Genetic potential for residual feed intake and diet fed during early- to mid-gestation influences post-natal DNA methylation of imprinted genes in muscle and liver tissues in beef cattle

Discovery of epigenetic modifications associated with feed efficiency or other economically important traits would increase our understanding of the molecular mechanisms underlying these traits. In combination with known genetic markers, this would provide opportunity to improve genomic selection accuracy in cattle breeding programs. It would also allow cattle to be managed to improve favorable gene expression. The objective of this study was to identify variation in DNA methylation between beef cattle of differential pre-natal nutrition and divergent genetic potential for residual feed intake (RFI). Purebred Angus offspring with the genetic potential for either high (HRFI) or low (LRFI) RFI were prenatally exposed to either a restricted maternal diet of 0.5 kg/d average daily gain (ADG) or a moderate maternal diet of 0.7 kg/d ADG from 30 to 150 d of gestation. We performed DNA methylation analysis of differentially methylated regions (DMR) of imprinted genes (Insulin-like growth factor 2 (IGF2) DMR2, IGF2/H19 imprinting control region (ICR) and IGF2 receptor (IGF2R) DMR2) using post-natal samples of longissimus dorsi (LD) muscle taken from male and female calves at birth and weaning, and of LD muscle, semimembranosus (SM) muscle, and liver samples collected from steers at slaughter (17 months of age). Interestingly, for all three DMR investigated in liver, LRFI steers had higher levels of methylation than HRFI steers. In LD muscle, IGF2/H19 ICR methylation differences for heifers at birth were due to pre-natal diet, while for steers at birth they were mostly the result of genetic potential for RFI with LRFI steers again having higher levels of methylation than HRFI steers. While results from repeated measures analysis of DNA methylation in steers grouped by RFI revealed few differences, in steers grouped by diet, we found higher methylation levels of IGF2 DMR2 and IGF2R DMR2 in LD muscle of restricted diet steers at weaning and slaughter than at birth, as well as increased methylation in LD muscle of restricted diet steers compared with moderate diet steers at weaning and/or slaughter. Our results suggest that differential pre-natal nutrition, and divergent genetic potential for RFI, induces tissue- and sex-specific alterations in post-natal IGF2 and IGF2R methylation patterns and that these patterns can vary with age in Angus beef cattle.     

Soil microbial communities resistant to changes in plant functional group composition

The soil community is an often ignored part of research which links plant biodiversity and ecosystem functioning despite their influence on numerous functions such as decomposition and nutrient cycling. Few consistent patterns have been detected that link plant and soil community composition. We used a removal experiment in a northern Canadian grassland to examine the effects of plant functional group identity on soil microbial community structure and function. Plant functional groups (graminoids, legumes and forbs) were removed independently from plots for five growing seasons (2003-2007) and in the fifth year effects on the soil microbial community were examined using substrate-induced respiration (SIR - a measure of metabolic diversity) and phospholipid fatty acid analysis (PLFA - a measure of microbial community composition). Removal treatments were also crossed with both a fertilizer treatment and a fungicide treatment to determine if effects of functional group identity on the soil community were context dependent. Plant functional group identity had almost no effect on the soil microbial community as measured by either SIR or PLFA. Likewise, soil properties including total carbon, pH, moisture and nutrients showed a limited response to plant removals in the fifth year after removals. We found a direct effect of fertilizer on the soil community, with fertilized plots having decreased metabolic diversity, with a decreased ability to metabolize amino acids and a phenolic acid, but there was no direct soil microbial response to fungicide. We show that in this northern Canadian grassland the soil microbial community is relatively insensitive to changes in plant functional group composition, and suggest that in northern ecosystems, where plant material is only slowly incorporated into the soil, five growing seasons may be insufficient to detect the impact of a changing plant community on the soil microbes.     

The lipid compositions of two isolates of Cladosporium cucumerinum do not explain their differences in sensitivity to fungicides which inhibit sterol biosynthesis

Isolate 840905 of Cladosporium cucumerinum, when grown on agar or in liqiud medium, was sensitive to triadimenol, HWG 1608 (tebuconazole), fenpropimorph and pimaricin but relatively resistant to terbinafine. Conversely, isolate 49628 was sensitive to terbinafine but relatively resistant to the other fungicides. Changes in sterol composition following treatment with the fungicides reflected the known modes of action of each fungicide. When individual enantiomers of triadimenol were tested against isolate 840905 the order of activity in reducing mycelial growth was 1 S, 2R > 1R, 2R > 1R, 2S ≈? 1S, 2S, and this was paralleled by the depletion of ergosterol and the appearance of 14α-methyl sterols. Isolate 49628 had a greater saturated:unsaturated fatty acid ratio than did isolate 840905 but no major changes in fatty acid composition of either isolate were induced by fungicide treatment. There appears to be no obvious explanation for the differences in fungicide sensitivity of the isolates in terms of their lipid compositions.     

Effects of pico-tesla electromagnetic field treatment on wound healing in rats

OBJECTIVE: To evaluate the effects of a pico-tesla electromagnetic field (PTEF) on healing of sutured and open skin wounds and clinicopathologic variables in rats. ANIMALS: 64 male Fischer-344 rats. PROCEDURE: An incision made in the dorsal aspect of the neck was sutured (n = 32) or left open to heal (32). In each group, 16 rats were not PTEF-treated (controls). Wound treatment consisted of exposure to a PTEF once daily. Rats in each group were euthanatized at days 2, 4, 7, and 14. Wounds were evaluated via tensiometry (sutured wounds), digital planimetry (open wounds), laser Doppler perfusion imaging, bacteriologic culture, and histologic examination. Blood samples were collected from all rats for analysis. RESULTS: At day 14, sutured wounds in PTEF-treated rats were stronger (ultimate stress) and tougher (strain energy) than were sutured wounds in control rats. Open wounds in PTEF-treated rats contracted more quickly at days 2 and 4 than did those in control rats. Compared with control wounds, histologic changes (indicative of improved healing) in sutured and open wounds in PTEF-treated rats were detected as early as day 4. Laser Doppler perfusion measurements, results of CBCs, serum biochemical analyses, and bacteriologic cultures were not different between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure to the PTEF caused no adverse effects on clinicopathologic, histologic, or bacteriologic variables tested in this study. It appears that PTEF is a safe form of adjuvant treatment for wounds and improves strength of sutured wounds and speeds contraction of open wounds.     

Postoperative integrity of veterinary surgical gloves

A multicenter, prospective study was performed to document the incidence of defective gloves postoperatively in veterinary surgery and to correlate defects with a variety of influencing factors. Gloves were collected after surgical procedures performed by the small animal clinical services at two veterinary teaching hospitals and one institution's student surgery laboratories. Gloves were evaluated for defects using electrical resistance testing. The overall incidence of glove defects was 23.3%. Significantly more defects occurred in gloves used for nonsoft-tissue procedures and in gloves worn on the nondominant hand. Eighty-four percent of all defects occurred in procedures lasting >60 minutes. No differences were detected in the brands of gloves used nor among surgeons of different experience levels. The individuals performing the surgery were not able to accurately predict the presence of a defect in their gloves. Surgeons should remain alert for possible glove defects and consider measures such as changing gloves every 60 minutes or double-gloving to minimize potential complications.     

Comparison of hematologic and biochemical values for blood samples obtained via jugular venipuncture and via vascular access ports in cats

OBJECTIVE: To determine whether hematologic and serum biochemical values for blood samples obtained from cats via vascular access ports (VAP) are comparable to those for samples obtained by direct venipuncture. DESIGN: Prospective study. ANIMALS: 14 healthy cats. PROCEDURE: A VAP was surgically implanted in a jugular vein in each cat. Blood samples were obtained from the VAP and by direct venipuncture of the contralateral jugular vein 10 weeks after VAP placement. Results of hematologic and serum biochemical analyses were compared by use of a paired t-test. The Pvalue to reject the null hypothesis was adjusted to account for multiple comparisons by using the Bonferroni procedure in which the nominal P-to-reject value is divided by the number of comparisons (0.05/24 = 0.002). RESULTS: Paired samples (VAP and venipuncture) obtained 10 weeks after VAP placement were evaluated for each cat. Of the 24 measured analytes, only potassium, total protein, and albumin concentrations differed significantly (P< 0.001 for all 3) between VAP and venipuncture samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that samples obtained from VAP are suitable for routine hematologic monitoring of feline cancer patients. Sample hemolysis may account for a slight increase in potassium, total protein, and albumin concentrations obtained from VAP samples. However, the values of variables most critical for monitoring of patients receiving chemotherapy (ie, mature neutrophil and platelet counts) are comparable. If proper techniques are used, VAP may be used for administration of chemotherapy as well as for blood collection in cats undergoing cancer treatment.     

Lack of agreement between thermodilution and echocardiographic determination of cardiac output during normovolemia and acute hemorrhage in clinically healthy, anesthetized dogs

Objective and hypothesis: To determine whether or not there is agreement between the thermodilution and echocardiographic measurement of cardiac output (CO) during normovolemia and acute hemorrhage. The hypothesis was that there will be agreement between echocardiographic measurement of CO (ECO) and thermodilution measurement of CO (TDCO) during normovolemia and acute hemorrhage. Design: CO was measured by both thermodilution and echocardiography during α‐chloralose anesthesia in dogs before and 15 and 30 minutes following acute arterial hemorrhage. Setting: Laboratory investigation. Animals: Eighteen clinically healthy dogs, weighing 20–25 kg, anesthetized with α‐chloralose. Interventions: Acute arterial hemorrhage of approximately 50% of the total blood volume. CO was measured by thermodilution and echocardiography before and 15 and 30 minutes following hemorrhage. Measurements and main results: Acute hemorrhage resulted in a significant decrease in CO. There was a lack of agreement between the 2 methods to measure CO at each time and at all anatomic points of measurement in the aorta and pulmonary artery. Conclusion: There is a lack of agreement between the 2 methods; thus, determination of CO by echocardiography may not be a clinically useful tool following hemorrhage in dogs.     

Linking Land-use, Water Body Type and Water Quality in Southern New Zealand

Land-use and vegetation cover have been linked to the nutrient levels (nitrogen, phosphorus) of surface waters in several countries. However, the links generally relate to streams and rivers, or to specific types of standing water, for example shallow lakes in a geologically defined region. We measured physical variables and nutrient chemistry of 45 water bodies representative of the wide range of lentic wetland environments (swamps, riverine wetlands, estuaries, reservoirs, shallow lakes, deep lakes) in Otago, New Zealand, and related these to catchment variables and land-use in order to assess the potential influence of catchment modification on water quality of these diverse wetlands. Catchment boundaries and land cover were derived from maps using ArcView GIS software. Our predictions that concentrations of nutrients and other components of water quality would correlate positively with the nature and intensity of catchment modification were confirmed in multivariate analyses. Physical and chemical measures were positively related to the extent of modification in the catchment (percentage of the catchment in pasture, planted forest, scrub and urban areas), and negatively related to lack of catchment modification (more of the catchment in bare ground, tussock grassland and indigenous forest). The strong negative correlations between nutrient concentrations, suspended sediment, water colour and the percentage of tussock cover in the catchment imply that increased conversion of the␣native tussock grassland to pastoral farming in␣Otago will increase nutrient concentrations and␣reduce water quality of the diverse lentic ecosystems.     

Survival in Dogs with Nasal Adenocarcinoma: 64 Cases (1981–1995)

Case records of 64 dogs with nasal adenocarcinoma were reviewed. The effects of age, gender, tumor stage, presence of metastatic lesions, and treatment method on survival time were examined. Surgery groups included rhinotomy (n = 9), transnasal curettage (n = 29), and no surgery (n = 26). Chemotherapy groups included fluorouracil-cyclophosphamide combination therapy (n = 15), mitoxantrone (n = 7), and no chemotherapy (n = 42). Fifty-three dogs received fractionated cobalt 60 radiation therapy. Surgical procedure, chemotherapy group, and stage of primary tumor were not significantly associated with survival time ( P > .05). Dogs that received radiation therapy had a significantly longer median survival time (424 days) than dogs that did not (126 days) ( P = .0001). The presence of either regional lymph node or pulmonary metastasis was associated with significantly shorter median survival time (109 days) when compared to dogs without metastases (393 days) ( P = .0125). When only dogs that had received radiation therapy were considered, neither surgical treatment nor chemotherapy group was associated with significant changes in median survival time. An alternate staging system emphasizing the presence or absence of metastases is proposed. Key words: Chemotherapy; Metastasis; Radiotherapy.     

Nitrogen mobilization, nitrogen uptake and growth of cuttings obtained from poplar stock plants grown in different N regimes and sprayed with urea in autumn

Nitrogen mobilization, nitrogen uptake and growth of cuttings obtained from poplar stock plants fertigated with different nitrogen (N) treatments and sprayed with urea in autumn were studied. Stock plants propagated from poplar cuttings were trained to a single shoot and fertigated with 0, 5, 10, 15 or 20 mmol l(-1) N during the first growing season. In October, a subset of stock plants from each N fertigation treatment was sprayed twice with either 3% urea or water, and overwintered outside. In March, total tree biomass and total N concentration and content of stems were estimated for stock plants in each treatment, and cuttings were taken from the middle of each stock plant and stored in plastic bags at 2 degrees C. In mid-April, cuttings were planted in 7.5-l pots containing N-free medium and grown outdoors with a weekly fertigation with nutrient solution containing 0 or 10 mmol l(-1) 15NH4 15NO3. In mid-July, cuttings were harvested, and new shoot (new stems and leaves), shank (old cutting stem) and roots were analyzed for new biomass growth and total N and 15N content. Growth of stock plants was positively related to N supply in the previous growing season. Foliar urea application in autumn had no effect on subsequent stock plant growth even though urea sprays increased both N concentration and content in stem tissues. Biomass growth of cuttings obtained from stock plants was closely related to their N content when the cuttings were grown in an N-free medium regardless of previous treatments applied to the stock plants. When N was supplied in the growth medium, the strength of the relationship between regrowth and N content of cuttings was significantly reduced. Cuttings from stock plants treated with foliar urea and grown in a N-free medium remobilized between 75 and 82% of their total N for new growth, whereas cuttings from plants receiving no urea spray remobilized only between 60 and 69% of their total N for new growth. Current N fertilization of the cuttings reduced the percentage of N remobilized. We conclude that new growth of poplar cuttings in spring was more dependent on currently applied N than on reserve N, and urea N applied as a spray in autumn was more easily remobilized than N taken up by roots during the previous season.