Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen

Dresden, Germany

Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen

Dresden, Germany

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Schirmeister F.,German Federal Institute for Risk Assessment | Wieczorek A.,German Federal Institute for Risk Assessment | Dieckmann R.,German Federal Institute for Risk Assessment | Taureck K.,Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen | Strauch E.,German Federal Institute for Risk Assessment
International journal of medical microbiology : IJMM | Year: 2014

Vibrio furnissii and Vibrio fluvialis are two closely related species which are regarded as emerging human pathogens. Human infections have been mainly associated with consumption of seafood or drinking of contaminated water. V. furnissii strains can be distinguished from V. fluvialis by their ability to produce gas from fermentation of carbohydrates. In this study, we compare two phenotypic (biochemical testing and matrix-assisted laser desorption/ionisation time of flight mass spectrometry, MALDI-TOF MS) and three genotypic techniques (rpoB sequencing, conventional PCR and real-time PCR) for determination of the two species. The methods were evaluated on a collection of 42 V. furnissii and 32 V. fluvialis strains, which were isolated from marine environments and from animals intended for food production. Four of the applied methods allowed the unambiguous discrimination of the two species, while the biochemical testing was the least reliable technique, due to a high variation in the phenotype of gas production from carbohydrates. In view of the One Health concept reliable diagnostic techniques are a prerequisite for preventive public health measurements, as pathogens isolated from animals can cross species borders and methods for detection of sources, reservoirs and ways of transmission of pathogenic bacteria are indispensable for the prevention of infectious diseases in humans and animals. Copyright © 2014 Elsevier GmbH. All rights reserved.


Stahnke H.,German Federal Institute for Risk Assessment | Kittlaus S.,Carl Zeiss GmbH | Kempe G.,Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen | Hemmerling C.,Landeslabor Berlin Brandenburg | Alder L.,German Federal Institute for Risk Assessment
Journal of Mass Spectrometry | Year: 2012

This study investigates to which extent the design of electrospray ion sources influences the susceptibility to matrix effects (MEs) in liquid chromatography-tandem mass spectrometry (LC-MS/MS). For this purpose, MEs were measured under comparable conditions (identical sample extracts, identical LC column, same chromatographic method and always positive ion mode) on four LC-MS/MS instrument platforms. The instruments were combined with five electrospray ion sources, viz. Turbo Ion Spray, Turbo VTM Source, Standard ESI, Jet Stream ESI and Standard Z-Spray Source. The comparison of MEs could be made at all retention times because the method of permanent postcolumn infusion was applied. The MEs ascertained for 45 pesticides showed for each electrospray ion source the same pattern, i.e. the same number of characteristic signal suppressions at equivalent retention times in the chromatogram. The Turbo Ion Spray (off-axis geometry), Turbo VTM Source (orthogonal geometry) and the Standard Z-Spray Source (double orthogonal geometry) did not differ much in their susceptibility to MEs. The Jet Stream ESI (orthogonal geometry) reaches a higher sensitivity by an additional heated sheath gas, but suffers at the same time from significantly stronger signal suppressions than the comparable Standard ESI (orthogonal geometry) without sheath gas. No relation between source geometry and extent of signal suppression was found in this study. Copyright © 2012 John Wiley & Sons, Ltd.


Otto P.H.,Friedrich Loeffler Institute | Rosenhain S.,Friedrich Loeffler Institute | Elschner M.C.,Friedrich Loeffler Institute | Hotzel H.,Friedrich Loeffler Institute | And 4 more authors.
Veterinary Microbiology | Year: 2015

Rotaviruses (RVs) are a major cause of neonatal diarrhoea in humans and animals worldwide. In this study, 425 faecal samples were collected between 1999 and 2013 from diarrhoeic livestock and companion animals at different locations in Germany and tested for RVs. A previously published real-time RT-PCR assay was optimized for detection of a larger variety of RV species A (RVA) strains, and real-time RT-PCR assays for detection of RV species B (RVB) and C (RVC) were newly developed. The detection limits of the assays were 1.54×102, 3.95×102 and 3.60×103 genome copies for RVA, RVB and RVC, respectively. RVA was identified in 85.2% of bovine samples, 51.2% of porcine samples, 50.0% of feline samples, 43.2% of equine samples and 39.7% of canine samples. RVB was found in 3.0% of bovine samples, 2.7% of equine samples and 1.6% of porcine samples. RVC was detected in 31.0% of porcine samples, 21.7% of feline samples, 9.0% of canine samples and 6.0% of bovine samples. For genotyping, 101 RVA-positive bovine samples were further analysed by semi-nested RT-PCR. Genotype combination G6P[5] was most frequently detected (67.3% of samples), followed by G6P[11] (13.9%), G10P[5] (4.0%), G8P[11] (3.0%), G6P[1] (1.0%), and G10P[11] (1.0%). Mixed RVA infections were detected in 5.9% of samples; no or incomplete typing was possible in 4.0% of the samples. This first overview on RV species and RVA genotypes in diarrhoeic livestock and companion animals from Germany indicates a broad circulation of a large variety of RVs. © 2015 Elsevier B.V.


Uphoff H.,Hessisches Landes Prufungs und Untersuchungsamt im Gesundheitswesen | an der Heiden M.,Robert Koch Institute | Schweiger B.,Robert Koch Institute | Campe H.,Bayerisches Landesamt fur Gesundheit und Lebensmittelsicherheit | And 11 more authors.
PLoS ONE | Year: 2011

During the autumn wave of the pandemic influenza virus A/(H1N1) 2009 (pIV) the German population was offered an AS03-adjuvanted vaccine. The authors compared results of two methods calculating the effectiveness of the vaccine (VE). The test-negative case-control method used data from virologic surveillance including influenza-positive and negative patients. An innovative case-series methodology explored data from all nationally reported laboratory-confirmed influenza cases. The proportion of reported cases occurring in vaccinees during an assumed unprotected phase after vaccination was compared with that occurring in vaccinees during their assumed protected phase. The test-negative case-control method included 1,749 pIV cases and 2,087 influenza test-negative individuals of whom 6 (0.3%) and 36 (1.7%), respectively, were vaccinated. The case series method included data from 73,280 cases. VE in the two methods was 79% (95% confidence interval (CI) = 35-93%; P = 0.007) and 87% (95% CI = 78-92%; P<0.001) for individuals less than 14 years of age and 70% (95% CI = -45%-94%, P = 0.13) and 74% (95% CI = 64-82%; P<0.001) for individuals above the age of 14. Both methods yielded similar VE in both age groups; and VE for the younger age group seemed to be higher. © 2011 Uphoff et al.


Goerigk D.,University of Leipzig | Theuss T.,University of Leipzig | Pfeffer M.,University of Leipzig | Konrath A.,Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen | And 5 more authors.
Tierarztliche Praxis Ausgabe G: Grosstiere - Nutztiere | Year: 2014

Orthopoxvirus infections appear to be rare in South American Camelids, because only a few cases have been reported in the literature. Based on a generalized infection with cowpox virus in an alpaca, the clinical symptoms, laboratory diagnostic findings and the pathological changes are described. The case history showed a long treatment because of chronic skin lesions. The main clinical symptom was miliary papules over the entire skin. Furthermore, a bilateral mucopurulent conjunctivitis occurred as well as excessive salivation due to a severe erosive-ulcerative stomatitis. Although the animal received intensive treatment, it died 8 days after admission to the clinic. During necropsy, an erosive-ulcerative laryngitis as well as a necrotising pneumonia and lymphadenitis were observed. Histopathological examination of representative organ samples led to the diagnosis of a suspected orthopoxvirus infection. Electron microscopy and quantitative polymerase chain reaction (qPCR) of tissue samples confirmed this diagnosis. The virus could be isolated in tissue culture and a PCR with subsequent nucleotide sequencing identified cowpox virus as the causative agent for this generalised infection. © Schattauer 2014.


Granzow H.,Institute of Infectology | Fichtner D.,Institute of Infectology | Schutze H.,Institute of Infectology | Lenk M.,Friedrich Loeffler Institute | And 3 more authors.
Journal of Fish Diseases | Year: 2014

Two isolates of a novel enveloped RNA virus were obtained from carp and koi carp with gill necrosis. Both isolates behaved identically and could be propagated in different cyprinid cell lines forming large syncytia. The virus was sensitive to lipid solvents and neither exhibited haemadsorption/haemagglutination nor reverse transcriptase activity. Mature virus particles displayed a spherical shape with diameter of 100-350 nm after negative staining and 100-300 nm in ultrathin sections, covered by short projections of 8-10 nm in length. Maturation of virus progeny was shown to occur by budding and envelopment of the filamentous helical nucleocapsids at the cell surface. A detailed comparison of ultrastructure and morphogenesis of the novel virus isolates with selected arena-, ortho- and paramyxoviruses as possible candidates for evaluation of taxonomic classification yielded no consistency in all phenotypic features. Thus, on the basis of ultrastructure the novel virus isolates could not be assigned unequivocally to any established virus family. © 2013 John Wiley & Sons Ltd.


Stahnke H.,German Federal Institute for Risk Assessment | Kittlaus S.,Carl Zeiss GmbH | Kempe G.,Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen | Alder L.,German Federal Institute for Risk Assessment
Analytical Chemistry | Year: 2012

In this study, the relationship between matrix concentration and suppression of electrospray ionization (matrix effects) was investigated. Ion suppression of pesticides present in QuEChERS extracts was used as an example. Residue-free extracts of four different commodities, avocado, black tea, orange, and rocket (arugula), were fortified with 39 pesticides each. For many of the resulting 156 pesticide/matrix combinations, considerable matrix effects were observed if the coextracted matrix of 8 mg of equivalent sample (in the case of tea: 1.6 mg) was injected with the undiluted extracts. The reduction of these matrix effects was measured at 10 levels of dilution up to 1000-fold. The results obtained indicate a linear correlation between matrix effects and the logarithm of matrix concentration (or dilution factor) until the zero-effect level of further dilution was reached. Using the logarithmic equations, it could be shown that a dilution of extracts by a factor of 25-40 reduces ion suppression to less than 20% if the initial suppression is ≤80%. For stronger matrix effects or complete elimination of suppression, higher dilution factors were needed. The observed correlation was independent from the two instrument platforms used, but the degree of matrix effects differed slightly between the two mass spectrometers in this study. © 2011 American Chemical Society.


PubMed | Friedrich Loeffler Institute, German Federal Institute for Risk Assessment and Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen
Type: Journal Article | Journal: Veterinary microbiology | Year: 2015

Rotaviruses (RVs) are a major cause of neonatal diarrhoea in humans and animals worldwide. In this study, 425 faecal samples were collected between 1999 and 2013 from diarrhoeic livestock and companion animals at different locations in Germany and tested for RVs. A previously published real-time RT-PCR assay was optimized for detection of a larger variety of RV species A (RVA) strains, and real-time RT-PCR assays for detection of RV species B (RVB) and C (RVC) were newly developed. The detection limits of the assays were 1.5410(2), 3.9510(2) and 3.6010(3) genome copies for RVA, RVB and RVC, respectively. RVA was identified in 85.2% of bovine samples, 51.2% of porcine samples, 50.0% of feline samples, 43.2% of equine samples and 39.7% of canine samples. RVB was found in 3.0% of bovine samples, 2.7% of equine samples and 1.6% of porcine samples. RVC was detected in 31.0% of porcine samples, 21.7% of feline samples, 9.0% of canine samples and 6.0% of bovine samples. For genotyping, 101 RVA-positive bovine samples were further analysed by semi-nested RT-PCR. Genotype combination G6P[5] was most frequently detected (67.3% of samples), followed by G6P[11] (13.9%), G10P[5] (4.0%), G8P[11] (3.0%), G6P[1] (1.0%), and G10P[11] (1.0%). Mixed RVA infections were detected in 5.9% of samples; no or incomplete typing was possible in 4.0% of the samples. This first overview on RV species and RVA genotypes in diarrhoeic livestock and companion animals from Germany indicates a broad circulation of a large variety of RVs.


PubMed | German Federal Institute for Risk Assessment and Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen
Type: Comparative Study | Journal: International journal of medical microbiology : IJMM | Year: 2014

Vibrio furnissii and Vibrio fluvialis are two closely related species which are regarded as emerging human pathogens. Human infections have been mainly associated with consumption of seafood or drinking of contaminated water. V. furnissii strains can be distinguished from V. fluvialis by their ability to produce gas from fermentation of carbohydrates. In this study, we compare two phenotypic (biochemical testing and matrix-assisted laser desorption/ionisation time of flight mass spectrometry, MALDI-TOF MS) and three genotypic techniques (rpoB sequencing, conventional PCR and real-time PCR) for determination of the two species. The methods were evaluated on a collection of 42 V. furnissii and 32 V. fluvialis strains, which were isolated from marine environments and from animals intended for food production. Four of the applied methods allowed the unambiguous discrimination of the two species, while the biochemical testing was the least reliable technique, due to a high variation in the phenotype of gas production from carbohydrates. In view of the One Health concept reliable diagnostic techniques are a prerequisite for preventive public health measurements, as pathogens isolated from animals can cross species borders and methods for detection of sources, reservoirs and ways of transmission of pathogenic bacteria are indispensable for the prevention of infectious diseases in humans and animals.


PubMed | Veterinary Practice Zettlitz, Landesuntersuchungsanstalt fur das Gesundheits und Veterinarwesen Sachsen and University of Gottingen
Type: Journal Article | Journal: PloS one | Year: 2015

Aristaless-like homeobox 4 (ALX4) gene is an important transcription regulator in skull and limb development. In humans and mice ALX4 mutations or loss of function result in a number of skeletal and organ malformations, including polydactyly, tibial hemimelia, omphalocele, biparietal foramina, impaired mammary epithelial morphogenesis, alopecia, coronal craniosynostosis, hypertelorism, depressed nasal bridge and ridge, bifid nasal tip, hypogonadism, and body agenesis. Here we show that a complex skeletal malformation of the hind limb in Galloway cattle together with other developmental anomalies is a recessive autosomal disorder most likely caused by a duplication of 20 bp in exon 2 of the bovine ALX4 gene. A second duplication of 34 bp in exon 4 of the same gene has no known effect, although both duplications result in a frameshift and premature stop codon leading to a truncated protein. Genotyping of 1,688 Black/Red/Belted/Riggit Galloway (GA) and 289 White Galloway (WGA) cattle showed that the duplication in exon 2 has allele frequencies of 1% in GA and 6% in WGA and the duplication in exon 4 has frequencies of 23% in GA and 38% in WGA. Both duplications were not detected in 876 randomly selected German Holstein Friesian and 86 cattle of 21 other breeds. Hence, we have identified a candidate causative mutation for tibial hemimelia syndrome in Galloway cattle and selection against this mutation can be used to eliminate the mutant allele from the breed.

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