Effect of release rate of the SF(6) tracer on methane emission estimates based on ruminal and breath gas samples

The release rate (RR) of sulphur hexafluoride (SF(6)) gas from permeation tube in the rumen appears to be positively related with methane (CH(4)) emissions calculated using the SF(6) tracer technique. Gas samples of breath and ruminal headspace were collected simultaneously in order to evaluate the hypothesis that transactions of SF(6) in the rumen are the source for this relationship. Six non-lactating dairy cows fitted with rumen cannulae were subdivided into two groups and randomly assigned to a two-period crossover design to permeation tubes with low RR (LRR = 1.577 mg/day) or two-times higher RR (HRR = 3.147 mg/day) RR. The cows were fed limited amounts of maize silage (80% ad libitum) split into two meals (40% at 0800 h and 60% at 1600 h). Each period consisted of 3-day gas sampling. Immediately before the morning feed and then each hour over 8 h, ruminal gas samples (50 ml) were withdrawn through the cannula fitted with stoppers to prevent opening. Simultaneously, 8-h integrated breath gas samples were collected over the same period. Ratios of concentration of CH(4)/SF(6), CO(2)/SF(6) and CO(2)/CH(4) and emission estimates of CH(4) and CO(2) were calculated for each sample source using the SF(6) tracer technique principles. The LRR treatment yielded higher (P < 0.001) ruminal CH(4)/SF(6) (by 1.79 times) and CO(2)/SF(6) (by 1.90 times) ratios than the HRR treatment; however, these differences were lower than the 2.0 times difference expected from the RR between the LRR and HRR. Consequently, the LRR treatment was associated with lower (P < 0.01) ruminal emissions of CH(4) over the 8-h collection period than with the HRR treatment (+11%), a difference also confirmed by the breath samples (+11%). RR treatments did not differ (P = 0.53) in ruminal or breath CO(2) emissions; however, our results confirm that the SF(6) tracer seems inappropriate for CO(2) emissions estimation in ruminants. Irrespective of the RR treatment, breath samples yielded 8% to 9% higher CH(4) emission estimates than the ruminal samples (P = 0.01). The relationship between rumen and breath sources for CH(4) emissions was better for LRR than for HRR treatment, suggesting that tracer performance decreases with the highest RR of SF(6) tested in our study (3.1 mg/day). A hypothesis is discussed with regard to the mechanism responsible for the relationship between RR and CH(4) emission estimates. The use of permeation tubes with small range in RR is recommended in animal experiments to decrease variability in CH(4) emission estimates using the SF(6) tracer technique.     

Effect of single nucleotide polymorphisms of CAPN1 and CAST genes on meat traits in Nellore beef cattle (Bos indicus) and in their crosses with Bos taurus

The objectives of this work were to study the segregation of single nucleotide polymorphisms of the calpain 1, large subunit ( CAPN1 ) and calpastatin ( CAST ) genes in Nellore ( Bos indicus ) and Nellore × Bos taurus beef cattle, as well as to evaluate their effects on meat traits. For this, 300 animals, including 114 Nellore, 67 Angus × Nellore, 44 Rubia Gallega × Nellore, 41 Canchim, 19 Brangus three-way crosses and 15 Braunvieh three-way crosses, were genotyped for the CAPN4751 [AF_248054.2:g.6545C>T (GenBank accession AF248054.2 )] and CAST/DdeI [AF_159246.1:g.2959A>G (GenBank accession AF159246.1 )] polymorphisms and phenotyped for Ribeye Area, Backfat Thickness, Intramuscular Fat, Shear Force (SF) and Myofibrillar Fragmentation Index (MFI). In relation to the CAPN4751 polymorphism, a frequency of 10.5% was observed for the C allele in the Nellore breed. In the total sample of studied animals, a significant association was found between genotypes and meat tenderness, assessed by SF ( P  = 0.005) and MFI ( P  = 0.008), with genotype CT being more favourable than TT. For the CAST/DdeI polymorphism, a frequency of 55.7% was found for the A allele in the Nellore breed. In the total sample, a significant association was observed between genotypes and meat tenderness – SF ( P  = 0.004) and MFI ( P  = 0.006), with genotype AA being more favourable than AG. The relationship between genotypes and aged meat tenderness in confluence with the distribution of favourable alleles shows great potential for application of the CAPN4751 and CAST/DdeI polymorphisms in the genetic improvement of the Nellore breed, whilst contributing to the validation, in this breed and in its crosses with B. taurus , of the association results previously described in the literature.     

Survival of Campylobacter jejuni in different gas mixtures

Campylobacter jejuni in fresh chilled chicken meat is known to be a major risk factor for human gastrointestinal disease. In the present study, the survival under chilled conditions of different C. jejuni strains exposed to different gas mixtures usually used for gas packaging of food was examined. Bolton broth and fresh, skinless chicken fillets were inoculated with six and four strains, respectively, and exposed to the gas mixtures 70/30% O(2)/CO(2), 70/30% N(2)/CO(2), and 100% N(2) (the latter only investigated in broth) at refrigeration temperature (4-5 degrees C). In broth culture, the strains survived significantly longer when exposed to 100% N(2) and 70/30% N(2)/CO(2) than in the oxygen-containing gas mixture, 70/30% O(2)/CO(2) (P<0.0001). For the two anaerobic gas mixtures, the reductions only reached 0.3-0.8 log(10) CFU mL(-1) within the same period. In the presence of oxygen, the numbers of C. jejuni were reduced by a minimum of 4.6 log(10) CFU mL(-1) over 21 days. When inoculated onto chicken fillets, the C. jejuni strains also died significantly faster in the oxygen-containing gas mixture, 70/30% O(2)/CO(2) (P<0.0001), reaching reductions of 2.0-2.6 log(10) CFU g(-1) after 8 days. In the gas mixture without oxygen (70/30% N(2)/CO(2)), no reductions were observed.     

Density-dependent reduction of nitric oxide diffusing capacity after pneumonectomy.

Airway lengthening after pneumonectomy (PNX) may increase diffusive resistance to gas mixing (1/D(G)); the effect is accentuated by increasing acinar gas density but is difficult to detect from lung CO-diffusing capacity (Dl(CO)). Because lung NO-diffusing capacity (Dl(NO)) is three- to fivefold that of Dl(CO), whereas 1/D(G) for NO and CO are similar, we hypothesized that a density-dependent fractional reduction would be greater for Dl(NO) than for Dl(CO). We measured Dl(NO) and Dl(CO) at two tidal volumes (Vt) and with three background gases [helium (He), nitrogen (N(2)), and sulfur hexafluoride (SF(6))] in immature dogs 3 and 9 mo after right PNX (5 and 11 mo of age). At maturity (11 mo), background gas density had no effect on Dl(NO), Dl(CO), or Dl(NO)-to-Dl(CO) ratio in sham controls. In PNX animals, Dl(NO) declined 25-50% in SF(6) relative to He and N(2), and Dl(NO)/Dl(CO) declined approximately 50% in SF(6) relative to He at a Vt of 15 ml/kg, consistent with a significant 1/D(G). At 5 mo of age, Dl(NO)/Dl(CO) declined 25-45% in SF(6) relative to He and N(2) in both groups, but Dl(CO) increased paradoxically in SF(6) relative to N(2) or He by 20-60%. Findings suggest that SF(6), besides increasing 1/D(G), may redistribute ventilation and/or enhance acinar penetration of the convective front.     

Effects of gaseous environment and temperature on the storage behaviour of Listeria monocytogenes on chicken breast meat

Portions of skinless chicken breast meat (pH 5.8) were inoculated with a strain of Listeria monocytogenes and stored at 1, 6 or 15 degrees C in (1) aerobic conditions; (2) 30% CO2 + air; (3) 30% CO2 + N2; and (4) 100% CO2. When samples were held at 1 degree C the organism failed to grow under any of the test conditions, despite marked differences between treatments in spoilage rate and ultimate microflora. At 6 degrees C counts of L. monocytogenes increased ca 10-fold in aerobic conditions before spoilage of the meat, but only when the inoculum culture was incubated at 1 degree C rather than 37 degrees C. In CO2 atmospheres growth of L. monocytogenes was inhibited on meat held at 6 degrees C, especially under 100% CO2. By contrast, storage at 15 degrees C led to spoilage of the meat within 2 d, in all gaseous environments, and listeria levels increased up to 100-fold. Differences in the behaviour of L. monocytogenes on poultry and red meats are discussed.     

Variation in meat quality characteristics between Sanga (Bos taurus africanus) and Sanga-derived cattle breeds and between Sanga and Brahman (Bos indicus)

Cattle breeds indigenous to Africa (Sanga) compare favourably to Bos indicus breeds with regard to adaptation to harsh environments. This study compared the meat quality of three Sanga breeds (Nguni, Tuli and Drakensberger), a Sanga-related breed (Bonsmara) and a B. indicus breed (Brahman) and supported these results with biochemical and histological measurements on the M. longissimus lumborum. Twelve young grain-fed steers of each breed were slaughtered and carcasses were electrically stimulated. All Sanga (and related) breeds, with the exception of the Tuli, had lower Warner-Bratzler shear force (SF) values at 2 and 21 days post mortem compared with the BR (P < 0.05). Measurements related to the calcium-dependent protease system and myofibrillar fragmentation explained the bulk of the variation among breeds, whereas variation in fibre type, sarcomere length and connective tissue properties gave less convincing support. With the exception of the Tuli, Sanga and Sanga-related breeds produced more tender (according to SF) meat than BR, mainly due to favourable calpain-to-calpastatin ratios. Small differences in colour, drip loss and cooking properties were found among breeds (P < 0.05).     

Effect of various gas atmospheres on the growth and survival of Campylobacter jejuni on beef

Pieces of fresh beef were inoculated with three strains of Campylobacter jejuni. The meat was then allocated to three treatments: (a) vacuum packaged, (b) packaged in an atmosphere of 20% CO2 + 80% N2, and (c) packaged into sterile Petri dishes in anaerobic cultivation boxes, which were filled with a gas mixture of 5% O2 + 10% CO2 + 85% N2. The packaging material in the first two treatments was PA 80/PE 100-PE 100/PA 80/PE 100. The survival of Campylobacter cells was followed at 37 degrees C, 20 degrees C and 4 degrees C for 48 h, 4 days and 25 days, respectively. At 37 degrees C the counts of two Campylobacter strains increased in each package treatment for 48 h. At 20 degrees C and at 4 degrees C the counts of the same two strains decreased by 1 to 2 log units and 0.5 to 1 log unit, respectively, during storage. The survival of the two strains was about the same in all package treatments. The third strain was the most sensitive of the strains studied. At 37 degrees C its numbers increased only in the optimal gas atmosphere; at 20 degrees C the strain was not detectable after 24 to 48 h storage and at 4 degrees C after 4 days storage. The aerobic plate counts were determined for all samples at the same time as Campylobacter counts. The high indigenous bacterial numbers of the meat samples did not appear to have a great effect on the survival or growth of campylobacters.     

Greenhouse gas emission during storage of pig manure on a pilot scale

The greenhouse gas emissions (CO2, CH4, N2O) from a 2 ton (4.4 m3) deep litter pig manure pile (storage time 113 days during winter season) were quantified by using a tent, which covered the whole pile during the measuring periods only. The emissions were calculated in CO2 equivalents per kilogram dry matter by. Additionally the retention time (use of tracer gas SF6) and the concentrations of the gases in different parts of the pile were determined. The average retention time of the gases in the pile was less than 2 h. CH4 is assumed to have been generated only in the centre of the pile, whereas CO2 was assumed to have been generated in a wider zone. The emissions of CH4, CO2 and N2O were at the highest in the beginning when nearly the whole pile had temperatures in the range of thermophilic microorganisms. This leads to the assumption that mainly thermophilic microorganisms formed the gases. The most important gas for global warming was found to be nitrous oxide.     

Genotypic effects of calpain 1 and calpastatin on the tenderness of cooked M. longissimus dorsi steaks from Jersey x Limousin, Angus and Hereford-cross cattle

Single nucleotide polymorphisms (SNPs) in the calpain 1 (CAPN1) and calpastatin (CAST) genes were studied to determine their effects on meat tenderness in Bos taurus cattle. Strip loins (M. longissimus dorsi) were removed from cattle in four resource populations after slaughter (n = 1042), aged under controlled conditions until fixed times after rigor mortis, cooked and measured using a tenderometer. Animals were genotyped for the CAPN1 SNP c.947C>G (p.Ala316Gly; AF252504) and for the CAST SNP c.2959A>G (AF159246). Frequencies of CAPN1 C alleles ranged from 23% to 68%, and CAST A alleles from 84% to 99.5%. From all data combined, the CAPN1 CC genotype (compared with the GG genotype) was associated with a 20.1 +/- 1.7% reduced average shear force at intermediate stages of ageing (P < 0.001) and with a 9.5 +/- 1.3% reduction near ultimate tenderness (P < 0.001). The heterozygote was intermediate. For CAST, corresponding values for AA compared with AG genotypes were reductions of 8.6 +/- 2.0% and 5.1 +/- 1.6% respectively (both P < 0.001), but there were too few GG genotypes for comparison. There were small interactions between the CAPN1 and CAST genotypes. For the CAPN1 and CAST genotypes combined, the maximal genotype effect in average shear force was 25.7 +/- 5.5% (P < 0.001) at intermediate stages and 15.2 +/- 4.8% near ultimate tenderness (P < 0.01).     

Combining Nitrous Oxide with Carbon Dioxide Decreases the Time to Loss of Consciousness during Euthanasia in Mice — Refinement of Animal Welfare?

Carbon dioxide (CO(2)) is the most commonly used euthanasia agent for rodents despite potentially causing pain and distress. Nitrous oxide is used in man to speed induction of anaesthesia with volatile anaesthetics, via a mechanism referred to as the "second gas" effect. We therefore evaluated the addition of Nitrous Oxide (N(2)O) to a rising CO(2) concentration could be used as a welfare refinement of the euthanasia process in mice, by shortening the duration of conscious exposure to CO2. Firstly, to assess the effect of N(2)O on the induction of anaesthesia in mice, 12 female C57Bl/6 mice were anaesthetized in a crossover protocol with the following combinations: Isoflurane (5%)+O(2) (95%); Isoflurane (5%)+N(2)O (75%)+O(2) (25%) and N(2)O (75%)+O(2) (25%) with a total flow rate of 3 l/min (into a 7 l induction chamber). The addition of N(2)O to isoflurane reduced the time to loss of the righting reflex by 17.6%. Secondly, 18 C57Bl/6 and 18 CD1 mice were individually euthanized by gradually filling the induction chamber with either: CO(2) (20% of the chamber volume.min-1); CO(2)+N(2)O (20 and 60% of the chamber volume.min(-1) respectively); or CO(2)+Nitrogen (N(2)) (20 and 60% of the chamber volume.min-1). Arterial partial pressure (P(a)) of O(2) and CO(2) were measured as well as blood pH and lactate. When compared to the gradually rising CO(2) euthanasia, addition of a high concentration of N(2)O to CO(2) lowered the time to loss of righting reflex by 10.3% (P<0.001), lead to a lower P(a)O(2) (12.55 ± 3.67 mmHg, P<0.001), a higher lactataemia (4.64 ± 1.04 mmol.l(-1), P = 0.026), without any behaviour indicative of distress. Nitrous oxide reduces the time of conscious exposure to gradually rising CO(2) during euthanasia and hence may reduce the duration of any stress or distress to which mice are exposed during euthanasia.     

Influence of cage or pen housing on carcass traits and meat quality of rabbit

Carcass traits and meat quality of rabbits reared in conventional cages (0.385 m2), small pens (0.662 m2) or large pens (4.052 m2) at a similar stock density of 15 rabbits/m2 were compared (n = 30 per group). Pens contained an elevated platform. Slaughter weight (SW; P < 0.01) and cold carcass weight (P < 0.05) decreased in the order of Cage < Small pen < Large pen groups. SW and cold carcass weight were 7% lower in rabbits housed in large pens than in cages. Dressing out and meat-to-bone ratio were not influenced by the housing system. Percentage of fat deposits was highest in caged rabbits (+0.15 point for scapular fat and +0.26 point for perirenal fat compared to rabbits reared in large pens P < 0.05). Rabbits housed in large pens had a bigger proportion of hind part (+1 point), and meat colour was shifted towards greater a* values (P < 0.01) compared to caged rabbits. Water holding capacity and shear test parameters in longissimus lumborum muscle, lipid content and shear test parameters of abductor cruralis cranialis, biceps femoris and semitendinosus muscles were not affected by the housing system. Tibia and femur bone moment of inertia increased in the order of Cage < Small pen < Large pen groups (P < 0.05), whereas elastic modulus, which is a measure of intrinsic stiffness, was highest in caged rabbits. This study showed that large pen housing altered carcass traits independently and increased meat redness and fracture resistance of tibia and femur.     

Effect of culture system on the yield and quality of bovine blastocysts as assessed by survival after vitrification

The aim of this study was to assess the effect of a bovine in vitro culture system on blastocyst yield and quality after vitrification. In Experiment 1, IVM/IVF zygotes were cultured in either synthetic oviduct fluid (SOF) in 5% CO2, 5% O2, 90% N2; or TCM199-granulosa cells (TCM199-GCM) in 5% CO2 in air. In vivo blastocysts were used as a control. Culture in SOF resulted in a significantly higher blastocyst yield on both Day 7 (31.3 vs 13.2%, P < 0.001) and 8 (36.8 vs 23.7%, P < 0.001) than did culture in TCM199-GCM. After vitrification, survival at 72 h of in vivo blastocysts was significantly higher than both in vitro groups, while significantly more blastocysts produced in TCM199-GCM survived compared to those produced in SOF (0, 43.5, 78.3% for SOF, TCM199-GCM and in vivo, respectively P < 0.01). In Experiment 2, SOF-GCM proved to be the best post-warming culture system of those tested and was adopted as the post-warming medium for all subsequent experiments. In Experiment 3, zygotes were cultured in SOF or SOF-GCM, in either 5% CO2 in air, or 5% CO2, 5% O2, 90% N2. In agreement with Experiment 1, culture in SOF in 5% O2 resulted in significantly more blastocysts at Day 7 (26.4 vs 17.3%, P < 0.01) and Day 8 (31.5 vs 23.2%, P < 0.01) than did culture in SOF-GCM. However, survival at 72 h post vitrification was significantly higher for SOF-GCM (44 vs 8.3%, P < 0.001). Increasing the O2 concentration to 20% significantly reduced the blastocyst eld from SOF (31.5 vs 17.3%, P < 0.001). In addition, the quality of blastocyst produced was reduced in terms of survival post vitrification (8.3 vs 0%, P < 0.05). In contrast, there was no difference in blastocyst yield (23.2 vs 25.2%) or survival (44.0 vs 36.9%) in SOF-GCM, irrespective of O2 concentration. Experiment 4 examined the duration of exposure to GCM necessary to acquire improved blastocyst quality. Zygotes were cultured in SOF; SOF until Day 3, followed by SOF-GCM for the remainder of the culture; SOF until Day 5, followed by SOF-GCM for the remainder of the culture; or SOF-GCM for the entire culture. Survival at 72 h post vitrification was significantly higher (P < 0.05) in Groups 2 (50.0%, 13/26) and 4 (55.3%, 26/47) than in Groups 1 (21.7%, 10/46) and 3 (10.8%, 4/37). In conclusion, culture system can affect blastocyst yield and quality and crytolerance is a useful indicator of blastocyst quality.     

In vitro development of preimplantation porcine nuclear transfer embryos cultured in different media and gas atmospheres

This study investigated the effect of culture media and gas atmospheres on the development of porcine nuclear transfer embryos. Oocytes derived from a local abattoir were matured for 42-44 h and enucleated. Fetal fibroblasts were prepared from a Day 35 porcine fetus. Confluent stage fetal fibroblasts were introduced into the perivitelline space of enucleated oocytes. Fusion and activation were induced simultaneously with two direct current (1.2 kV/cm for 30 micros) in 0.3 M mannitol medium. For parthenogenetic activation, the same pulses were used. In Experiment 1, parthenogenetically activated oocytes were cultured in North Carolina State University-23 (NCSU-23), Porcine Zygote Medium-3 (PZM-3), or Beltsville Embryo Culture Medium-3 (BECM-3). Parthenogenetically activated oocytes cultured in PZM-3 had a higher (P < 0.05) developmental rate to the blastocyst stage (15.2% versus 3.7-9.6%) as compared to BECM-3 or NCSU-23. The number of nuclei in Day 6 blastocysts was higher (P < 0.05) in PZM-3 (23.6) and NCSU-23 (21.4) than BECM-3 (14.2). In Experiment 2, parthenogenetically activated oocytes were cultured in NCSU-23 under a gas atmosphere of 5% CO(2) in air for 6 days (T1), 5% CO(2), 5% O(2), 90% N(2) for 6 days (T2), 5% CO(2) in air for 3 days, then 5% CO(2), 5% O(2), 90% N(2) for 3 days (T3), or 5% CO(2), 5% O(2), 90% N(2) for 3 days, then 5% CO(2) in air for 3 days (T4). Blastocyst formation rates were not different among treatments (12.9 =/-3.6 %, 13.5 +/- 4.2%, 10.8+/-2.4%, and 12.6+/-2.7%, respectively). However, T2 (36.7+/-2.9) and T3 (33.8+/-3.0) resulted in more nuclei per blastocyst than T1 (23.2+/-2.1) or T4 (26.0+/-2.1 ). In Experiment 3, reconstructed porcine nuclear transfer (NT) embryos were cultured in NCSU-23 or PZM-3 under a gas atmosphere of 5% CO(2) in air or 5% CO(2), 5% O(2), 90% N(2). Developmental rates to blastocyst stage for porcine NT embryos cultured in NCSU-23 under a gas atmosphere of 5% CO(2) in air or 5% CO(2), 5% O(2), 90% N(2) were 7.2+/-1.4% and 12.3+/-1.4%, and the number of nuclei was 12.2=/-0.8% and 19.4+/-1.0, respectively. NT embryos cultured in PZM-3 under a gas atmosphere of 5% CO(2) in air or 5% CO(2), 5% O(2), 90% N(2) had developmental rates to blastocyst stage of 18.8+/-1.9 %, and 17.8+/-3.8% the nuclei number was 20.9 +/- 1.9 and 21.9+/-3.3, respectively. NT embryos cultured in NCSU-23 had a higher developmental rate to the blastocyst stage in 5% CO(2), 5% O(2), 90% N(2) than in 5% CO(2) in air (P < 0.05). Regardless of gas atmospheres, NT embryos cultured in PZM-3 had a higher developmental rate (18.3 =/- 1.7% versus 16.9 +/- 1.2%) and nuclei number (21.4 +/-1.8 versus 16.9 +/- 1.2) than in NCSU-23 (P < 0.05). In conclusion, a gas atmosphere of 5% CO(2), 5% O(2), 90% N(2) supported a higher development rate of porcine NT embryos than 5% CO(2) in air when the porcine NT embryos were cultured in NCSU-23. Furthermore, regardless of atmosphere, PZM-3 supported a higher development rate of porcine nuclear transfer embryos than NCSU-23.     

Stress reactivity and its relationship to beef quality in Nguni steers supplemented with Acacia karroo leaves

The objective of this study was to determine the levels of catecholamines and their relationship to beef quality in Nguni steers fed on Acacia karroo leaves. A total of 30 19-month-old steers were randomly assigned to A. karroo leaves (AK), sunflower cake (SF) and the control with no supplement (CN) diets. The AK and SF diets provided the steers with an additional 150 g of protein per day for 60 days. Catecholamine levels were determined from urine samples collected from each steer before and after slaughter. The Musculus longissimus thoracis et lumborum was sampled for selected meat quality measurements. Nguni steers on the CN diet had higher (P < 0.05) concentrations of post-mortem urinary norepinephrine and dopamine compared with those that received the AK and SF diets. Norepinephrine was negatively linearly related (P < 0.05) to the Warner-Bratzler shear force value of meat aged for 21 days and cooking loss of meat aged for 2 days (CL2) in steers that were given the SF diet. Meat pH and drip loss values were inversely related (P < 0.05) to epinephrine concentration in steers that received the AK diet. Dopamine concentration was negatively linearly related (P < 0.05) to water holding capacity and CL2 for steers on the CN diet. For steers on the CN diet, lightness (L*) values increased (P < 0.05) with increase in dopamine concentration. It was concluded that stress responsiveness and its relationship to certain beef quality attributes could be positively manipulated by supplementation with A. karroo leaves.     

Comparison of various atmospheric conditions for isolation and subcultivation of Mycoplasma hyorhinis from cell cultures

The efficiency of aerobic incubation was compared with incubation under various oxygen and carbon dioxide conditions for the isolation and subcultivation of three strains of Mycoplasma hyorhinis from VERO-cell cultures and subcultivation of three laboratory strains. Under anaerobic conditions with a low oxidation-reduction potential (at or below -115 mV) as obtained in jars, with catalysts, containing mixtures of 5%-10% CO2 in H2, very poor or no growth of any of the six M. hyorhinis strains was observed. When traces of oxygen were present (that is, under conditions with higher oxidation-reduction potentials, e.g. when omitting the catalyst in the above gas mixtures or in 5% CO2 + 95% N2) isolation from cell cultures was successful in most tests, but subcultivation of these primary isolates was seldom possible under these semi-anaerobic conditions. However, in most cases these primary isolates could be subcultivated aerobically, although aerobic conditions were unsatisfactory for isolation in about half of the experiments. Isolation of M. hyorhinis was optimal in 5% O2 + 95% N2, under which condition the isolates could also always be subcultivated. Isolation failed occasionally when 5% O2 + 5% CO2 + 90% N2 was used, thus indicating that 5% CO2 was slightly inhibitory. 5% CO2 in air and 10% CO2 either in air, H2 or N2 were also inadequate for isolation from cell cultures. In contrast to the findings with these cell culture-adapted M. hyorhinis strains, the laboratory strains could be subcultivated easily under all conditions tested except those with an oxidation-reduction potential at or below -115 mV; 100% CO2 was inhibitory for all 6 strains. Our findings may partly explain why M. hyorhinis is often considered "non-cultivable" on artificial media once adapted to cell cultures. The findings emphasize the need to employ also a micro-aerophilic condition (5% O2 in 95% N2) in the examination of cell cultures for mycoplasma.     

Stunning pigs with nitrogen and carbon dioxide mixtures: effects on animal welfare and meat quality

The aim of this study was to assess the effect of exposure to the gas mixtures of 70% nitrogen (N₂) and 30% carbon dioxide (CO₂; 70N30C), 80% N₂ and 20% CO₂ (80N20C) and 85% N₂ and 15% CO₂ (85N15C) on aversion, stunning effectiveness and carcass, as well as meat quality in pigs, and to compare them with the commercial stunning of 90% CO₂ (90C). A total of 68 female pigs were divided into four groups and stunned with one of the gas mixtures. During the exposure to the gas, behavioural variables (retreat attempts, escape attempts, gasping, loss of balance, muscular excitation and vocalizations) were recorded, and at the end of the stunning, corneal reflex and rhythmic breathing were assessed. After slaughter, meat quality parameters such as pH at 45 min post mortem (pH45) and at 24 h post mortem (pHu), electrical conductivity, drip loss and colour, in the Longissimus thoracis (LT) and Semimembranosus (SM) muscles were measured, and the presence of ecchymosis on the hams was noted. The PROC MIXED and the PROC GENMOD of SAS® were used to analyse the parametric and binomial variables, respectively. The ‘gas mixture’ was always considered a fixed effect and the ‘live weight’ as a covariate. To assess the correlation between meat quality and behaviour measures, PROC CORR was used. Pigs exposed to 90C showed a higher percentage of escape attempts and gasping, a lower percentage of vocalization and shorter muscular excitation phase than pigs exposed to the other N₂ and CO₂ mixtures (P < 0.05). After stunning, no pig exposed to 90C showed corneal reflex or rhythmic breathing, whereas 85% and 92% of the animals exposed to N₂ and CO₂ mixtures showed corneal reflex and rhythmic breathing, respectively. Animals stunned with 80N20C and 85N15C had a lower pH45 (P < 0.01) than animals exposed to 90C. Electrical conductivity in the SM muscle was lower (P < 0.001) in 90C and 70N30C pigs than in 80N20C and 85N15C pigs, whereas in LT, it was lower (P < 0.05) in 90C pigs than in 85N15C. As the CO₂ concentration of the gas mixture was decreased, the prevalence of exudative pork increased. Twenty-five percent of animals exposed to N₂ and CO₂ mixtures (n = 68) had ecchymosis in their carcasses, whereas no animal stunned with 90C had ecchymosis. In conclusion, although N₂ and CO₂ stunning exhibit fewer signs of aversion than 90C, their induction time to unconsciousness is longer, and this may negatively affect meat and carcass quality.     

How does barley supplementation in lambs grazing alfalfa affect meat sensory quality and authentication?

Excessive flavour in lamb meat is undesirable for consumers and can prompt purchase resistance. Volatile indoles responsible for off-flavours accumulate more in the fat of lambs on pasture than on grain and are enhanced when lambs graze alfalfa. Here, we investigated whether barley supplementation of lambs grazing alfalfa influences meat sensory quality. Using three groups of 12 male Romane lambs, we compared three feeding regimes: alfalfa grazing (AG), alfalfa grazing + daily supplementation with barley (29 g/kg live weight^0.75, AGS) and stall feeding with concentrate and hay (SF). As some of the compounds involved in meat sensory traits may act as dietary biomarkers, we also investigated potential implications for meat authentication. Although barley represented 38% of the diet in AGS lambs, it did not offer any advantage for animal average daily gain or parasitism level. Animal performance, carcass weight and fatness did not differ between feeding regimes. Dorsal fat firmness tended to be greater in AG than AGS and greater in AGS than SF. Skatole and indole concentrations in perirenal and dorsal fat were lower in SF lambs than in AG and AGS lambs (P<0.01 to P<0.0001), but did not differ between AG and AGS lambs. Yellowness, chroma and hue angle of perirenal fat were lower in SF lambs than in AG and AGS lambs (P<0.001), but did not differ between AG and AGS lambs. Absolute value of the mean integral for both perirenal fat and subcutaneous caudal fat (AVMI_PF and AVMI_SC), quantifying the intensity of light absorption by carotenoids in perirenal and subcutaneous caudal fat, respectively, were lower in SF lambs than in AG and AGS lambs (P<0.0001 for both comparisons), but did not differ between AG and AGS lambs. Meat colour was unaffected by the treatment. We confirm that lambs grazing alfalfa accumulate high levels of volatile indoles in their fat, but we show that barley supplementation to lambs grazing alfalfa is not effective in reducing fat volatile indoles concentration and excessive odour/flavour in the meat. We also confirm that both perirenal fat skatole concentration and AVMI_PF are of interest for discriminating lambs that grazed alfalfa from lambs that were stall-fed, and we show that they are not effective for discriminating supplemented from non-supplemented grazing lambs.     

红豆草与土壤氮含量对大气二氧化碳浓度升高的响应

在封闭的植物培养箱中,通过盆栽实验,研究了红豆草和土壤氮含量对CO₂浓度增加的响应.结果表明,与正常CO₂浓度(355～370 μmol·mol^-1)相比,CO₂浓度升高(700 μmol·mol^-1),植物生物量增加25.1%(P＜0.01),但植物体氮浓度降低25.3%(P＜0.001),植物全氮没有显著的变化.经3个月盆栽实验后,与原始土壤相比,两种CO₂浓度处理土壤全N、NO₃^--N和NH₄⁺-N都有所降低,而土壤微生物氮则显著增加,这可能与植物生长有关.不同CO₂浓度处理土壤NH₄⁺-N浓度基本一致,但在高CO₂浓度下,土壤NO₃^--N浓度显著降低,而微生物生物氮显著增加.对整个土壤-植物系统而言,盆栽实验后,整个系统全氮有少量增加,但变化不显著,特别是在高CO₂浓度条件下,土壤-植物系统全氮最大,这可能与培养材料红豆草为豆科植物,而且在高CO₂浓度下生物量增加,导致氮的固定量增加有关.