Epidemiological analysis of the dynamic and diversity of Salmonella spp. in five German pig production clusters using pheno- and genotyping methods: an exploratory study.

An exploratory study in five conventional pig production clusters was carried out to investigate the dynamic and diversity of Salmonella spp. within different production stages and sample site categories (pooled feces, direct and non-direct environment). Observing two production cycles per production cluster, a total of 1276 samples were collected along the pig production chain. Following a microbiological examination via culture, 2246 subcultures were generated out of 285 Salmonella positive samples and analysed by pheno- and genotyping methods. Based on a combination of serotyping, MLVA (multiple-locus variable-number tandem repeat (VNTR) analysis), PFGE (pulse-field gel electrophoresis) and MLST (multilocus sequence typing), an amount of 22.3% Salmonella positive samples were characterized in clonal lineages and its variants. Within each production cluster, one main clonal lineage could be identified and persisted over both production cycles with a large diversity of variants and a wide distribution in sample site categories and production stages. Results underline the importance of biosecurity with emphasis on the environment to prevent persistence and circulation of Salmonella within herds. Furthermore, the combined implementation of MLVA, PFGE and MLST with conventional culture techniques for isolate classification could be successfully applied as an effective and valuable tool for identifying similar pattern of Salmonella occurrence within pig production clusters.

Statistical tools to improve assessing agreement between several observers.

In the context of assessing the impact of management and environmental factors on animal health, behaviour or performance it has become increasingly important to conduct (epidemiological) studies in the field. Hence, the number of investigated farms per study is considerably high so that numerous observers are needed for investigation. In order to maintain the quality and validity of study results calibration meetings where observers are trained and the current level of agreement is assessed have to be conducted to minimise the observer effect. When study animals were rated independently by the same observers by a categorical variable the exclusion test can be performed to identify disagreeing observers. This statistical test compares for each variable and each observer the observer-specific agreement with the overall agreement among all observers based on kappa coefficients. It accounts for two major challenges, namely the absence of a gold-standard observer and different data type comprising ordinal, nominal and binary data. The presented methods are applied on a reliability study to assess the agreement among eight observers rating welfare parameters of laying hens. The degree to which the observers agreed depended on the investigated item (global weighted kappa coefficients: 0.37 to 0.94). The proposed method and graphical description served to assess the direction and degree to which an observer deviates from the others. It is suggested to further improve studies with numerous observers by conducting calibration meetings and accounting for observer bias.

Associations between host characteristics and antimicrobial resistance of Salmonella typhimurium.

A collection of Salmonella Typhimurium isolates obtained from sporadic salmonellosis cases in humans from Lower Saxony, Germany between June 2008 and May 2010 was used to perform an exploratory risk-factor analysis on antimicrobial resistance (AMR) using comprehensive host information on sociodemographic attributes, medical history, food habits and animal contact. Multivariate resistance profiles of minimum inhibitory concentrations for 13 antimicrobial agents were analysed using a non-parametric approach with multifactorial models adjusted for phage types. Statistically significant associations were observed for consumption of antimicrobial agents, region type and three factors on egg-purchasing behaviour, indicating that besides antimicrobial use the proximity to other community members, health consciousness and other lifestyle-related attributes may play a role in the dissemination of resistances. Furthermore, a statistically significant increase in AMR from the first study year to the second year was observed.

A quantitative approach to analyse linkages between antimicrobial resistance properties in Salmonella Typhimurium isolates.

This study used statistical methods to investigate linkages in phenotypic resistance profiles in a population sample of 321 Salmonella Typhimurium isolates from sporadic salmonellosis cases in Lower Saxony, Germany, collected during 2008-2010. A resistance index was applied to calculate the conditional probability of resistance to one antimicrobial agent given the resistance to one or more other antimicrobial agent(s). A susceptibility index was defined analogously. A contingency plot, which visualizes the association between resistances to two antimicrobial agents, facilitated the interpretation. Linkages between minimum inhibitory concentrations (MIC) were analysed using Spearman's rank correlation coefficient and jittered scatter plots. Applying these methods provided a compact description of multi-resistance and linkages between resistance properties in large datasets. Moreover, this approach will improve monitoring of antimicrobial resistance dynamics of bacteria in human or animal populations by identifying linked resistance to antimicrobial agents (cross- or co-resistance) with a non-molecular method.