approaches that leverage next-generation sequencing to rapidly quantify transposon mutant phenotypes are ushering in a new wave of biological discovery. Here, we have adapted a genome-scale approach, random barcoded transposon mutant sequencing (RB-TnSeq), to discover fitness factors that promote growth of three related bacterial strains in a common niche, tomato xylem sap. Fitness of the wild type and mutants show that Ralstonia spp. are adapted to grow well in xylem sap from their natural host plant, tomato. Our screen identified multiple sap-specific fitness factors with roles in maintaining the bacterial envelope. These factors include putative adaptations to resist plant defenses that may include antimicrobial proteins and specialized metabolites that damage bacterial membranes.

Implementation of OIE international standards: challenges and opportunities for monitoring.

The World Organisation for Animal Health (OIE) is a major actor in international cooperation to improve animal health and welfare throughout the world. The OIE sets international standards to support Member Countries in their efforts to prevent and control animal diseases, strengthen Veterinary Services and Aquatic Animal Health Services, and facilitate safe international trade. Member Countries face many challenges in the implementation of OIE standards. Poor governance and a lack of resources and technical capacity are often major constraints. Trade concerns raised at the World Trade Organization (WTO) can also be a signal that countries are experiencing difficulties in implementing international standards. In May 2018, the World Assembly of OIE Delegates adopted a resolution recommending the establishment of an observatory to monitor the implementation of OIE standards. This monitoring mechanism will help the OIE to improve its international standard-setting process and identify the capacity-building needs of Member Countries. Monitoring implementation will be challenging as the OIE does not prescribe a specific procedure for implementing OIE standards. World Organisation for Animal Health Member Countries use a range of approaches to implement OIE standards, because of differences in animal health situations, legal frameworks and procedures, trade profiles, and acceptable levels of risk. Given this complexity, this article proposes a 'cross-over' approach to monitoring implementation that would require the collection of information from various trusted sources, such as the World Animal Health Information System (WAHIS), the OIE Performance of Veterinary Services (PVS) Pathway mission reports and the WTO Sanitary and Phytosanitary Information Management System database. This approach aims to document what is currently happening and to identify potential patterns in Member Country practices when implementing OIE standards.

L'Organisation mondiale de la santé animale (OIE) est un acteur majeur de la coopération internationale oeuvrant à l'amélioration de la santé et du bien-être animal dans le monde. L'OIE élabore des normes internationales visant à soutenir les efforts déployés par ses Membres pour prévenir et lutter contre les maladies animales, renforcer les Services vétérinaires et les Services chargés de la santé des animaux aquatiques, et faciliter un commerce international sûr. Les Membres rencontrent de nombreuses difficultés lors de la mise en oeuvre des normes de l'OIE. Une mauvaise gouvernance et l'insuffisance des ressources et des capacités techniques constituent souvent des contraintes majeures. Les préoccupations commerciales soulevées au sein de l'Organisation mondiale du commerce (OMC) sont également révélatrices des difficultés rencontrées par les pays pour mettre en oeuvre les normes internationales. En mai 2018, l'Assemblée mondiale des Délégués de l'OIE a adopté une résolution recommandant la création d'un observatoire destiné à assurer le suivi de la mise en oeuvre des normes de l'OIE. Grâce à ce mécanisme de suivi, l'OIE sera à même d'améliorer le processus d'élaboration de ses normes internationales et d'identifier les besoins de ses Membres en matière de renforcement des capacités. Le suivi de la mise en oeuvre ne sera pas une tâche facile, dans la mesure où l'OIE ne prescrit pas de procédure spécifique pour appliquer ses normes. Les Membres de l'OIE recourent à diverses approches pour mettre en oeuvre les normes de l'OIE, dictées par des différences dans les situations zoosanitaires, les cadres et procédures juridiques, les profils commerciaux et les niveaux de risque considérés comme acceptables. Face à une telle complexité, les auteurs proposent une méthode transversale de suivi de la mise en oeuvre, qui passe par la collecte d'informations auprès de diverses sources fiables telles que le Système mondial d'information sanitaire de l'OIE (WAHIS), les rapports de mission du Processus de l'OIE pour évaluer la Performance des Services vétérinaires (Processus PVS) et la base de données de l'OMC de gestion des renseignements sanitaires et phytosanitaires. Cette approche vise à documenter ce qui se fait actuellement et à identifier d'éventuelles tendances dans les pratiques des Membres en matière de mise en oeuvre des normes de l'OIE.

La Organización Mundial de Sanidad Animal (OIE) es uno de los actores principales de la cooperación internacional para mejorar la sanidad y el bienestar animales en todo el mundo. La OIE elabora normas internacionales destinadas a apoyar el esfuerzo de los Países Miembros por prevenir y combatir las enfermedades animales, fortalecer los Servicios Veterinarios y los Servicios de sanidad de los animales acuáticos y facilitar un comercio internacional seguro. Los Países Miembros afrontan numerosos desafíos a la hora de implementar las normas de la OIE. La insuficiencia de mecanismos de gobernanza y la falta de recursos y capacidad técnica constituyen a menudo obstáculos de gran calado. Las preocupaciones comerciales expresadas ante la Organización Mundial del Comercio (OMC) también pueden ser una señal de las dificultades que afrontan los países para implementar las normas internacionales. En mayo de 2018, la Asamblea Mundial de Delegados de la OIE aprobó una resolución en la que recomendaba la creación de un observatorio para el seguimiento de la implementación de las normas de la OIE. Este mecanismo ayudará a la OIE a mejorar su proceso de elaboración de normas internacionales y a identificar las necesidades de capacidades específicas de los Países Miembros. El seguimiento de la implementación no será fácil, pues la OIE no prescribe ningún procedimiento específico para poner en práctica sus normas. Los Países Miembros utilizan modalidades distintas debido a las diferencias en cuanto a situaciones zoosanitarias, ordenamiento y procedimientos jurídicos, perfiles comerciales y niveles de riesgo aceptables. Teniendo en cuenta esta compleja situación, los autores proponen aquí un enfoque «transversal¼ de seguimiento de la implementación de las normas, que requeriría reunir información de diversas fuentes fidedignas, como el Sistema Mundial de Información Zoosanitaria (WAHIS), los informes de las misiones de evaluación de las prestaciones de los Servicios Veterinarios (Proceso PVS) de la OIE y la base de datos del sistema de la OMC de gestión de información relativa a las medidas sanitarias y fitosanitarias. Tal enfoque tiene por objetivo documentar lo que está ocurriendo actualmente e identificar posibles patrones en las prácticas de los Países Miembros al implementar las normas de la OIE.

Evolution of satellite plasmids can prolong the maintenance of newly acquired accessory genes in bacteria.

Transmissible plasmids spread genes encoding antibiotic resistance and other traits to new bacterial species. Here we report that laboratory populations of Escherichia coli with a newly acquired IncQ plasmid often evolve 'satellite plasmids' with deletions of accessory genes and genes required for plasmid replication. Satellite plasmids are molecular parasites: their presence reduces the copy number of the full-length plasmid on which they rely for their continued replication. Cells with satellite plasmids gain an immediate fitness advantage from reducing burdensome expression of accessory genes. Yet, they maintain copies of these genes and the complete plasmid, which potentially enables them to benefit from and transmit the traits they encode in the future. Evolution of satellite plasmids is transient. Cells that entirely lose accessory gene function or plasmid mobility dominate in the long run. Satellite plasmids also evolve in Snodgrassella alvi colonizing the honey bee gut, suggesting that this mechanism may broadly contribute to the importance of IncQ plasmids as agents of bacterial gene transfer in nature.

One Health operations: a critical component in the International Health Regulations Monitoring and Evaluation Framework.

Under the International Health Regulations (IHR, 2005), a legally binding document adopted by 196 States Parties, countries are required to develop their capacity to rapidly detect, assess, notify and respond to unusual health events of potential international concern. To support countries in monitoring and enhancing their capacities and complying with the IHR (2005), the World Health Organization (WHO) developed the IHR Monitoring and Evaluation Framework (IHR MEF). This framework comprises four complementary components: the State Party Annual Report, the Joint External Evaluation, after-action reviews and simulation exercises. The first two are used to review capacities and the second two to help to explore their functionality. The contribution of different disciplines, sectors, and areas of work, joining forces through a One Health approach, is essential for the implementation of the IHR (2005). Therefore, WHO, in partnership with the Food and Agriculture Organization of the United Nations (FAO), the World Organisation for Animal Health (OIE), and other international and national partners, has actively worked on facilitating the inclusion of the relevant sectors, in particular the animal health sector, in each of the four components of the IHR MEF. Other tools complement the IHR MEF, such as the WHO/OIE IHR-PVS [Performance of Veterinary Services] National Bridging Workshops, which facilitate the optimal use of the results of the IHR MEF and the OIE Performance of Veterinary Services Pathway and create an opportunity for stakeholders from animal health and human health services to work on the coordination of their efforts. The results of these various tools are used in countries' planning processes and are incorporated in their National Action Plan for Health Security to accelerate the implementation of IHR core capacities. The present article describes how One Health is incorporated in all components of the IHR MEF.

En vertu du Règlement sanitaire international (RSI, 2005), instrument juridique ayant force obligatoire pour les 196 États Parties dans le monde, les pays s'engagent à renforcer leurs capacités de détection, d'évaluation, de notification et de réaction en cas d'événements sanitaires inhabituels ou présentant une dimension internationale inquiétante. Le Cadre de suivi et d'évaluation du RSI (2005) a été élaboré par l'Organisation mondiale de la santé (OMS) afin de soutenir les pays souhaitant évaluer et améliorer leurs capacités et leur niveau de conformité avec le RSI (2005). Ce cadre comprend quatre composantes complémentaires : le rapport annuel de l'État Partie, l'Évaluation extérieure conjointe, les examens « après action¼ et les exercices de simulation. Les deux premières composantes permettent de faire le point sur les capacités tandis que les deux dernières visent une connaissance détaillée de leur fonctionnement. La mise en oeuvre du RSI (2005) demande aux différentes disciplines, secteurs et domaines d'activités de fédérer leurs forces dans une approche Une seule santé. Par conséquent, en partenariat avec l'Organisation des Nations Unies pour l'alimentation et l'agriculture (FAO), avec l'Organisation mondiale de la santé animale (OIE) et avec d'autres partenaires internationaux et nationaux, l'OMS a fait en sorte de faciliter l'intégration de tous les secteurs concernés, en particulier celui de la santé animale, dans les diverses composantes du Cadre d'évaluation du RSI. D'autres outils complètent celui-ci, en particulier les ateliers de liaison nationaux OMS/OIE sur le RSI et le Processus d'évaluation des performances des Services vétérinaires (PVS), dont le but est de faciliter l'utilisation optimale des résultats du Cadre d'évaluation du RSI et du Processus PVS de l'OIE et de fournir aux acteurs des services de santé animale et de santé publique la possibilité de se concerter sur les modalités d'une synergie de leur action. Les résultats de ces outils sont ensuite pris en compte par les pays lors des procédures de planification et intégrés dans les Plans d'action nationaux pour la sécurité sanitaire afin d'accélérer la mise en oeuvre des capacités fondamentales décrites dans le RSI. Les auteurs décrivent l'intégration du concept Une seule santé dans chacune des composantes du Cadre d'évaluation du RSI.

Según lo dispuesto en el Reglamento Sanitario Internacional (RSI, 2005), documento jurídicamente vinculante suscrito por 196 Estados Partes, los países están obligados a dotarse de la capacidad necesaria para detectar, evaluar, notificar y afrontar con rapidez todo evento sanitario inusual que pueda revestir importancia internacional. Para ayudar a los países a dotarse de mejores capacidades, a seguir de cerca su evolución al respecto y a dar cumplimiento al RSI (2005), la Organización Mundial de la Salud (OMS) elaboró el marco de seguimiento y evaluación del RSI, que consta de cuatro elementos complementarios: el informe anual que debe presentar cada Estado Parte; la evaluación externa conjunta; exámenes posteriores a las intervenciones; y ejercicios de simulación. Los dos primeros sirven para examinar las capacidades, y los dos segundos para ayudar a estudiar su funcionalidad. Para la aplicación del RSI (2005) es fundamental la contribución de diferentes disciplinas, sectores y ámbitos de trabajo, que aúnen esfuerzos actuando desde los postulados de Una sola salud. Por ello la OMS, en colaboración con la Organización de las Naciones Unidas para la Alimentación y la Agricultura (FAO), la Organización Mundial de Sanidad Animal (OIE) y otros asociados internacionales y nacionales, ha trabajado activamente para facilitar la integración de los sectores pertinentes, en particular el de la sanidad animal, en cada uno de los cuatro componentes del marco de seguimiento y evaluación del RSI. Hay otros dispositivos que vienen a complementar este marco, por ejemplo los talleres nacionales dedicados a la creación de nexos entre el RSI y el proceso PVS (Prestaciones de los Servicios Veterinarios) de la OIE, organizados conjuntamente por la OMS y la OIE, que facilitan un uso idóneo de los resultados del marco de seguimiento y evaluación del RSI y del proceso PVS y brindan a las partes interesadas de los servicios sanitarios y zoosanitarios la oportunidad de trabajar sobre la coordinación de sus respectivas actividades. Los resultados de estas diversas herramientas alimentan después los procesos de planificación de los países y son incorporados a su Plan de acción nacional de seguridad sanitaria para acelerar la implantación de las capacidades básicas prescritas en el RSI. Los autores explican cómo se incorpora la filosofía de Una sola salud a todos los componentes del marco de seguimiento y evaluación del RSI.

Performance of Veterinary Services Pathway evolution and One Health aspects.

Animals, and the health systems which ensure their protection, play a vital role in the security and economic and social well-being of humanity, and are therefore a key component of the One Health concept. For global and national health security, prevention is better than cure, and targeting 'risk at source' in animal populations is a vital strategy in safeguarding the planet from risks of emerging zoonoses and antimicrobial resistance (AMR). Neglected zoonoses - such as rabies and brucellosis - continue to have a significant global impact on human health and are also best managed at their animal source. The World Organisation for Animal Health (OIE) has built international consensus on the principles of good governance and the quality of Veterinary Services, which are incorporated within its international standards. The OIE has a proven track record in the provision of Member Country support based on these standards, especially since the advent of its flagship Performance of Veterinary Services (PVS) Pathway programme in 2006-2007. To date, approximately 140 countries have benefited from the structured and sustainable process of animal health systems evaluation and planning afforded by the PVS Pathway. The PVS Tool, the basic methodology upon which the PVS Pathway is based, addresses One Health by evaluating the Veterinary Authority's ability to coordinate with other Competent Authorities that have a role to play in One Health, most notably public health, food safety, and environmental authorities. Despite the undoubted success of the PVS Pathway, the OIE felt that it was time to consider how the programme might be developed to adapt to new challenges. Consequently, during 2017-2018, the OIE embarked on a process of PVS evolution, during which it carried out extensive consultation and further tailored the PVS Pathway to a changing global context. These improvements, which include both fundamental adaptations to the PVS Pathway methods and the development of new PVS Pathway activities targeting topics such as multisectoral collaboration, rabies and AMR, have further strengthened and embedded the One Health approach within the PVS Pathway.

Parce qu'ils jouent un rôle crucial pour la sécurité et le bien-être économique et social de l'humanité, les animaux et les systèmes sanitaires en charge de leur protection sont une composante clé du concept Une seule santé. En matière de sécurité sanitaire à l'échelle du monde ou d'un pays, il vaut toujours mieux prévenir que guérir ; c'est pourquoi la stratégie consistant à cibler le risque à sa source est la seule qui puisse protéger la planète contre les zoonoses émergentes et le développement de l'antibiorésistance. L'impact sur la santé publique des zoonoses négligées comme la rage et la brucellose reste important et c'est également à leur source animale que les interventions visant à les contrôler sont les plus efficaces. L'Organisation mondiale de la santé animale (OIE) a forgé un consensus international autour des principes de bonne gouvernance et de qualité des Services vétérinaires et les a inscrites au coeur de ses normes internationales. L'OIE a démontré sa capacité à apporter aux Pays membres un soutien basé sur ces normes, en particulier depuis la création en 2006­2007 du Processus sur les Performances des Services vétérinaires (PVS), son programme phare. À ce jour, près de 140 pays ont bénéficié d'une procédure structurée d'évaluation et de planification durable de leurs systèmes de santé animale, grâce au Processus PVS. L'Outil PVS, instrument méthodologique du Processus PVS, couvre certains aspects relevant de l'approche Une seule santé en évaluant les capacités de concertation des Autorités vétérinaires avec d'autres autorités compétentes ayant un rôle à jouer dans ce contexte, en particulier celles en charge de la santé publique, de la sécurité sanitaire des aliments et de la protection de l'environnement. En dépit de la réussite incontestée du Processus PVS, l'OIE a estimé que le temps était venu d'envisager l'évolution de ce programme afin de l'adapter aux nouveaux défis. En conséquence, l'OIE a lancé en 2017­2018 la phase d'Évolution du Processus OIE à travers de larges consultations visant à adapter le Processus PVS aux mutations du contexte mondial. Les améliorations apportées, qui portent à la fois sur les fondements méthodologiques et sur la conception de nouvelles activités du Processus PVS dédiées à des sujets tels que la collaboration multisectorielle, la rage et la résistance aux agents antimicrobiens ont renforcé l'approche Une seule santé ainsi que son ancrage dans le Processus PVS.

Los animales y los sistemas sanitarios que velan por su protección cumplen una función vital para la seguridad y el bienestar económico y social de la humanidad, razón por la cual constituyen un elemento básico del concepto de Una sola salud. Desde el punto de vista de la seguridad sanitaria del mundo y de los países, más vale prevenir que curar, y el hecho de ir a combatir un riesgo en las poblaciones animales en las que tiene su origen es una estrategia indispensable para salvaguardar al planeta de los peligros que entrañan las zoonosis emergentes y la resistencia a los antimicrobianos. La mejor forma de luchar contra zoonosis desatendidas como la rabia o la brucelosis, que en todo el mundo siguen repercutiendo sensiblemente en la salud humana, pasa por atacarlas en su origen animal. La Organización Mundial de Sanidad Animal (OIE) ha sabido suscitar un consenso internacional en torno a los principios de buen gobierno y calidad de los Servicios Veterinarios, integrados ahora en sus normas internacionales. La OIE goza de contrastada experiencia en la prestación de apoyo a los Países Miembros basándose en estas normas, especialmente desde la instauración en 2006­2007 de su emblemático programa llamado Proceso PVS (Prestaciones de los Servicios Veterinarios). Hasta la fecha, alrededor de 140 países han podido beneficiarse del procedimiento estructurado y sostenible de evaluación y planificación de los sistemas de sanidad animal que se propone a través del Proceso PVS. La Herramienta PVS aporta la metodología básica en que descansa el Proceso PVS: con ella se trabaja en clave de Una sola salud evaluando la capacidad de la Autoridad Veterinaria del país para coordinarse con las demás autoridades competentes que cumplen alguna función relacionada con Una sola salud, sobre todo las de salud pública, seguridad sanitaria de los alimentos y medio ambiente. Pese al indiscutible éxito cosechado por el Proceso PVS, la OIE estimó llegado el momento de plantearse hacia dónde hacer evolucionar el programa para adaptarlo a nuevas problemáticas. Obrando en consecuencia, en 2017 y 2018 la OIE se embarcó en un proceso de «evolución del PVS¼ durante el cual celebró vastas consultas y adaptó aún más el Proceso PVS a un panorama mundial en constante evolución, incorporándole mejoras que incluyen a la vez una serie de ajustes básicos en los métodos del Proceso PVS y la creación de nuevas actividades encuadradas en él sobre temas como la colaboración multisectorial, la rabia o la resistencia a los antimicrobianos, mejoras que a la postre han servido para potenciar la filosofía de Una sola salud e integrarla aún más en el Proceso PVS.

Are Village Animal Health Workers Able to Assist in Strengthening Transboundary Animal Disease Control in Cambodia?

A cross-sectional survey of 445 Village Animal Health Workers (VAHWs) from 19 provinces in Cambodia was undertaken. The aim was to establish their levels of training, farm visit frequency, reasons for visits and disease reporting practices, enabling the strengths and weaknesses of the VAHW system in Cambodia to be determined, in providing both a fee-based smallholder livestock clinical service and a government partnership in transboundary animal disease (TAD) surveillance and control. The study used 'guided group interviews' and identified that VAHWs had good contact with farmers with 61.5% making more than one farm visit daily. However, incomes from services remained low, with 45% VAHWs obtaining between 20 and 40% of their household income from VAHW activities. VAHWs recorded relatively high rates of disease reporting, with 72% claiming they report diseases immediately and 74% undertaking monthly reporting to veterinary authorities. Logistic regression analysis revealed VAHW contact frequency with district and/or provincial officers was associated with more VAHW farm visits, and frequency of VAHW visits to smallholder farms was positively associated with average monthly expenditure on animal medication and equipment. This suggests that increased veterinary extension to VAHWs and access to veterinary equipment, vaccines and drugs may further increase VAHW-farmer engagement. VAHWs provide an accessible, market-based, animal health 'treatment and reporting' service linked to livestock smallholders across Cambodia. However, for improved TAD prevention and more efficient control of outbreaks, research that assesses provision of an animal health 'preventive-based' business model is urgently needed to reduce both the costs to farmers and the risks to the economy due to foot-and-mouth disease and other TADs in Cambodia.

A four-year pathology review of the near total face transplant.

In December of 2008, our institution performed a near total face transplant. The patient was monitored for signs of rejection assessed by paired skin and mucosa biopsies. The results of histological review of 120 biopsies collected during the first 4 years posttransplant are discussed. All biopsies were stained with hematoxylin and eosin, periodic acid-Schiff, immunohistochemical and TUNEL assays and graded using the Banff 2007 classification. Grade III rejection was diagnosed clinically at weeks 45 and 66, posttransplant; week 45 was determined as folliculitis while the erythema episode at week 66 confirmed an acute rejection (AR) that required hospitalization. The mucosa frequently showed interface inflammation without clinical signs of rejection and was not present in skin biopsies. In all, 34 of the 45 mucosal biopsies (75%) showed these interface changes. Clinical symptoms concurred with skin pathology in two grade III rejections. The mucosa showed histologic signs of rejection more frequently, which may indicate: increased mucosal sensitivity to rejection, a different type or subtype of AR that is specific to the mucosa, or a nonspecific process such as a drug effect. With more data and world experience, the diagnosis of face transplant rejection will be better defined and the Banff classification enhanced.

Printing ink compounds in foods: UK survey results.

Three hundred and fifty foodstuffs packaged in printed paper/board were purchased from UK retail outlets. Solvent extracts of all foods and associated quality assurance samples were analysed by gas chromatography-mass spectrometry (GC-MS) to determine the presence and concentrations of 20 printing ink compounds: benzophenone, 4-methylbenzophenone, 2-methylbenzophenone, 3-methylbenzophenone, 4-hydroxybenzophenone, 2-hydroxybenzophenone, 4-phenylbenzophenone, methyl-2-benzoylbenzoate, 1-hydroxycyclohexyl phenyl ketone, 2-isopropylthioxanthone, 4-isopropylthioxanthone, 2,4-diethyl-9H-thioxanthen-9-one, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-4'-(methylthio)-2-morpholinopropiophenone, 4-(4-methylphenylthio)benzophenone, ethyl-4-dimethylaminobenzoate, 2-ethylhexyl-4-(dimethylamino)benzoate, N-ethyl-p-toluene-sulphonamide, triphenyl phosphate and di-(2-ethylhexyl) fumarate. The presence of one or more of the compounds benzophenone, 4-phenylbenzophenone, methyl-2-benzoylbenzoate, 1-hydroxycyclohexyl phenyl ketone, 2,2-dimethoxy-2-phenylacetophenone, 4-(4-methylphenylthio)benzophenone, ethyl-4-dimethylaminobenzoate, 2-ethylhexyl-4-dimethylaminobenzoate and triphenyl phosphate was confirmed in some food samples. Analysis of the associated packaging material was also carried out to confirm whether or not it was likely that the occurrence of these compounds in the foods was due to migration from the printed paper/board packaging. With the exception of triphenyl phosphate, detected in one foodstuff, all the packaging material contained the substance(s) found in the food.

Income disparities in life expectancy in the City of Toronto and Region of Peel, Ontario.

INTRODUCTION: To understand the lack of a gradient in mortality by neighbourhood income in a previous study, we used individual-level data from the 1991-2001 Canadian census mortality follow-up study to examine income-related disparities in life expectancy and probability of survival to age 75 years in the City of Toronto and Region of Peel. METHODS: We calculated period life tables for each sex and income adequacy quintile, overall and separately for immigrants and non-immigrants. RESULTS: For all cohort members of both sexes, including both immigrants and non-immigrants, there was a clear gradient across the income quintiles, with higher life expectancy in each successively richer quintile. However, the disparities by income were much greater when the analysis was restricted to non-immigrants. The lesser gradient for immigrants appeared to reflect the higher proportion of recent immigrants in the lower income quintiles. CONCLUSION: These findings highlight the importance of using individual-level ascertainment of income whenever possible, and of including immigrant status and period of immigration in assessments of health outcomes, especially for areas with a high proportion of immigrants.

Survey of raw milk cheeses for microbiological quality and prevalence of foodborne pathogens.

Cheese may be manufactured in the United States using raw milk, provided the cheese is aged for at least 60 days at temperatures not less than 35°F (1.7°C). There is now increased concern among regulators regarding the safety of raw milk cheese due to the potential ability of foodborne pathogens to survive the manufacturing and aging processes. In this study, 41 raw milk cheeses were obtained from retail specialty shops, farmers' markets, and on-line sources. The cheeses were then analyzed for the presence of Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, Staphylococcus aureus, and Campylobacter. Aerobic plate counts (APC), coliform and yeast/mold counts were also performed. The results revealed that none of the enteric pathogens were detected in any of the samples tested. Five samples contained coliforms; two of those contained E. coli at less than 10(2) cfu/g. Three other cheese samples contained S. aureus. The APC and yeast-mold counts were within expected ranges. Based on the results obtained from these 41 raw milk cheeses, the 60-day aging rule for unpasteurized milk cheeses appears adequate for producing microbiologically safe products.

Malolactic fermentation as a technique for the deacidification of hard apple cider.

Malolactic fermentation (MLF), the conversion of malate to lactate, is an important process leading to the deacidification of hard apple cider. MLF is dependent on the levels of inhibitory factors such as sulfur dioxide and ethanol. To assess the effect of these 2 factors on MLF, hard apple cider was produced from pasteurized, unfiltered apple cider (Malus domestica cvs Red Delicious, Golden Delicious, Braeburn, and Fuji). Apple cider was treated with 2 levels of sulfur dioxide (50 and 80 ppm) and then fermented using Saccharomyces cerevisiae montrachet. After the primary fermentation, 1 set of the samples remained unadjusted and 100% ethyl alcohol was used to adjust other sets of samples to 7%, 9%, or 11% (v/v) ethanol. Following the ethanol adjustment, Oenococcus oeni MCW was used to initiate the MLF in half of the samples. Cider parameters monitored throughout the fermentations included organic acid content, titratable acidity, pH, ethanol production, and sugar content. Since samples containing either sulfur dioxide level had similar sugar utilization rates and ethanol production it was concluded that sulfur dioxide had no effect on the primary fermentation. Sulfur dioxide content was shown to have an impact on MLF. There was no difference in the rate of malic acid consumption, but lactic acid production was faster in the 50-ppm sulfur dioxide samples. MLF was not inhibited by ethanol content.

Investigation of the performance of safety systems for protection of the elderly.

This study investigates injury occurrence for belted occupants as a function of age. An analysis of NASS/CDS 1997-2003 data was conducted to determine crash involvement rates and injury rates for front seat occupants versus mean occupant age. In frontal and near-side crashes, the average age of MAIS 3+ belted front seat occupants injured in crashes less severe than 15 mph is of the order of 50 years. The average age of the population exposed to crashes less severe than 15 mph is under 40 years old. The crash exposure and frequency if injuries to the elderly were both found to be the highest in low severity crashes. The chest is the most frequent body region injured for the elderly. These findings suggest the need for more benign safety systems to protect the elderly in low severity crashes. Design of safety systems for the elderly should give priority to reducing the chest loading in low severity frontal and near-side crashes.

Validation of the urgency algorithm for near-side crashes.

The URGENCY algorithm uses vehicle crash sensor data in Automatic Crash Notification (ACN) systems to assist in instantly identifying crashes that are most likely to have time critical injuries. The algorithm also provides the capability of improving injury identification, using data obtained from the scene. The prime purpose of the algorithm is to automatically provide emergency medical responders with objective information on crash severity to assist in detecting the approximately 1% of crashes with serious injuries needing the most urgent medical care. The algorithm calculates the risk of a MAIS 3+ injury being present in the crashed vehicle, instantly at the time of the crash. The prediction can be subsequently updated as more information becomes available. The algorithm was based on a multiple regression analysis using data from the National Accident Sampling System/Crashworthiness Data System, (NASS/CDS) years 1988-95. In this paper, the accuracy of the algorithm was evaluated for near side crashes by applying it retrospectively to the population of injured occupants in NASS 1997-2000. URGENCY was applied to the population of injured occupants in near side crashes. Using an injury risk criterion of 50%, URGENCY identified 69% of the crashes with MAIS 3+ injuries. By lowering injury risk criterion to 40%, URGENCY identified 78% of the crashes with MAIS 3+ injuries. Vehicle side intrusion was found to be a highly influential variable. By changing side intrusion from a binary to a continuous variable, the correctly identified crashes increased from 69% to 81%. Examination of the consequence of missing variables found that unknown values of occupant height and weight had a negligible effect on the ability to capture the MAIS 3+ injured. However, lack of knowledge of these variables did increase the magnitude of the false positives.

Cytogenetic-clinicopathologic correlations in rhabdomyosarcoma: a report of five cases.

Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children younger than the age of 15 years. Histologically, RMS can be subdivided into two major subtypes; embryonal (E-RMS) and alveolar (A-RMS) rhabdomyosarcoma, with E-RMS being the more common. Although cytogenetic and molecular genetic findings have been reported extensively for RMS, clinicopathologic-genetic correlations among these tumors have not been reported in detail. In this report, we correlate the cytogenetic findings, including fluorescence in situ hybridization and spectral karyotyping, with pathologic findings and outcome for five RMS, including two A-RMS, one E-RMS, one botryoid RMS, and one anaplastic nonclassified RMS (N-RMS). The findings in A-RMS and E-RMS generally were consistent with previous reports; however, gain of chromosome 7 in A-RMS and gain of chromosome 9 segments in E-RMS observed here have seldom been reported in the literature. Importantly, the botryoid RMS had a cytogenetic profile similar to other types of E-RMS. An add(11)(q21) observed in this tumor, together with a t(8;11)(q12 approximately 13;q21) reported previously, indicates that 11q21 rearrangements may be nonrandomly related to botryoid RMS. In addition, the N-RMS expressed a cytogenetic pattern similar to that observed in E-RMS, thus providing genetic evidence that anaplastic N-RMS is a variant of E-RMS. Finally, these cases provide cogent evidence for the diagnostic and prognostic significance of the pathologic-genetic classification of RMS.

A descriptive analysis of giardiasis cases reported in Ontario, 1990-1998.

Cases of giardiasis in Ontario were described using notifiable disease data from the Ontario Ministry of Health for the years 1990-1998 inclusive. The mean annual age- and sex-adjusted incidence rate was 25.77 cases per 100,000 population for the 25,289 cases reported. Children under five years of age had the highest incidence of disease. Males had a higher mean annual incidence in all age groups. Four deaths occurred among cases. The most frequently reported symptoms were loose stools or watery diarrhea (50.1%). A seasonal pattern was noted, peaking in late summer and early autumn. The most frequently reported probable risk settings were the home (40.1%) and travel (39.1%). The study findings suggest that a high proportion of cases occur in urban areas and spatial analysis showed the highest incidence around Lake Huron and Georgian Bay. Unfiltered water and person-to-person contact are believed to be important sources of infection.

Validation of methods used to recover Escherichia coli O157:H7 and Salmonella spp. subjected to stress conditionst.

Escherichia coli O157:H7, Salmonella spp., and Salmonella Typhimurium DT104 were stressed with lactic acid and cell-free supernatants from lactic acid bacteria and plated on three different media to determine if injured cells were recovered. A comparison of the susceptibility and recovery of antibiotic-resistant strains of the pathogens and nonresistant strains was also made. Acid stress conditions were created by adjusting the pH of a cocktail mixture (two to four strains) of the pathogen to 3.50 with lactic acid and holding for 18 h. The pathogen cocktail was also stressed with a cell-free supernatant of Lactobacillus lactis (pH 3.90) in a 4:6 ratio. Both nonstressed and stressed cocktail cultures were plated on Trypticase soy agar (TSA) and violet red bile agar (VRBA) for E. coli and xylose lysine tergitol4 (XLT4) for Salmonella. Repair of injured cells was evaluated by pour plating the stressed cells on a 5-ml thin layer of TSA and allowing a 2-h room temperature incubation followed by overlaying with VRBA or XLT4. There were significant reductions in the populations of both pathogens under both stress conditions when plating was done on nonselective media. Injured E. coli O157:H7 was not recovered on recovery or selective media compared with TSA. Numbers of cells of supernatant-stressed Salmonella spp. plated on selective and recovery media were similar to those on TSA. Acid-stressed cells for all Salmonella spp. were not recovered on TSA, selective, or recovery media at levels comparable to recovery on TSA. Antibiotic-resistant strains showed similar recovery patterns on all media evaluated. However, the antibiotic-resistant strains were less sensitive to both stress conditions. The use of antibiotic-resistant strains resulted in a greater recovery of stressed pathogens than the use of recovery media.