Molecular characterization of Listeria monocytogenes and the significance for food hygiene

Research on the pathogen Listeria monocytogenes is a key issue both for the clinical and the food microbiologist owing to the unique pathway of infection and the exposure of humans via contaminated foods. Although, in Austria, the incidence of listeriosis is about 870-fold lower than the incidence for Salmonella infection, the food law manages both foodborne pathogens with a comparable stringency. The current risk management is based on the assumption that environmental L. monocytogenes isolates, from which the pool of "foodborne" isolates is recruited, are of similar pathogenicity compared to clinical and outbreak isolates. This verdict became doubted in the recent years. Characterization of L. monocytogenes by virulence gene sequencing, virulence studies in vivo and in vitro and by molecular typing was considerably stimulating the discussion on virulence variability in L. monocytogenes. This article provides insights in the value of epidemiological follow-up studies by presenting a typing study on 15 cases of listeriosis observed in a district hospital in Turkey. Furthermore results from typing L. monocytogenes either by virulence gene sequencing, mismatch amplification mutation assay or by pulsed field gel electrophoresis are discussed. The close interaction of molecular microbiology with food microbiology both in applied and basic science is currently creating a new discipline of molecular food microbiology. We are convinced that veterinary medicine will contribute to this exiting development in a fruitful way.     

Variation in clinical and parasitological traits in Pietrain and Meishan pigs infected with Sarcocystis miescheriana

Future prophylaxis needs new concepts, including natural disease resistance of hosts against infectious agents. Genomic approaches to detect and improve disease resistance in farm animals and the molecular mechanisms involved in host-parasite interactions depend to a high degree on the trait differences between founder breeds, i.e. on the animal model. The present study evaluates differences in susceptibility/resistance against Sarcocystis miescheriana in the European Pietrain (PI) and the Chinese Meishan (ME) pig breeds, based on 25 individuals, infected orally with 5x10(4) sporocysts of S. miescheriana. Significant differences appeared in clinical, serological, haematological and parasitological findings. The major discriminating period post infection (p.i.) was between days 42 and 45. Severity of signs was negatively correlated with specific immunoglobulin titres during the first 3 weeks p.i. and positively with the load of bradyzoites in muscle tissues of the pigs. Loads of bradyzoites in muscle tissues were 20 times higher in PI than in ME. Sarcocystis-specific differences between the two breeds were in the range of 1-2 standard deviations. The study lays the foundation for further experiments to analyse chromosomal regions, candidate genes, and thus the molecular basis of Sarcocystis susceptibility/resistance as a model for host-parasite interaction in protozoan infectious disease.     

Intraspecific Relationship of Plasmopara halstedii Isolates Differing in Pathogenicity and Geographic Origin Based on ITS Sequence Data

Sequence parts of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA were analysed to screen for the intraspecific variability of a non-coding genomic region in 15 Plasmopara halstedii populations of different pathotype and geographic origin. Samples revealed uniformity in a ca. 790 Bp fragment comprising of the ITS-1, 5.8S and front parts of the ITS-2. In contrast, clear differences were found in a ca. 810 Bp fragment of the ITS-2 thus allowing differentiation between populations of pathotype 100, 310 and 330 and a group of populations representing pathotypes 700, 701, 703, 710 and 730. Samples of pathotypes 700 to730 originated from Slovakia, France, and Germany, but were uniform in both ITS sequence parts, thus indicating very recent origin of these highly aggressive physiological races. The potential use of ITS sequences for pathotype differentiation and phylogenetic studies in P. halstedii is discussed.     

Metabolic labeling of plant cell cultures with K15NO3 as a tool for quantitative analysis of proteins and metabolites

Strategies for robust quantitative comparison between different biological samples are of high importance in experiments that address biological questions beyond the establishment of protein lists. Here, we propose the use of ¹⁵N-KNO₃ as the only nitrogen source in Arabidopsis cell cultures in order to achieve a metabolically fully labeled cell population. Proteins from such metabolically labeled culture are distinguishable from unlabeled protein populations by a characteristic mass shift that depends on the amino acid composition of the tryptic peptide analyzed. In addition, the metabolically labeled cell extracts are also suitable for comparative quantitative analysis of nitrogen-containing cellular metabolic complement. Protein extracts from unlabeled and from standardized ¹⁵N-labeled cells were combined into one sample for joined analytical processing. This has the advantage of (i) reduced experimental variability and (ii) immediate relative quantitation at the level of single extracted peptide and metabolite spectra. Together ease and accuracy of relative quantitation for profiling experiments is substantially improved. The metabolic labeling strategy has been validated by mixtures of protein extracts and metabolite extracts from the same cell cultures in known ratios of labeled to unlabeled extracts (1:1, 1:4, and 4:1). We conclude that saturating metabolic ¹⁵N-labeling provides a robust and affordable integrative strategy to answer questions in quantitative proteomics and nitrogen focused metabolomics.     

颗粒料和破碎料饲喂小虾的效果

虾料的外在物理特性变化范围很大 ,但在是否为小虾生长所必需这一问题上 ,还存在着疑问。相关的试验结果表明使用颗粒状开食料的斑节对虾 ( Penaeus monodon)幼苗的生长性能要显著好于饲喂破碎料的。饲喂 2种粒度碎粒状开食料的罗氏沼虾 ( Macrobrachium rosenbergii)间的生长性能无差异 ,因此得出结论 :幼虾阶段使用 2种粒度的破碎料是没有必要的。试验结果还表明颗粒料的长度 (长或短 )对斑节对虾的生长没有影响。目前 ,小虾的商业化养殖仍是一个新兴行业 ,相关的知识还很缺乏。因此 ,生产实践中往往按主观判断行事 ,特别是决定饲料的物…     

The "Friedrich-Loeffler-Institut": past, present and future of research in infectious diseases of animals

The Friedrich-Loeffler-Institut, founded in 1910 by Friedrich Loeffler, the discoverer of the first animal virus, foot-and-mouth disease virus, is the oldest virological research facility in the world. Beyond viruses, its area of competence has significantly expanded since its foundation and now also covers bacterial, parasitic and prion diseases of livestock, poultry and aquatic animals. Presently located at four sites within Germany (Insel Riems, Jena,Tübingen,Wusterhausen) the tasks of the institute as delineated in the Animal Disease Act encompass research on infectious animal diseases including zoonoses, import/export examinations, epidemiological studies in case of outbreaks of notifiable animal diseases, acting as reference laboratory for notifiable animal diseases and nationwide quality management of diagnosis of notifiable animal diseases. It is obliged to publish and maintain up-to-date diagnostic regimes for notifiable animal diseases, and it publishes a yearly report on animal health in Germany. With the increasing importance of infectious diseases of animals, in particular those potentially harmful to man (zoonoses), the Friedrich-Loeffler-Institut will be moving into new facilities including laboratories and animal facilities up to the highest biosafety level at its main site Insel Riems on the occasion of its 100th anniversary.     

Susceptibility of cocooned pupae and adults of the parasitoid <Emphasis Type="Italic">Microplitis mediator</Emphasis> to selected insecticides

The braconid parasitoid Microplitis mediator (Haliday) is a key natural enemy of the cabbage moth, Mamestra brassicae (L.), in Europe. In the context of an Integrated Pest Management approach, the use of selective insecticides is essential for the conservation of naturally occurring beneficial arthropods. The present laboratory study investigated the side effects of six insecticides applied at recommended field rates on adults and cocooned pupae of M. mediator. Male and female parasitoids were paired in drum cells contaminated with dry residues of insecticides. Besides lethal effects after 24 h, parasitization capacity and longevity of the surviving parasitoids was evaluated. Lethal effects on cocooned pupae were also investigated by assessing adult emergence from treated cocoons. Pirimicarb caused 100% adult mortality after 24 h, whereas the other tested insecticides caused no direct toxic effects. However, sub-lethal effects in terms of reduced parasitization activity, percentage of parasitism or female longevity were found for flonicamid, pymetrozine, spinosad and thiacloprid. Spirotetramat shortened only male longevity. Adult emergence from treated cocoons was reduced only by flonicamid and pymetrozine.