Animal welfare in cross-ventilated, compost-bedded pack, and naturally ventilated dairy barns in the upper Midwest.

The objective of this cohort study was to investigate animal welfare in 2 newer dairy housing options in the upper Midwest, cross-ventilated freestall barns (CV) and compost-bedded-pack barns (CB), compared with conventional, naturally ventilated freestall barns (NV). The study was conducted on 18 commercial dairy farms, 6 of each housing type, in Minnesota and eastern South Dakota. The primary breed in all farms was Holstein; 1 CV and 1 NV herd had approximately 30% Jersey-Holstein crossbreds. All freestall herds used sand for bedding. Farms were visited 4 times (once in each season) between January and November 2008, and approximately 93% of all animals in each pen were visually scored on each visit. Outcome-based measurements of welfare (locomotion, hock lesions, body condition score, hygiene, respiration rates, mortality, and mastitis prevalence) were collected on each farm. Lameness prevalence (proportion of cows with locomotion score ≥3 on a 1 to 5 scale, where 1=normal and 5=severely lame) in CB barns (4.4%) was lower than that in NV (15.9%) and CV (13.1%) barns. Lameness prevalence was similar between CV and NV barns. Hock lesion prevalence (proportion of cows with a lesion score ≥2 on a 1 to 3 scale, where 1=normal, 2=hair loss, and 3=swelling) was lower in CB barns (3.8%) than in CV (31.2%) and NV barns (23.9%). Hygiene scores (1 to 5 scale, where 1=clean and 5=very dirty) were higher for CB (3.18) than CV (2.83) and NV (2.77) barns, with no differences between CV and NV barns. Body condition scores, respiration rates, mastitis prevalence, culling, and mortality rates did not differ among housing systems. The CV and NV barns were evaluated using the cow comfort index (proportion of cows lying down in a stall divided by all animals touching a stall) and the stall usage index (proportion of cows lying divided by all animals in the pen not eating). The CV barns tended to have greater cow comfort index (85.9%) than the NV barns (81.4%) and had greater stall usage index (76.8% and 71.5%, respectively). Dairy cattle housed in CB barns had reduced lameness and hock lesions compared with those housed in freestall barns and had no adverse associations with body condition, respiration rates, mastitis prevalence, culling, or mortality. When comparing the 2 freestall housing options, CV barns had improved cow comfort indices compared with NV barns. Although cows in CB barns had better feet and leg health, as indicated by the reduced lameness and hock lesion prevalence, acquiring bedding and managing the bedded pack could limit their use.

Towards a recombinant vaccine against diphtheria toxin.

Two recombinant fragments of diphtheria toxin (DT) were fused to an engineered tandem repeat of the immunoglobulin (Ig) binding domain of protein A, called ZZ. These fragments are (i) the receptor binding domain (DTR), which comprises amino acids 382 to 535 of DT, and (ii) a linear peptide (DT(168-220)) which comprises residues 168 to 220 of the loop between fragment A and fragment B of DT. The fusion proteins were produced in Escherichia coli and purified by affinity chromatography. In vitro experiments showed that the DTR domain is responsible for the capacity of ZZ-DTR to bind to Vero cells and is capable of inhibiting the cytotoxicity of DT for these cells. These findings suggest that DTR binds to the cell surface receptors of DT and hence adopts a conformation that is similar to that of the receptor binding domain of DT. We compared the capacities of ZZ-DTR, ZZ-DT(168-220), and a chemically detoxified form of DT currently used for vaccination to elicit antibodies in rabbits. The toxoid was more immunogenic than ZZ-DT(168-220), which in turn was more immunogenic than ZZ-DTR. However, ZZ-DT(168-220) antiserum was poorly efficient at neutralizing DT cytotoxicity on Vero cells, whereas ZZ-DTR antiserum was only 15-fold less potent than anti-DT antisera.

Subunit composition, crystallization and preliminary crystallographic studies of the Desulfovibrio gigas aldehyde oxidoreductase containing molybdenum and [2Fe-2S] centers.

The Desulfovibrio gigas aldehyde oxidoreductase contains molybdenum bound to a pterin cofactor and [2Fe-2S] centers. The enzyme was characterized by SDS/PAGE, gel-filtration and analytical ultracentrifugation experiments. It was crystallized at 4 degrees C, pH 7.2, using isopropanol and MgCl2 as precipitants. The crystals diffract beyond 0.3-nm (3.0-A) resolution and belong to space group P6(1)22 or its enantiomorph, with cell dimensions a = b = 14.45 nm and c = 16.32 nm. There is one subunit/asymmetric unit which gives a packing density of 2.5 x 10(-3) nm3/Da (2.5 A3/Da), consistent with the experimental crystal density, rho = 1.14 g/cm3. One dimer (approximately 2 x 100 kDa) is located on a crystallographic twofold axis.

Isolation, crystallization, crystal structure analysis and refinement of B-phycoerythrin from the red alga Porphyridium sordidum at 2.2 A resolution.

The light-harvesting pigment-protein complex B-phycoerythrin from the red alga Porphyridium sordidum has been isolated and crystallized. B-Phycoerythrin consists of three different subunits forming an (alpha beta)6 gamma aggregate. The three-dimensional structure of the (alpha beta)6 hexamer was solved by Patterson search techniques using the molecular model of C-phycocyanin from Fremyella diplosiphon. The asymmetric unit of the crystal cell (space group P3, with a = b = 111.2 A, c = 59.9 A, alpha = beta = 90 degrees, gamma = 120 degrees) contains two (alpha beta) monomers related by a local dyad. Three asymmetric units are arranged around the crystallographic 3-fold axis building an (alpha beta)6 hexamer, as in C-phycocyanin. The crystal structure has been refined by energy-restrained crystallographic refinement and model building. The conventional R-factor of the final model was 18.9% with data to 2.2 A resolution. The molecular structures of the alpha and beta-subunits resemble those of C-phycocyanin. Major changes in comparison to phycocyanin are caused by deletion or insertion of segments involved in protein-chromophore interactions. The singly linked phycoerythrobilin chromophores alpha-84, alpha-140a, beta-84 and beta-155 are each covalently bound to a cysteine by ring A. The doubly linked chromophore beta-50/beta-61 is attached at cysteine beta-50 through ring A and at cysteine beta-61 through ring D. B-Phycoerythrin contains additionally a 30 kDa gamma-subunit, which is presumably located in the central cavity of the hexamer. It is disordered, as a consequence of crystal and local symmetry averaging.