Evaluation of a new sandwich enzyme-linked immunosorbent assay for detection of bovine viral diarrhea virus in unprocessed fetal bovine serum.

A new sandwich enzyme-linked immunosorbent assay (S-ELISA) kit that uses raw (unprocessed) fetal bovine serum (FBS) as the testing sample was evaluated for upstream bovine viral diarrhea virus (BVDV) testing. Pooled FBS samples (n = 84) were tested using the S-ELISA. Thirty serum samples originating from persistently infected (PI) calves that had been confirmed by virus isolation (VI) as BVDV positive and another 30 samples previously confirmed by VI as BVDV negative were also evaluated. Of the 84 field samples, the S-ELISA detected 13 (15.5%) BVDV-positive specimens. When these 13 positive samples were tested by VI and immunofluorescent assay, 11 (84.6%) were positive and 2 (15.4%) were negative. The S-ELISA was positive for all 30 PI samples (100%) and negative for all 30 negative samples (100%). These data indicate that the new kit is a relatively reliable diagnostic tool and can be considered for upstream detection of BVDV-contaminated raw FBS pools.

The effect of omitted milking on the behaviour of cows in the context of cluster attachment failure during automatic milking.

In robotic milking there is always a slight chance of failure to attach the milking cluster. Attachment failure is most likely for cows whose udder conformation is less convenient for robot attachment. In general, after milking failure cows try to revisit the milking robot if they are not sent to a separate area. Since it is difficult to estimate the effect of milking failure on such a cow and her welfare in conditions of robotic milking, a specific 16-day trial was conducted on 12 cows. These cows were milked in a milking parlour with six milking stalls. Each afternoon milking, three cows were not milked. All the cows were closely observed in the cubicle house for 1 h after the afternoon milking. Thereafter, all cows were brought to the milking parlour the third time and the three unmilked cows were milked. In total, each cow was observed 12 times after milking and four times after omitted milking. The following behavioural traits were registered: time budget for the 1 h, occurrence and time until eating, drinking, lying, urination and defecation, and aggressive interactions. Milking order was defined on the basis of how often a cow came to the milking parlour in the first batch of six cows. Moreover, the data related to the milk yield and the use of the automatic feeding installation with the complete diet were analysed. After omitted milking, only the cows from the first batch stood longer in cubicles (14.2 min of 1 h) and lay less (5.4 min of 1 h) than milked cows of the same batch (respectively 7.0 min and 16.3 min for standing and lying in cubicles) (P<0.01). After omitted milking, cows urinated earlier and more frequently (64.5%) than milked cows (36.3%) (P<0.002) (both batches). There were no statistically significant differences in eating time and feed intake after milking and omitted milking. Milk yield per cow averaged 24.9 kg during days with omitted (delayed by 1 h) milking and 25.3 kg during the days without omitted milking (P<0.05). It was concluded that cows show some signs of discomfort after omitted milking (urination); this discomfort seemed to be greater in cows coming earlier to the milking parlour (afterwards they preferred to stand rather than to lie). The 60% of cases of milk leakage found after omitted milking indicates that failed cluster attachment can be accompanied by an extra risk factor for the occurrence of mastitis. However, omitted milking as a treatment did not influence feeding and aggressive behaviour or milking order when unmilked cows were brought to the milking parlour the third time together with the milked cows. Our methods and results can be useful for estimating the effects of robot milking failures on a cow. Future studies should pay particular attention to high-yielding cows and to longer periods of delayed milking.

Microstructural characteristics of calcium hydroxyapatite/poly-L-lactide based composites.

Besides its high osteoinductive properties, hydroxyapatite (HAp) exhibits a relatively low mechanical strength. In order to improve the mechanical properties and reliability of HAp based composites, the addition of selected polymers is highly recommended. The main objective of this work is to study the microstructural characteristics of HAp/poly-L-lactide (PLLA) composites obtained by cold or hot processing. The composites were prepared from a mixture of a chloroform solution of poly-L-lactide with granulated HAp. After elimination of chloroform by vacuum evaporation, dense compacts were obtained by cold or hot pressing. The pressing pressure ranged from 98.10 to 294.3 MPa for both cold and hot pressing. The hot pressing was performed in the temperature region 293-457 K for a time period of 15-60 min. Depending on the PLLA amount and the pressing procedure it is possible to obtain highly porous or nearly fully dense composites. The scanning electron microscopy examination of fracture as well as of free surfaces revealed that the final porosity and wetting are affected to a great extent by the synthesis conditions and amount of polymer added. An increase in temperature to 457 K for a longer period of time results in fully dense compacts. The formation of a nearly continuous polymer network that leads to the hardening of HAp has also been observed. However, it should be pointed out that some layers of HAp may be free of polymer film since PLLA penetrates more deeply into the porous HAp.

Synthesis and properties of hydroxyapatite/poly-L-lactide composite biomaterials.

Calcium hydroxyapatite (HAp) and poly-L-lactide (PLLA) were synthesized chemically. The obtained HAp was of high purity and, after special thermal treatment, of high crystallinity as well. Synthesis of PLLA was performed using L-lactide as a monomer and nontoxic initiator. In this way a polymer of large molar weight (about 400,000) was obtained. The HAp and PLLA obtained were used as constituents of the HAp/PLLA composite biomaterial, a potential material for implants. The composite was obtained by mixing completely dissolved PLLA with granules of HAp. The composite was compacted by cold and hot pressing at pressures of 49.0-490.5 MPa and temperatures of 20-184 degrees C. The material obtained at optimum process parameters had a density of 99.6% and compressive strength of 93.2 MPa.