Fate of recombinant DNA and Cry1Ab protein after ingestion and dispersal of genetically modified maize in comparison to rapeseed by fallow deer ( Dama dama )

The fate of recombinant DNA in fallow deer (Dama dama) was investigated by feeding a diet of isogenic or genetically modified (GM) maize expressing Cry1Ab protein against the European corn borer (Ostrinia nubilalis). To study the degradability of ingested DNA, polymerase chain reaction (PCR) assays were introduced to detect fragments of the endogenous, highly abundant chloroplast-specific rubisco gene, the maize-specific zein gene and the recombinant cry1Ab gene. PCR analysis revealed that small chloroplast- and maize-specific DNA fragments were detectable in contents of rumen, abomasums, jejunum, caecum and colon and occasionally in visceral tissues. In contrast, no fragments of the recombinant cry1Ab gene were detectable in gastrointestinal (GI) contents. The Cry1Ab protein was analysed using an enzyme-linked immunosorbent assay (ELISA) and immunoblotting technique. Neither ELISA nor immunoblotting yielded positive signals of immunoactive Cry1Ab protein in GI contents and tissues of fallow deer fed with GM maize. In conclusion, after uptake of GM maize, neither cry1Ab-specific gene fragments nor Cry1Ab protein were detected in the GI tract of fallow deer, indicating complete digestion of the GM maize. Additional investigations on the germination capacity of conventional rapeseed and maize seed after ingestion by fallow deer and faecal excretion (endozoochory) were performed to draw conclusions regarding a potential spreading of germinable GM crop seed by deer. Germination tests revealed that germinable rapeseed kernels were detectable in faeces; in contrast, no intact maize seeds were found in faece

Degradation of Cry1Ab protein from genetically modified maize (MON810) in relation to total dietary feed proteins in dairy cow digestion

To investigate the relative degradation and fragmentation pattern of the recombinant Cry1Ab protein from genetically modified (GM) maize MON810 throughout the gastrointestinal tract (GIT) of dairy cows, a 25 months GM maize feeding study was conducted on 36 lactating Bavarian Fleckvieh cows allocated into two groups (18 cows per group) fed diets containing either GM maize MON810 or nearly isogenic non-GM maize as the respective diet components. All cows were fed a partial total mixed ration (pTMR). During the feeding trial, 8 feed (4 transgenic (T) and 4 non-transgenic (NT) pTMR) and 42 feces (26 T and 18 NT) samples from the subset of cows fed T and NT diets, and at the end of the feeding trial, digesta contents of rumen, abomasum, small intestine, large intestine and cecum were collected after the slaughter of six cows of each feeding group. Samples were analyzed for Cry1Ab protein and total protein using Cry1Ab specific ELISA and bicinchoninic acid assay, respectively. Immunoblot analyses were performed to evaluate the integrity of Cry1Ab protein in feed, digesta and feces samples. A decrease to 44% in Cry1Ab protein concentration from T pTMR to the voided feces (9.40 versus 4.18 μg/g of total proteins) was recorded. Concentrations of Cry1Ab protein in GIT digesta of cows fed T diets varied between the lowest 0.38 μg/g of total proteins in abomasum to the highest 3.84 μg/g of total proteins in rumen. Immunoblot analysis revealed the extensive degradation of recombinant Cry1Ab protein into a smaller fragment of around 34 kDa in GIT. The results of the present study indicate that the recombinant Cry1Ab protein from MON810 is increasingly degraded into a small fragment during dairy cow digestion

Trans-Atlantic exchanges have shaped the population structure of the Lyme disease agent Borrelia burgdorferi sensu stricto

The origin and population structure of Borrelia burgdorferi sensu stricto (s.s.), the agent of Lyme disease, remain obscure. This tick-transmitted bacterial species occurs in both North America and Europe. We sequenced 17 European isolates (representing the most frequently found sequence types in Europe) and compared these with 17 North American strains. We show that trans-Atlantic exchanges have occurred in the evolutionary history of this species and that a European origin of B. burgdorferi s. s. is marginally more likely than a USA origin. The data further suggest that some European human patients may have acquired their infection in North America. We found three distinct genetically differentiated groups: i) the outgroup species Borrelia bissettii, ii) two divergent strains from Europe, and iii) a group composed of strains from both the USA and Europe. Phylogenetic analysis indicated that different genotypes were likely to have been introduced several times into the same area. Our results demonstrate that irrespective of whether B. burgdorferi s. s. originated in Europe or the USA, later trans-Atlantic exchange(s) have occurred and have shaped the population structure of this genospecies. This study clearly shows the utility of next generation sequencing to obtain a better understanding of the phylogeography of this bacterial species

Fate of recombinant DNA and Cry1Ab protein after ingestion and dispersal of genetically modified maize in comparison to rapeseed by fallow deer (Dama dama

Abstract The fate of recombinant DNA in fallow deer (Dama dama) was investigated by feeding a diet of isogenic or genetically modified (GM) maize expressing Cry1Ab protein against the European corn borer (Ostrinia nubilalis). To study the degradability of ingested DNA, polymerase chain reaction (PCR) assays were introduced to detect fragments of the endogenous, highly abundant chloroplast-specific rubisco gene, the maize-specific zein gene and the recombinant cry1Ab gene. PCR analysis revealed that small chloroplast-and maize-specific DNA fragments were detectable in contents of rumen, abomasums, jejunum, caecum and colon and occasionally in visceral tissues. In contrast, no fragments of the recombinant cry1Ab gene were detectable in gastrointestinal (GI) contents. The Cry1Ab protein was analysed using an enzyme-linked immunosorbent assay (ELISA) and immunoblotting technique. Neither ELISA nor immunoblotting yielded positive signals of immunoactive Cry1Ab protein in GI contents and tissues of fallow deer fed with GM maize. In conclusion, after uptake of GM maize, neither cry1Ab-specific gene fragments nor Cry1Ab protein were detected in the GI tract of fallow deer, indicating complete digestion of the GM maize. Additional investigations on the germination capacity of conventional rapeseed and maize seed after ingestion by fallow deer and faecal excretion (endozoochory) were performed to draw conclusions regarding a potential spreading of germinable GM crop seed by deer. Germination tests revealed that germinable rapeseed kernels were detectable in faeces; in contrast, no intact maize seeds were found in faeces

The origin of mid-infrared emission in massive young stellar objects: multi-baseline VLTI observations of W33A

The circumstellar structure on 100 AU scales of the massive young stellar object W33A is probed using the VLTI and the MIDI instrument. N-band visibilities on 4 baselines are presented which are inconsistent with a spherically symmetric geometry. The visibility spectra and SED are simultaneously compared to 2D axi-symmetric dust radiative transfer models with a geometry including a rotationally flattened envelope and outflow cavities. We assume an O7.5 ZAMS star as the central source, consistent with the observed bolometric luminosity. The observations are also compared to models with and without (dusty and gaseous) accretion disks. A satisfactory model is constructed which reproduces the visibility spectra for each (u,v) point. It fits the silicate absorption, the mid-IR slope, the far-infrared peak, and the (sub)mm of the SED. It produces a 350 micron morphology consistent with observations. The 10 micron emission on 100 AU scales is dominated by the irradiated walls of the cavity sculpted by the outflow. The visibilities rule out the presence of dust disks with total (gas and dust) masses more than 0.01 Msun. However, optically thick accretion disks, interior to the dust sublimation radius, are allowed to accrete at rates equalling the envelope's mass infall rate (up to 10^(-3) Msun/yr) without substantially affecting the visibilities due to the extinction by the extremely massive envelope of W33A.Comment: Accepted for publication in A&

Probing the envelopes of massive young stellar objects with diffraction limited mid-infrared imaging

Massive stars form whilst they are still embedded in dense envelopes. As a result, the roles of rotation, mass loss and accretion in massive star formation are not well understood. This study evaluates the source of the Q-band, lambda=19.5 microns, emission of massive young stellar objects (MYSOs). This allows us to determine the relative importance of rotation and outflow activity in shaping the circumstellar environments of MYSOs on 1000 AU scales. We obtained diffraction limited mid-infrared images of a sample of 20 MYSOs using the VLT/VISIR and Subaru/COMICS instruments. For these 8 m class telescopes and the sample selected, the diffraction limit, ~0.6", corresponds to approximately 1000 AU. We compare the images and the spectral energy distributions (SEDs) observed to a 2D, axis-symmetric dust radiative transfer model that reproduces VLTI/MIDI observations of the MYSO W33A. We vary the inclination, mass infall rate, and outflow opening angle to simultaneously recreate the behaviour of the sample of MYSOs in the spatial and spectral domains. The mid-IR emission of 70 percent of the MYSOs is spatially resolved. In the majority of cases, the spatial extent of their emission and their SEDs can be reproduced by the W33A model featuring an in-falling, rotating dusty envelope with outflow cavities. There is independent evidence that most of the sources which are not fit by the model are associated with ultracompact HII regions and are thus more evolved. We find that, in general, the diverse 20 micron morphology of MYSOs can be attributed to warm dust in the walls of outflow cavities seen at different inclinations. This implies that the warm dust in the outflow cavity walls dominates the Q-band emission of MYSOs. In turn, this emphasises that outflows are an ubiquitous feature of massive star formation.Comment: Accepted for publication in A&A. The images in this version have been compressed. A high resolution version is available on reques

The Physical Conditions for Massive Star Formation: Dust Continuum Maps and Modeling

Fifty-one dense cores associated with water masers were mapped at 350 micron. These cores are very luminous, 10^3 < Lbol/Lsun < 10^6, indicative of the formation of massive stars. Dust continuum contour maps and photometry are presented for these sources. The spectral energy distributions and normalized radial profiles of dust continuum emission were modeled for 31 sources using a one-dimensional dust radiative transfer code, assuming a power law density distribution in the envelope, n = n_f (r/r_f)^{-p}. The best fit density power law exponent, p, ranged from 0.75 to 2.5 with = 1.8 +/- 0.4. The mean value of p is comparable to that found in regions forming only low mass stars. The mean p is incompatible with a logatropic sphere (p = 1), but other star formation models cannot be ruled out. Different mass estimates are compared and mean masses of gas and dust are reported within a half-power radius determined from the dust emission and within a radius where the total density exceeds 10^4 cm^3. Evolutionary indicators commonly used for low mass star formation may have some utility for regions forming massive stars. For comparison with extragalactic star formation studies, the luminosity to dust mass ratio is calculated for these sources with a method most parallel to that used in studies of distant galaxies and is found to be similar to that seen in high redshift starburst galaxies.Comment: 45 pages, 20 figures, accepted to ApJ Supplemen

Resolved 24.5 micron emission from massive young stellar objects

Massive young stellar objects (MYSO) are surrounded by massive dusty envelopes. Our aim is to establish their density structure on scales of ~1000 AU, i.e. a factor 10 increase in angular resolution compared to similar studies performed in the (sub)mm. We have obtained diffraction-limited (0.6") 24.5 micron images of 14 well-known massive star formation regions with Subaru/COMICS. The images reveal the presence of discrete MYSO sources which are resolved on arcsecond scales. For many sources, radiative transfer models are capable of satisfactorily reproducing the observations. They are described by density powerlaw distributions (n(r) ~ r^(-p)) with p = 1.0 +/-0.25. Such distributions are shallower than those found on larger scales probed with single-dish (sub)mm studies. Other sources have density laws that are shallower/steeper than p = 1.0 and there is evidence that these MYSOs are viewed near edge-on or near face-on, respectively. The images also reveal a diffuse component tracing somewhat larger scale structures, particularly visible in the regions S140, AFGL 2136, IRAS 20126+4104, Mon R2, and Cep A. We thus find a flattening of the MYSO envelope density law going from ~10 000 AU down to scales of ~1000 AU. We propose that this may be evidence of rotational support of the envelope (abridged).Comment: 21 pages, accepted for A&

Effects of Feeding Bt MON810 Maize to Pigs for 110 Days on Peripheral Immune Response and Digestive Fate of the cry1Ab Gene and Truncated Bt Toxin

peer-reviewedBackground: The objective of this study was to evaluate potential long-term (110 days) and age-specific effects of feeding genetically modified Bt maize on peripheral immune response in pigs and to determine the digestive fate of the cry1Ab gene and truncated Bt toxin. Methodology/Principal Findings: Forty day old pigs (n = 40) were fed one of the following treatments: 1) isogenic maize-based diet for 110 days (isogenic); 2) Bt maize-based diet (MON810) for 110 days (Bt); 3) Isogenic maize-based diet for 30 days followed by Bt maize-based diet for 80 days (isogenic/Bt); and 4) Bt maize-based diet (MON810) for 30 days followed by isogenic maize-based diet for 80 days (Bt/isogenic). Blood samples were collected during the study for haematological analysis, measurement of cytokine and Cry1Ab-specific antibody production, immune cell phenotyping and cry1Ab gene and truncated Bt toxin detection. Pigs were sacrificed on day 110 and digesta and organ samples were taken for detection of the cry1Ab gene and the truncated Bt toxin. On day 100, lymphocyte counts were higher (P<0.05) in pigs fed Bt/isogenic than pigs fed Bt or isogenic. Erythrocyte counts on day 100 were lower in pigs fed Bt or isogenic/Bt than pigs fed Bt/isogenic (P<0.05). Neither the truncated Bt toxin nor the cry1Ab gene were detected in the organs or blood of pigs fed Bt maize. The cry1Ab gene was detected in stomach digesta and at low frequency in the ileum but not in the distal gastrointestinal tract (GIT), while the Bt toxin fragments were detected at all sites in the GIT. Conclusions/Significance: Perturbations in peripheral immune response were thought not to be age-specific and were not indicative of Th 2 type allergenic or Th 1 type inflammatory responses. There was no evidence of cry1Ab gene or Bt toxin translocation to organs or blood following long-term feeding.The research leading to these results has received funding from the European Union’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 211820 and the Teagasc Walsh Fellowship programme