Identification of Mycobacterium avium ssp. paratuberculosis infected dairy herds by environmental sampling

In herds with known prevalence (P) use of environmental sampling (ES) to detect Mycobacterium avium ssp. paratuberculosis (MAP) infected cattle herds was proofed in relation to P. In 31 MAP-infected free stall dairy herds and 15 non-infected herds P was defined by annually repeated whole herd testing by fecal culture (34 877 individual samples). Eight infected herds had a very low (> 0-2%), 14 a low (> 2-5%), four a medium (> 5-10%), and five a high P (> 10%). A mean number of nine environmental samples per herd were collected from the floor of lactating cows, milking, calving and sick cow areas and the crossover to the calf area. After twelve weeks cultivation on HEYM-medium with and without mycobactin positive samples were further characterized by PCR. All non-infected herds (100%) showed negative and 22 (71%) of the infected herds positive results in ES. Nine infected herds with negative ES results had a low P (0.04-4,04%). Proportion of positive ES depended on P and on sampling areas with 53.3% positive results in lactating cow areas and 45.2% in milking areas. For P > 5%, ES in these two areas caused a positive herd status; herds with P < 5% required sampling in the other areas too. The ES method has a herd sensitivity of 87% for dairy herds with P > 2% and provides an efficient tool to determine MAP infection status or herd prevalence.     

Tularemia in Alaska, 1938 - 2010

Tularemia is a serious, potentially life threatening zoonotic disease. The causative agent, Francisella tularensis, is ubiquitous in the Northern hemisphere, including Alaska, where it was first isolated from a rabbit tick (Haemophysalis leporis-palustris) in 1938. Since then, F. tularensis has been isolated from wildlife and humans throughout the state. Serologic surveys have found measurable antibodies with prevalence ranging from < 1% to 50% and 4% to 18% for selected populations of wildlife species and humans, respectively. We reviewed and summarized known literature on tularemia surveillance in Alaska and summarized the epidemiological information on human cases reported to public health officials. Additionally, available F. tularensis isolates from Alaska were analyzed using canonical SNPs and a multi-locus variable-number tandem repeats (VNTR) analysis (MLVA) system. The results show that both F. t. tularensis and F. t. holarctica are present in Alaska and that subtype A.I, the most virulent type, is responsible for most recently reported human clinical cases in the state.     

Toxicokinetics of mercury in blood compartments and hair of fish-fed sled dogs

Background

Understanding mercury (Hg) distribution in blood and the importance of hair as an excretory pathway is critical for evaluating risk from long term dietary Hg exposure. The major objective of this study was to characterize changes in total Hg concentrations in specific blood compartments and hair over time due to long term piscivory.

Methods

Eight sled dogs (Canis lupus familiaris) were fed either a fish and kibble diet (n = 4), or a fish-free control diet (n = 4) for 12 weeks. Concentrations of Hg were monitored throughout the exposure period, and for 10 weeks post exposure, until Hg concentrations in all blood compartments of one of the exposed dogs dropped below detection limit. Additionally, foreleg hair was sampled during acclimation and weeks 0 and 12.

Results

Hg was detected primarily in whole blood and packed cells, although it was sporadically detected at low concentrations in plasma and serum in two of the fish fed dogs. Dogs ingested an estimated average of 13.4 ± 0.58 μg Hg per kg body weight per day. Hg was detectable in whole blood and packed cells within a week of exposure. Detected concentrations continued to rise until plateauing at approximately 3-6 weeks of exposure at a mean of 9.2 ± 1.97 ng/g (ppb) in whole blood. Hg concentration decreased post exposure following 1st order elimination. The mean half-life (t_1/2) in whole blood for Hg was 7 weeks. Mean Hg in hair for the fish-fed dogs at week 12 was 540 ± 111 ppb and was significantly greater (about 7-fold) than the Hg hair concentration for the control dogs. The hair to blood ratio for Hg in fish-fed dogs was 59.0 ± 7.6:1.

Conclusions

This study found the sled dog model to be an effective method for investigating and characterizing blood Hg distribution (whole blood, serum, plasma, packed cells) and toxicokinetics associated with a piscivorous diet, especially for Hg-exposed fur bearing mammals (such as polar bears). Although hair excretion and hair to blood Hg ratios were not similar to human concentrations and ratios, the sled dog toxicokinetics of Hg in blood, was more similar to that of humans than traditional laboratory animals (such as the rat).     

Initial stage of cheese production: a molecular modeling study of bovine and camel chymosin complexed with peptides from the chymosin-sensitive region of κ-casein

Bovine chymosin has long been the preferred enzyme used to coagulate cow's milk, in the initial stage of cheese production, during which it cleaves a specific bond in the milk protein κ-casein. Recently, camel chymosin has been shown to have a 70% higher clotting activity toward cow's milk and, moreover, to cleave κ-casein more selectively. Bovine chymosin, on the other hand, is a poor clotting agent toward camel's milk. This paper reports a molecular modeling study aimed at understanding this disparity, based on homology modeling and molecular dynamics simulations using peptide fragments of κ-casein from cow and camel in both bovine and camel chymosin. The results show that the complex between bovine chymosin and the fragment of camel κ-casein is indeed less stable in the binding pocket. The results also indicate that this in part may be due to charge repulsion between a lysine residue in bovine chymosin and an arginine residue in the P4 position of camel κ-casein.     

Effects of storage atmosphere and heme state on the color and visible reflectance spectra of salmon ( Salmo salar ) fillets

It has previously been observed that the color of mackerel muscle is dependent on the status of heme as myoglobin and hemoglobin and hence the storage atmosphere. This study gives strong indications of this being the case also in salmon. Three different storage conditions were used to promote the oxidized, reduced, and carbon monoxide (CO) bound forms of heme in salmon and mackerel fillets. Color determination (instrumental color analysis, imaging, and sensory evaluation) and spectroscopic measurements were performed to study how spectral changes corresponded to color variations. Storage in CO significantly increased the redness in mackerel. This was also seen in salmon to such a degree that it was visible over normal levels of salmon carotenoids. Air storage increased the yellowness and reduced the redness in mackerel, but this effect was partly concealed in salmon by the astaxanthin absorption. The spectral differences due to storage condition could be ascribed to the spectral features characterizing heme of different oxidation states and bound to different ligands. The status of heme should therefore always be considered when experiments related to salmon color are performed. The findings could help in the understanding, control, and prediction of color loss in salmon during processing, storage, and transport.     

Recovery of infectious virus from full-length cowpox virus (CPXV) DNA cloned as a bacterial artificial chromosome (BAC)

ABSTRACT: Transmission from pet rats and cats to humans as well as severe infection in felids and other animal species have recently drawn increasing attention to cowpox virus (CPXV). We report the cloning of the entire genome of cowpox virus strain Brighton Red (BR) as a bacterial artificial chromosome (BAC) in Escherichia coli and the recovery of infectious virus from cloned DNA. Generation of a full-length CPXV DNA clone was achieved by first introducing a mini-F vector, which allows maintenance of large circular DNA in E. coli, into the thymidine kinase locus of CPXV by homologous recombination. Circular replication intermediates were then electroporated into E. coli DH10B cells. Upon successful establishment of the infectious BR clone, we modified the full-length clone such that recombination-mediated excision of bacterial sequences can occur upon transfection in eukaryotic cells. This self-excision of the bacterial replicon is made possible by a sequence duplication within mini-F sequences and allows recovery of recombinant virus progeny without remaining marker or vector sequences. The in vitro growth properties of viruses derived from both BAC clones were determined and found to be virtually indistinguishable from those of parental, wild-type BR. Finally, the complete genomic sequence of the infectious clone was determined and the cloned viral genome was shown to be identical to that of the parental virus. In summary, the generated infectious clone will greatly facilitate studies on individual genes and pathogenesis of CPXV. Moreover, the vector potential of CPXV can now be more systematically explored using this newly generated tool.     

Swine infection with Trichinella spiralis: Comparative analysis of the mucosal intestinal and systemic immune responses

The immune protective response developed by swine against Trichinella spiralis is not yet fully understood, particularly at the mucosal level. This study aimed to characterise intestinal immunity to T. spiralis by comparison with the systemic response in specifically pathogen-free pigs. For this purpose, the kinetics of cytokine and antibody production were assessed in the intestinal mucosa and serum of swine infected with T. spiralis for up to 60 days post-infection (dpi). An ex vivo model of jejunum mucosa culture was used to collect the supernatant as a source of antibodies (Abs). Mucosal antibodies were observed by Western blot from 15 dpi, while serum antibodies were expressed from 20 dpi. Both sources of antibodies initially recognized a 110 kDa protein, followed by the identification of 35, 43/46 and 55/59 kDa proteins. IgG1 and IgA antibodies were strongly expressed within the mucosa. The expression levels of Type 1 (IFN-gamma, IL-12), Type 2 (IL-4, IL-6), pro-inflammatory (TNF-alpha) and regulatory (IL-10, TGF-beta) cytokines were assessed by RT-PCR in the intestinal mucosa and spleen. Both IL-10 and IFN-gamma mRNA levels were increased in mucosa, whereas IL-6 and IL-12 mRNA were expressed in spleen. Taken together, these results demonstrated a mixed Type 1/Type 2 profile, the Type 2 profile being dominant in the mucosa.     

Electrical anisotropy below slow- and fast-moving plates: paleoflow in the upper mantle?

Upper mantle electrical conductivities can be explained by hydrogen diffusivity in hydrous olivine. Diffusivity enhances the conductivity of olivine anisotropically, making the a axis the most conductive of the three axes. Therefore, the hypothesis that plate motion induces lattice-preferred orientation of olivine can be tested with the use of long-period electromagnetic array measurements. Here, we compared electrical anisotropies below the slow-moving Fennoscandian and fast-moving Australian plates. The degree of olivine alignment is greater in the mantle below the Fennoscandian plate than below the Australian plate. This finding may indicate that convection rather than plate motion is the dominant deformation mechanism.     

Metabolomic responses in grain,ear, and straw of winter wheat under increasing sulfur treatment

Only little is known about the effect of a varying sulfur (S) nutrition on the pattern of metabolites in different organs of the ears of winter wheat (Tritcum aestivum L.) at final maturity. More insights into the metabolome as influenced by increasing S‐fertilizer rates would, however, be of particular interest in order to unravel S‐dependent physiological processes related to grain filling in wheat. We have therefore investigated the effects of varying sulfur nutrition on metabolite composition and distribution in the organs of the wheat ear and vegetative organs at final maturity. Gas chromatography–mass spectrometry–based metabolite profiles revealed that S deficiency decreased the bulk of metabolites in the straw in favor of an increasing metabolite concentration in the husk, rachis, and grains. Surprisingly, only four out of 109 detectable metabolites, namely N‐acetyl glucosamine, lysine, ferulic acid, and β‐aminoisobutyric acid were most responsible for organ‐specific differences in the metabolite profiles. Under S‐deficient conditions, N‐acetyl‐glucosamine, lysine, and β‐aminoisobutyric were increasingly transferred from source tissues into the ears and grains.     

Redox control on carbon mineralization and dissolved organic matter along a chronosequence of paddy soils

Paddy soils are subjected to periodically changing redox conditions. In order to understand better the redox control on long‐term carbon turnover, we assessed carbon mineralization and dissolved organic carbon (DOC) of paddy topsoils sampled along a chronosequence spanning 2000 years of rice cultivation. Non‐paddy soils were used as references. We exposed soils to alternating redox conditions for 12 weeks in incubation experiments. Carbon mineralization of paddy soils was independent of redox conditions. Anoxic conditions caused increasing DOC concentrations for paddy soils, probably because of desorption induced by increasing pH. We assume desorption released older, previously stabilized carbon, which then was respired by a microbial community well adapted to anoxic conditions. This assumption is supported by the ¹⁴C signatures of respired CO₂, indicating larger mineralization of older carbon under anoxic than under oxic conditions. The increasing DOC concentrations under anoxic conditions did not result in an equivalent increase in carbon mineralization, possibly because of little reducible iron oxide. Therefore, net DOC and CO₂ production were not positively related under anoxic conditions. The overall 20–75% smaller carbon mineralization of paddy soils than of non‐paddy soils resulted from less respiration under oxic conditions. We conclude that carbon accumulation in paddy as well as in other wetland soils results from a microbial community well adapted to anoxic conditions, but less efficient in mineralizing carbon during transient oxic periods. Carbon accumulation might be even larger when mineralization under anoxic conditions is restricted by a lack of alternative electron acceptors.