The aim of this study was to determine the prevalence, major management systems, and fresh cow clinical conditions associated with ketosis in western European dairy herds. A total of 131 dairies were enrolled in Germany, France, Italy, the Netherlands, and the United Kingdom during 2011 to 2012. A milk-based test for ketones (Keto-Test; Sanwa Kagaku Kenkyusho Co. Ltd., Nagoya, Japan; distributed by Elanco Animal Health, Antwerp, Belgium) was used for screening cows between d 7 and 21 after calving and ketosis was defined as a Keto-Test ≥100µmol/L. Study cows were observed for clinical disease up to 35d postcalving. Multivariate analysis (generalized estimating equation logistic regression) was performed to determine country, farm, management, feed, and cow factors associated with ketosis and to determine associations between ketosis and fresh cow diseases. Thirty-nine percent of the cows were classified as having ketosis. The herd average of ketosis was 43% in Germany, 53% in France, 31% in Italy, 46% in the Netherlands, and 31% in the United Kingdom. Of the 131 farms, 112 (85%) had 25% or more of their fresh cows resulting as positive for ketosis. Clinical ketosis was not reported in most farms and the highest level of clinical ketosis reported was 23%. The risks of ketosis were significantly lower in Italy and the United Kingdom compared with France, the Netherlands, and Germany. Larger herd size was associated with a decreased risk of ketosis. The farms that fed partially mixed rations had 1.5 times higher odds of ketosis than those that fed total mixed rations. Cows that calved in April to June had the highest odds of ketosis, with about twice as high odds compared with cows that calved in July to September. The cows that calved in January to March tended to have 1.5 times higher risk of ketosis compared with cows that calved in July to September. The odds of ketosis in parity 2 and parity 3 to 7 was significantly higher (1.5 and 2.8 times higher, respectively) than the odds of ketosis in parity 1. The odds of ketosis was significantly smaller in parity 2 compared with parity 3 to 7. Ketosis was associated with significantly higher odds of all common fresh cow conditions: metritis, mastitis, displaced abomasum, clinical ketosis, lameness, and gastrointestinal disorders. Odds of ketosis in cows having had twins or dystocia were not increased, whereas higher odds of ketosis were observed in cows with milk fever or retained placenta.

Ketosis is associated with many transition cow diseases and the subclinical form has been found to be a common condition in high-producing dairy cows. The objectives of this field study in the Netherlands were (1) to determine risk factors for subclinical ketosis [SCK; 1.2-2.9mmol of β-hydroxybutyrate (BHBA)/L of serum] and clinical ketosis (CK: ≥3.0mmol of BHBA/L of serum) at 7 to 14 d in milk and (2) to assess the association of SCK and CK with production parameters at the first dairy herd improvement (DHI) testing. Twenty-three dairies were enrolled by a local veterinary practice from 2009 to 2010, and 1,715 cows were screened for ketosis by measuring serum BHBA concentrations at 7 to 14 d in milk. Overall, 47.2% of cows had SCK and 11.6% had CK. Mixed generalized logit models with a random effect of herd were used to evaluate cow level factors associated with SCK and CK. The associations of SCK and CK with milk production parameters were tested using mixed linear models with a random effect of herd. Cows at a moderate (3.25-3.75) or fat (≥4) body condition score before calving were more likely to develop SCK and CK than thin (body condition score≤3.0) cows. The risk for developing SCK was higher in parity 2 and older cows compared with heifers, whereas for CK only, parity ≥3 cows had a higher risk. The quarter of the year in which a cow calved was associated with the risk for SCK and CK. For SCK quarter 1 (January-March) and quarter 2 (April-June), and for CK quarter 1, quarter 2, and quarter 3 (July-September) all increased the risk of development of the condition compared with quarter 4 (October-December). An increased yield of colostrum at first milking was associated with increasing risk for SCK and CK. Prolonged previous lactation length and dry period length were both associated with increased odds for SCK and CK. Subclinical ketosis and CK were associated with a higher milk yield, a higher milk fat percentage, and a lower milk protein percentage at first DHI test day. Overall the study reinforces previous findings that the major risk factors for both SCK and CK are increasing parity, overconditioning of animals prepartum, season of calving, and dry period length. In addition, previous lactation length and liters of colostrum have been identified as additional risk factors for the development of ketosis.

In situ forming bone substitute materials are attractive candidates for filling irregularly shaped defects. In this study, a chemically modified form of the Pluronic F127 hydrogel was used. Similar to the parent form, this derivative underwent a sol-gel transition in the body and additional radical curing resulted in a stable three-dimensional network gel with a controllable degradation rate. An extra cell source of autologous bone marrow-derived mesenchymal stem cells was mixed with the hydrogel to increase the ossification process, when implanted in noncritical size unicortical tibia defects. These cells were cultured and predifferentiated on two types of cell carrier systems, that is, gelatin CultiSpher-S microcarriers and hydroxyapatite tubular carriers. Radiographic and histological evaluation revealed that bone regeneration was comparable in the defects with the bone substitute compositions and the untreated control defects at 2 and 4 weeks postimplantation and that newly formed bone originated from the cells on the CultiSpher-S carriers. This resulted, 6 and 8 weeks postimplantation, in faster bone repair in the defects filled with the hydrogel plus CultiSpher-S carriers in comparison to the control defects. Surprisingly, there was no formation of new bone originating from the hydroxyapatite carriers. The hydrogel by itself seemed to stimulate the natural repair process.

An in situ crosslinkable, biodegradable, methacrylate-endcapped poly(D,L-lactide-co-e-caprolactone) in which crosslinkage is achieved by photoinitiators was developed for bone tissue regeneration. Different combinations of the polymer with bone marrow-derived mesenchymal stem cells (BMSCs) and a-tricalcium phosphate (a-TCP) were tested in a unicortical tibial defect model in eight goats. The polymers were randomly applied in one of three defects (6.0 mm diameter) using a fourth unfilled defect as control. Biocompatibility and bone-healing characteristics were evaluated by serial radiographies, histology, histomorphometry, and immunohistochemistry. The results demonstrated cell survival and proliferation in the polymer-substituted bone defects. The addition of a-TCP was associated with less expansion and growth of the BMSCs than other polymer composites.

Pluronic® F127 is a biocompatible, injectable, and thermoresponsive polymer with promising biomedical applications. In this study, a chemically modified form, i.e., Pluronic ALA-L with tailored degradation rate, was tested as an encapsulation vehicle for osteoblastic cells. UV cross-linking of the modified polymer results in a stable hydrogel with a slower degradation rate. Toxicological screening showed no adverse effects of the modified Pluronic ALA-L on the cell viability. Moreover, high viability of embedded cells in the cross-linked Pluronic ALA-L was observed with life/death fluorescent staining during a 7-day-culture period. Cells were also cultured on macroporous, cross-linked gelatin microbeads, called CultiSpher-S® carriers, and encapsulated into the modified cross-linked hydrogel. Also, in this situation, good cell proliferation and migration could be observed in vitro. Preliminary in vivo tests have shown the formation of new bone starting from the injected pre-loaded CultiSpher-S® carriers.