Federal Research Institute for Animal Health

Greifswald, Germany

Federal Research Institute for Animal Health

Greifswald, Germany
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Herrmann K.,German Institute of Human Nutrition | Engst W.,German Institute of Human Nutrition | Meinl W.,German Institute of Human Nutrition | Florian S.,German Institute of Human Nutrition | And 8 more authors.
Carcinogenesis | Year: 2014

Methyleugenol-a natural constituent of herbs and spices-is hepatocarcinogenic in rodent models. It can form DNA adducts after side-chain hydroxylation and sulfation. We previously demonstrated that human sulfotransferases (SULTs) 1A1 and 1A2 as well as mouse Sult1a1, expressed in Salmonella target strains, are able to activate 1′-hydroxymethyleugenol (1′-OH-ME) and 3′-hydroxymethylisoeugenol (3′-OH-MIE) to mutagens. Now we investigated the role of these enzymes in the formation of hepatic DNA adducts by methyleugenol in the mouse in vivo. We used FVB/N mice [wild-type (wt)] and genetically modified strains in this background: Sult1a1 knockout (ko), transgenic for human SULT1A1/2 (tg) and the combination of both modifications (ko-tg). Methyleugenol (50mg/kg body mass) formed 23, 735, 3770 and 4500 N2-(trans-methylisoeugenol-3′-yl)-2′-deoxyguanosine adducts per 108 2′-deoxyribonucleosides (dN) in ko, wt, ko-tg and tg mice, respectively. The corresponding values for an equimolar dose of 1′-OH-ME were 12, 1490, 12 400 and 13 300 per 108 dN. Similar relative levels were observed for the minor adduct, N6-(trans-methylisoeugenol-3′-yl)-2′-deoxyadenosine. Thus, the adduct formation by both compounds was nearly completely dependent on the presence of SULT1A enzymes, with human SULT1A1/2 producing stronger effects than mouse Sult1a1. Moreover, a dose of 0.05mg/kg methyleugenol (one-fourth of the estimated average daily exposure of humans) was sufficient to form detectable adducts in humanized (ko-tg) mice. Although 3′-OH-MIE was equally mutagenic to 1′-OH-ME in Salmonella strains expressing human SULT1A1 or 1A2, it only formed 0.14% of hepatic adducts in ko-tg mice compared with an equimolar dose of 1′-OH-ME, suggesting an important role of detoxifying pathways for this isomer in vivo. © The Author 2013. Published by Oxford University Press. All rights reserved.

Wadsworth J.D.F.,University College London | Joiner S.,University College London | Linehan J.M.,University College London | Balkema-Buschmann A.,Federal Research Institute for Animal Health | And 8 more authors.
Emerging Infectious Diseases | Year: 2013

Public and animal health controls to limit human exposure to animal prions are focused on bovine spongiform encephalopathy (BSE), but other prion strains in ruminants may also have zoonotic potential. One example is atypical/Nor98 scrapie, which evaded statutory diagnostic methods worldwide until the early 2000s. To investigate whether sheep infected with scrapie prions could be another source of infection, we inoculated transgenic mice that overexpressed human prion protein with brain tissue from sheep with natural field cases of classical and atypical scrapie, sheep with experimental BSE, and cattle with BSE. We found that these mice were susceptible to BSE prions, but disease did not develop after prolonged postinoculation periods when mice were inoculated with classical or atypical scrapie prions. These data are consistent with the conclusion that prion disease is less likely to develop in humans after exposure to naturally occurring prions of sheep than after exposure to epizootic BSE prions of ruminants.

Future livestock production is likely to be affected by both rising ambient temperatures and indirect effects mediated by modified growth conditions of feed plants such as increased atmospheric CO2 concentrations and drought. Corn was grown at elevated CO2 concentrations of 550 ppm and drought stress using free air carbon dioxide enrichment technology. Whole plant silages were generated and fed to sheep kept at three climatic treatments. Differential blood count was performed. Plasma DON and de-epoxy-DON concentration were measured. Warmer environment increased rectal and skin temperatures and respiration rates (p < 0.001 each) but did not affect blood parameters and the almost complete metabolization of DON into de-epoxy-DON. Altered growth conditions of the corn fed did not have single effects on sheep body temperature measures and differential blood count. Though the thermoregulatory activity of sheep was influenced by the thermal environment, the investigated cultivation factors did not indicate considerable impacts on the analysed parameters.

Danicke S.,Federal Research Institute for Animal Health
Mycotoxin research | Year: 2012

The aim of this study was to examine the effects of a control diet (CON, 0.25 mg DON/kg diet) or a Fusarium toxin-contaminated diet (FUS, 4.49 mg DON/kg diet) without and with humic substances (HS) (CON-HS and FUS-HS, 0.23 and 4.56 mg DON/kg diet, respectively) on piglets during a 5-week growth trial starting after weaning (6.7 ± 0.9 kg live weight, n = 20/group). Feed intake was significantly reduced by feeding the FUS containing diets by approximately 21% compared with the CON diet irrespective of HS supplementation. The decrease in live weight gain paralleled the feed intake depression and amounted to approximately 26%. Feeding the FUS diet was clearly reflected by the DON levels in blood. While only traces of DON with median concentrations of 3 ng/ml (2-5 ng/ml) and 2 ng/ml (0-3 ng/ml) were detected in piglets fed the CON and CON-HS diets, respectively, significantly higher levels of 22.5 ng/ml (7-30 ng/ml) and 23.5 ng/ml (15-32 ng/ml) were found in piglets fed the FUS and FUS-HS diet, respectively. The urinary excretion of DON and its metabolite de-epoxy-DON as percentage of DON intake was not significantly influenced by HS supplementation and amounted to 24.1 and 20.2% for groups FUS and FUS-HS, respectively. In conclusion, the tested HS preparation cannot be recommended as a DON inactivating feed supplement for pigs.

Danicke S.,Federal Research Institute for Animal Health
Mycotoxin research | Year: 2012

Both hydrothermal treatment and wet preservation of mainly deoxynivalenol (DON)-containing, Fusarium toxin (FUS)-contaminated cereal grains with sodium metabisulfite (Na2S2O5 [SBS]) were successfully demonstrated to reduce the DON contamination through formation of the sulfonated derivative of DON, termed as DON sulfonate (DONS). The wet preservation is particularly interesting from a practical viewpoint as it can be easily performed at the farm level where the cereal grains are harvested and utilized in pig feeding. This review compiles the literature with regard to the chemical characterization and the detection of DONS, technical procedures and their efficacies, toxicological aspects and toxic effects of DON, DONS and SBS, and detection of DONS, DON and further metabolites in physiological specimens of pigs.

Ramp K.,Institute of Molecular Biology | Topfstedt E.,Institute of Molecular Biology | Wackerlin R.,Institute of Diagnostic Virology | Hoper D.,Institute of Diagnostic Virology | And 4 more authors.
Avian Diseases | Year: 2012

Even though Newcastle disease virus (NDV) live vaccine strains can be applied to 1-day-old chickens, they are pathogenic to chicken embryos when given in ovo 3 days before hatch. Based on the reverse genetics system, we modified recombinant NDV (rNDV) established from lentogenic vaccine strain Clone 30 by introducing specific mutations within the fusion (F) and hemagglutinin- neuraminidase (HN) proteins, which have recently been suggested as being responsible for attenuation of selected vaccine variants (Mast et al. Vaccine 24:17561765, 2006) resulting in rNDV49. Another recombinant (rNDVGu) was generated to correct sequence differences between rNDV and vaccine strain NDV Clone 30. Recombinant viruses rNDV, rNDV49, and rNDVGu have reduced virulence compared with NDV Clone 30, represented by lower intracerebral pathogenicity indices and elevated mean death time. After in ovo inoculation, hatchability was comparable for all infected groups. However, only one chicken from the NDV Clone 30 group survived a 21-day observation period; whereas, the survival rate of hatched chicks from groups receiving recombinant NDV was between 40% and 80%, with rNDVGu being the most pathogenic virus. Furthermore, recombinant viruses induced protection against challenge infection with virulent NDV 21 days post hatch. Differences in antibody response of recombinant viruses indicate that immunogenicity is correlated to virulence. In summary, our data show that point mutations can reduce virulence of NDV. However, alteration of specific amino acids in F and HN proteins of rNDV did not lead to further attenuation as indicated by their pathogenicity for chicken after in ovo inoculation. © 2012 American Association of Avian Pathologists.

Maan S.,Institute for Animal Health | Maan N.S.,Institute for Animal Health | van Rijn P.A.,Central Veterinary Institute of Wageningen UR | van Gennip R.G.P.,Central Veterinary Institute of Wageningen UR | And 9 more authors.
PLoS ONE | Year: 2010

In mid September 2008, clinical signs of bluetongue (particularly coronitis) were observed in cows on three different farms in eastern Netherlands (Luttenberg, Heeten, and Barchem), two of which had been vaccinated with an inactivated BTV-8 vaccine (during May-June 2008). Bluetongue virus (BTV) infection was also detected on a fourth farm (Oldenzaal) in the same area while testing for export. BTV RNA was subsequently identified by real time RT-PCR targeting genome-segment (Seg-) 10, in blood samples from each farm. The virus was isolated from the Heeten sample (IAH "dsRNA virus reference collection" [dsRNA-VRC] isolate number NET2008/05) and typed as BTV-6 by RT-PCR targeting Seg-2. Sequencing confirmed the virus type, showing an identical Seg-2 sequence to that of the South African BTV-6 live-vaccine-strain. Although most of the other genome segments also showed very high levels of identity to the BTV-6 vaccine (99.7 to 100%), Seg-10 showed greatest identity (98.4%) to the BTV-2 vaccine (RSAvvv2/02), indicating that NET2008/05 had acquired a different Seg-10 by reassortment. Although Seg-7 from NET2008/05 was also most closely related to the BTV-6 vaccine (99.7/100% nt/aa identity), the Seg-7 sequence derived from the blood sample of the same animal (NET2008/06) was identical to that of the Netherlands BTV-8 (NET2006/04 and NET2007/01). This indicates that the blood contained two different Seg-7 sequences, one of which (from the BTV-6 vaccine) was selected during virus isolation in cell-culture. The predominance of the BTV-8 Seg-7 in the blood sample suggests that the virus was in the process of reassorting with the northern field strain of BTV-8. Two genome segments of the virus showed significant differences from the BTV-6 vaccine, indicating that they had been acquired by reassortment event with BTV-8, and another unknown parental-strain. However, the route by which BTV-6 and BTV-8 entered northern Europe was not established. © 2010 Maan et al.

Reiche S.,Federal Research Institute for Animal Health | Dwai Y.,University of Leipzig | Dwai Y.,Tishreen University | Bussmann B.M.,University of Leipzig | And 3 more authors.
PLoS ONE | Year: 2015

The diversity of virus-specific antibodies and of B cells among different individuals is unknown. Using single-cell cloning of antibody genes, we generated recombinant human monoclonal antibodies from influenza nucleoprotein-specific memory B cells in four adult humans with and without preceding influenza vaccination. We examined the diversity of the antibody repertoires and found that NP-specific B cells used numerous immunoglobulin genes. The heavy chains (HCs) originated from 26 and the kappa light chains (LCs) from 19 different germ line genes. Matching HC and LC chains gave rise to 43 genetically distinct antibodies that bound influenza NP. The median lengths of the CDR3 of the HC, kappa and lambda LC were 14, 9 and 11 amino acids, respectively. We identified changes at 13.6% of the amino acid positions in the V gene of the antibody heavy chain, at 8.4% in the kappa and at 10.6 %in the lambda V gene. We identified somatic insertions or deletions in 8.1% of the variable genes.We also found several small groups of clonal relatives that were highly diversified. Our findings demonstrate broadly diverse memory B cell repertoires for the influenza nucleoprotein.We found extensive variation within individuals with a high number of point mutations, insertions, and deletions, and extensive clonal diversification. Thus, structurally conserved proteins can elicit broadly diverse and highly mutated B-cell responses. Copyright: © 2015 Reiche et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Scheuch M.,Federal Research Institute for Animal Health | Hoper D.,Federal Research Institute for Animal Health | Beer M.,Federal Research Institute for Animal Health
BMC Bioinformatics | Year: 2015

Background: Fuelled by the advent and subsequent development of next generation sequencing technologies, metagenomics became a powerful tool for the analysis of microbial communities both scientifically and diagnostically. The biggest challenge is the extraction of relevant information from the huge sequence datasets generated for metagenomics studies. Although a plethora of tools are available, data analysis is still a bottleneck. Results: To overcome the bottleneck of data analysis, we developed an automated computational workflow called RIEMS - Reliable Information Extraction from Metagenomic Sequence datasets. RIEMS assigns every individual read sequence within a dataset taxonomically by cascading different sequence analyses with decreasing stringency of the assignments using various software applications. After completion of the analyses, the results are summarised in a clearly structured result protocol organised taxonomically. The high accuracy and performance of RIEMS analyses were proven in comparison with other tools for metagenomics data analysis using simulated sequencing read datasets. Conclusions: RIEMS has the potential to fill the gap that still exists with regard to data analysis for metagenomics studies. The usefulness and power of RIEMS for the analysis of genuine sequencing datasets was demonstrated with an early version of RIEMS in 2011 when it was used to detect the orthobunyavirus sequences leading to the discovery of Schmallenberg virus. © 2015 Scheuch et al.; licensee BioMed Central.

PubMed | Federal Research Institute for Animal Health, Ghent University, Institute of Tropical Medicine, Bangladesh Agricultural University and 3 more.
Type: | Journal: Zoonoses and public health | Year: 2017

To determine the role of different Brucella (B.) spp. in Bangladesh, 62 animal samples and 500 human sera were tested. Animal samples from cattle, goats and sheep (including milk, bull semen, vaginal swabs and placentas) were cultured for Brucella spp. Three test-positive human sera and all animal samples were screened by Brucella genus-specific real-time PCR (RT-PCR), and positive samples were then tested by IS711 RT-PCR to detect B.abortus and B.melitensis DNA. Only B.abortus DNA was amplified from 13 human and six animal samples. This is the first report describing B.abortus as the aetiological agent of brucellosis in occupationally exposed humans in Bangladesh. Of note is failure to detect B.melitensis DNA, the species most often associated with human brucellosis worldwide. Further studies are required to explore the occurrence of Brucella melitensis in Bangladesh.

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