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Broeders S.,Scientific Institute of Public Health WIV ISP | Huber I.,Bavarian Health and Food Safety Authority LGL | Grohmann L.,Federal Office of Consumer Protection and Food Safety BVL | Berben G.,Walloon Agricultural Research Center | And 5 more authors.
Trends in Food Science and Technology | Year: 2014

As for many areas of molecular testing, detection of Genetically Modified Organisms (GMO) relies on the real-time Polymerase Chain Reaction (qPCR) technology. Due to the increasing number of GMO, a screening approach using qualitative screening methods has become an integrated part of GMO detection. However, specific guidelines for the validation of these methods are lacking. Here, a pragmatic approach to conduct in-house and inter-laboratory validation studies for GMO screening methods, is proposed. Such guidelines could be adapted to other areas where qualitative qPCR methods are used for molecular testing allowing to implement easily a more reliable screening phase where necessary. © 2014 Elsevier Ltd.


Huang J.-H.,ETH Zurich | Fecher P.,Bavarian Health and Food Safety Authority LGL | Ilgen G.,University of Bayreuth | Hu K.-N.,U.S. National Institute of Diabetes and Digestive and Kidney Diseases | Yang J.,University of Sichuan
Food Chemistry | Year: 2012

Arsenite and arsenate speciation was performed in 121 commercially purchased samples of 12 rice types to understand their relative relevance in rice grain. General effectiveness of a recently developed extraction protocol based on 0.28 M nitric acid at 95 °C was verified by checking the recovery of total grain arsenic and by comparing arsenic speciation in NIST-CRM-1568a, NMIJ-CRM-7503a and IMEP-107 with certificated and literature values. The arsenic speciation highlights the predominance of arsenite in 115 and dimethylarsinic acid in six samples and common minor components including arsenate, monomethylarsonic acid and two unknown arsenical species. Our data also indicate potential influences of other trace elements on As speciation in rice grain. Averagely, arsenite accounts for 90% of inorganic grain arsenic regardless of geographic origin, rice type, grain size, cultural practice and polish treatment. The high arsenite predominance indicates an elevated toxic effect of arsenic in rice than is perceived to date. © 2011 Elsevier Ltd. All rights reserved.


Huang J.-H.,ETH Zurich | Ilgen G.,University of Bayreuth | Fecher P.,Bavarian Health and Food Safety Authority LGL
Journal of Analytical Atomic Spectrometry | Year: 2010

Extraction using 0.28 M nitric acid at 95 °C for 90 min recovers not only arsenic quantitatively from rice grains independent of types but also preserves arsenic speciation completely. It is simple, economic, effective and reliable and is especially suitable for routine analysis of large amounts of samples. © The Royal Society of Chemistry 2010.


Bottcher S.,Robert Koch Institute | Neubauer K.,Robert Koch Institute | Baillot A.,Governmental Institute of Public Health of Lower Saxony NLGA | Rieder G.,Bavarian Health and Food Safety Authority LGL | And 2 more authors.
International Journal of Medical Microbiology | Year: 2015

Germany is a partner of the Global Polio Eradication Initiative. Assurance of polio free status is based on enterovirus surveillance, which focuses on patients with signs of acute flaccid paralysis or aseptic meningitis/encephalitis, representing the key symptoms of poliovirus infection. In response to the wild poliovirus outbreak in Syria 2013 and high number of refugees coming from Syria to Germany, stool samples from 629 Syrian refugees/asylum seekers aged <3 years were screened for wild poliovirus between November 2013 and April 2014. Ninety-three samples (14.8%) were positive in an enterovirus specific PCR. Of these, 12 contained Sabin-like polioviruses. The remaining 81 samples were characterized as non-polio enteroviruses representing several members of groups A-C as well as rhinovirus. Wild-type poliovirus was not detected via stool screening involving molecular and virological methods, indicating a very low risk for the importation by Syrian refugees and asylum seekers at that time. © 2015 Elsevier GmbH.


Takla A.,Robert Koch Institute | Wichmann O.,Robert Koch Institute | Klinc C.,Bavarian Health and Food Safety Authority LGL | Hautmann W.,Bavarian Health and Food Safety Authority LGL | And 3 more authors.
Eurosurveillance | Year: 2013

In Germany, mumps has been notifiable until 2013 only in the five Eastern federal states (EFS) of former East Germany. Due to different immunisation policies until 1990 and varying vaccination coverages thereafter, mumps incidences cannot be extrapolated to the 11 Western federal states (WFS). We studied mumps-related International Classification of Diseases (ICD-10) code diagnoses claimed through statutory health insurances between 2007 and 2011 to estimate countrywide mumps incidences in the outpatient sector, and compared them with case numbers from ambulatory notification data. Overall, 32,330 outpatient mumps cases were claimed. Annual incidence ranged between 9.3/100,000 and 11.8/100,000 and showed a significant decreasing trend. Compared with EFS, mumps incidence in WFS was higher and indicated a shift towards older age groups. Notified outpatient case numbers in EFS were 13-fold lower and from voluntary surveillance during an outbreak in the WFS Bavaria 8-fold lower than from insurance data (n=316 versus n=4,217 and n=238 versus 1,995, respectively). Of all notified cases with available information, 75.4% (EFS) and 57.6% (Bavaria) were unvaccinated; 6.8% (EFS) and 19.3% (Bavaria) required hospitalisation. In Germany, mumps is still endemic despite decades of vaccination, with considerable underreporting in the established notification systems. © 2007-2013. All rights reserved.


Schabauer L.,University of Veterinary Medicine Vienna | Wenning M.,TU Munich | Huber I.,Bavarian Health and Food Safety Authority LGL | Ehling-Schulz M.,University of Veterinary Medicine Vienna
BMC Veterinary Research | Year: 2014

Background: The routine diagnosis of Streptococcus spp. and other mastitis associated gram-positive, catalase-negative cocci is still based upon biochemical tests and serological methods, which frequently provide ambiguous identification results. We therefore aimed to establish an accurate identification system for differential diagnosis of mastitis associated Streptococcus spp. and related species using biophysical techniques such as Fourier-transform infrared (FTIR) spectroscopy and MALDI - TOF/MS.Results: Based on a panel of 210 isolates from cases of bovine mastitis, an unsupervised FTIR spectral reference library was established and an artificial neural network (ANN) - assisted identification system was developed. All bacterial isolates were previously identified by species-specific PCR and/or 16S rRNA gene sequence analysis. An overall identification rate of 100% at species level for 173 strains unknown to the ANN and the library was achieved by combining ANN and the spectral database, thus demonstrating the suitability of our FTIR identification system for routine diagnosis. In addition, we investigated the potential of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of mastitis associated Streptococcus spp. and related bacteria. Using the Microflex LT System, MALDI Biotyper software™ (V3.3) we achieved an accuracy rate of 95.2%. A blind study, including 21 clinical samples from dairy cows, revealed a 100% correct species identification rate for FTIR and 90.5% for MALDI-TOF MS, indicating that these techniques are valuable tools for diagnosis.Conclusions: This study clearly demonstrates that FTIR spectroscopy as well as MALDI-TOF MS can significantly improve and facilitate the identification and differentiation of mastitis associated Streptococcus spp. and related species. Although the FTIR identification system turned out being slightly superior to MALDI-TOF MS in terms of identification on species level, both methods offer interesting alternatives to conventional methods currently used in mastitis diagnosis as both of them provide high accuracy at low operating costs once the instrument is acquired. © 2014 Schabauer et al.; licensee BioMed Central Ltd.


von Rosenberg S.J.,Ludwig Maximilians University of Munich | Wehr U.A.,Ludwig Maximilians University of Munich | Wehr U.A.,Bavarian Health and Food Safety Authority LGL
Journal of Animal Physiology and Animal Nutrition | Year: 2012

Summary: Two different lanthanum salts, lanthanum carbonate (LaCO3) and Lancer®, a lanthanide citrate mixture, were tested for their effects on bone metabolism in a small animal model for post-menopausal osteoporosis. Forty female outbred Wistar Han rats, sham-operated (SHAM, positive control, n=10) or ovariectomized (OVX, n=30) at 4months of age, were allotted into following groups (n=10/group): (i) SHAM, (ii) OVX control (negative control), (iii) OVX+LaCO3 (1.74g/kg feed) and (iv) OVX+Lancer® (8g/kg feed). Effects on bone were investigated by bone markers [osteocalcin (Oc) in serum and excretion of pyridinoline (PYD) in urine] and by physical parameters of bone structure and bone composition (bone mass, calcium, phosphorus and magnesium content in bone crude ash). Bone micro-architecture and bone mineral density were evaluated by peripheral quantitative computed tomography and micro-computed tomography (μCT). The animal model could be validated by differences between OVX control and SHAM. Body mass and feed intake were the same among the four groups. Oc was clearly increased in the two experimental groups (p<0.001) vs. SHAM and OVX control. Bone mass and calcium content in bone ash were significantly higher than in OVX control. The Ca/P ratio in bone ash of the two lanthanide groups did not differ from SHAM. Bone-protecting effects of lanthanides were clearly demonstrated by an increased trabecular density which is the region of interest for osteoporotic bone loss. A 3D imaging of bone micro-architecture by μCT visualized descriptively the positive effects of lanthanides on bone formation. The results of this study demonstrate an improvement of bone formation and bone-protecting effects of lanthanides in the OVX rat. Thus, lanthanum salts suggest a prevention of post-menopausal bone loss and may be of benefit in experimental osteopenia following ovariectomy. © 2012 Blackwell Verlag GmbH.


Gerdes L.,Bavarian Health and Food Safety Authority LGL | Busch U.,Bavarian Health and Food Safety Authority LGL | Pecoraro S.,Bavarian Health and Food Safety Authority LGL
European Food Research and Technology | Year: 2012

Identification of specific material derived from genetically modified organisms (GMO) present in food, feed or seed samples screened positive for the presence of genetic modification(s) is mandatory for the official food and feed control in the European Union. Since the introduction of regulation (EC) No 1829/2003 in 2004, the number of maize GMO events either approved in the EU or with a pending application grew constantly. By the sheer multitude of events and crossed events (stacks), maize poses a special challenge on official food and feed control. We developed a modular qualitative detection system for the parallel identification of maize GMO events to cope with the increasing number of GMO potentially present in routine samples. This system is based on validated real-time PCR assays in a microtitre plate format grouped modularly by crop species. The maize module identifies in parallel, i. e. simultaneously, 15 maize events and RoundupReady soy in a single analytical run of approximately 2 h. Maize modules can be conveniently prepared in advance and stored at -20 °C until use. Ready-to-use reference DNA mixtures serve as positive controls. The modular approach is flexible as it allows easy change or addition of individual detection reactions, if necessary, e. g. when new validated methods become available. 23 food, 14 feed and 8 seed samples were successfully analysed with the maize module. The parallel detection of nine different GMO maize and soy events in single routine samples demonstrated the usefulness of the parallelised modular approach for routine GMO analysis. © 2011 Springer-Verlag.


Huber I.,Bavarian Health and Food Safety Authority LGL | Campe H.,Bavarian Health and Food Safety Authority LGL | Sebah D.,Bavarian Health and Food Safety Authority LGL | Hartberger C.,Bavarian Health and Food Safety Authority LGL | And 4 more authors.
Eurosurveillance | Year: 2011

For surveillance purposes real-time PCR assays for influenza viruses had to be adapted to the pandemic influenza A(H1N1)2009 strain. We combined published primers and probes for influenza A, influenza B and an internal amplification control with a detection system for influenza A(H1N1)2009 to set up a rapid, reliable, simple and cost-effective high-throughput multiplex one-step real-time RT-PCR. The workflow also includes automated sample preparation for high-throughput screening. The lower limit of detection of the multiplex assay was 3.5×102 RNA copies per PCR reaction. The diagnostic sensitivity of the multiplex assay was 87.7%, but increased to 99.4% for influenza-positive samples yielding Ct values of less than 34 cycles in the respective diagnostic assay. High specificity was confirmed by sequencing and correct detection of 15 reference samples from two quality assurance studies. The multiplex PCR was introduced for surveillance of samples from a network of general practitioners and paediatricians in Bavaria, Germany during the influenza pandemic of 2009. Comparison with surveillance data from reported cases proved the reliability of the multiplex assay for influenza surveillance programmes.


Gerdes L.,Bavarian Health and Food Safety Authority LGL | Busch U.,Bavarian Health and Food Safety Authority LGL | Pecoraro S.,Bavarian Health and Food Safety Authority LGL
Food Analytical Methods | Year: 2012

The development and cultivation of genetically modified crops is still increasing globally. Food and feed imports from outside the European Union (EU) will subsequently require more effort from the responsible authorities in monitoring the compliance with effective labelling regulations. The aim of this study was the development of the GMOfinder, a database for collection and interpretation of information related to the screening for genetically modified organisms (GMOs). Different genetic elements (e. g. promoters, terminators, structural genes) are artificially introduced into plants to establish new genetic modifications. The introduced elements may vary between GMO events, depending on the intended trait(s). Screening for such inserted elements with (real-time) polymerase chain reaction is a common first step to analyse samples for the presence of any genetical modification. From the pattern of detectable and nondetectable elements, valuable conclusions about the identity of putative present GMO event(s) can be drawn with the GMOfinder. Information about selected genetic elements from the literature, applications for authorisation and other (web) sources were systematically integrated in a tabular matrix format. Special care was taken to additionally record the sources of the information, thus facilitating evaluation of screening results, and tracing of possible errors in the matrix. The GMOfinder accesses data from the element matrix with implemented algorithms and facilitates to interpret the outcome of screenings. Such a preselection helps to systematically narrow down the candidates for subsequent identification reactions. Optional display of events with potentially masked elements completes the included features. © 2012 Springer Science+Business Media, LLC.

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