Comparison of greenhouse gas emissions from sheep measured using both respiration and portable accumulation chambers.

Methane (CH4) is a potent gas produced by ruminants, and new measurement techniques are required to generate large datasets suitable for genetic analysis. One such technique are portable accumulation chambers (PAC), a short-term sampling method. The objectives of the current study were to explore the relationship between CH4 and carbon dioxide (CO2) output measured using both PAC and respiration chambers (RC) in growing lambs, and separately investigate the relationship among CH4, CO2 and measured ad libitum DM intake (DMI). Methane, CO2 and DMI were measured on 30 Suffolk and 30 Texel ewe lambs (age 253 ± 12 days) using the RC and PAC sequentially. The experiment was conducted over a 14-day period, with DMI measured from days 1 to 14; measurements in RC were conducted from days 10 to 12, while measurements in PAC were taken twice, the day immediately prior to the lambs entering the RC (day 9; PAC Pre-RC) and on the day lambs exited the RC (day 13; PAC Post-RC). Greater CH4 and CO2 output was measured in the RC than in the PAC (P < 0.01); similarly mean CH4 yield was greater when measured in the RC (15.39 ± 0.452 g CH4/kg DMI) compared to PAC (8.01 ± 0.767 g CH4/kg DMI). A moderate correlation of 0.37 was found between CH4 output measured in PAC Pre-RC and the RC, the corresponding regression coefficient of CH4 output measured in the RC regressed on CH4 output measured in PAC Pre-RC was close to unity (0.74; SE 0.224). The variance of CH4 and CO2 output within the measurement technique did not differ from each other (P > 0.05). Moderate to strong correlations were found between CH4 and CO2 per kg of live weight and CH4 and CO2 yield. Results from this study highlight the suitability of PAC as a ranking tool to rank animals based on their gaseous output when compared to the RC. However, repeated measurements separated by several days may be beneficial if precise rankings are required. Given the close to unity regression coefficient of CH4 output measured in the RC regressed on CH4 output measured in PAC Pre-RC suggests that PAC could also be potentially used to estimate absolute CH4 output; however, further research is required to substantiate this claim. When DMI is unknown, CH4 and CO2 per kg of live weight are a suitable alternative to the measurement of CH4 and CO2 yield.

The effect of calf jacket usage on performance, behaviour and physiological responses of group-housed dairy calves.

Poor performance and ill-health of calves in the pre-wean period can affect future productivity. Increasing numbers of producers are opting to use calf jackets as a means of mitigating the potential negative effects of low ambient temperatures, wind speed and precipitation on growth and health. This study aimed to use a range of noninvasive monitoring technologies to investigate the effects of using calf jackets in the first 3 weeks of life on calf performance and behavioural and physiological parameters. Ninety Holstein-Friesian calves were allocated to one of the two treatments: (i) Jacketed until 21 days of age and (J; n = 44) ii. Nonjacketed (NJ; n = 46). Calves were group housed and fed milk replacer (MR) and concentrate solid feed via automatic feeders. Calves were weaned at day 56, and the experiment was completed at day 63. Health assessments were conducted on a daily basis throughout the experiment using predefined faecal and respiratory scoring protocols. A range of novel, noninvasive monitoring technologies were used to examine the activity, heart rate and thermal profiles of calves on an individual basis throughout the experimental period. There were no differences in calf live weight (LWT), average daily gain (ADG) or feed conversion efficiency (FCE) in J and NJ calves between days 5 to 20. However, NJ calves consumed more MR and had more unrewarded visits to the milk feeder than J calves during this period. Although calf LWT was comparable across treatments in the week following jacket removal (days 21 to 28), both ADG and FCE tended to be greater in NJ calves. There were no treatment differences in calf LWT at the end of the study (d63). When measured over a period of 24 h and at a mean ambient temperature of 7.7°C, skin surface temperature was 6.37°C higher in J calves. Core body temperature was higher in J calves between days 5 to 20; however, there were no differences in IR eye or IR rectal temperature. No differences in lying behaviour occurred, with calves spending 18 and 17 h/day lying between days 5 to 20 and days 21 to 28, respectively. Under the climatic and management conditions described, no significant benefits to calf performance were found as a result of the provision of calf jackets to group-housed calves in the first 3 weeks of life. The higher frequency of unrewarded visits to the milk feeder in NJ calves during the first 3 weeks of life could be suggestive of a lack of satiety in these calves.

Evaluation of factors associated with immunoglobulin G, fat, protein, and lactose concentrations in bovine colostrum and colostrum management practices in grassland-based dairy systems in Northern Ireland.

The objectives of this study were to investigate colostrum feeding practices and colostrum quality on commercial grassland-based dairy farms, and to identify factors associated with colostrum quality that could help inform the development of colostrum management protocols. Over 1 yr, background information associated with dairy calvings and colostrum management practices were recorded on 21 commercial dairy farms. Colostrum samples (n = 1,239) were analyzed for fat, protein, lactose, and IgG concentration. A subset was analyzed for somatic cell count and total viable bacteria count. Factors associated with nutritional and IgG concentrations were determined using both univariate and multivariate models. This study found that 51% of calves were administered their first feed of colostrum via esophageal tube, and the majority of calves (80%) were fed >2 L of colostrum at their first feed (mean = 2.9 L, SD = 0.79), at a mean time of 3.2 h (SD 4.36) after birth, but this ranged across farms. The mean colostral fat, protein, and lactose percentages and IgG concentrations were 6.4%, 14%, 2.7%, and 55 mg/mL, respectively. The mean somatic cell count and total viable count were 6.3 log10 and 6.1 log10, respectively. Overall, 44% of colostrum samples contained <50 mg/mL IgG, and almost 81% were in excess of industry guidelines (<100,000 cfu/mL) for bacterial contamination. In the multivariate model, IgG concentration was associated with parity and time from parturition to colostrum collection. The nutritional properties of colostrum were associated with parity, prepartum vaccination, season of calving, and dry cow nutrition. The large variation in colostrum quality found in the current study highlights the importance of routine colostrum testing, and now that factors associated with lower-quality colostrum on grassland-based dairy farms have been identified, producers and advisers are better informed and able to develop risk-based colostrum management protocols.

Effect of concentrate supplementation during the dry period on colostrum quality and effect of colostrum feeding regimen on passive transfer of immunity, calf health, and performance.

The objectives were to evaluate the effect of (1) supplementing concentrates to multiparous Holstein cows during the dry period on colostral and milk immunoglobulin G (IgG) concentration; and (2) feeding calves colostrum at either 5 or 10% of their body weight (BW) on passive transfer of immunity, health, and performance. Holstein multiparous cows (n=37) were assigned to 1 of 2 nutritional treatments during an 8-wk dry period: (1) offered ad libitum grass silage only (GS) or (2) offered ad libitum access to the same grass silage plus concentrate [total mixed ration in a 75:25 dry matter (DM) ratio], providing a mean concentrate DM intake of 3.0kg/cow per day (GSC). Both treatment groups were offered identical levels of mineral and vitamin supplementation. Calves from these cows were weighed immediately after birth and fed either 5% (5BW) or 10% (10BW) of their BW in colostrum from their own dams within 2.5h of birth. Calves in the 10BW group received their second feed of colostrum from first-milking colostrum. Concentrate supplementation during the dry period had no effect on colostral IgG concentration, first-milking IgG yield, or fat, protein, and lactose contents. However, cows in GSC produced a greater mean milk yield over the first 8 milkings compared with cows in the GS group. Concentrate supplementation had no effect on calf BW or BW gain, serum IgG, or apparent efficiency of absorption (AEA) at 24h after birth. However, offspring from the GSC group had fewer cases of enteritis during the first 56d of life compared with offspring from the GS group. Calves in the 10BW group had greater mean serum IgG concentration for the first 3d following birth; however, at 24h after birth, we observed no treatment effect on AEA. The rate of enteritis was greater for calves in the 5BW treatment compared with 10BW. The colostrum-feeding regimen had no effect on BW gain or on the incidence of pneumonia among calf treatment groups. In conclusion, concentrate supplementation regimens offered during the dry period had a positive effect on colostrum yield, and offspring from the GSC group had a reduced rate of enteritis. Feeding 10% of BW of colostrum versus 5% of BW resulted in a greater serum IgG concentration for the first 3d postpartum, and 10BW calves had a reduced rate of enteritis. Overall, to achieve successful passive transfer, decrease the rate of enteritis, and increase efficiency in the dairy calf, we recommend that dairy calves be fed 10% of their BW in colostrum as soon as possible after birth.

Metabolic regulation of stem cell function.

Stem cell function is regulated by intrinsic mechanisms, such as transcriptional and epigenetic regulators, as well as extrinsic mechanisms, such as short-range signals from the niche and long-range humoral signals. Interactions between these regulatory mechanisms and cellular metabolism are just beginning to be identified. In multiple systems, differentiation is accompanied by changes in glycolysis, oxidative phosphorylation and the levels of reactive oxygen species. Indeed, metabolic pathways regulate proliferation and differentiation by regulating energy production and the generation of substrates for biosynthetic pathways. Some metabolic pathways appear to function differently in stem cells as compared with restricted progenitors and differentiated cells. They also appear to influence stem cell function by regulating signal transduction, epigenetic marks and oxidative stress. Studies to date illustrate the importance of metabolism in the regulation of stem cell function and suggest complex cross-regulation likely exists between metabolism and other stem cell regulatory mechanisms.

The effect of calf nutrition on the performance of dairy herd replacements.

Sixty-five Holstein-Friesian calves were randomly allocated to one of eight nutritional treatments at 4 days of age. In this factorial design study, the treatments comprised of four levels of milk replacer (MR) mixed in 6 l of water (500, 750, 1000 and 1250 g/day) × two crude protein (CP) concentrations (230 and 270 g CP/kg dry matter (DM)). MR was fed via automatic teat feeders and concentrates were offered via automated dispensers during the pre-wean period. MR and calf starter concentrate intake were recorded until weaning with live weight and body measurements recorded throughout the rearing period until heifers entered the dairy herd at a targeted 24 months of age. There was no effect of MR protein concentration on concentrate or MR intake, and no effect on body size or live weight at any stage of development. During the pre-weaning period, for every 100 g increase in MR allowance, concentrate consumption was reduced by 39 g/day. While, for every 100 g increase in the amount of MR offered, live weight at days 28 and 270 increased by 0.76 and 2.61 kg, respectively (P < 0.05). Increasing MR feed levels increased (P < 0.05) heart girth and body condition score at recordings during the first year of life, but these effects disappeared thereafter. Increasing MR feeding level tended to reduce both age at first observed oestrus and age at first service but no significant effect on age at first calving was observed. Neither MR feeding level nor MR CP content affected post-calving live weight or subsequent milk production. Balance measurements conducted using 44 male calves during the pre-weaning period showed that increasing milk allowance increased energy and nitrogen (N) intake, diet DM digestibility, true N digestibility and the biological value of the dietary protein. Increasing the MR protein content had no significant effect on the apparent digestibility of N or DM.

Tracer method to determine residence time in a permeable reactive barrier.

A method is presented to evaluate ground water residence time in a zero-valent iron (ZVI) permeable reactive barrier (PRB) using radon-222 ((222)Rn) as a radioactive tracer. Residence time is a useful indicator of PRB hydraulic performance, with application to estimating the volumetric rate of ground water flow through a PRB, identifying flow heterogeneity, and characterizing flow conditions over time as a PRB matures. The tracer method relies on monitoring the decay of naturally occurring aqueous (222)Rn as ground water flows through a PRB. Application of the method at a PRB site near Monticello, Utah, shows that after 8 years of operation, residence times in the ZVI range from 80 to 486 h and correlate well with chemical parameters (pH, Ca, SO(4), and Fe) that indicate the relative residence time. Residence times in this case study are determined directly from the first-order decay equation because we show no significant emanation of (222)Rn within the PRB and no measurable loss of (222)Rn other than by radioactive decay.

Effects of feeding level and protein content of milk replacer on the performance of dairy herd replacements.

It has been suggested that United Kingdom recommendations for feeding the neonatal calf (500 g milk replacer (MR)/day; 200-230 g CP/kg milk powder) are inadequate to sustain optimal growth rates in early life. The current study was undertaken with 153 high genetic merit, male and female Holstein-Friesian calves (PIN2000 = £48) born between September and March, with heifers reared and bred to calve at 24 months of age. Calves were allocated to one of four pre-weaning dietary treatments arranged in a 2 MR feeding level (5 v. 10 l/day) × 2 MR protein content (210 v. 270 g CP/kg dry matter (DM)) factorial design. MR was reconstituted at a rate of 120 g/l of water, throughout, and was offered via computerised automated milk feeders. Calves were introduced to pre-weaning diets at 5 days of age and weaned at day 56. During the first 56 days of life, calves offered 10 l MR/day had significantly higher liveweight gains (P < 0.001) than calves fed 5 l MR/day. No significant differences in liveweight gain were found between calves fed 210 g CP/kg DM MR and those fed 270 g CP/kg DM MR from birth to day 56. Differences in live weight and body size due to feeding level disappeared by day 90. Neither MR feeding level nor MR CP content affected age at first service or age at successful service, and with no milk production effects, the results indicate no post-weaning benefits of increased nutrition during the milk-feeding period in dairy heifers.

Stem cells use distinct self-renewal programs at different ages.

Stem cells expand in number during development and persist throughout life by undergoing self-renewing divisions. The question of how stem cells self-renew throughout life is a fundamental problem in cell biology, with broad implications for understanding development, tissue regeneration, cancer, and aging. Recent insights demonstrate that self-renewal programs depend on key transcriptional regulators that are often shared among stem cells in different tissues but that often change between stem cells at different stages of life: Embryonic, fetal, young adult, and old adult stem cells are maintained by different self-renewal programs. Self-renewal programs change over time to contend with changes in tissue growth and repair demands as well as the increasing risk of malignant transformation during aging. The downstream mechanisms by which these programs regulate the cell cycle, developmental potential, and timing of differentiation are just starting to be elucidated. One key requirement for self-renewal is repression of the p16(Ink4a) and p19(Arf) tumor suppressors. This is accomplished by overlapping transcriptional regulators whose expression and function change with age, so as to maintain self-renewal potential throughout life while allowing increased expression of p16(Ink4a) and p19(Arf) in aging stem cells. This reduces stem cell function in aging tissues but also reduces cancer incidence.

Stem cell self-renewal and cancer cell proliferation are regulated by common networks that balance the activation of proto-oncogenes and tumor suppressors.

Networks of proto-oncogenes and tumor suppressors that control cancer cell proliferation also regulate stem cell self-renewal and possibly stem cell aging. Proto-oncogenes promote regenerative capacity by promoting stem cell function but must be balanced with tumor suppressor activity to avoid neoplastic proliferation. Conversely, tumor suppressors inhibit regenerative capacity by promoting cell death or senescence in stem cells. For example, the polycomb family proto-oncogene, Bmi-1, is consistently required for the self-renewal of diverse adult stem cells, as well as for the proliferation of cancer cells in the same tissues. Bmi-1 promotes stem cell self-renewal partly by repressing the expression of Ink4a and Arf, tumor suppressor genes that are commonly deleted in cancer. Despite ongoing Bmi-1 expression, Ink4a expression increases with age, potentially reducing stem cell frequency and function. Increased tumor suppressor activity during aging therefore may partly account for age-related declines in stem cell function. Thus, networks of proto-oncogenes and tumor suppressors have evolved to coordinately regulate stem cell function throughout life. Imbalances within such networks cause cancer or premature declines in stem cell activity that resemble accelerated aging.

Stem cells, cancer, and cancer stem cells.

Stem cell biology has come of age. Unequivocal proof that stem cells exist in the haematopoietic system has given way to the prospective isolation of several tissue-specific stem and progenitor cells, the initial delineation of their properties and expressed genetic programmes, and the beginnings of their utility in regenerative medicine. Perhaps the most important and useful property of stem cells is that of self-renewal. Through this property, striking parallels can be found between stem cells and cancer cells: tumours may often originate from the transformation of normal stem cells, similar signalling pathways may regulate self-renewal in stem cells and cancer cells, and cancer cells may include 'cancer stem cells' - rare cells with indefinite potential for self-renewal that drive tumorigenesis.

Neuronal potential and lineage determination by neural stem cells.

How do neural stem cells ensure that they give rise to the right number and type of neurons at the right time? Over the past year several regulatory mechanisms have been identified, including promotion of neurogenesis by proneural bHLH genes, instruction of gliogenesis by Notch, and cell-intrinsic changes in the neurogenic capacity of stem cells in culture and in vivo.

Uranium precipitation in a permeable reactive barrier by progressive irreversible dissolution of zerovalent iron.

A permeable reactive barrier (PRB) containing zerovalent iron [Fe(O)] was installed at a former uranium milling site in Monticello, UT. A large-scale column experiment was conducted at the site to test the feasibility of Fe(O) to treat U prior to installing the PRB. Effluents from the field column experiment had pH values near 7.34, moderate decreases in C(IV) and Ca concentrations, and an elevated Fe concentration (27.1 mg/L). In contrast, groundwater exiting the PRB had a pH value of 9.82, decreases in C(IV) and Ca concentrations, and a low concentration of Fe (0.17 mg/L). A geochemical model was used to explain the chemical changes that occurred in both the field column experiment and the PRB. The model simulated the systems by the progressive irreversible dissolution of Fe(O). Modeling results indicated that a longer residence time in the PRB compared with the shorter residence time in the column contributed to the disparate effluent qualities. Prior to modeling, a controlled laboratory column experiment was conducted to help evaluate the dominant chemical mechanisms by which Fe(O) removes U from aqueous solutions. Results of the laboratory column experiment indicated that only a small amount of U could be adsorbed to ferric minerals, and, therefore, this mechanism was not considered in the model.

Neuronal differentiation: proneural genes inhibit gliogenesis.

Proneural basic helix-loop-helix transcription factors such as Neurogenin are activators of neuronal gene expression. Recent studies show they reinforce neuronal differentiation by also inhibiting the expression of glial genes, and are required in vivo to prevent premature and excessive gliogenesis.

Neural crest stem cells undergo cell-intrinsic developmental changes in sensitivity to instructive differentiation signals.

Rat neural crest stem cells (NCSCs) prospectively isolated from uncultured E14.5 sciatic nerve and transplanted into chick embryos generate fewer neurons than do NCSCs isolated from E10.5 neural tube explants. In addition, they differentiate primarily to cholinergic parasympathetic neurons, although in culture they can also generate noradrenergic sympathetic neurons. This in vivo behavior can be explained, at least in part, by a reduced sensitivity of sciatic nerve-derived NCSCs to the neurogenic signal BMP2 and by the observation that cholinergic neurons differentiate at a lower BMP2 concentration than do noradrenergic neurons in vitro. These results demonstrate that neural stem cells can undergo cell-intrinsic changes in their sensitivity to instructive signals, while maintaining multipotency and self-renewal capacity. They also suggest that the choice between sympathetic and parasympathetic fates may be determined by the local concentration of BMP2.

Stem cell potential: can anything make anything?

Recent results suggest that stem cells from one tissue can give rise to cells from developmentally unrelated tissues. These results strongly support the idea that certain progenitors retain much broader developmental potentials than expected, and other progenitors may be able to acquire broader potentials in culture.

Culture in reduced levels of oxygen promotes clonogenic sympathoadrenal differentiation by isolated neural crest stem cells.

Isolated neural crest stem cells (NCSCs) differentiate to autonomic neurons in response to bone morphogenetic protein 2 (BMP2) in clonal cultures, but these neurons do not express sympathoadrenal (SA) lineage markers. Whether this reflects a developmental restriction in NCSCs or simply inappropriate culture conditions was not clear. We tested the growth and differentiation potential of NCSCs at approximately 5% O(2), which more closely approximates physiological oxygen levels. Eighty-three percent of p75(+)P(0-) cells isolated from embryonic day 14.5 sciatic nerve behaved as stem cells under these conditions, suggesting that this is a nearly pure population. Furthermore, addition of BMP2 plus forskolin in decreased oxygen cultures elicited differentiation of thousands of cells expressing tyrosine hydroxylase, dopamine-beta-hydroxylase, and the SA lineage marker SA-1 in nearly all colonies. Such cells also synthesized and released dopamine and norepinephrine. These data demonstrate that isolated mammalian NCSCs uniformly possess SA lineage capacity and further suggest that oxygen levels can influence cell fate. Parallel results indicating that reduced oxygen levels can also promote the survival, proliferation, and catecholaminergic differentiation of CNS stem cells (Studer et al., 2000) suggests that neural stem cells may exhibit a conserved response to reduced oxygen levels.

Transient Notch activation initiates an irreversible switch from neurogenesis to gliogenesis by neural crest stem cells.

The genesis of vertebrate peripheral ganglia poses the problem of how multipotent neural crest stem cells (NCSCs) can sequentially generate neurons and then glia in a local environment containing strong instructive neurogenic factors, such as BMP2. Here we show that Notch ligands, which are normally expressed on differentiating neuroblasts, can inhibit neurogenesis in NCSCs in a manner that is completely dominant to BMP2. Contrary to expectation, Notch activation did not maintain these stem cells in an uncommitted state or promote their self-renewal. Rather, even a transient activation of Notch was sufficient to cause a rapid and irreversible loss of neurogenic capacity accompanied by accelerated glial differentiation. These data suggest that Notch ligands expressed by neuroblasts may act positively to instruct a cell-heritable switch to gliogenesis in neighboring stem cells.

In vivo proliferation and cell cycle kinetics of long-term self-renewing hematopoietic stem cells.

A rare set of hematopoietic stem cells (HSC) must undergo a massive expansion to produce mature blood cells. The phenotypic isolation of HSC from mice offers the opportunity to determine directly their proliferation kinetics. We analyzed the proliferation and cell cycle kinetics of long-term self-renewing HSC (LT-HSC) in normal adult mice. At any one time, approximately 5% of LT-HSC were in S/G2/M phases of the cell cycle and another 20% were in G1 phase. BrdUrd incorporation was used to determine the rate at which different cohorts of HSC entered the cell cycle over time. About 50% of LT-HSC incorporated BrdUrd by 6 days and >90% incorporated BrdUrd by 30 days. By 6 months, 99% of LT-HSC had incorporated BrdUrd. We calculated that approximately 8% of LT-HSC asynchronously entered the cell cycle per day. Nested reverse transcription-PCR analysis revealed cyclin D2 expression in a high proportion of LT-HSC. Although approximately 75% of LT-HSC are quiescent in G0 at any one time, all HSC are recruited into cycle regularly such that 99% of LT-HSC divide on average every 57 days.