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.     

Quantification of reservoir bags as airway pressure-limiting devices in a nonrebreathing system

ObjectiveTo evaluate the influence of reservoir bag types, volumes and previous use on the peak pressures (Pmax) and the times to develop 30 cmH₂O pressure (P30) within a nonrebreathing system with a closed adjustable pressure-limiting (APL) valve.Study designIn vitro study using three-way factorial design with repeated measure on one factor.SubjectsA total of 75 new anesthesia reservoir bags (five types, three volumes, five bags from each type × volume). The bag types were reusable latex (RL), disposable latex (DL) and three disposable neoprene (DN-1, DN-2 and DN-3).MethodsEach bag was tested three times (treatments): new, after prestretching and 1 week later. The bags were attached to a Bain system and anesthesia machine with closed APL valve and patient port with O₂ flow 2 L minute^–1 until Pmax was reached. The Pmax and time to reach P30 values were determined from recorded pressure traces. General linear mixed model analysis was used to examine the effects of bag type, volume and treatment. One-sided 95% upper prediction limits of Pmax were calculated to test the null hypothesis that predicted Pmax of new bags would be ≥ 50 cmH₂O for each factor combination.ResultsRL bags were the least and DN-3 bags were the most compliant. Prestretching increased compliance in all bag types. Smaller bags of RL, DL and DN-1 were less compliant than larger ones. The predicted Pmax values were < 50 cmH₂O only for DN-3 bags after prestretching. The time to reach P30 was critically low when using 0.5 L bags (median 17 seconds).Conclusions and clinical relevanceTo minimize the risk of barotrauma, highly compliant reservoir bags (e.g. DN-3) are recommended and reusable bags should be avoided. Bags should be prestretched before first use, 0.5 L bags should be avoided and fresh gas flow minimized.     

Investigation of a possible chain of Streptococcus suis infection in a pig breeding community using PCR-based genotyping

Streptococcus (S.) suis plays an important role in pig breeding causing invasive diseases such as meningitis and polyarthritis. Herd problems with Streptococcus suis are common in many pig farms and are frequently characterised by disease outbreaks. In this context, it is often important to identify chains of infection, e.g. between a farrow-to-wean and a grower farm. In the following case report a possible chain of infection among the different farms of a farrow-to-finish system was investigated. In two grower units herd problems with S. suis were present; the mortality was higher than 5 %. It appeared likely, that the streptococci causing these problems were transmitted from a single farrow-to-wean unit to the two different grower farms. In the respective farms swabs were taken from different healthy animals and, in the case of the grower farms, also from the infected animals. Genotypic profiling of strains by a multiplex-PCR and AFLP typing method revealed that two different S. suis pathotypes were responsible for the herd problems. Both pathotypes could not be detected in the farrow-to-wean farm.Thus, a chain of infection originating from the farrow-to-wean farm appeared unlikely. The multiplex PCR was in this case sufficient to elucidate the described problem.     

Combining agent-based and stock-flow modelling approaches in a participative analysis of the integrated land system in Reichraming, Austria

The integrated modelling of coupled socio-ecological systems in land-change science requires innovative model concepts capable of grasping the interrelations between socioeconomic and natural components. Here, we discuss the integrated socio-ecological model SERD (Simulation of Ecological Compatibility of Regional Development) that was developed for the municipality of Reichraming in Upper Austria in a participative 2-year process involving local stakeholders. SERD includes three main components: (1) an agent-based actors module that simulates decisions of farmsteads, the municipal administration and other important actors; (2) a spatially explicit (GIS based) land-use module that simulates land-use change at the level of individual parcels of land and (3) an integrated socio-ecological stock-flow module that simulates carbon and nitrogen flows through both socioeconomic and ecological system compartments. We report on outcomes of a scenario analysis that outlines possible future trajectories depending on both external (e.g. agricultural subsidies and prices) and internal (e.g. innovation, willingness to co-operate) factors. We find that both external and internal factors can affect the behaviour of the integrated system considerably. Local and regional policies are found to be able to counteract adverse global socioeconomic conditions to some extent, but not to reverse the trend altogether. We also find strong interdependencies between socioeconomic and ecological components of the system. Fully evaluating these interdependencies is, however, not possible at the local scale alone and will require explicit consideration of higher-level effects in future research.     

On-line gas chromatography combustion/pyrolysis isotope ratio mass spectrometry (HRGC-C/P-IRMS) of pineapple (Ananas comosus L. Merr.) volatiles

By use of extracts prepared by liquid-liquid separation of the volatiles from self-prepared juices of pineapple fruits (Ananas comosus) (n = 14) as well as commercial pineapple recovery aromas/water phases (n = 3), on-line capillary gas chromatography-isotope ratio mass spectrometry was employed in the combustion (C) and the pyrolysis (P) modes (HRGC-C/P-IRMS) to determine the delta(13)C(VPDB) and delta(2)H(VSMOW) values of selected pineapple flavor constituents. In addition to methyl 2-methylbutanoate 1, ethyl 2-methylbutanoate 2, methyl hexanoate 3, ethyl hexanoate 4, and 2,5-dimethyl-4-methoxy-3[2H]-furanone 5, each originating from the fruit, the delta(13)C(VPDB) and delta(2)H(VSMOW) data of commercial synthetic 1-5 and "natural" (biotechnologically derived) 1-4 were determined. With delta(13)C(VPDB) data of pineapple volatiles 1-4 varying from -12.8 to -24.4 per thousand, the range expected for CAM metabolism was observed. Compound 5 showed higher depletion from -20.9 to -28.6 per thousand. A similar situation was given for the delta(2)H(VSMOW) values of 3-5 from pineapple ranging from -118 to -191 per thousand, whereas 1 and 2 showed higher depleted values from -184 to -263 per thousand. In nearly all cases, analytical differentiation of 1-5 from pineapple and natural as well as synthetic origin was possible. In general, natural and synthetic 1-5 exhibited delta(13)C(VPDB) data ranging from -11.8 to -32.2 per thousand and -22.7 to -35.9 per thousand, respectively. Their delta(2)H(VSMOW) data were in the range from -242 to -323 per thousand and -49 to -163 per thousand, respectively.     

Prevalence of herd specific factors and limb disorders,and their associations in intensive swine production

A longitudinal observational study in 180 pig breeding herds was performed to calculate prevalences of herd specific factors as well as typical limb disorders and to estimate their associations in a 2-step regression analysis. Regarding herd size, genetics, feeding and weight gain herds were distributed almost equal. The population density and the hygiene status were considered proper in most herds. In the farrowing units partially slatted floors of metal or plastic with slats > 9 mm, in the weaning units fully slatted floors of plastic, and in the rearing units fully slatted floors of concrete were most common. Less than 6% of the farms housed their pigs on solid concrete with straw bedding. Herd prevalences of fault floors varied between 18 and 43%. As a herd health problem (morbidity > 25%) claw hematomas and limb abrasions in just 1-week old piglets, overgrown claws and bursa swellings in weaned pigs, and bursa swellings in rearing pigs were wide spread. Leg deformations by osteopathy or arthritis occurred only sporadically. In the risk analysis claw hematomas of piglets were associated with slatted floors, particulary with slats < 10 mm. Abrasions were associated with concrete and rough floor surfaces at all. Overgrown claws and bursa swellings in weaned and in rearing pigs were associated with damaged, slippery or rough floor surfaces. Other associations were not detected. The quality of floor might be more important than the type of housing.     

Suitability of SSCP-PCR analysis for molecular detection of quinolone resistance in Campylobacter jejuni

Foodborne infections with Campylobacter spp. are increasing, especially antibiotic resistant strains are emerging. Quinolone resistant isolates can cause failure of therapy in severe clinical infections. Molecular characterisation is needed for the detection of resistant variants of C. jejuni. Therefore 23 isolates from poultry and human medicine as well as three control strains were tested for their minimal inhibitory concentration, their Single-Strand-Conformation-Polymorphism (SSCP)-PCR pattern (a method for the detection of resistance determining point mutations), and their sequence of the quinolone resistance determining region (QRDR). Six different SSCP types could be identified: two types for quinolone resistant isolates and other types containing so called silent mutations without influence on the resistance. A genotypic monitoring of the quinolone resistance in C. jejuni can be useful for the early detection of new resistance variants. As a screening method for detection of point mutations in the QRDR the SSCP-PCR can be applied. Compared to other genotypic methods the SSCP-PCR is less time and cost consuming and needs only standard technical equipment.     

Report on Salmonella isolates submitted to the German National Veterinary Salmonella Reference Laboratory in the year 1999

The National Salmonella Reference Laboratory (NRL-Salm) of the Federal Institute for Health Protection of Consumers and Veterinary Medicine receives putative Salmonella isolates originating from animals, food, feed and the environment for typing. This report summarises the results of the sero- and phagetyping studies. In livestock S. Typhimurium DT 104 with 24% and S. Enteritidis PT 4 with 9% of all isolations predominate like in the human isolates. These sero- and phagetypes are frequently isolated from food as well and consequently can reach the consumer via the food chain.