Ocular refraction: heritability and genome-wide search for eye morphometry traits in an isolated Sardinian population

No genes influencing oculometric phenotypes have yet been identified, despite it being well known that eye morphometry is involved in refraction and that genetics may play an important role. We have therefore performed a heritability analysis and genome-wide search (GWS) of biometric ocular traits in an isolated Sardinian population, assessing the genetic contribution and identifying the associated genetic loci. A complete eye examination including refraction and ocular biometry measurements such as axial length (AL), anterior chamber depth (ACD) and corneal curvature (CC), was performed on 789 subjects. Heritability analysis was carried out by means of parent–offspring regression and variance component models. Univariate and bivariate linkage analysis was performed by using 654 microsatellite markers spanning the genome. CC showed a mean heritability of 57%. AL and ACD were found to have significantly different variances (P<0.01) in males and females, so that heritability was calculated separately for each sex. AL had an estimated heritability in females of 31% and in males of 60%, whereas ACD had an estimated heritability of 47% in females and of 44% in males. In the GWS, the most suggestive evidence of linkage was identified on chromosome 2 for AL (LOD 2.64), on chromosome 1 for ACD (LOD 2.32) and on chromosomes 7, 2 and 3 for CC (LOD 2.50, 2.44 and 2.34, respectively). High heritability of eye morphometry traits was thus revealed. The identified loci are the first linkage signals available in ocular biometry. Notably, the observed significant differences in parental transmission deserve further study.The authors Ginevra Biino and Maria Antonietta Palmas contributed equally to this work     

Phenol Biodegradation and Metal Removal by a Mixed Bacterial Consortium

This paper reports the tolerance and biodegradation of phenol by a heavy metal–adapted environmental bacterial consortium, known as consortium culture (CC). At the highest tolerable phenol concentration of 1200 mg/L, CC displayed specific growth rate of 0.04 h^?1, phenol degradation rate of 6.11 mg L^?1 h^?1 and biomass of 8.45 ± 0.35 (log₁₀ colony-forming units [CFU]/ml) at the end of incubation. Phenol was degraded via the ortho-cleavage pathway catalyzed by cathechol-1,2-dioxygenase with specific activity of 0.083 (µmol min^?1 mg^?1 protein). The different constituent bacterial isolates of CC preferentially grow on benzene, toluene, xylene, ethylbenzene, cresol, and catechol, suggesting a synergistic mechanism involved in the degradation process. Microtox assay showed that phenol degradation was achieved without producing toxic dead-end metabolites. Moreover, lead (Pb) and cadmium (Cd) at the highest tested concentration of 1.0 and 0.1 mg/L, respectively, did not inhibit phenol degradation by CC. Simultaneous metal removal during phenol degradation was achieved using CC. These findings confirmed the dual function of CC to degrade phenol and to remove heavy metals from a mixed-pollutant medium.     

Variability for the in vitro culture response in tomato recombinant inbred lines

The aim of this work was to estimate genetic variability for in vitro culture response of recombinant inbred lines (RILs) of the genus Lycopersicon. The callus percentage (C), the regeneration percentage (R) and the productivity rate (PR) were evaluated 45 d after culture initiation in a set of 16 elite tomato RILs and their parents. The narrow sense heritability (h ²) values were 0.38 ± 0.04 for C, 0.46 ± 0.04 for R, and 0.28 ± 0.03 for R, while the genetic correlation (r _g ) values were −0.96 ± 0.07 between C and R, 0.81 ± 0.14 between PR and R, and −0.79 ± 0.16 between PR and C. Three AFLP markers associated to the in vitro traits were identified.     

Embryonic and larval traits of the temperate damselfish Chromis crusma reveal important similarities with other Pomacentridae throughout the family's thermal range

Embryonic development and larval morphology of Chromis crusma was described from five nests sampled between 21 and 25 m depth in central Chile (33°S). From each nest, a set of c. 100 randomly selected eggs were hand-collected and transported in seawater to the laboratory. Subsets of c. 30 eggs per nest were maintained in 50 ml glass containers at a constant ambient temperature of c. 12°C (range 11.5–12.9°C). Egg length (L) and width (W) and larval notochordal length (L_N) were measured from photographs. Geometric morphometric analyses were performed in newly hatched and 1 week old larvae to quantify shape changes. Ellipsoid eggs had an average (mean ± SE) size of 1.12 ± 0.05 mm L and 0.67 ± 0.02 mm W, with volume being similar throughout 15 developmental stages (i.e., ellipsoid-shaped; 0.27 mm³). Planktonic larvae hatched between 5 and 11 days at 12°C and had a mean L_N of 3.13 ± 0.25 mm, a yolk sack volume of 0.03 mm³ and an oil droplet volume of 0.005 mm³. Morphological traits at hatching included: (a) lack of paired fins and jaws; (b) single medial fin fold; (c) lack of eye pigmentation; (d) yolk sac present near anterior tip; (e) melanophores distributed along ventral surface with one pair over the forehead. In order to generate an up-to-date summary of developmental traits within Pomacentridae, we reviewed literature on egg development (e.g., shape and number of oil droplets), hatching and larval traits (e.g., morphology, pigmentation patterns). Thirty-two publications accounting for 35 species were selected, where eggs, embryonic development, hatching and larval traits were found for 26, 21, 24 and 34 species, respectively. In order to evaluate potential phylogenetic and environmental relationships within the early stages of Pomacentridae, cluster analyses (Bray Curtis similarity, group average) were also performed on egg and larval traits of 22 species divided by subfamily (Stegastinae, Chrominae, Abudefdufinae, Pomacentrinae) and thermal ranges (i.e., low: 16.5°C (range: 12–21°C), medium: 24.5°C (range:21–28°C) and high: 27°C (range: 26–28°C)), suggesting that early developmental patterns can be segregated by both temperature and phylogenetic relationships.     

Estimation of social genetic effects on feeding behaviour and production traits in pigs

Pigs are housed in groups during the test period. Social effects between pen mates may affect average daily gain (ADG), backfat thickness (BF), feed conversion rate (FCR), and the feeding behaviour traits of pigs sharing the same pen. The aim of our study was to estimate the genetic parameters of feeding behaviour and production traits with statistical models that include social genetic effects (SGEs). The data contained 3075 Finnish Yorkshire, 3351 Finnish Landrace, and 968 F1-crossbred pigs. Feeding behaviour traits were measured as the number of visits per day (NVD), time spent in feeding per day (TPD), daily feed intake (DFI), time spent in feeding per visit (TPV), feed intake per visit (FPV), and feed intake rate (FR). The test period was divided into five periods of 20 days. The number of pigs per pen varied from 8 to 12. Two model approaches were tested, i.e. a fixed group size model and a variable group size model. For the fixed group size model, eight random pigs per pen were included in the analysis, while all pigs in a pen were included for the variable group size model. The linear mixed-effects model included sex, breed, and herd*year*season as fixed effects and group (batch*pen), litter, the animal itself (direct genetic effect (DGE)), and pen mates (SGEs) as random effects. For feeding behaviour traits, estimates of the total heritable variation (T² ± SE) and classical heritability (h² ± SE, values given in brackets) from the variable group size model (e.g. period 1) were 0.34 ± 0.13 (0.30 ± 0.04) for NVD, 0.41 ± 0.10 (0.37 ± 0.04) for TPD, 0.40 ± 0.15 (0.14 ± 0.03) for DFI, 0.53 ± 0.15 (0.28 ± 0.04) for TPV, 0.66 ± 0.17 (0.28 ± 0.04) for FPV, and 0.29 ± 0.13 (0.22 ± 0.03) for FR. The effect of social interaction was minimal for ADG (T² = 0.29 ± 0.11 and h² = 0.29 ± 0.04), BF (T² = 0.48 ± 0.12 and h² = 0.38 ± 0.07), and FCR (T² = 0.37 ± 0.12 and h² = 0.29 ± 0.04) using the variable group size model. In conclusion, the results indicate that social interactions have a considerable indirect genetic effect on the feeding behaviour and FCR of pigs but not on ADG and BF.     

Evidence of Shared Genetic Effects Between Pre‐ and Postobesity Epidemic BMI Levels

United States has experienced a widespread obesity epidemic. However, it is unclear whether the obesogenic environment has uncovered genes previously unimportant in adiposity or whether genes influencing obesity are the same before and after the obesity epidemic. The objective of this study was to test whether BMI pre‐ and postobesity epidemic would be controlled by shared genetic effects. A 25–30‐year follow‐up of parents and children who participated in the National Institutes of Health–National Heart, Lung, and Blood Institute Lipid Research Clinics (LRC) Princeton School Study, 1973–1976, were followed up in 1999–2004 in the Princeton Follow‐up Study (PFS). Heritability of BMI and genetic correlations between pre‐epidemic BMI and BMI z‐scores in adolescents and postobesity epidemic BMI were calculated. Even though they had similar ages, offspring had higher BMI in PFS than their parents in LRC (28.5 ± 6.6 vs. 26.1 ± 4.4, P < 0.0001). BMI measurements in offspring were strongly heritable (BMI_LRC: h² = 0.78 ± 0.17; BMI z‐score_LRC: h² = 0.61 ± 0.16; BMI_PFS: h² = 0.64 ± 0.16, all P ≤ 0.0001). Further, the change of BMI exhibited a high heritability (h² = 0.51 ± 0.18, P = 0.003). Bivariate analysis of BMI in LRC and PFS showed significant genetic correlation (0.70 ± 0.16, P = 0.005), whereas the environmental correlation was not significant (0.36 ± 0.17). Although the obesogenic environment may have changed between the 1970s and 2000s, many of the same genes are likely to be involved in establishing genetic susceptibility to obesity. Furthermore, shared genetic effects survive the period of the transition from adolescence to adulthood.     

Glutathione and transferrin in rhesus monkeys1

Transferrin types and glutathione (GSH) levels were determined for 38 rhesus monkeys (Macaca mulatto). Eight transferrin alleles were found: their gene frequencies were: Tf^A 0.04, Tf^c 0.50, Tf^D 0.03, Tf^D 0.09, TF 0.11, TF 0.05, Tf^G 0.11, and Tf ^H' 0.08. The 16 phenotypes found and the number of each was: Tf AC 3, Tf CC 9, Tf CD‘ 2, Tf CD 2, Tf CF 5, Tf CF 3, Tf CH’ 3, Tf CG 2, Tf D'D‘ 1, Tf D'F 1, Tf D'G 1, TfD'H’ l, TfFG2, TfF'G ‘l.TfCG l.and Tf H'H’ 1. Mean GSH level was 310.6 mg/100 ml red cells. The Tf CC group had significantly lower GSH levels than Tf^c heterozygotes or non-Tf^c homozygotes. The Tf^c heterozygotes had significantly lower GSH levels than non-Tf^c homozygotes. Tf^c, Tf^D' and TF alleles were associated with lower GSH levels. Glutathione levels were higher in older animals and were higher in September than in December. Sex differences in glutathione levels were not found.     

Heritability of anti‐predatory traits: vigilance and locomotor performance in marmots

Animals must allocate some proportion of their time to detecting predators. In birds and mammals, such anti‐predator vigilance has been well studied, and we know that it may be influenced by a variety of intrinsic and extrinsic factors. Despite hundreds of studies focusing on vigilance and suggestions that there are individual differences in vigilance, there have been no prior studies examining its heritability in the field. Here, we present one of the first reports of (additive) genetic variation in vigilance. Using a restricted maximum likelihood procedure, we found that, in yellow‐bellied marmots (Marmota flaviventris), the heritability of locomotor ability (h² = 0.21), and especially vigilance (h² = 0.08), is low. These modest heritability estimates suggest great environmental variation or a history of directional selection eliminating genetic variation in these traits. We also found a significant phenotypic (r_P = ?0.09 ± 0.04, P = 0.024) and a substantial, but not significant, genetic correlation (r_A = ?0.57 ± 0.28, P = 0.082) between the two traits (slower animals are less vigilant while foraging). We found no evidence of differential survival or longevity associated with particular phenotypes of either trait. The genetic correlation may persist because of environmental heterogeneity and genotype‐by‐environment interactions maintaining the correlation, or because there are two ways to solve the problem of foraging in exposed areas: be very vigilant and rely on early detection coupled with speed to escape, or reduce vigilance to minimize time spent in an exposed location. Both strategies seem to be equally successful, and this ‘locomotor ability‐wariness’ syndrome may therefore allow slow animals to compensate behaviourally for their impaired locomotor ability.     

Prostaglandin Endoperoxide H Synthase-2 (PGHS-2) Variants and Risk of Obesity and Microvascular Dysfunction Among Tunisians: Relevance of rs5277 (306G/C) and rs5275 (8473T/C) Genetic Markers

The purpose of this study was to determine the impact of six PGHS-2 genetic variants on obesity development and microvascular dysfunction. The study included 305 Tunisian subjects (186 normal weights, 35 overweights and 84 obeses). PCR analyses were used for allelic discrimination between polymorphisms. Prostaglandin (PGE2, PGI2), leptin, and matrix metalloproteinase (MMP1, 2, 3, 9) levels were evaluated by ELISA. Fatty acid composition was performed by gas chromatography–mass spectrometry. Our results revealed that subjects carrying the PGHS-2 306CC (rs5277) and 8473CC (rs5275) genotypes present higher anthropometric values compared to wild-type genotypes (306GG, BMI (Kg/m²): 27.11?±?0.58; WC (cm): 93.09?±?1.58; 306CC, BMI: 33.83?±?2.46; WC: 109.93?±?5.41; 8473TT, BMI: 27.75?±?0.68; WC: 93.96?±?1.75; 8473CC, BMI: 33.72?±?2.2; WC: 117.89?±?2.94). A reduced microvascular reactivity and a higher PGE2 level were also found in individuals with the 306CC and 8473CC genotypes in comparison to 306GG and 8473TT carriers (306GG, Peak Ach-CVC (PU/mmHg): 0.46?±?0.03; PGE2 (pg/ml): 7933.1?±?702; 306CC, Peak Ach-CVC: 0.24?±?0.01; PGE2: 13,380.3?±?966.2; 8473TT, Peak Ach-CVC: 0.48?±?0.05; PGE2: 7086.41?±?700.31; 8473CC, Peak Ach-CVC: 0.23?±?0.01; PGE2: 13,175.7?±?1165.8). Fatty acid analysis showed a significant increase of palmitic acid (PA) (34.2?±?2.09 vs. 16.82%?±?1.76, P?<?0.001), stearic acid (SA) (25.76?±?3.29 vs. 9.05%?±?2.53, P?<?0.001), and linoleic acid (LA) (5.25?±?1.18 vs. 0.5%?±?0.09, P?<?0.001) levels in individuals carrying the PGHS-2 306CC genotype when compared to GG genotype individuals. Subjects with the 8473CC genotype showed also a significant increase of PA, SA ,and LA levels when compared to TT genotype carriers (PA: 38.02?±?1.51 vs. 12.65%?±?1.54, P?<?0.001; SA: 32.96?±?1.87 vs. 1.38%?±?0.56, P?<?0.001; LA: 26.84?±?2.09 vs. 3.7%?±?1.54, P?<?0.001). Logistic regression analysis revealed that PGHS-2 306CC and 8473CC variants are significantly associated with obesity status (OR 6.25, CI (1.8–21.6), P?=?0.004; OR 3.01, CI (1.13–8.52), P?=?0.03, respectively). Haplotypes containing the C₃₀₆:T₈₄₇₃ (OR 2.91; P?=?0.01) and G_306:C₈₄₇₃ (OR 5.25; P?=?0.002) combinations were associated with an enhanced risk for obesity development in the studied population. In conclusion, our results highlight that PGHS-2 306G/C and 8473T/C variants could be useful indicators of obesity development, inflammation, and microvascular dysfunction among Tunisians.

     

Quantitative Genetics of Response to Novelty and Other Stimuli by Infant Rhesus Macaques (Macaca mulatta) Across Three Behavioral Assessments

Primate behavior is influenced by both heritable factors and environmental experience during development. Previous studies of rhesus macaques (Macaca mulatta) examined the effects of genetic variation on expressed behavior and related neurobiological traits (heritability and/or genetic association) using a variety of study designs. Most of these prior studies examined genetic effects on the behavior of adults or adolescent rhesus macaques, not in young macaques early in development. To assess environmental and additive genetic variation in behavioral reactivity and response to novelty among infants, we investigated a range of behavioral traits in a large number (N?=?428) of pedigreed infants born and housed in large outdoor corrals at the Oregon National Primate Research Center (ONPRC). We recorded the behavior of each subject during a series of brief tests, involving exposure of each infant to a novel environment, to a social threat without the mother present, and to a novel environment with its mother present but sedated. We found significant heritability (h ² ) for willingness to move away from the mother and explore a novel environment (h ² ?=?0.25?±?0.13; P?=?0.003). The infants also exhibited a range of heritable behavioral reactions to separation stress or to threat when the mother was not present (h ² ?=?0.23?±?0.13–0.24?±?0.15, P?<?0.01). We observed no evidence of maternal environmental effects on these traits. Our results extend knowledge of genetic influences on temperament and reactivity in nonhuman primates by demonstrating that several measures of behavioral reactivity among infant rhesus macaques are heritable.     

The Essential Role of Cytosolic Cl− in Ca2+ Regulation of an Amiloride-Sensitive Channel in Fetal Rat Pneumocyte

An amiloride-sensitive, Ca²⁺-activated nonselective cation (NSC) channel in the apical membrane of fetal rat alveolar epithelium plays an important role in stimulation of Na⁺ transport by a beta adrenergic agonist (beta agonist). We studied whether Ca²⁺ has an essential role in the stimulation of the NSC channel by beta agonists. In cell-attached patches formed on the epithelium, terbutaline, a beta agonist, increased the open probability (P _o ) of the NSC channel to 0.62 ± 0.07 from 0.03 ± 0.01 (mean ±se; n= 8) 30 min after application of terbutaline in a solution containing 1 mm Ca²⁺. The P _o of the terbutaline-stimulated NSC channel was diminished in the absence of extracellular Ca²⁺ to 0.26 ± 0.05 (n= 8). The cytosolic Ca²⁺ concentration ([Ca²⁺] _c ) in the presence and absence of extracellular Ca²⁺ was, respectively, 100 ± 6 and 20 ± 2 nm (n= 7) 30 min after application of terbutaline. The cytosolic Cl⁻ concentration ([Cl⁻] _c ) in the presence and absence of extracellular Ca²⁺ was, respectively, 20 ± 1 and 40 ± 2 mm (n= 7) 30 min after application of terbutaline. The diminution of [Ca²⁺] _c from 100 to 20 nm itself had no significant effects on the P _o if the [Cl⁻] _c was reduced to 20 mm; the P _o was 0.58 ± 0.10 at 100 nm [Ca²⁺] _c and 0.55 ± 0.09 at 20 nm [Ca²⁺] _c (n= 8) with 20 mm [Cl⁻] _c in inside-out patches. On the other hand, the P _o (0.28 ± 0.10) at 20 nm [Ca²⁺] _c with 40 mm [Cl⁻] _c was significantly lower than that (0.58 ± 0.10; P < 0.01; n= 8) at 100 nm [Ca²⁺] _c with 20 mm [Cl⁻] _c , suggesting that reduction of [Cl⁻] _c is an important factor stimulating the NSC channel. These observations indicate that the extracellular Ca²⁺ plays an important role in the stimulatory action of beta agonist on the NSC channel via reduction of [Cl⁻] _c . Received: 11 August 2000/Revised: 4 December 2000     

Effects of UCP2 and UCP3 Variants on the Manifestation of Overweight in Korean Children

To investigate the associations of uncoupling protein (UCP)2 and UCP3 gene variants with overweight and related traits, we genotyped UCP2−866G>A, UCP2Ala55Val, and UCP3−55C>T in 737 Korean children and 732 adults and collected data regarding anthropometric status and blood biochemistry. Information concerning the children's lifestyles and dietary habits was collected. The UCP2−866G>A and UCP3−55C>T gene variants showed significant associations with BMI level, waist circumference, and body weight in the children but not in the adults. Compared with −866GG carriers, the −866GA and AA carriers showed a strong decreasing trend in the risk for overweight (odds ratio (OR), 0.67; 95% confidence interval (CI), 0.45–1.01; P = 0.053). In comparison with UCP3−55CC carriers, children carrying −55CT and TT showed a significant reduction in the risk of overweight (OR, 0.67; 95% CI, 0.46–0.98; P = 0.039). There was also evidence of interactions between the effects of the combined UCP2−UCP3 genotype and obesity‐related metabolic traits. The greatest protective effect against overweight was seen in those with the combined genotype non‐UCP2−866GG and non‐UCP3−55CC, as compared with those carrying both UCP2−866GG and UCP3−55CC (OR, 0.60; 95% CI, 0.38–0.95; P = 0.030). In the subgroup with a low level of physical activity, UCP3−55CC carriers had higher BMI values than UCP3−55T carriers (16.6 ± 2.3 kg/m² vs. 16.1 ± 1.9 kg/m², P = 0.016). Low physical activity may aggravate the susceptibility to overweight in UCP2−866GG and UCP3−55CC carriers.     

Genetic parameter estimates for pre-weaning performance and reproduction traits in Markhoz goats

Genetic parameters for growth, mortality and reproductive performances of Markhoz goats were estimated from data collected during 1993–2010 at Markhoz goat Performance Testing Station in Sanandaj, Iran. For kid performance traits 3763 records were available for birth weight (BW), 2931 for weaning weight (WW), average daily gain (ADG) and Kleiber ratio (KR) (approximated as ADW/WW^0.75) and 3032 for pre-weaning mortality (PWM). For doe reproductive performance traits there were 2920 records available for litter size at birth (LSB), litter size at weaning (LSW), total litter weight at birth (TLWB) and litter mean weight per kid born (LMWKB), and 2182 for total litter weight at weaned (TLWW) and litter mean weight per kid weaned (LMWKW). Genetic parameters were estimated with univariate and bivariate models using restricted maximum likelihood (REML) procedures. Random effects were explored by fitting additive direct genetic effects, maternal additive genetic effects, maternal permanent environmental effects, the covariance between direct and maternal genetic effects, and common litter effects in different models for pre-weaning traits of kids. Also, in addition to an animal model, sire and threshold models, using a logit link function, were used for analyses of PWM. Models for LSB, LSW, TLWB, TLWW, LMWKB, and LMWKW included direct additive genetic effects, permanent environmental effects due to the animal as well as service sire effects. Estimated direct heritabilities were moderate for pre-weaning traits (0.22 for BW, 0.16 for WW, 0.21 for ADG, and 0.27 for KR and 0.29 for PWM), and low for reproduction traits (0.01 for LSB, 0.01 for LSW, 0.02 for TLWB, 0.03 for TLWW, 0.07 for LMWKB, and 0.06 for LMWKW). The estimates for the maternal additive genetic variance ratios were lower than direct heritability for BW (0.07) and KR (0.04). The estimate for the maternal permanent environmental variance ratios (c²) varied from 0.01 for KR to 0.07 for WW and ADG. The magnitude of common litter variance ratios (l²) was more substantial for BW (0.46) than the PWM (0.19) and KR (0.16). The estimate for the permanent environmental variance due to the animal (c²) ranged from 0.03 for LMWKB to 0.07 for TLWB and LMWKW, whereas service sire effects (s²) ranged from 0.02 to 0.04. The correlation between direct and maternal genetic effects were negative and high for BW (?0.51) and KR (?0.62). The genetic correlations between pre-weaning growth traits were positive and moderate to strong, as were genetic correlations between reproductive traits. Between BW and PWM the correlation was ?0.35. Phenotypic and environmental correlations for all traits were generally lower than genetic correlations.     

Genetic associations between maternal traits and aggressive behaviour in Large White sows

The present study examined the possibilities and consequences of selecting pigs for reduced aggression and desirable maternal behaviour. Data were recorded from 798 purebred Large White gilts, with an age of 217±17.7 (mean±SD) days, which were observed at mixing with unfamiliar conspecifics. The reaction of the sows towards separation from their litter was assessed for 2022 litters from 848 Large White sows. Sows’ performance during their time in the farrowing unit was scored based on the traits farrowing behaviour (i.e. need of birth assistance), rearing performance (i.e. litter quality at day 10 postpartum (pp)), usability (i.e. additional labour input during lactation period e.g. for treatments) and udder quality of the sow (i.e. udder attachment). For agonistic behaviour, traits heritabilities of h²=0.11±0.04 to h²=0.28±0.06 were estimated. For the sow’s reaction towards separation from her litter low heritabilities were found (h²=0.03±0.03 for separation test on day 1 pp and h²=0.02±0.03 for separation test on day 10 pp). Heritabilities for lactating sow’s performance (farrowing behaviour, rearing performance, usability of the sow and udder quality) in the farrowing unit ranged from h²=0.03±0.02 to h²=0.19±0.03. Due to these results it can be assumed that selection for these traits, for example, for udder quality or reduced aggression, is possible. Antagonistic associations were found between separation test on day 1 pp and different measures of aggressiveness (r_g=−0.22±0.26 aggressive attack and r_g=−0.41±0.33 reciprocal fighting). Future studies should determine economic as well as welfare-related values of these traits in order to decide whether selection for these traits will be reasonable.     

Oxidation of NADH by plant mitochondria: Kinetics and effects of calcium ions

The kinetics of NADH oxidation by the outer membrane electron transport system of intact beetroot (Beta vulgaris L.) mitochondria were investigated. Very different values for V_max and the K_m for NADH were obtained when either antimycin A-insensitive NADH-cytochrome c activity (V_max= 31 ± 2.5 nmol cytochrome c (mg protein)^?1 min^?1; K_m= 3.1 ± 0.8 μM) or antimycin A-insensitive NADH-ferricyanide activity (V_max= 1.7 ± 0.7 μmol ferricyanide (mg protein)^?1 min^?1; K_m= 83 ± 20 μM) were measured. As ferricyanide is believed to accept electrons closer to the NADH binding site than cytochrome c, it was concluded that 83 ± 20 μM NADH represented a more accurate estimate of the binding affinity of the outer membrane dehydrogenase for NADH. The low K_m determined with NADH-cytochrome c activity may be due to a limitation in electron flow through the components of the outer membrane electron transport chain. The K_m for NADH of the externally-facing inner membrane NADH dehydrogenase of pea leaf (Pisum sativum L. cv. Massey Gem) mitochondria was 26.7 ± 4.3 μM when oxygen was the electron acceptor. At an NADH concentration at which the inner membrane dehydrogenase should predominate, the Ca²⁺ chelator, ethyleneglycol-(β-aminoethylether)-N,N,-tetraacetic acid (EGTA), inhibited the oxidation of NADH through to oxygen and to the ubiquinone-10 analogues, duroquinone and ubiquinone-1, but had no effect on the antimycin A-insensitive ferricyanide reduction. It is concluded that the site of action of Ca²⁺ involves the interaction of the enzyme with ubiquinone and not with NADH.     

The quantitative genetics of sexually selected traits,preferred traits and preference: a review and analysis of the data

The maintenance of variation in sexually selected traits is a puzzle that has received increasing attention in the past several decades. Traits that are related to fitness, such as life‐history or sexually selected traits, are expected to have low additive genetic variance (and hence, heritability) due to the rapid fixation of advantageous alleles. However, previous analyses have suggested that the heritabilities of sexually selected traits are on average higher than nonsexually selected traits. We show that the heritabilities of sexually selected traits are not significantly different from those of nonsexually selected traits overall or when separated into the three trait categories: behavioural, morphological and physiological. In contrast with previous findings, the heritability of preference is quite low (h² = 0.25 ± 0.06) and is in the same range as life‐history traits. We distinguish preferred traits as a category of sexually selected traits and find that the heritability of the former is not significantly different than sexually selected traits overall (0.48 ± 0.04 vs. 0.46 ± 0.03). We test the hypothesis that the heritability of sexually selected traits is negatively correlated with the strength of sexual selection. As predicted, there is a significant negative correlation between the heritabilities of sexually selected traits and the strength of selection. This suggests that heritabilities do indeed decrease as sexual selection increases but sexual selection is not strong enough to cause heritabilities of sexually selected traits to deviate from the same type of nonsexually selected traits.     

Comparison of ecosystem water-use efficiency among Douglas-fir forest, aspen forest and grassland using eddy covariance and carbon isotope techniques

Comparisons were made among Douglas‐fir forest, aspen (broad leaf deciduous) forest and wheatgrass (C₃) grassland for ecosystem‐level water‐use efficiency (WUE). WUE was defined as the ratio of photosynthetic CO₂ assimilation rate and evapotranspiration (ET) rate. The ET data measured by eddy covariance were screened so that they overwhelmingly represented transpiration. The three sites used in this comparison spanned a range of vegetation (plant functional) types and environmental conditions within western Canada. When compared in the relative order Douglas‐fir (located on Vancouver Island, BC), aspen (northern Saskatchewan), grassland (southern Alberta), the sites demonstrated a progressive decline in precipitation and a general increase in maximum air temperature and atmospheric saturation deficit (D_max) during the mid‐summer. The average (±SD) WUE at the grassland site was 2.6±0.7 mmol mol^?1, which was much lower than the average values observed for the two other sites (aspen: 5.4±2.3, Douglas‐fir: 8.1±2.4). The differences in WUE among sites were primarily because of variation in ET. The highest maximum ET rates were approximately 5, 3.2 and 2.7 mm day^?1 for the grassland, aspen and Douglas‐fir sites, respectively. There was a strong negative correlation between WUE and D_max for all sites. We also made seasonal measurements of the carbon isotope ratio of ecosystem respired CO₂ (δ_R) in order to test for the expected correlation between shifts in environmental conditions and changes to the ecosystem‐integrated ratio of leaf intercellular to ambient CO₂ concentration (c_i/c_a). There was a consistent increase in δ_R values in the grassland, aspen forest and Douglas‐fir forest associated with a seasonal reduction in soil moisture. Comparisons were made between WUE measured using eddy covariance with that calculated based on D and δ_R measurements. There was excellent agreement between WUE values calculated using the two techniques. Our δ_R measurements indicated that c_i/c_a values were quite similar among the Douglas‐fir, aspen and grassland sites, despite large variation in environmental conditions among sites. This implied that the shorter‐lived grass species had relatively high c_i/c_a values for the D of their habitat. By contrast, the longer‐lived Douglas‐fir trees were more conservative in water‐use with lower c_i/c_a values relative to their habitat D. This illustrates the interaction between biological and environmental characteristics influencing ecosystem‐level WUE. The strong correlation we observed between the two independent measurements of WUE, indicates that the stable isotope composition of respired CO₂ is a useful ecosystem‐scale tool to help study constraints to photosynthesis and acclimation of ecosystems to environmental stress.     

A single amino acid substitution in the MurF UDP‐MurNAc‐pentapeptide synthetase renders Streptococcus pneumoniae dependent on CO2 and temperature

The respiratory tract pathogen Streptococcus pneumoniae encounters different levels of environmental CO₂ during transmission, host colonization and disease. About 8% of all pneumococcal isolates are capnophiles that require CO₂‐enriched growth conditions. The underlying molecular mechanism for caphnophilic behaviour, as well as its biological function is unknown. Here, we found that capnophilic S. pneumoniae isolates from clonal complex (CC) 156 (i.e. Spain^9V‐3 ancestry) and CC344 (i.e. Norway^NT‐42 ancestry) have a valine at position 179 in the MurF UDP‐MurNAc‐pentapeptide synthetase. At ≤ 30°C, the growth characteristics of capnophilic and non‐capnophilic CC156 strains were equal, but at > 30°C growth and survival of MurF^V179 strains was dependent on > 0.1% CO₂‐enriched conditions. Expression of MurF^V179 in S. pneumoniae R6 and G54 rendered these, otherwise non‐capnophilic strains, capnophilic. Time‐lapse microscopy revealed that a capnophilic CC156 strain undergoes rapid autolysis upon exposure to CO₂‐poor conditions at 37°C, and staining with fluorescently labelled vancomycin showed a defect in de novo cell wall synthesis. In summary, in capnophilic S. pneumoniae strains from CC156 and CC344 cell wall synthesis is placed under control of environmental CO₂ levels and temperature. This mechanism might represent a novel strategy of the pneumococcus to rapidly adapt and colonize its host under changing environmental conditions.     

Morphometric characterization of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa using computer-assisted spermatozoa analysis (ASMA)

As part of a larger study on sperm quality and cryopreservation methods, the present study characterized the head morphometry of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa, using both scanning electron microscopy (SEM) and computer‐assisted morphology analysis (ASMA). The latter method has been used rarely in fish and this is its first application on sharpsnout sea bream and gilthead sea bream spermatozoa. Results obtained using SEM are expensive and time‐consuming, while ASMA provides a faster and automated evaluation of morphometric parameters of spermatozoa head. For sharpsnout sea bream spermatozoa, similar head measurement values were obtained using both ASMA and SEM, having a mean ± standard error length of 2.57 ± 0.01 μm vs 2.54 ± 0.02 μm, width of 2.22 ± 0.02 μm vs 2.26 ± 0.04 μm, surface area of 4.44 ± 0.02 μm² vs 4.50 ± 0.04 μm² and perimeter of 7.70 ± 0.02 μm vs 7.73 ± 0.04 μm using ASMA and SEM, respectively. Although gilthead sea bream spermatozoa were found to be smaller than those of sharpsnout sea bream, spermatozoal head morphometry parameters were also found to be similar regardless of evaluation method, having a mean head length of 1.97 ± 0.01 μm vs 1.94 ± 0.02 μm, head width of 1.80 ± 0.01 μm vs 1.78 ± 0.02 μm, surface area of 3.16 ± 0.03 μm² vs 3.18 ± 0.06 μm² and perimeter of 6.52 ± 0.04 μm vs 6.56 ± 0.08 μm using ASMA and SEM, respectively. The results demonstrate that ASMA can be considered as a reliable technique for spermatozoal morphology analysis, and can be a useful tool for studies on fish spermatozoa, providing quick and objective results.     

Ex vitro experimental study on concentration polari-zation of macromolecules (LDL) at an arterial stenosis

To verify the previous theoretical prediction that the disturbed flow distal to a stenosis enhances lipid accumulation at the blood/arterial wall interface, we designed a canine carotid arterial stenosis model and measured ex vitro the luminal surface concentration of bovine serum albumin (as a tracer macromolecule) by directly taking liquid samples from the luminal surface of the artery. The experimental results showed that due to the presence of a filtration flow, the luminal surface albumin concentration cw was higher than the bulk concentration co as predicted by our theory. The measurement revealed that the luminal surface concentration of macromolecules was indeed enhanced significantly in regions of the disturbed flow. At Re = 50, the relative luminal surface concentration cw/co was 1.66±0.10 in the vortex region, while the cw/co was 1.37±0.06 in the laminar flow region. When Re increased to 100,the cw/co in the vortex flow region and the laminar flow region reduced to 1.39±0.07 and 1.24±0.04,respectively. The effect of the filtration rate, vw, on the luminal surface concentration of albumin was remarkably apparent. At Re=50 and 100, when vw=8.9±1.7×10-6 cm/s, cw in the vortex region was 77% and 52% higher than co respectively, meanwhile when vw = 4.8±0.6×10-6 cm/s, cw in the vortex region was only 66% and 39% higher than co respectively. In summary, the present study has provided further experimental evidence that concentration polarization can occur in the arterial system and fluid layer with highly concentrated lipids in the area of flow separation point may be responsible for the formation and development of atherosclerosis.