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The objective of this study was to investigate the effects of replacing barley and soybean meal with increasing levels of by-products on production, digestive, and metabolic parameters in early-mid lactation dairy cows offered perennial ryegrass-based pasture. Forty-eight (32 multiparous and 16 primiparous) dairy cows that were 64 ± 24 d in milk were assigned to 1 of 4 pasture-based dietary treatments (n = 12) in a randomized block design experiment that ran for 70 d. Treatments consisted of a perennial ryegrass-based pasture and 1 of 4 supplementary concentrates: BP35, BP55, BP75, and BP95 containing 35, 55, 75, and 95% by-products, respectively, in the concentrate on a dry matter basis. The by-products used were soyhulls, dried distillers grains, and palm kernel extract in equal proportions. Barley and soybean meal were replaced as by-product inclusion level increased. In this study, intakes of pasture dry matter (15.7 kg) and total dry matter (21.1 kg) were not affected by treatment. Similarly, milk production parameters (milk yield, milk composition, somatic cell count, and urea) were not different between treatments. Unsaturated fatty acids were lower in the milk of cows offered BP35 and BP55 compared with those offered BP75 and BP95. Concentrations of β-hydroxybutyrate, nonesterified fatty acids, and other blood metabolites were within normal range and did not differ between treatments, and cow body condition score and body weight were also not different. Equally, N was unaffected by diet. Blood urea N was lower in the BP75 group compared with BP35. This study demonstrated that barley and soybean meal can be replaced with soyhulls, dried distillers grains, and palm kernel extract without affecting milk production, digestive, or metabolic parameters in dairy cows offered a pasture-based diet., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Relations between early child care and adolescent functioning at the end of high school (EOHS; M age = 18.3 years) were examined in a prospective longitudinal study of 1,214 children. Controlling for extensive measures of family background, early child care was associated with academic standing and behavioral adjustment at the EOHS. More experience in center-type care was linked to higher class rank and admission to more selective colleges, and for females to less risk taking and greater impulse control. Higher quality child care predicted higher academic grades and admission to more selective colleges. Fewer hours in child care was related to admission to more selective colleges. These findings suggest long-term benefits of higher quality child care, center-type care, and lower child-care hours for measures of academic standing at the EOHS. (PsycINFO Database Record, ((c) 2016 APA, all rights reserved).)

Fatty acid alkyl esters (FAAEs) were determined at 10-100mg/L in biodiesel and blends with petrodiesel without sample pre-treatment using gas chromatography with a tandem differential mobility detector. Selectivity was provided through chromatographic separations and atmospheric pressure chemical ionization reactions in the detector with mobility characterization of gas ions. Limits of detection were ~0.5ng with an average of 2.98% RSD for peak area precision, ≤1.3% RSD for retention time precision, and ≤9.2% RSD for compensation voltage precision. Biodiesel blends were classified using principal component analysis (PCA) and hierarchical cluster analysis (HCA). Unsupervised cluster analysis captured 52.72% of variance in a single PC while supervised analysis captured 71.64% of variance using Fisher ratio feature selection. Test set predictions showed successful clustering according to source or feedstock when regressed onto the training set model. Detection of the regulated substance methyl linolenate (C18:3 me) was achieved in 6-10s with a 1m long capillary column using dual ion filtering in the tandem differential mobility detector., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The productivity of grazing systems is primarily limited by the scale and efficiency of systems applied to the grazable land platform adjacent to the milking parlor. The objective of this study was to compare forage production, utilization and quality, milk production, and requirement for supplementary feeds for 2 different grazing platform stocking rate (GPSR) treatments over 4 yr. Animals were randomly allocated to 1 of 2 GPSR treatments: high-closed (HC; 3.1 cows/ha) and high-open (HO; 4.5 cows/ha), which were designed to represent alternative GPSR in a post-European Union milk quota, spring calving, pasture-based milk production system. Animal production data were analyzed using Proc MIXED of SAS with GPSR, year, and parity included as fixed effects in the final model. Within a seasonal spring calving grazing system, at high GPSR and offering moderate amounts of additional supplements based on pasture supply deficits, both systems produced more milk and fat plus protein per hectare in comparison with Irish commercial dairy farms. Although requiring additional supplementation, increased GPSR resulted in increased milk production per hectare but also in an increased requirement for concentrate and forage supplementation during lactation. No significant influence of GPSR was found on body weight and body condition score or reproductive performance during the 4-yr study period. In addition, GPSR also had no effect on pasture production, utilization, or quality during the study period. The strategic use of additional supplements with restricted pasture availability at higher GPSR maintained milk production per cow and significantly increased milk production per hectare., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The production and utilization of increased quantities of high quality pasture is of paramount importance in pasture-based milk production systems. The objective of this study was to evaluate the cumulative effects of alternative integrated grazing strategies, incorporating alternative stocking rate (SR) and grazing severities, on pasture productivity and grazing efficiency over multiple years within farm systems using perennial ryegrass dominant pastures. Three whole-farm SR treatments were compared over 4 complete grazing seasons (2009 to 2012 inclusive): low (2.51 cows/ha; LSR), medium (2.92 cows/ha; MSR), and high (3.28 cows/ha; HSR). Each system had its own farmlet containing 18 paddocks and remained on the same treatment for the duration of the study. Stocking rate had a significant effect on all grazing variables with the exception of soil fertility status and sward density. Increased SR resulted in increased total annual net pasture accumulation, improved sward nutritive value, and increased grazed pasture utilization. Total annual net pasture accumulation was greatest in HSR [15,410kg of dry matter (DM)/ha], intermediate for MSR (14,992kg of DM/ha), and least for LSR (14,479kg of DM/ha) during the 4-yr study period. A linear effect of SR on net pasture accumulation was detected with an increase in net pasture accumulation of 1,164.4 (SE=432.7) kg of DM/ha for each 1 cow/ha increase in SR. Pregrazing pasture mass and height and postgrazing residual pasture mass and height were greatest for LSR, intermediate for the MSR, and lowest for the HSR. In comparison with the LSR, the imposition of a consistently increased grazing severity coupled with increased whole farm SR in MSR and HSR treatments arrested the decline in sward nutritive value, typically observed during mid-season. Incorporating the individual beneficial effects of SR on pasture accumulation, nutritive value, and utilization efficiency, total proportional energy (unité fourragère lait) utilization per hectare increased significantly with increasing SR (+0.026 and +0.081 for MSR and HSR, respectively). These results quantify the significant effect of grazing management practices on the feed production capability of modern perennial ryegrass pastures for intensive grazing dairy production systems. Furthermore, these results highlight the importance of consistently imposing grazing treatments over multiple years, and within integrated whole farm systems, to accurately assess the longer term effects of alternate grazing management practices on pasture productivity., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Saxitoxins (STXs) are paralytic alkaloids produced by marine dinoflagellates in response to biotic and abiotic stressors yielding harmful algal blooms. Because STX impacts coastal, near-shore communities to a greater extent than would be predicted by its relative abundance, it has been referred to as a "molecule of keystone significance" in reference to Robert Paine's Keystone Species Concept. Pisaster ochraceus, the predator upon which Paine's concept was founded, inhabits waters regularly plagued by harmful algal blooms, but the effects of STX on Pisaster have not yet been investigated. Here, we used laboratory and field experiments to examine the potential consequences of exposure to STX on sea stars' feeding, attachment to the substrate, and success in fertilization. Pisaster exhibited similar feeding behaviors when offered non-toxic prey, STX-containing prey, or a combination of the two. Although feeding behavior is unaffected, consumption of STX poses a physiological tradeoff. Sea stars in the laboratory and field had significantly lower thresholds of the force needed to detach them from their substrates after either being exposed to, or consuming, STX. High pressure (or high performance) liquid chromatography analysis indicated an accumulation of STX (and structural analogues) in sea stars' viscera, likely due to trophic transfer from toxic prey. Incidence of fertilization tended to decrease when gametes were exposed to high, yet ecologically relevant, STX concentrations of STX. These findings suggest that the molecule of keystone significance, STX, produced during harmful algal blooms extends its impacts to rocky intertidal communities by way of the keystone predator P. ochraceus., (© The Author 2015. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.)

Economically viable and productive farming systems are required to meet the growing worldwide need for agricultural produce while at the same time reducing environmental impact. Within grazing systems of animal production, increasing concern exists as to the effect of intensive farming on potential N losses to ground and surface waters, which demands an appraisal of N flows within complete grass-based dairy farming systems. A 3-yr (2011 to 2013) whole-farm system study was conducted on a free-draining soil type that is highly susceptible to N loss under temperate maritime conditions. Soil solution concentrations of N from 3 spring-calving, grass-based systems designed to represent 3 alternative whole-farm stocking rate (SR) treatments in a post-milk quota situation in the European Union were compared: low (2.51 cows/ha), medium (2.92 cows/ha), and high SR (3.28 cows/ha). Each SR had its own farmlet containing 18 paddocks and 23 cows. Nitrogen loss from each treatment was measured using ceramic cups installed to a depth of 1m to sample the soil water. The annual and monthly average nitrate, nitrite, ammonia, and total N concentrations in soil solution collected were analyzed for each year using a repeated measures analysis. Subsequently, and based on the biological data collated from each farm system treatment within each year, the efficiency of N use was evaluated using an N balance model. Based on similar N inputs, increasing SR resulted in increased grazing efficiency and milk production per hectare. Stocking rate had no significant effect on soil solution concentrations of nitrate, nitrite, ammonia, or total N (26.0, 0.2, 2.4, and 32.3 mg/L, respectively). An N balance model evaluation of each treatment incorporating input and output data indicated that the increased grass utilization and milk production per hectare at higher SR resulted in a reduction in N surplus and increased N use efficiency. The results highlight the possibility for the sustainable intensification of grass-based dairy systems and suggest that, at the same level of N inputs, increasing SR has little effect on N loss in pastoral systems with limited imported feed. These results suggest that greater emphasis should be attributed to increased grass production and utilization under grazing to further improve the environmental impact of grazing systems., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Disparities between nutrient intake and demand often result in a state of negative energy balance (EB) in the early-lactation dairy cow. Reducing dietary crude protein (CP) content and providing glucogenic nutrients may overcome this issue. This study evaluates whether or not offering a diet lower in CP and higher in nonfiber carbohydrates (LP-NFC) can improve EB and the metabolic status of the early-lactation dairy cow compared with a diet higher in CP and fibrous carbohydrates (HP-FC). Twenty Holstein-Friesian dairy cows were assigned to 1 of 2 dietary treatments in a randomized block design. Diets were isoenergetic (6.57 MJ of net energy for lactation) and formulated to contain 15% CP and 6% starch (HP-FC), or 12% CP and 28% starch (LP-NFC) and were offered for the first 63 d of lactation. Intake and milk yield were determined daily, whereas milk and blood samples, weights, and body condition scores were collected weekly. Intakes (mean ± standard errors of the mean, SEM) of dry matter (17.4 ± 0.6 kg/d) and energy (113.0 ± 4.6 MJ of net energy for lactation) were not different between treatments. However, the HP-FC group had a higher milk yield (31.8 vs. 28.9 ± 1.4 kg/d) and a lower EB compared with the LP-NFC group. Blood urea N concentration (3.5 vs. 1.8 ± 0.2 mmol/L) was higher, whereas bilirubin (6.0 vs. 6.7 ± 0.2 mmol/L) and β-hydroxybutyrate concentrations (0.7 vs. 0.8 ± 0.05 mmol/L) were lower in the HP-FC group compared with the LP-NFC group. These data suggest that EB can be improved during early lactation through the manipulation of milk output by offering a lower CP, higher NFC diet., (Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The primary objective of the study was to quantify the effect of stocking rate (SR) and calving date (CD) on milk production, dry matter intake (DMI), energy balance (EB), and milk production efficiency over 4 consecutive years (2009 to 2012). Two groups of Holstein-Friesian dairy cows with different mean CD were established from within the existing research herd at Moorepark (Teagasc, Ireland). Animals were assigned to either an early calving (mean CD February 14) treatment or a late calving (mean CD March 2) treatment. Animals within each CD treatment were randomly allocated to 1 of 3 whole-farm SR treatments: low (LSR; 2.51 cows/ha), medium (MSR; 2.92 cows/ha), and high (HSR; 3.28 cows/ha), and animals remained on the same farmlet for the duration of the study. Individual animal DMI was estimated 3 times per year at grass using the n-alkane technique in March (spring), May (summer), and September (autumn), corresponding to, on average, 45, 132, and 258 d in milk, respectively. A total of 138 spring-calving dairy cows were used during each year of the study. The effects of SR, CD, season, and their interaction were studied using mixed models. Individual animal milk production, body weight, body condition score, and the efficiency of milk production were significantly decreased as SR increased due to a reduction in herbage availability. The existence of CD × SR × season interactions for production, DMI, and EB indicate that delaying the herd mean CD can be an effective strategy to minimize the reduction in animal performance, particularly in spring at higher SR. This study further confirms the benefits of a new approach to the evaluation of herbage allowance known as the individual herbage allowance, which encompasses the 3 main factors restricting DMI in rotational grazing; namely, the average daily herbage allowance of the group, the intake capacity of the individual animal within the group, and the relative intake capacity of the animal within the competing herd., (Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Objective: The purpose of this case series is to report how the symptom section of the Sport Concussion Assessment Tool 2 (SCAT2) was used to manage athletes with concussions in a high school training room setting and to address the need for SCAT2 baseline measurements., Clinical Features: During a 4-month period, 3 doctors of chiropractic with certification from the American Chiropractic Board of Sports Physicians managed 15 high school athletes with concussions in a multidisciplinary setting. Fourteen athletes were male American football players, and one was a female volleyball player., Intervention and Outcome: Of the 15 athletes, 3 athletes had baseline SCAT2 documentation. Athletes were evaluated and returned to play with a graded return to play protocol using the SCAT2 symptoms and serial physical examinations. Once participants were asymptomatic, they began a graded return to play process. A total of 47 SCAT2 tests were performed on the 15 athletes, averaging 3.13 SCAT2 evaluations per patient. Of the 15 athletes evaluated, 6 were managed and cleared for return to play; 2 of the athletes sustained concussions in the last week of the season, thus ending their season; and 3 athletes were cleared by medical doctors. None of the athletes under care reported an adverse event., Conclusion: The utilization of the SCAT2 with serial physical examinations provided objective measures for athlete's injuries, allowing the practitioners to evaluate concussions. More efforts are needed to collect baseline SCAT2 to compare these scores with subsequent SCAT2 scores following athletic injuries.

The objective of this study was to determine the effect of pregrazing pasture herbage mass (HM) on CH4 emissions, ruminal fermentation, and ADG of grazing beef heifers at 2 stages of the grazing season. Thirty Limousin cross heifers were allocated to 1 of 2 target pregrazing HM treatments [a low HM (LHM) or high HM (HHM) treatment] for 126 d in a randomized block design experiment. Pasture herbage and heifer rumen fluid samples were collected, and enteric CH4 emissions were determined using an SF6 tracer technique during two 5-d measurement periods [MP; MP 1 (25 to 29 May) and MP 2 (6 to 10 September)]. Both DMI and GE intake (GEI) were measured during MP 2, and ADG of the heifers was measured every 14 d throughout the 126-d grazing period. Mean HM for the LHM and HHM treatments were 1,300 and 2,000 kg DM/ha, respectively, during MP 1 and 2,800 and 3,200 kg DM/ha, respectively, during MP 2. The CP concentration of the offered herbage was greater (P < 0.01) for the LHM treatment during MP 1 and tended (P < 0.1) to be greater for the LHM herbage during MP 2. No difference (P > 0.10) in the NDF concentration of the herbage was found between the HM treatments during MP 1 or 2. There was no effect (P > 0.10) of HM treatment on total CH4 emissions (g/d) for either MP [mean value across HM treatments of 121 (SED 5.4) g/d during MP 1 and 132 (8.8) g/d during MP 2], but CH4 emissions (g) per kilogram of ADG were reduced (P < 0.05) from heifers fed the LHM treatment during MP 1 and 2 [mean values for LHM and HHM of 135 and 163 (SED 9.5) g/kg, respectively, during MP 1 and corresponding values of 150 and 194 (9.9) g/kg during MP 2]. Heifers fed the LHM treatment had greater (P < 0.001) ADG throughout the grazing period [mean value across the 126-d grazing period of 0.88 (SEM 0.032) kg/d] than those fed the HHM treatment [corresponding value of 0.73 (0.034)]. For MP 2, CH4 emissions per kilogram of DMI (g CH4/kg DMI) and per megajoule of GEI (MJ CH4/MJ GEI) tended (P ≤ 0.08) to be less for heifers fed the LHM [19.3 (0.08) g/kg and 0.056 (0.0020) MJ/MJ, respectively] than for the HHM (21.1 g/kg and 0.061 MJ/MJ) treatment, and there were no differences (P > 0.10) in DMI or GEI of the heifers between the HM treatments. The results of this study suggest that offering a low pregrazing HM sward will reduce enteric CH4 emissions relative to ADG throughout the grazing season because of increased ADG.

A stochastic whole-farm simulation model was used to examine alternative strategies for new entrant dairy farmers to grow and develop dairy farm businesses in the context of European Union (EU) milk quota abolition in 2015. Six alternative strategies were compared: remain static, natural growth expansion, waiting until after EU milk quota abolition to expand, a full-scale expansion strategy without milk quotas and not incurring super levy penalties, a full-scale expansion strategy with milk quotas and incurring super levy penalties, and once-a-day milking until EU milk quota abolition, followed by full-scale expansion. Each discrete whole farm investment strategy was evaluated over a 15-yr period (2013-2027) using multiple financial stability and risk indicators, including overall discounted farm business profitability, net worth change, return on investment, and financial risk. The results of this study indicate that, although associated with increased risk, dairy farm expansion will ensure the future profitability of the farm business. Within the context of EU milk quotas until 2015, the most attractive expansion strategy is to increase cow numbers while avoiding super levy fines using once-a-day milking techniques, increasing to the full capacity of the dairy farm once milk quotas are removed. In contrast, the results also indicate that dairy farms that remain static will experience a significant reduction in farm profitability in the coming year due to production cost inflation. Cash flow deficits were observed during the initial year of expansion and, therefore, rapidly expanding dairy farm businesses require a significant cash reserve to alleviate business risk during the initial year of expansion. The results of this analysis also indicate that dairy farm businesses that expand using lower cost capital investments and avoid milk quota super levy fines significantly reduce the financial risks associated with expansion., (Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Dietary change, an inconsistent nutrient intake and high levels of milk production make the early post-partum period (PP) a challenging time for the lactating dairy cow. This experiment investigates the effects of two early PP nutritional management strategies (NM): abrupt introduction to pasture (AP) or a total mixed ration (TMR) for 21 days followed by a gradual introduction to pasture over 7 days (GP), with (Y) or without (C) live yeast (YS) on milk production, energy balance (EB) and selected metabolic and reproductive variables. Forty multiparous dairy cows were assigned to one of four dietary treatments in a two (AP vs. GP) by two (Y vs. C) factorial, randomized block design. The experiment was conducted from days 1 to 70 PP. Blood samples were taken on day 1, day 5 and every 10 days until day 45 to determine metabolites, whilst intake (DMI), and EB were determined during week 6 PP. Milk was sampled weekly for fat, protein and lactose. Trans-rectal scanning for reproductive variables commenced on day 10 PP. Animals in the GP group had a higher DMI (p = 0.04), higher fat yield (p = 0.08) and fewer days to first ovulation (p = 0.09) vs. those in the AP group. EB (-3.5 ± 0.67 units of energy for milk production) and body condition score loss (0.70 ± 0.09) were not affected by NM. However, non-esterified fatty acids (NEFA) (p < 0.01) were higher, and glucose (p = 0.02) was lower in the AP vs. the GP group. Supplementary YS tended to improve EB (p = 0.09) and reduced NEFA (p < 0.01) vs. non-supplemented animals. These data suggest that offering animals a nutritionally balanced TMR during the first 3 weeks PP followed by a gradual introduction to pasture can improve DMI vs. pasture-based diets. Additionally, the blood metabolic profile suggests a more favourable energy status in the GP group or where YS was supplemented during the early PP period., (Journal of Animal Physiology and Animal Nutrition © 2013 Blackwell Verlag GmbH.)

β-Glucans have been identified as natural biomolecules with immunomodulatory activity. The first objective of the present study was to compare the effects of purified β-glucans derived from Laminaria digitata, L. hyperborea and Saccharomyces cerevisiae on piglet performance, selected bacterial populations and intestinal volatile fatty acid (VFA) production. The second aim was to compare the gene expression profiles of the markers of pro- and anti-inflammation in both unchallenged and lipopolysaccharide (LPS)-challenged ileal and colonic tissues. β-Glucans were included at 250 mg/kg in the diets. The β-glucans derived from L. hyperborea, L. digitata and S. cerevisiae all reduced the Enterobacteriaceae population (P<0·05) without influencing the lactobacilli and bifidobacteria populations (P>0·05) in the ileum and colon. There was a significant interaction between gastrointestinal region and β-glucan source in the expression of cytokine markers, IL-1α (<0·001), IL-10 (P<0·05), TNF-α (P<0·05) and IL-17A (P<0·001). β-Glucans did not stimulate any pro- or anti-inflammatory cytokine markers in the ileal epithelial cells. In contrast, the expression of a panel of pro- and anti-inflammatory cytokines (IL-1α, IL-10, TNF-α and IL-17A) was down-regulated in the colon following exposure to β-glucans from all the three sources. However, the data suggest that the soluble β-glucans derived from L. digitata may be acting via a different mechanism from the insoluble β-glucans derived from L. hyperborea and S. cerevisiae, as the VFA profile was different in the L. digitata-treated animals. There was an increase in IL-8 gene expression (P<0·05) in the gastrointestinal tract from the animals exposed to L. digitata following an LPS ex vivo challenge that was not evident in the other two treatment groups. In conclusion, β-glucans from both seaweed and yeast sources reduce Enterobacteriaceae counts and pro-inflammatory markers in the colon, though the mechanisms of action may be different between the soluble and insoluble fibre sources.

Comprehensive two-dimensional (2D) separations, such as comprehensive 2D gas chromatography (GC×GC), liquid chromatography (LC×LC), and related instrumental techniques, provide very large and complex data sets. It is often up to the software to assist the analyst in transforming these complex data sets into useful information, and that is precisely where the field of chemometric data analysis plays a pivotal role. Chemometric tools for comprehensive 2D separations are continually being developed and applied as researchers make significant advances in novel state-of-the-art algorithms and software, and as the commercial sector continues to provide user friendly chemometric software. In this review, we build upon previous reviews of this topic, by focusing primarily on advances that have been reported in the past five years. Most of the reports focus on instrumental platforms using GC×GC with either flame ionization detection (FID) or time-of-flight mass spectrometry (TOFMS) detection, or LC×LC with diode array absorbance detection (DAD). The review covers the following general topics: data preprocessing techniques, target analyte techniques, comprehensive nontarget analysis techniques, and software for chemometrics in multidimensional separations., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Forty-four early-lactation dairy cows of mixed parity were used to examine the effect of 4 supplemental concentrate types (n=11) on milk production and metabolic status. Animals were blocked by parity and calving date, and blocks were balanced for previous milk yield and milk protein yield. Cows received grazed pasture plus 5.17 kg of DM/d of 1 of the following isoenergetic (1.1 units of energy for lactation) concentrates: 1) high crude protein (CP) with rolled barley (HP, 19% CP); b) low CP with rolled barley (LP, 15% CP); c) low CP with barley and a supplemental methionine hydroxy analog (HMBi; LP + HMBi, 15% CP); and d) low CP with ground corn (LP-corn, 15% CP). Milk yield was recorded from d 1 to 100 postpartum, with weekly milk sampling, body weight, and body condition score (BCS) measurements. Blood and rumen sampling were conducted weekly from wk 2 to 6 postpartum. Milk yield was lower for cows in the LP treatment compared with those offered other concentrate types (25.2 vs. 27.5 ± 0.39 kg/d). Animals in the HP group had a higher milk yield than those in the LP + HMBi group (28.2 vs. 26.8 ± 0.39 kg/d). Milk fat yield was lower from animals in the LP-corn group compared with those in the LP + HMBi group (0.94 vs. 1.03 ± 0.03 kg/d). Milk protein yield was lower in the LP group compared with those in the HP group (0.88 vs. 0.97 ± 0.02 kg/d). Animal body weight, BCS, and BCS loss were not affected by concentrate type. However, nonesterified fatty acids were higher from animals in the HP group than for those in the LP + HMBi group (0.41 vs. 0.33 ± 0.03 mmol/L), and β-hydroxy butyric acid was higher from animals in the HP group than for those in the other treatments (0.71 vs. 0.59 ± 0.03 mmol/L). Glucose was higher from animals in the LP-corn group than for those in the HP and LP groups (3.3 vs. 3.2 ± 0.05 mmol/L). Blood urea-N was higher from animals offered HP compared with those offered the other treatments (5.49.6 vs. 4.21 ± 0.44 mmol/L). However, rumen NH(3)-N and volatile fatty acid concentration in the rumen were not affected by supplemental concentrate type. Reducing supplemental concentrate CP reduced milk yield. However, milk fat production and energy-corrected milk were not different, reducing the likelihood of an improved energy balance or a more favorable blood metabolic profile in early-lactation dairy cows grazing perennial ryegrass. Offering HMBi with low-CP concentrates or replacing rolled barley with ground maize improves milk production relative to low-CP concentrates and metabolic status relative to high-CP concentrates., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Forty-four early lactation (64 ± 20 d in milk) dairy cows of mixed parity were used to assess the effect of 4 supplementary concentrate types (n=11) on N partitioning. Animals were blocked on parity and calving date, and blocks were balanced for previous milk yield and milk protein yield. Cows received grazed pasture plus 5.17 kg of dry matter (DM)/d of one of the following isoenergetic concentrates: high crude protein (CP) with rolled barley (HP, 19% CP); low CP with rolled barley (LP, 15% CP); low CP with barley and supplementary 2-hydroxy-4-methylthio butanoic acid (HMBi; LP+HMBi, 15% CP); and low CP with ground corn (LP Corn, 15% CP). Nitrogen partitioning studies were conducted at wk 6 and 10 postpartum by using the n-alkane technique to determine pasture dry matter intake (DMI). Pasture DMI (13.3kg of DM/d) and dietary digestibility of DM were not affected by concentrate type. Milk yield was lower for LP compared with other concentrate types (25.4 vs. 28.3 kg/d). Yields of milk protein and milk casein were not affected by concentrate type. However, milk solid yield and milk fat yield were higher for LP+HMBi (1.97 and 0.92 kg/d) compared with LP (1.72 and 0.87 kg/d). Concentrations of fat, protein, lactose, and casein were not affected by concentrate type. Dietary N intake was higher for HP compared with other treatments (0.545 vs. 0.482 kg/d, HP vs. average of the 3 LP treatments). Dietary N intakes were not different among low CP concentrates. Fecal N excretion was not affected by concentrate type. However, urinary N excretion was related to N intake and was higher for HP compared with other treatments (0.261 vs. 0.195 kg/d, HP vs. average of the 3 LP treatments). Urinary N excretion was not different among low CP concentrates. Milk N output was higher for HP (0.139 kg/d) compared with LP (0.12 kg/d) but not LP+HMBi (0.137 kg/d) or LP Corn (0.138 kg/d). The portion of feed N excreted as feces N was lower for HP compared with other treatments (0.272 vs. 0.327, HP vs. average of the 3 LP treatmentsHowever, the portion of feed N excreted as urine N was higher for HP (0.466) compared with LP+HMBi (0.408) and LP Corn (0.366) but not compared with LP. The portion of feed N excreted as milk N was higher for LP Corn (0.282) compared with HP (0.257) but not LP+HMBi or LP. Dietary reformulation to reduce N excretion in pasture-based dairy production systems is possible. However, maintenance of milk yield and milk N when concentrate CP was reduced (19 vs. 15%) required the use of either protected AA (HMBi) or ground corn., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Plant-derived hydrolysates are widely used in mammalian cell culture media to increase yields of recombinant proteins and monoclonal antibodies (mAbs). However, these chemically varied and undefined raw materials can have negative impact on yield and/or product quality in large-scale cell culture processes. Traditional methods that rely on fractionation of hydrolysates yielded little success in improving hydrolysate quality. We took a holistic approach to develop an efficient and reliable method to screen intact soy hydrolysate lots for commercial recombinant mAb manufacturing. Combined high-resolution (1) H nuclear magnetic resonance (NMR) spectroscopy and partial least squares (PLS) analysis led to a prediction model between product titer and NMR fingerprinting of soy hydrolysate with cross-validated correlation coefficient R(2) of 0.87 and root-mean-squared-error of cross-validation RMSECV% of 11.2%. This approach screens for high performance hydrolysate lots, therefore ensuring process consistency and product quality in the mAb manufacturing process. Furthermore, PLS analysis was successful in discerning multiple markers (DL-lactate, soy saccharides, citrate and succinate) among hydrolysate components that positively and negatively correlate with titer. Interestingly, these markers correlate to the metabolic characteristics of some strains of taxonomically diverse lactic acid bacteria (LAB). Thus our findings indicate that LAB strains may exist during hydrolysate manufacturing steps and their biochemical activities may attribute to the titer enhancement effect of soy hydrolysates., (Copyright © 2012 American Institute of Chemical Engineers (AIChE).)

The aim of this study was to assess the influence of age, body weight (BW) and body condition score (BCS) of maiden Holstein-Friesian heifers before mating start date (MSD) on the rate of puberty, subsequent production and longevity and their implications with regard to farm profitability. Data were available on 871 Holstein-Friesian heifers from 48 herds. BW was recorded electronically and BCS was recorded by a single operator on a scale of 1 to 5. Heifer age was calculated as the number of days from birth to the day of visit. All of the independent variables of interest were grouped into three or four categories. Three age categories (thirtiles), four BW categories (quartiles) and four BCS categories (≤ 2.75, 3.00, 3.25 and ≥ 3.50) were formed. Heifers with an identifiable corpus lutuem (CL) in the presence or absence of large follicles and peri-ovulatory signs and with a plasma progesterone (P4) concentration ≥ 1 ng/ml were classified as pubertal. In addition, heifers without an identifiable CL in the presence or absence of large follicles and peri-ovulatory signs but with a P4 concentration ≥ 1 ng/ml were also classified as pubertal. Age, BW and BCS at MSD were all found to be significantly associated with pubertal rate (P < 0.05). Age was shown to have no practical implications on subsequent cow performance. BW at MSD was favourably associated with subsequent calving date (P < 0.05), subsequent cow BW (P < 0.001) and potential (305 days) milk fat plus protein yield (P < 0.001). BCS at MSD was found to be favourably associated with milk fat plus protein yield potential (P < 0.05) and BCS (P < 0.001) during lactation. The economic analysis undertaken indicated that larger, well-grown heifers will be more profitable because of superior production potential, all else being equal. However, because of the finding of poorer reproductive efficiency in heifers grown to more than 343 kg at MSD, heifers at ∼330 kg at MSD are deemed optimal. This will correspond to mature cow BW of ∼550 kg.

The two main goals of the analytical method described herein were to (1) use principal component analysis (PCA), hierarchical clustering (HCA) and K-nearest neighbors (KNN) to determine the feedstock source of blends of biodiesel and conventional diesel (feedstocks were two sources of soy, two strains of jatropha, and a local feedstock) and (2) use a partial least squares (PLS) model built specifically for each feedstock to determine the percent composition of the blend. The chemometric models were built using training sets composed of total ion current chromatograms from gas chromatography-quadrupole mass spectrometry (GC-qMS) using a polar column. The models were used to semi-automatically determine feedstock and blend percent composition of independent test set samples. The PLS predictions for jatropha blends had RMSEC=0.6, RMSECV=1.2, and RMSEP=1.4. The PLS predictions for soy blends had RMSEC=0.5, RMSECV=0.8, and RMSEP=1.2. The average relative error in predicted test set sample compositions was 5% for jatropha blends and 4% for soy blends., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This study examined gender differences in the level and developmental course of relational aggression in middle childhood, as well as early predictors and outcomes of relational aggression, after controlling for concurrent physical aggression. Relational (RAgg) and Physical aggression (PAgg) scores for 558 boys and 545 girls at ages 8-11 in the NICHD Study of Early Child Care and Youth Development (SECCYD) were created by combining the highest rating for each item across mother and teacher reports. Longitudinal analyses were conducted using Latent Curve Models of RAgg with PAgg as a time-varying covariate, with all parameters allowed to vary by gender. Boys and girls had different growth parameters of RAgg. Girls' RAgg intercept was higher and the slope was not different from zero; boys' RAgg intercept was lower and the slope declined. Mother-child conflict in early childhood predicted RAgg intercept for both boys and girls, but maternal harsh control and sensitivity were also uniquely predictive for girls, whereas center care was uniquely predictive for boys. RAgg intercept predicted adolescent self-reports of depression for girls and delinquency and risk-taking for both boys and girls; the magnitude of the association with risk-taking was significantly greater for boys.

Two groups of Holstein-Friesian dairy cows with different mean calving dates (CD) were established from within the existing research herd at Moorepark (Teagasc, Ireland). Animals were assigned to either an early calving (mean CD February 12) treatment or a late calving (mean CD February 25) treatment. Animals within each CD treatment were randomly allocated to 1 of 3 stocking rate (SR) treatments, low (2.51 cows/ha), medium (2.92 cows/ha), or high (3.28 cows/ha), which were designed to represent 3 alternative whole-farm SR in a spring-calving, grass-based milk production system following abolition of the European Union milk quotas. A total of 138 spring-calving dairy cows, comprising 2 strains of Holstein-Friesian, North American (NA) and New Zealand (NZ), were used in each year (2009 and 2010). The effects of CD, SR treatment, genetic strain, and their interactions on reproductive performance, body weight, body condition score, blood metabolites, hormone and immunological parameters, and health status were analyzed. Stocking rate and CD had no effect on pregnancy rates, immunological parameters, or health status, although a tendency was observed for more reproductive intervention as SR increased. Earlier calving and increased SR also resulted in reduced body weight, body condition score, and metabolic status in early lactation. Strain of Holstein-Friesian also affected reproductive performance. The NZ strain tended to have a higher submission rate and 42 d pregnancy rate compared with the NA strain, and a strain by SR interaction was observed for pregnancy rate to first service. Earlier calving and increased SR can be achieved without adverse effects on overall pregnancy rates. The existence of a SR by strain interaction for several reproductive variables suggests that the smaller NZ strain is better adapted to increased SR systems., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to compare the biological and economic efficiencies of 2 likely future pasture-based systems of milk production differing in overall stocking rate and concentrate supplementation level on a wetland drumlin soil in the Border Midlands Western region of Ireland. Physical performance data were obtained from a 3-yr systems comparison at Ballyhaise College, Co. Cavan, comparing 2 production systems: a high grass (HG) system (578 kg of concentrate/cow at 2.45 livestock units per hectare) and a high intensity (HI) system (1,365 kg of concentrate/cow at 2.92 livestock units/ha). Animal production data were analyzed using a mixed model, with feed system, year, and parity included as fixed effects in the final model. Feed system had a significant effect on all yield variables with higher yields in the HI system. Production system had no significant influence on reproductive performance. The Moorepark Dairy Systems Model, a stochastic budgetary simulation model, was used to simulate a model farm integrating biological data from each feed system to identify the economic effect of each system at 3 future milk prices of 22, 27, and 33 euro cents per liter (€c/L). Two economic scenarios were investigated within the model: scenario 1 (S1) assumed fixed cow numbers (n=55 cows) and scenario 2 (S2) assumed fixed land area (n=40 ha). At a milk price of 27 or 33 €c/L, profit per cow, per kilogram of milk solids, and per hectare were similar for HG and HI in S1 and higher for HI in S2. At a milk price of 22 €c/L, all systems were unprofitable, with increased losses realized in the HI system (both S1 and S2) compared with the HG system. Pasture-based systems of milk production in the northern region of Ireland are capable of highly efficient and profitable milk production. Moreover, the efficacy of increased supplementation to remove the constraints of pasture seasonality will depend on the cost of supplementation and the price paid for additional milk produced., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Four primiparous and 4 multiparous midlactation dairy cows were stratified by pre-experimental milk yield (23.5 ± 2.3 kg/d), protein yield (0.75 ± 0.066 kg/d), parity, and days in lactation (121 ± 10 d) into 4 groups of 2 in a 2 × 2 factorial, Latin square design (n = 8) to assess the effect of forage source and a supplementary methionine hydroxy analog on nitrogen (N) balance where low crude protein (CP) diets (13.3%) are offered. Diets contained either predominantly grass silage [GS (G- and G+)] or corn silage [CS (C- and C+)] as the forage source and were offered with (G+ and C+) or without (G- and C-) the isopropyl ester of 2-hydroxy-4 methylthio butanoic acid (HMBi). The G- and G+ contained 46% GS and 10% CS in the dry matter (DM), whereas C- and C+ contained 12% GS and 52% CS in the DM. Supplementary HMBi was included at a rate of 0.2% of DM in G+ and C+ diets. Diets were isonitrogenous (9.8 ± 0.4% protein truly digested in the small intestine) and isoenergetic (0.96 ± 0.01 units of energy for lactation; kg/DM). Each of the 4 experimental periods lasted 24 d: 14 d for dietary adaptation, followed by 10 d of housing in individual metabolism stalls; N balance was conducted on the last 5 d of each experimental period. Intake of DM was higher for CS-based vs. GS-based diets (20.23 vs. 18.41 kg/d). No effect of dietary treatment was found on milk yield or yields of milk fat, protein, and lactose. Supplementing with HMBi tended to improve milk solids yield (1.69 vs. 1.59 kg/d), casein yield (0.59 vs. 0.55 kg/d), and concentrations of casein (2.89 vs. 2.73%) and protein (3.58 vs. 3.49%) in the milk. Dietary N intake was higher for CS-based vs. GS-based diets (0.460 vs. 0.422 kg/d). However, forage source or supplementary HMBi had no effect on N excretion in the feces, urine, or milk. Excretion of urinary urea was positively related to N intake. Concentrations of urea N in the plasma (2.34 vs. 1.72 mmol/L), milk (2.54 vs. 2.24 mmol/L), and urine (123.32 vs. 88.79 mmol/L), and total excretion of urinary urea N (40.23 vs. 35.09 g/d) were higher for animals offered CS-based vs. GS-based diets. Corn silage improved N intake through improved DM intake. However, neither forage source nor HMBi supplementation affects N output in the feces, urine, or milk., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study is to quantify the milk production response per cow and per hectare (ha) for an incremental stocking rate (SR) change, based on a meta-analysis of published research papers. Suitable experiments for inclusion in the database required a comparison of at least two SRs under the same experimental conditions in addition to details on experimental length and milk production results per cow and per ha. Each additional increased SR treatment was also described in terms of the relative milk production change per cow and per ha compared to the lower base SR (b&#95;SR). A database containing 109 experiments of various lengths with 131 comparisons of SR was sub-divided into Type I experiments (common experimental lengths) and Type II experiments (variable experimental lengths). Actual and proportional changes in milk production according to SR change were analysed using linear mixed model procedures with study included as a random effect in the model. Low residual standard errors indicated a good precision of the predictive equations with the exception of proportional change in milk production per cow. For all milk yield variables analysed, the results illustrate that while production per cow is reduced, a strong positive relationship exists between SR and milk production per ha. An SR increase of one cow/ha resulted in a decrease in daily milk yield per cow of 7.4% and 8.7% for Type I and Type II data, respectively, whereas milk yield per ha increased by 20.1% and 19.6%, respectively. Within the Type II data set, a one cow/ha increase in SR also resulted in a 15.1% reduction in lactation length (equivalent to 42 days). The low predictability of proportional change in milk production per cow according to the classical SR definition of cows per ha over a defined period suggests that SR may be more appropriately defined in terms of the change in available feed offered per animal within each treatment.

The percent composition of blends of biodiesel and conventional diesel from a variety of retail sources were modeled and predicted using partial least squares (PLS) analysis applied to gas chromatography-total-ion-current mass spectrometry (GC-TIC), gas chromatography-mass spectrometry (GC-MS), comprehensive two-dimensional gas chromatography-total-ion-current mass spectrometry (GCxGC-TIC) and comprehensive two-dimensional gas chromatography-mass spectrometry (GCxGC-MS) separations of the blends. In all four cases, the PLS predictions for a test set of chromatograms were plotted versus the actual blend percent composition. The GC-TIC plot produced a best-fit line with slope=0.773 and y-intercept=2.89, and the average percent error of prediction was 12.0%. The GC-MS plot produced a best-fit line with slope=0.864 and y-intercept=1.72, and the average percent error of prediction was improved to 6.89%. The GCxGC-TIC plot produced a best-fit line with slope=0.983 and y-intercept=0.680, and the average percent error was slightly improved to 6.16%. The GCxGC-MS plot produced a best-fit line with slope=0.980 and y-intercept=0.620, and the average percent error was 6.12%. The GCxGC models performed best presumably due to the multidimensional advantage of higher dimensional instrumentation providing more chemical selectivity. All the PLS models used 3 latent variables. The chemical components that differentiate the blend percent compositions are reported., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Two experiments, a performance experiment and a mineral balance study, were conducted on grower-finisher pigs (42 to 101 kg live weight) to investigate the effects of Peniophora lycii phytase enzyme and 25-hydroxyvitamin D3 (25-OHD3) on growth performance, carcass characteristics, nutrient retention and excretion, and bone and blood parameters. The two experiments were designed as a 2 × 2 factorial (two levels of phytase and two levels of 25-OHD3). The four diets were T1, low-phosphorous diet; T2, T1 + phytase; T3, T1 + 25-OHD3 and T4, T1 + phytase + 25-OHD3 diet. In all, 25 μg of 25-OHD3 was used to replace 1000 IU of vitamin D3 in diets T3 and T4. Diets were pelleted (70°C) and formulated to contain similar concentrations of energy (13.8 MJ DE/kg), lysine (9.5 g/kg) and digestible phosphorus (P; 1.8 g/kg). Neither the inclusion of phytase nor 25-OHD3 in the diet had any effect on pig performance. There was an interaction between phytase and 25-OHD3 on calcium (Ca) and P retention (P < 0.01) and on the apparent digestibility of ash (P < 0.01), P (P < 0.001) and Ca (P < 0.001). Pigs offered phytase diets only, had a higher retention of Ca and P and digestibility of ash (P < 0.01), P (P < 0.001) and Ca (P < 0.01) compared with pigs offered unsupplemented diets. However, when the combination of phytase and 25-OHD3 were offered, no effects were detected compared with 25-OHD3 diets only. Pigs fed phytase diets had higher bone ash (P < 0.01), bone P (P < 0.01) and bone Ca (P < 0.05) concentrations compared with pigs offered non-phytase diets. In conclusion, pigs offered phytase diets had a significantly increased bone ash, Ca and P than pigs offered unsupplemented phytase diets. However, there was no advantage to offering a combination of phytase and 25-OHD3 on either bone strength or mineral status compared to offering these feed additives separately.

The primary objective of the study was to quantify the effect of genetic improvement using the Irish total merit index (Economic Breeding Index) on dry matter intake and feed efficiency across lactation and to quantify the variation in performance among alternative definitions of feed efficiency. Three genotypes of Holstein-Friesian dairy cattle were established from within the Moorepark dairy research herd: 1) low Economic Breeding Index North American Holstein-Friesian representative of the Irish national average dairy cow, 2) high genetic merit North American Holstein-Friesian, and 3) high genetic merit New Zealand Holstein-Friesian. Animals from within each genotype were randomly allocated to 1 of 2 possible intensive pasture-based feed systems: 1) the Moorepark pasture system (2.64 cows/ha and 500 kg of concentrate supplement per cow per lactation) and 2) a high output per hectare pasture system (2.85 cows/ha and 1,200 kg of concentrate supplement per cow per lactation). A total of 128 and 140 spring-calving dairy cows were used during the years 2007 and 2008, respectively. Each group had an individual farmlet of 17 paddocks, and all groups were managed similarly throughout the study. The effects of genotype, feed system, and the interaction between genotype and feed system on dry matter intake, milk production, body weight, body condition score, and different definitions of feed efficiency were studied using mixed models with factorial arrangements of genotypes and feed systems accounting for the repeated cow records across years. No significant genotype-by-feed-system interactions were observed for any of the variables measured. Results showed that aggressive selection using the Irish Economic Breeding Index had no effect on dry matter intake across lactation when managed on intensive pasture-based systems of milk production, although the ranking of genotypes for feed efficiency differed depending on the definition of feed efficiency used. Performance of animals grouped on alternative definitions of feed efficiency showed that conventional definitions such as feed conversion efficiency or residual feed intake may be inappropriate measures of efficiency for lactating dairy cows. An alternative definition, residual solids production, is proposed. This definition of feed efficiency identifies animals that produce greater volumes of milk solids at similar levels of feed intake without excessive body tissue mobilization and with improved fertility performance. The results also suggest that although there are differences in feed efficiency between strains of Holstein-Friesian, there is also variation within genotypes so that improvements in feed efficiency can be realized if the appropriate definition of feed efficiency is incorporated into breeding programs., (Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of the study was to quantify the effect of genetic improvement using the Irish total merit index, the Economic Breeding Index (EBI), on overall performance and lactation profiles for milk, milk solids, body weight (BW), and body condition score (BCS) within 2 pasture-based systems of milk production likely to be used in the future, following abolition of the European Union's milk quota system. Three genotypes of Holstein-Friesian dairy cattle were established from within the Moorepark dairy research herd: LowNA, indicative of animals with North American origin and average or lower genetic merit at the time of the study; HighNA, North American Holstein-Friesians of high genetic merit; and HighNZ, New Zealand Holstein-Friesians of high genetic merit. Animals from within each genotype were randomly allocated to 1 of 2 possible pasture-based feeding systems (FS): 1) The Moorepark pasture (MP) system (2.64 cows/ha and 344 kg of concentrate supplement per cow per lactation) and 2) a high output per hectare (HC) system (2.85 cows/ha and 1,056 kg of concentrate supplement per cow per lactation). Pasture was allocated to achieve similar postgrazing residual sward heights for both treatments. A total of 126, 128, and 140 spring-calving dairy cows were used during the years 2006, 2007, and 2008, respectively. Each group had an individual farmlet of 17 paddocks and all groups were managed similarly throughout the study. The effects of genotype, FS, and the interaction between genotype and FS on milk production, BW, and BCS across lactation were studied using mixed models with factorial arrangements of genotype and FS accounting for the repeated cow records across years. No significant genotype by FS interaction was observed for any of the variables measured. Results show that milk solids production of the national average dairy cow can be increased across lactation through increased EBI. High EBI genotypes (HighNA and HighNZ) produced more milk solids per cow and per hectare than the LowNA genotype (2.7 and 4.1%, respectively). The results also suggest that when concentrate supplementation is used to facilitate increased stocking rates, increased herbage utilization and decreased substitution of concentrate for herbage can be achieved. When implemented, the HC FS could increase the overall productivity of pasture-fed dairy farming systems where land availability is the primary limiting factor of production., (Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The key objectives of this study were to investigate differences in milking characteristics and udder health between Holstein-Friesian (HF), Jersey (J), and Jersey x Holstein-Friesian (F(1)) cows and to determine possible associations between milking characteristics and udder health. Records were available from 329 lactations (162 cows): 65 HF, 48J, and 49F(1). Data included lactation mean milk yield, somatic cell score (SCS), incidence of mastitis, average milk flow (AMF), peak milk flow (PMF), and milking duration (MD). Breed group had a significant effect on milk yield and was higher with the HF cows (18.0 kg/d) compared with the J cows (14.2 kg/d). Udder health (SCS and incidence of mastitis at least once during lactation) were similar across the breed groups. Average milk flow was greater with the HF cows (1.36 kg/min) compared with the J cows (1.09 kg/min). Peak milk flow also tended to greater with the HF cows. No difference in MD was observed between the breed groups. The performance of the F(1) cows tended to be similar to the mid-parent (breed) mean for udder health and MD, but heterosis was evident for milk yield, AMF, and PMF. Correlations examined showed that phenotypic milk yield was negatively associated with SCS. Increased milk yield was synonymous with increased AMF, PMF, and MD. Correlations between SCS and milking characteristics were weak. Correlations also showed that cows with low AMF and PMF had extended MD. Therefore, no difference in udder health was observed between HF, J, or F(1) cows. The fact that higher yielding animals exhibit faster milking speeds was confirmed; however, no difference in MD was observed between the breed groups. Such findings indicate that regularity in the milking process will be maintained within mixed-breed herds., (2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The transition into school occurs at the intersection of multiple environmental settings. This study applied growth curve modeling to a sample of 1,364 American children, followed from birth through age 6, who had been categorized by their exposure to cognitive stimulation at home and in preschool child care and 1st-grade classrooms. Of special interest was the unique and combined contribution to early learning of these 3 settings. Net of socioeconomic selection into different settings, children had higher math achievement when they were consistently stimulated in all 3, and they had higher reading achievement when consistently stimulated at home and in child care. The observed benefits of consistent environmental stimulation tended to be more pronounced for low-income children.

The objectives of this study were to investigate differences in grazing behavior among Holstein-Friesian (HF), Jersey (JE), and Jersey x Holstein-Friesian (F(1)) cows under an intensive, seasonal, grass-based environment and to determine whether associations exist among grazing behavior, intake capacity, and production efficiency. Data from a total of 108 animals (37 HF, 34 JE, and 37F(1)) were available for analysis. Measurements included milk production, body weight (BW), intake, and grazing behavior. Breed group had a significant effect on all of the production, grass dry matter intake, and efficiency parameters investigated. No differences were observed among the breeds for grazing time, number of grazing bouts, grazing bout duration, and total number of bites. Grazing mastications were higher for the JE cows compared with the HF cows. Grass dry matter intake per bite and rate of intake per minute were higher for the HF cows compared with the JE cows. Large differences between the breeds were apparent when grazing behavior measurements were expressed per unit of BW and per unit of intake. In absolute terms, the HF cows spent more time ruminating and had more mastications during rumination than the JE cows. However, when expressed per unit of BW, ruminating time was greater for the JE cows and they tended to have more ruminating mastications compared with the HF cows. Despite these differences, ruminating time and ruminating mastications per unit of intake were similar for the 2 breeds. For the most part, the F(1) cows tended to be similar to the mid-parent mean, but results showed an increase in biting rate, lower grazing duration per bout, and a tendency to achieve a high intake per bite compared with the average of the parent breeds. The results obtained also indicate that inherent grazing and ruminating differences exist between cows varying in intake capacity and production efficiency. Cows with higher intake capacities have increased grazing time and rate of intake per unit of BW. Increased production efficiency, on the other hand, appears to be aided, in particular by improvements in mastication behavior during grazing., (Copyright 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Multidimensional analysis of instant coffee and barley beverage samples using size exclusion chromatography (SEC) combined with a dynamic surface tension detector (DSTD) and a UV-vis absorbance detector (UV) is reported. A unique finding of this study was the action of the tetrabutylammonium (TBA) cation as a modifying agent (with bromide as the counter anion) that substantially increased the surface pressure signal and sensitivity of many of the proteins in the chromatographically separated samples. The tetrabutylammonium bromide (TBAB) enhancement of the surface pressure signal was further investigated by studying the response of 12 commercial standard proteins (alpha-lactalbumin, beta-lactoglobulin, human serum albumin (HSA), albumin from chicken egg white (OVA), bovine serum albumin (BSA), hemoglobin, alpha-chymotrypsinogen A, cytochrome C, myoglobin, RNase A, carbonic anhydrase, and lysozyme) in buffer performed using flow injection analysis (FIA) coupled with the DSTD with and without various concentrations of TBAB. The FIA-DSTD data show that 1mM TBAB enhances sensitivity of HSA detection, by lowering the limit of detection (LOD) from 2mg/mL to 0.1mg/mL. Similarly, the LOD for BSA was reduced from 1mg/mL to 0.2mg/mL. These FIA-DSTD experiments allowed the detection conditions to be optimized for further SEC-UV/DSTD experiments. Thus, the SEC-UV/DSTD system has been optimized and successfully applied to the selective analysis of surface-active protein fractions in a commercial instant coffee sample and in a soluble barley sample. The complementary selectivity of using the DSTD relative to an absorbance detector is also demonstrated.

The objectives of this study were 1) to investigate production and energetic efficiencies among lactating dairy Holstein-Friesian (HF), Jersey (J), and Jersey x Holstein-Friesian (F(1)) cows over a total lactation at pasture and 2) to measure the associations among efficiency variables and performance traits. Data from 110 cows were available (37 HF, 36 J, and 37 F(1)). Breed groups were not balanced for parity; 16 HF, 10 J, and 9 F(1) were in parity 1, whereas the remainder were in parity 2. Milk production, body weight (BW), body condition score (BCS), and estimates of dry matter intake (DMI) corresponding to 51, 108, 149, 198, and 233 d in milk were available. Breed group had a significant effect on all the production parameters investigated: milk yield, solids-corrected milk (SCM), milk fat, protein and lactose concentrations, and milk solids (MLKS; fat + protein yield). Daily MLKS yield was similar for HF and J (1.33 and 1.28 kg/d, respectively). There was a tendency for F(1) (1.41 kg/d) to produce more MLKS compared with HF. The HF breed had higher BW throughout the study compared with F(1) and J. Mean BCS was higher for F(1) (3.00) and J (2.93) compared with HF (2.76). Mean DMI was similar with HF (16.9 kg) and F(1) (16.2 kg) and was lowest with J (14.7 kg). Breed group had a significant effect on all the efficiency parameters investigated: total DMI per 100 kg of BW, SCM per 100 kg of BW, MLKS per 100 kg of BW, and MLKS per total DMI, which tended to be highest for J. Production efficiency based on net energy intake per MLKS was most favorable for F(1) and J compared with HF [12.5, 13.0, and 14.1 UFL, respectively, where 1 UFL is defined as the net energy content of 1 kg of standard barley for milk production (O'Mara, 2000)]. Significant estimates of hybrid vigor were evidenced for milk yield, milk lactose content, SCM, MLKS, net energy for lactation, BW, BCS, and net energy intake per MLKS. The correlations examined indicated that production efficiency was positively associated with MLKS yield.

Three genetic groups of Holstein-Friesian dairy cows were established from within the Moorepark (Teagasc, Ireland) dairy research herd: LowNA, indicative of the Irish national average-genetic-merit North American Holstein-Friesian; HighNA, high-genetic-merit North American Holstein-Friesian; HighNZ, high-genetic-merit New Zealand Holstein-Friesian. Genetic merit in this study was based on the Irish total merit index, the Economic Breeding Index. Animals from within each genetic group were randomly allocated to 1 of 2 possible post-European Union-milk-quota pasture-based feeding systems (FS): 1) The Moorepark (MP) pasture system (2.64 cows/ha and 500 kg of concentrate supplement per cow per lactation) and 2) a high output per hectare (HC) pasture system (2.85 cows/ha and 1,200 kg of concentrate supplement per cow per lactation). A total of 126, 128, and 140 spring-calving dairy cows were used during the years 2006, 2007, and 2008, respectively. Each group had an individual farmlet of 17 paddocks, and all groups were managed similarly throughout the study. The effects of genetic group, FS, and the interaction between genetic group and FS on reproductive performance, body weight, body condition score, and blood metabolite concentrations were studied using mixed models with factorial arrangements of genetic groups and FS. Odds ratios were used in the analysis of binary fertility traits, and survival analysis was used in the analysis of survival after first calving. When treatment means were compared, the HighNA and HighNZ genotypes (with greater genetic merit for fertility performance) had greater first-service pregnancy rates and had a greater proportion of cows pregnant after 42 d of the breeding season than the LowNA group. Both HighNA and HighNZ genotypes were submitted for artificial insemination earlier in the breeding season and had greater survival than the LowNA genotype. There was no significant FS or genotype by FS interactions for any of the reproductive, blood metabolite, body weight, or body condition score measures. The results demonstrate that increased genetic merit for fertility traits resulted in improved reproductive performance and that the poor reproductive capacity of inferior-genetic-merit animals for fertility was not improved through concentrate supplementation at pasture.

The objective of this study was to investigate potential differences in udder health and immune response traits among Holstein-Friesian (HF), Norwegian Red (NR), and NR x HF (NRX) cows on 30 commercial Irish dairy farms. A total of 648 second-lactation cows (HF n = 274, NR n = 207, and NRX n = 167) were immunized with hen egg white lysozyme (HEWL) to induce antibody-mediated immune response (AMIR). Candida albicans was used to induce a cell-mediated immune response, with in vivo delayed-type hypersensitivity used as the indicator. Antibody response to HEWL was measured by ELISA. Udder health defined as mean somatic cell score (SCS) over the lactation, mean SCS within 30 d of the beginning of the immunization, peak SCS for each individual cow during lactation, and incidence of mastitis were statistically superior for the NR. The NR had a greater primary AMIR, producing greater concentrations of anti-HEWL immunoglobulin G on d 14 compared with HF and NRX. No difference was observed among the breed groups in the magnitude of secondary AMIR (d 21 post-immunization) or cell-mediated immune response. The proportion of high and low responders was similar across breed groups. Cows with high AMIR and high cell-mediated immune response had significantly lower mean SCS within 30 d of the start of the immunization, but greater occurrence of clinical mastitis, recorded as a binary trait over the course of the lactation. Otherwise, no significant difference in udder health was evident between cows designated as high and low responders. Although differences in mean breed group SCS values were in line with group mean AMIR values, no association was found among the traits when correlated on an individual cow basis. Results highlight the superiority of the NR with regard to udder health and suggest that improvements to udder health may result from crossbreeding with the NR. However, the immune response traits investigated proved to be inconsistent indicators of udder health.

An experiment (complete randomised design) was conducted to investigate the effects of Laminaria hyperborea and Laminaria digitata seaweed extract inclusion on gut morphology, selected intestinal microbiota populations, volatile fatty acid concentrations and the immune status of the weaned pig. Twenty-eight piglets (24 days of age, 6.5 ± 1.4 kg live weight) were assigned to one of four dietary treatments for 7 days and then sacrificed: (T1) basal diet (control); (T2) basal diet and 1.5 g/kg L. hyperborea seaweed extract; (T3) basal diet and 1.5 g/kg L. digitata seaweed extract; and (T4) basal diet and 1.5 g/kg of a combination of L. hyperborea and L. digitata seaweed extract. The seaweed extract contained both laminarin and fucoidan. Digesta samples were taken from the caecum and colon to measure the enterobacteria, bifidobacteria and lactobacilli populations and for volatile fatty acid analysis. Tissue samples were taken from the duodenum, jejunum and ileum for morphological examination. Blood samples were taken to determine the cytokine gene expression profile and to measure the phagocytotic capacity of the blood. Pigs offered diets containing L. hyperborea seaweed extract had less bifidobacteria in the colon (P < 0.05) and lactobacilli in the caecum (P < 0.05) and colon (P < 0.001). The inclusion of L. digitata seaweed extract resulted in lower populations of enterobacteria in the caecum and colon (P < 0.01), bifidobacteria in the caecum (P < 0.05), and lactobacilli in the caecum (P < 0.05) and colon (P < 0.001). Pigs offered the combination of L. hyperborea and L. digitata seaweed extracts had less enterobacteria (P < 0.05) and lactobacilli (P < 0.01) in the caecum and colon. Pigs offered the L. digitata-supplemented diet had a reduced villous height in the duodenum and jejunum (P < 0.05). The inclusion of the L. digitata seaweed extract increased the molar proportion of butyric acid in the colon (P < 0.05). There was a significant reduction in the ammonia concentration in the colon with the inclusion of L. hyperborea (P < 0.01) and L. digitata (P < 0.05) seaweed extracts. An increase in the expression of the Interleukin-8 mRNA was observed on day 6 with the supplementation of the combination of L. hyperborea and L. digitata seaweed extract (P < 0.05). The inclusion of L. hyperborea seaweed extract resulted in an increase in total monocyte number (P < 0.05). In conclusion, the supplementation of L. hyperborea and L. digitata seaweed extract alone and in combination reduced the enterobacteria, bifidobacteria and lactobacilli populations in the caecum and colon, while only marginal effects on the immune response was observed.

Four bacteria, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus warneri, and Micrococcus luteus, were grown at temperatures of 23, 30, and 37 degrees C and were characterized by pyrolysis-gas chromatography/differential mobility spectrometry (Py-GC/DMS) providing, with replicates, 120 data sets of retention time, compensation voltage, and ion intensity, each for negative and positive polarity. Principal component analysis (PCA) for 96 of these data sets exhibited clusters by temperature of culture growth and not by genus. Analysis of variance was used to isolate the constituents with dependences on growth temperature. When these were subtracted from the data sets, Fisher ratios with PCA resulted in four clusters according to genus at all temperatures for ions in each polarity. Comparable results were obtained from unsupervised PCA with 24 of the original data sets. The ions with taxonomic features were reconstructed into 3D plots of retention time, compensation voltage, and Fisher ratio and were matched, through GC-mass spectrometry (MS), with chemical standards attributed to the thermal decomposition of proteins and lipid A. Results for negative ions provided simpler data sets than from positive ions, as anticipated from selectivity of gas phase ion-molecule reactions in air at ambient pressure.

Comprehensive two-dimensional (2D) separations provide the analyst with a tremendous amount of complex data. In order to glean useful information from this complex data, advancements in commercially available software that implement chemometrics are currently available and continue to evolve. Future advancements will no doubt involve commercializing (or adapting) specialized, in-house chemometric techniques that are currently found only in the hands of technical experts and researchers in industry, government, and academia. In order to make timely advancements, future commercialization of novel chemometric techniques should involve collaborations among instrument software manufacturers, professional programmers, technical experts, and researchers. During the last decade, this field has seen a steady advancement from single analyte target analysis to comprehensive non-target analysis of entire multidimensional sample profiles (involving sample classification and/or data mining for discovery-based sample comparisons). The advancements in instrumentation and chemometric software tools have a tremendous impact in various applications: fuels, food, environmental, pharmaceuticals, metabolomics, etc. Most of the development has been for software to apply with gas chromatography-based instrumentation, such as comprehensive two-dimensional gas chromatography (GC x GC) and comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC x GC-TOF-MS). More recently there have been notable advancements in liquid-phase instrumentation as well.

Pyrolysis gas chromatography-differential mobility spectrometry (py-GC-DMS) analysis of E. coli, P. aeruginosa, S. warneri and M. luteus, grown at temperatures of 23, 30, and 37 degrees C, provided data sets of ion intensity, retention time, and compensation voltage for principal component analysis. Misaligned chromatographic axes were treated using piecewise alignment, the impact on the degree of class separation (DCS) of clusters was minor. The DCS, however, was improved between 21 to 527% by analysis of variance with Fisher ratios to remove chemical components independent of growth temperature. The temperature dependent components comprised 84% of all peaks in the py-GC-DMS analysis of E. coli and were attributed to the pyrolytic decomposition of proteins rather than lipids, as anticipated. Components were also isolated in other bacteria at differing amounts: 41% for M. luteus, 14% for P. aeruginosa, and 4% for S. warneri, and differing patterns suggested characteristic dependence on temperature of growth for these bacteria. These components are anticipated to have masses from 100 to 200 Da by inference from differential mobility spectra.

The first extensive study of yeast metabolite GC x GC-TOFMS data from cells grown under fermenting, R, and respiring, DR, conditions is reported. In this study, recently developed chemometric software for use with three-dimensional instrumentation data was implemented, using a statistically-based Fisher ratio method. The Fisher ratio method is fully automated and will rapidly reduce the data to pinpoint two-dimensional chromatographic peaks differentiating sample types while utilizing all the mass channels. The effect of lowering the Fisher ratio threshold on peak identification was studied. At the lowest threshold (just above the noise level), 73 metabolite peaks were identified, nearly three-fold greater than the number of previously reported metabolite peaks identified (26). In addition to the 73 identified metabolites, 81 unknown metabolites were also located. A Parallel Factor Analysis graphical user interface (PARAFAC GUI) was applied to selected mass channels to obtain a concentration ratio, for each metabolite under the two growth conditions. Of the 73 known metabolites identified by the Fisher ratio method, 54 were statistically changing to the 95% confidence limit between the DR and R conditions according to the rigorous Student's t-test. PARAFAC determined the concentration ratio and provided a fully-deconvoluted (i.e. mathematically resolved) mass spectrum for each of the metabolites. The combination of the Fisher ratio method with the PARAFAC GUI provides high-throughput software for discovery-based metabolomics research, and is novel for GC x GC-TOFMS data due to the use of the entire data set in the analysis (640 MB x 70 runs, double precision floating point).

Simulated chromatographic separations were used to study the performance of piecewise retention time alignment and to demonstrate automated unsupervised (without a training set) parameter optimization. The average correlation coefficient between the target chromatogram and all remaining chromatograms in the data set was used to optimize the alignment parameters. This approach frees the user from providing class information and makes the alignment algorithm applicable to classifying completely unknown data sets. The average peak in the raw simulated data set was shifted up to two peak-widths-at-base (average relative shift=2.0) and after alignment the average relative shift was improved to 0.3. Piecewise alignment was applied to severely shifted GC separations of gasolines and reformate distillation fraction samples. The average relative shifts in the raw gasolines and reformates data were 4.7 and 1.5, respectively, but after alignment improved to 0.5 and 0.4, respectively. The effect of piecewise alignment on peak heights and peak areas is also reported. The average relative difference in peak height was -0.20%. The average absolute relative difference in area was 0.15%.

Comprehensive two-dimensional gas chromatography combined with time-of-flight mass spectrometry (GCxGC-TOFMS) provides high resolution separations of complex samples with a mass spectrum at every point in the separation space. The large volumes of multidimensional data obtained by GCxGC-TOFMS analysis are analyzed using a principal component analysis (PCA) method described herein to quickly and objectively discover differences between complex samples. In this work, we submitted 54 chromatograms to PCA to automatically compare the metabolite profiles of three different species of plants, namely basil (Ocimum basilicum), peppermint (Mentha piperita), and sweet herb stevia (Stevia rebaudiana), where there were 18 chromatograms for each type of plant. The 54 scores of the m/z 73 data set clustered in three groups according to the three types of plants. Principal component 1 (PC 1) separated the stevia cluster from the basil and peppermint clusters, capturing 61.84% of the total variance. Principal component 2 (PC 2) separated the basil cluster from the peppermint cluster, capturing 16.78% of the total variance. The PCA method revealed that relative abundances of amino acids, carboxylic acids, and carbohydrates were responsible for differentiating the three plants. A brief list of the 16 most significant metabolites is reported. After PCA, the 54 scores of the m/z 217 data set clustered in three groups according to the three types of plants, as well, yielding highly loaded variables corresponding with chemical differences between plants that were complementary to the m/z 73 information. The PCA data mining method is applicable to all of the monitored selective mass channels, utilizing all of the collected data, to discover unknown differences in complex sample profiles.

A useful methodology is introduced for the analysis of data obtained via gas chromatography with mass spectrometry (GC-MS) utilizing a complete mass spectrum at each retention time interval in which a mass spectrum was collected. Principal component analysis (PCA) with preprocessing by both piecewise retention time alignment and analysis of variance (ANOVA) feature selection is applied to all mass channels collected. The methodology involves concatenating all concurrently measured individual m/z chromatograms from m/z 20 to 120 for each GC-MS separation into a row vector. All of the sample row vectors are incorporated into a matrix where each row is a sample vector. This matrix is piecewise aligned and reduced by ANOVA feature selection. Application of the preprocessing steps (retention time alignment and feature selection) to all mass channels collected during the chromatographic separation allows considerably more selective chemical information to be incorporated in the PCA classification, and is the primary novelty of the report. This methodology is objective and requires no knowledge of the specific analytes of interest, as in selective ion monitoring (SIM), and does not restrict the mass spectral data used, as in both SIM and total ion current (TIC) methods. Significantly, the methodology allows for the classification of data with low resolution in the chromatographic dimension because of the added selectivity from the complete mass spectral dimension. This allows for the successful classification of data over significantly decreased chromatographic separation times, since high-speed separations can be employed. The methodology is demonstrated through the analysis of a set of four differing gasoline samples that serve as model complex samples. For comparison, the gasoline samples are analyzed by GC-MS over both 10-min and 10-s separation times. The successfully classified 10-min GC-MS TIC data served as the benchmark analysis to compare to the 10-s data. When only alignment and feature selection was applied to the 10-s gasoline separations using GC-MS TIC data, PCA failed. PCA was successful for 10-s gasoline separations when the methodology was applied with all the m/z information. With ANOVA feature selection, chromatographic regions with Fisher ratios greater than 1500 were retained in a new matrix and subjected to PCA yielding successful classification for the 10-s separations.

This report is about applying a Fisher ratio method to entire four dimensional (4D) data sets from third-order instrumentation data. The Fisher ratio method uses a novel indexing scheme to discover the unknown chemical differences among known classes of complex samples. This is the first report of a Fisher ratio analysis procedure applied to entire 4D data sets of third-order separation data, which, in this case, is comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry analyses of metabolite extracts using all of the collected mass channels. Current analysis methods for third-order separation data use only user-defined subsets of the 4D data set. First, in a validation study, the Fisher ratio method was demonstrated to objectively evaluate and determine the chemical differences between three controlled urine samples that differed by known spiked chemical components. It was determined that, out of more than 600 recognizable chemical components in a single sample, the six spiked components, along with only two other matrix components, differed most significantly in concentration among the control samples. In a second study, the Fisher ratio method was used in a novel application to discover the unknown chemical differences between urine metabolite samples from pregnant women and nonpregnant women. A brief list of the top 11 components that were most significantly different in concentration between the pregnant and nonpregnant samples was generated. Because the Fisher ratio calculation statistically differentiates regions of the chromatogram with large class-to-class variations from regions containing large within-class variations, the Fisher ratio method should generally be robust against biological diversity in a sample population. Indeed, application of principal component analysis in this second study failed due to biological diversity of the samples.

A comprehensive two-dimensional (2D) retention time alignment algorithm was developed using a novel indexing scheme. The algorithm is termed comprehensive because it functions to correct the entire chromatogram in both dimensions and it preserves the separation information in both dimensions. Although the algorithm is demonstrated by correcting comprehensive two-dimensional gas chromatography (GC x GC) data, the algorithm is designed to correct shifting in all forms of 2D separations, such as LC x LC, LC x CE, CE x CE, and LC x GC. This 2D alignment algorithm was applied to three different data sets composed of replicate GC x GC separations of (1) three 22-component control mixtures, (2) three gasoline samples, and (3) three diesel samples. The three data sets were collected using slightly different temperature or pressure programs to engender significant retention time shifting in the raw data and then demonstrate subsequent corrections of that shifting upon comprehensive 2D alignment of the data sets. Thirty 12-min GC x GC separations from three 22-component control mixtures were used to evaluate the 2D alignment performance (10 runs/mixture). The average standard deviation of first column retention time improved 5-fold from 0.020 min (before alignment) to 0.004 min (after alignment). Concurrently, the average standard deviation of second column retention time improved 4-fold from 3.5 ms (before alignment) to 0.8 ms (after alignment). Alignment of the 30 control mixture chromatograms took 20 min. The quantitative integrity of the GC x GC data following 2D alignment was also investigated. The mean integrated signal was determined for all components in the three 22-component mixtures for all 30 replicates. The average percent difference in the integrated signal for each component before and after alignment was 2.6%. Singular value decomposition (SVD) was applied to the 22-component control mixture data before and after alignment to show the restoration of trilinearity to the data, since trilinearity benefits chemometric analysis. By applying comprehensive 2D retention time alignment to all three data sets (control mixtures, gasoline samples, and diesel samples), classification by principal component analysis (PCA) substantially improved, resulting in 100% accurate scores clustering.

A fast and objective chemometric classification method is developed and applied to the analysis of gas chromatography (GC) data from five commercial gasoline samples. The gasoline samples serve as model mixtures, whereas the focus is on the development and demonstration of the classification method. The method is based on objective retention time alignment (referred to as piecewise alignment) coupled with analysis of variance (ANOVA) feature selection prior to classification by principal component analysis (PCA) using optimal parameters. The degree-of-class-separation is used as a metric to objectively optimize the alignment and feature selection parameters using a suitable training set thereby reducing user subjectivity, as well as to indicate the success of the PCA clustering and classification. The degree-of-class-separation is calculated using Euclidean distances between the PCA scores of a subset of the replicate runs from two of the five fuel types, i.e., the training set. The unaligned training set that was directly submitted to PCA had a low degree-of-class-separation (0.4), and the PCA scores plot for the raw training set combined with the raw test set failed to correctly cluster the five sample types. After submitting the training set to piecewise alignment, the degree-of-class-separation increased (1.2), but when the same alignment parameters were applied to the training set combined with the test set, the scores plot clustering still did not yield five distinct groups. Applying feature selection to the unaligned training set increased the degree-of-class-separation (4.8), but chemical variations were still obscured by retention time variation and when the same feature selection conditions were used for the training set combined with the test set, only one of the five fuels was clustered correctly. However, piecewise alignment coupled with feature selection yielded a reasonably optimal degree-of-class-separation for the training set (9.2), and when the same alignment and ANOVA parameters were applied to the training set combined with the test set, the PCA scores plot correctly classified the gasoline fingerprints into five distinct clusters.

Experiences that are deeply engaging and enjoyable, engender full concentration, and present a balance between challenge and skill promote children's development. This chapter describes a study that sought to identify the kinds of settings and activities that foster engagement and, by extension, positive youth development. The after-school experiences of 191 ethnically diverse youth living in three states, some of whom participated in after-school programs and some of whom did not, were studied. Youth were equipped with logbooks and watches that were programmed to signal at random times. When signaled, youth recorded their location, social partners, activity, and feelings. The study found pervasive differences in the experiences at programs and elsewhere. Youth spent more time in academic and arts enrichment, organized sports and physical activities, community service, and homework at programs versus elsewhere, and they spent less time eating and watching TV at programs. They also reported higher levels of motivation, engagement, and positive affect at programs. At the same time, there were few differences in activities, emotions, effort, or motivation of program participants and nonparticipants when both groups were elsewhere. The similarities in these experiences while elsewhere suggest that the program context, not differences in youth characteristics or interests, was responsible for the feelings of engagement that were reported at programs.

Since the 1990s, there has been a growing recognition of the importance of the out-of-school context for children and adolescents. Fueled in part by family demographics that include substantial numbers of employed mothers and single mothers, in part by concerns about poor academic performance and problem behaviors, and in part by intensified efforts to find ways to promote positive youth development, researchers and practitioners have focused their attention on two particular out-of-school settings: after-school programs and structured activities. The research findings pertaining to full-time (i.e., 5 days a week) after-school programs are mixed, which may reflect the substantial heterogeneity of the programs in terms of children being served, the types of activities offered, and the training and background of the staff. The federal funding of the 21st Century CLCs and various state and local initiatives has increased the numbers of low-income and English-learning students participating in after-school programs. A substantial number of programs are becoming more school-like. The available research suggests that (under some conditions) attending after-school programs is linked to improved social and academic outcomes. Children are more likely to show academic and social benefits when staff-child relationships are positive and nonconflictual, when programs offer a variety of age-appropriate activities from which children can select those of interest, and when children attend on a regular basis. The research findings about voluntary structured activities are more straightforward. Participation in these activities has been consistently linked to positive academic and social developmental outcomes in numerous studies. What appears to be key is that the activities are voluntary, are characterized by sustained engagement and effort, and provide opportunities to build or develop skills. Although the available research has begun to inform our understanding of the out-of-school context, further research is sorely needed. First, there is a need for research to identify the social, cognitive, and linguistic processes by which participation in programs and structured activities influences child and youth developmental outcomes. For example, researchers need to consider the competitiveness of sport activities in relation to children's social and emotional functioning. Researchers also might examine after-school experiences as settings in which complex thought processes can develop. Heath (1999) has conducted initial work in the area of language development by obtaining language samples during voluntary structured activities and analyzing their content. In the initial samples, students engaged in few sustained conversations on a topic and they frequently changed topics. After 3-4 weeks at the program, however, Heath noted substantial changes in the students' conversations and language. The use of conditionals (should, would, could) increased. She also noted increases in strategies to obtain clarifications from others and increases in the use of shifted registers and genres. Heath's (1999) linguistic analyses in conjunction with research that considers social and motivational processes underscore the broader point that the out-of-school context is complex and multi-layered and likely to be of substantial importance in the lives of children and youth. Research is needed to identify other important developmental processes in programs and structured activities. A promising procedure for identifying these processes is experience sampling (Csikszentmihalyi & Larson, 1987; Larson, 1989). Experience sampling methodology allows researchers to collect systematic data about an individual's activities, thoughts, and affective states by obtaining reports from participants at multiple randomly sampled points in time. Participants are signaled to provide a report in a variety of ways, such as with beepers or alarm watches. This record of experiences is not usually captured by other data collection methods. For example, program observations provide data on observed activities, interactions, and program climate, but do not offer insights into students' feelings and experiences within the after-school environment. Questionnaire and survey data are retrospective, asking respondents to recall past experiences and feelings regarding their after-school activities. Experience sampling could be used to examine any number of processes in after-school programs and structured activities. A better understanding of the effects of program content also is needed. Whether after-school programs should focus exclusively on enrichment activities or exclusively on academic activities, or include both enrichment and academic components, is the subject of heated debate. Some after-school scholars (Halpern, 1999; Heath, 1999; Eccles, in press) have argued forcefully that a focus on academics undermines the unique strengths and role of programs, and that programs should emphasize extracurricular enrichment activities. Others (Noam, 2004) have supported the move by policy makers and educators to make programs more academic, with an emphasis on homework help, tutoring, and preparation for academic achievement tests. The effects of different approaches to after-school programming have not been evaluated systematically. Research that describes, compares, and then tests effects of different program content models is needed to determine which types of programs are successful in attracting and keeping students (a necessary condition for programs to effect change), and to determine whether different types of programs are differentially associated with improvements in student outcomes such as school attendance, academic achievement, social competencies, and behavioral adjustment. A related question is whether structured activities that are obligatory or required have the same effects as voluntary structured activities do. Researchers also should further examine the impact of different attendance patterns on child developmental outcomes. We do not have solid information about optimal intensity and duration of attendance in terms of outcomes. There are suggestions in the literature that long-term, frequent attendance at programs is associated with positive outcomes for low-income children. Research needs to examine whether these results hold for middle-income children and youth as well. Finally, experimental studies should be conducted in which children and adolescents are randomly assigned to after-school programs and structured activities. All of the research to date on structured activities, and most of the research on after-school programs, has been nonexperimental, so questions about selection bias remain. Experimental studies in which children and adolescents are randomly assigned to participation in programs and activities would be a valuable next step in understanding relations between participation and child and youth outcomes. Such research should not be conducted until we have more information about the components of high-quality programming in terms of program content and developmental processes, however.

Purpose: Tooth decay is one of the more common diseases of childhood. Slightly >40% of US children are already affected by the time they reach kindergarten. Primary care physicians can play an important role in prevention and control of this disease because of their ready access to this population. Unlike dentists, they see a large percentage of children during their infant and toddler years. However, few studies have been conducted on oral screenings and referrals by primary care physicians or the effectiveness of their oral health preventive activities. The purpose of this study was to determine the accuracy of pediatric primary care providers' screening and referral for Early Childhood Caries., Methods: We sought to compare independent, blinded oral screening results and referral recommendations made by primary care providers with those of a pediatric dentist, considered for purposes of the study to be the reference gold standard. The study was conducted at a private pediatric group practice in North Carolina. The practice was selected because it serves a large volume of Medicaid patients and includes a large number of pediatric primary care providers (11 pediatricians and 1 nurse practitioner). Study participants included Medicaid-eligible children younger than 36 months of age with erupted teeth. The pediatric primary care providers in this practice received 2 hours of training in infant oral health. The training consisted of a review of the study methods and clinical slides illustrating dental caries in various stages of progression. Specific instructions were given to the providers on how to recognize a cavitated carious lesion and how to determine when a dental referral is needed. Providers were instructed to refer any child with 1 or more cavitated carious lesions, soft tissue pathology, or evidence of trauma to the teeth or mouth. Before commencing the study, calibration and a comparative analysis were performed to establish reliability and validity of the examinations performed by the pediatric dentist. Both a pediatric dentist and a pediatric primary care provider conducted a dental screening on each child and recorded carious teeth and whether a dental referral was needed. Sensitivity and specificity were calculated to compare the pediatric primary care providers' screenings to the gold standard (pediatric dentist) in 3 categories: caries at the tooth level, caries at the patient level (1 or more affected teeth), and need for referral., Results: The final study sample consisted of 258 preschool-aged children (122 males and 136 females) with a mean age of 21.2 months (standard deviation [SD]: 9.13). One hundred eighty-four (71.3%) of the participants were white, 58 (22.5%) were black, and 16 (6.2%) were Hispanic. Tooth-Level Analysis: The pediatric dentist reported an average of 0.30 (SD: 0.005) cavitated teeth per child, whereas the pediatric primary care providers reported a mean of 0.25 (SD: 0.004). This difference was not statistically significant (t test). The pediatric dentist identified 80 (2.4%) teeth with cavitated carious lesions, whereas the pediatric primary care providers identified 64 (1.9%), 25 of which were false-positives. Their screening results include 41 false-negative teeth. Thus, the primary care providers tended to under-count the number of teeth with carious lesions. They achieved a sensitivity of 0.49 (95% confidence interval [CI]: 0.47-0.51) and a specificity of 0.99 (95% CI: 0.99-1.0) when their screening results for individual teeth were compared with the gold standard. Patient-Level Analysis: At the patient level, the pediatric dentist identified 25 (9.7%) children with 1 or more teeth affected by cavitated lesions. The pediatric primary care providers collectively identified 30 (11.6%) children who had cavitated lesions. They achieved a sensitivity of 0.76 (95% CI: 0.71-0.81) and a specificity of 0.95 (95% CI: 0.93-0.98) in identifying those children with cavitated carious lesions. There were 6 false-negatives and 11 false-positives when the pediatric primary care providers' findings were compared with the gold standard. At the patient-level, the positive predictive value of the dental screening was 0.63 and the negative predictive value was 0.97. Dental Referral: The pediatric dentist referred a total of 27 (10.5%) children to a dentist. Two of these children were referred for trauma and the other 25 were referred for cavities. The pediatric primary care providers referred a total of 23 (8.9%) children to a dentist. Two referrals were made because the provider was concerned about stains on the teeth, whereas the remaining 21 were referred for cavities. The pediatric primary care providers achieved a sensitivity of 0.63 (95% CI: 0.57-0.69) and a specificity of 0.98 (95% CI: 0.96-0.99) when their recommendations for referral were compared with the gold standard. The number of children receiving a referral from a pediatric primary care provider for cavities (N = 21) was less than the number of children they identified as having cavities (N = 30). The providers as a whole tended to under-refer, and only 70% of children with evidence of dental disease received a referral., Conclusions: After 2 hours of training in infant oral health, the pediatric primary care providers in this study achieved an adequate level of accuracy in identifying children with cavitated carious lesions. Additional training and research would be needed to optimize pediatric primary care providers' identification of carious teeth if that were the goal of screening. However, the purpose of screening by nondental personnel generally is to accurately identify those in need of referral, which does not require a tooth-by-tooth identification of cavities. Additional research is also needed to determine how to improve dental referrals by pediatric primary care providers. Results of our study suggest that dental screenings can easily be incorporated into a busy pediatrics practice and that pediatric primary care providers can significantly contribute to the overall oral health of young children by the identification of those children who need to be seen by a dentist.