Influences of a Supplemental Blend of Essential Oils Plus 25-Hydroxy-Vit-D3 and Zilpaterol Hydrochloride (ß2 Agonist) on Growth Performance and Carcass Measures of Feedlot Lambs Finished under Conditions of High Ambient Temperature.

Forty-eight Pelibuey × Katahdin male intact lambs (25.12 ± 3.79 kg LW) were used in a 70-d growing-finishing trial. Dietary treatments consisted of total mixed corn-based diet supplemented with: (1) no feed additives (Control); (2) 150 mg of essential oils blend plus 0.10 mg of 25-hydroxy-Vit-D3/kg diet offered throughout the 70-d experimental period (EOD3); (3) Control diet fed during the first 35 days and zilpaterol hydrochloride (ZH) supplementation at 6 mg/kg diet offered during the final 35 days of the experiment (32 days with ZH with a withdrawal 3-d before harvest), and (4) basal diet supplemented with EOD3 during first 35 days finishing, and EOD3 in combination with ZH (EOD3 + ZH) during the subsequent 32-days with ZH withdrawal 3 days before harvest. The temperature-humidity index during the experiment averaged 80.4 ± 3.2. There were no treatment interactions (p > 0.20) on growth performance and carcass measures. Supplemental EOD3 did not affect (p = 0.43) dry matter intake (DMI), but increased (p < 0.01) carcass adjusted average daily gain (ADG, 9.2%), gain efficiency (GF, 6.7%), and observed vs. expected dietary net energy for maintenance (NEm, 4.8%) and for gain (NEg, 6.4%). Supplemental ZH did not affect dry matter intake (DMI, p = 0.50) but increased (p < 0.01) carcass adjusted ADG (14.5%), GF (13%) and observed vs. expected dietary NEm (9%) and NEg (11.7%). Compared to control lambs, the combination of both additives increased ADG (24.9%), GF (21.2%), and observed vs. expected dietary NEm and NEg (14.2% and 18.9%, respectively). There were no treatment interactions on carcass characteristics, visceral organ mass, or on gene expression of IGF1, IGF2 and mTOR in longissimus muscle (LM). Supplemental EOD3 increased hot carcass weight (HCW; 4.0%, p < 0.01) but did not affect other carcass measures. Supplemental EOD3 decreased (3%, p = 0.03) intestine mass weight (g intestine/kg empty body weight). Supplemental ZH increased HCW (6%, p < 0.01), dressing percentage (1.7%, p = 0.04), and LM area (9.7%, p < 0.01), and decreased kidney-pelvic-fat percentage (16.2%, p < 0.01), fat thickness (14.7%, p = 0.03), and visceral fat. Compared to controls, the combination of EOD3 with ZH increased HCW (10.2%). It is concluded that growth performance responses to supplemental EOD3 and ZH are additive. Both supplements can be fed in combination without detrimental effects on expected benefits when fed separately. In addition, ZH supplementation improves carcass traits.

Essential Oils Combined with Vitamin D3 or with Probiotic as an Alternative to the Ionophore Monensin Supplemented in High-Energy Diets for Lambs Long-Term Finished under Subtropical Climate.

Supplementation with natural additives such as essential oils (EO) or probiotics has resulted in comparable growth performance to that of supplemental monensin in fattening lambs in hot environments. Supra-supplementation levels of vitamin D3 improved the carcass weight and dressing percentage of steers fattened under tropical conditions. We hypothesized that certain combinations of these natural additives could be complementary. For this reason, a feeding trial was carried out using 48 Pelibuey × Katahdin non-castrated male lambs (107 ± 14 d age; 17.9 ± 2.51 kg LW). Lambs were fed an 88:12 concentrate to forage ratio basal diet supplemented (dry matter basis, DMI) with: (1) no additive (CON); (2) 28 mg monensin/kg diet (MON); (3) 150 mg of essential oils containing a combination of thymol, eugenol, vanillin, guaiac, and limonene plus 0.12 mg vitamin D3 (EO + D3)/kg diet; and (4) 300 mg of essential oils containing a combination of carvacrol and cynamaldehyde plus 2 g probiotic (2.2 × 108 CFU of bacillus subtilis/kg diet, EO + BS). Lambs were grouped by initial weight and assigned within six weight groupings to 24 pens (2 lambs/pen, 6 replicas per treatment) in a randomized complete block design. The experiment lasted 121 days. Daily maximal THI exceeded the 80 "danger or "emergency" range for 119 days of the 121 days of the trial. Lambs supplemented with MON had similar DMI, growth performance, and dietary energetics to those of CON lambs. Lambs supplemented with EO + BS had a greater (9.2%, p ≤ 0.05) average daily gain (ADG) than the CON and MON groups due to enhanced (10.2%, p ≤ 0.05) dry matter intake. Thus, gain efficiency (GF) and estimated dietary energy were similar for CON, MON, and EO + BS. Lambs receiving EO + D3 had similar (0.254 vs. 0.262 kg/d) ADG but a lower DMI (8%, p < 0.05) compared with EO + BS lambs. Consequently, GF and estimated dietary net energy were greater (4.9 and 3.7%, respectively; p ≤ 0.05) for EO + D3 lambs. Even when ambient heat load was elevated, the efficiency of utilization of dietary energy (observed-to-expected dietary net energy) was close to 1.00 (0.992) expected for EO + D3 lambs. In contrast, efficiency of energy utilization was depressed by -4.4% for lambs on the other treatments. Compared with the other treatments, lambs receiving EO + D3 had greater longissimus muscle area (5.6%, p < 0.05) and lower kidney pelvic fat (21.8%, p ≤ 0.05). There were no treatment effects on shoulder tissue composition or whole cuts (expressed as % of cold carcass weight). Compared to CON, lambs that were fed with natural additives showed 3.5% lower (p ≤ 0.05) intestine mass. All supplemental additives decreased visceral fat mass, which was minimal with EO + D3 treatment. Combinations of essential oils with vitamins or probiotics were superior to antibiotic monensin in finishing diets for feedlot lambs. Combining EO with probiotics promoted DM intake and gain but not gain efficiency, while combining EO with vitamin D3 supra-supplementation increased dietary energy efficiency and improved some carcass characteristics in lambs fattening under high ambient heat loads.

Effect of Combining the Ionophore Monensin with Natural Antimicrobials Supplemented in the Last Phase of Finishing of Lambs: Growth Performance, Dietary Energetics, and Carcass Characteristics.

With the aim of evaluating the effect of combining an antibiotic ionophore with plant extracts and probiotics on the productive efficiency (performance and carcass) during the last phase of lamb fattening, 24 Pelibuey × Katahdin male lambs (38.47 ± 3.92 kg, initial weight) were fed with a high-energy diet during for 56 days, and assigned, under a complete randomized block design experiment to one of the following supplement treatments: (1) 28 mg of monensin/kg diet DM supplemented alone (MON), (2) combination of MON plus 2 g/kg diet of a product contained Bacillus subtilis 2.2 × 108 CFU kg diet DM (MON + BS), (3) combination of MON + BS plus 300 mg essential oils/kg diet DM (MON + BS + EO), and (4) BS alone. At the end of the feeding trial (56-d), lambs were slaughtered and carcass variables were measured. Compared to the rest of the treatments, combining MON with BS improved dietary NE by 3.4% and the efficiency of utilization of dietary energy consumed. Inclusion of EO in the MON + BS combination resulted in a similar average daily weight gain (ADG) and feed efficiency (GF) when compared with MON + BS, but showed a lower dietary net energy (NE), hot carcass weight, and dressing percentage. Lambs receiving BS alone showed greater average ADG and dry matter intake (DMI) than lambs receiving MON + BS + EO, but similar feed GF and dietary NE. There were no treatment effects on tissue composition, whole cut, or visceral organ mass. It was concluded that combining probiotics with the ionophore monensin can improve the efficiency of dietary energy utilization in the last phase of finishing. Probiotics supplemented alone result in greater ADG without a difference in dietary energy efficiency when compared with MON alone. Inclusion of EO in the MON + BS combination did not show advantages; on the contrary, it reduced carcass weight and dressing percentage. It is necessary to further research the potential complementary effects of combining diverse sources of natural additives with synthetic antibiotics.

Linking glucocorticoid variations to monthly and daily behavior in a wild endangered neotropical primate.

Identifying the factors swaying physiological stress levels in wild animals can help depict how they cope with environmental and social stressors, shedding light on their feeding ecology, behavioral plasticity, and adaptability. Here, we used noninvasive methods to explore the link between glucocorticoid levels and behavior in an endangered neotropical primate facing habitat fragmentation pressure, the black lion tamarin (Leontopithecus chrysopygus). We investigated monthly and day-to-day glucocorticoid variations independently to attempt to disentangle the complex nature of the adrenocortical activity. Between May 2019 to March 2020, we followed two groups of black lion tamarins in two different areas, a continuous forest and a small fragment, and gathered behavioral data (over 95 days in total; 8.6 ± 3.9 days/month) and fecal samples (Nsamples = 468; 4.93 ± 3.5 samples/day) simultaneously. Preliminary analyses enabled us to identify circadian variations linked to the biological rhythm, which were taken into account in subsequent models. Monthly analyses revealed that black lion tamarin fecal glucocorticoid metabolite levels vary according to changes in activity budget associated with the fruit consumption, movement, and resting time of the groups. At a day-to-day level, while intergroup encounters led to increases in fecal glucocorticoid metabolite concentrations, we found that changes in food intake or activity level did not trigger physiological stress responses. These findings suggest that diet and ranging patterns, driven by food availability and distribution, influence physiological stress at a seasonal scale, while acute stressors such as interspecific competition trigger short-term stress responses. Exploring fecal glucocorticoid metabolite variations over different timescales can help uncover the predictive and reactive facets of physiological stress in wild species. Moreover, having a comprehensive understanding of the physiological state of species is a valuable conservation tool for evaluating how they cope in changing environments.

Acute effects of sodium bicarbonate ingestion on cycling time-trial performance: A systematic review and meta-analysis of randomized controlled trials.

This study aimed to investigate the isolated effects of NaHCO3 on cycling time-trial performance. Furthermore, we investigated whether the ingestion time of NaHCO3, standardized or individualized based on time to peak, could be effective in improving cycling time-trial performance. A systematic review was carried out on randomized placebo-controlled studies. A random-effects meta-analysis assessed the standardized mean difference (SMD) between NaHCO3 and placebo conditions. Eighteen studies were qualitatively (systematic review) and quantitatively (meta-analysis) analysed concerning mean power output (Wmean) (n = 182) and time performance (n = 201). The reviewed studies showed a low risk of bias and homogenous results for Wmean (I2 = 0%) and performance time (I2 = 0%). Overall, when compared to placebo, the NaHCO3 ingestion improved the Wmean (SMD: 0.42; 95% CI: 0.21-0.63; P = 0.001) and performance time (SMD: 0.22; 95% CI: 0.02-0.43; P = 0.03). Similarly, the NaHCO3 ingestion using a time-to-peak strategy improved the Wmean (SMD: 0.39; 95% CI: 0.03-0.75; P = 0.04; I2 = 15%) and performance time (SMD: 0.34; 95% CI: 0.07-0.61, P = 0.01, I2 = 0%). The present findings reveal that NaHCO3 ingestion has the potential to increase the overall performance time and Wmean in cycling time trials.HighlightsNaHCO3 is an effective strategy to increase cycling time-trial performance.The standardized protocol did not improve the cycling time-trial performance parameters.The individualized time-to-peak NaHCO3 ingestion has a positive effect on time and Wmean during cycling time-trial performance.

Energy supply per neuron is constrained by capillary density in the mouse brain.

Neuronal densities vary enormously across sites within a brain. Does the density of the capillary bed vary accompanying the presumably larger energy requirement of sites with more neurons, or with larger neurons, or is energy supply constrained by a mostly homogeneous capillary bed? Here we find evidence for the latter, with a capillary bed that represents typically between 0.7 and 1.5% of the volume of the parenchyma across various sites in the mouse brain, whereas neuronal densities vary by at least 100-fold. As a result, the ratio of capillary cells per neuron decreases uniformly with increasing neuronal density and therefore with smaller average neuronal size across sites. Thus, given the relatively constant capillary density compared to neuronal density in the brain, blood and energy availability per neuron is presumably dependent on how many neurons compete for the limited supply provided by a mostly homogeneous capillary bed. Additionally, we find that local capillary density is not correlated with local synapse densities, although there is a small but significant correlation between lower neuronal density (and therefore larger neuronal size) and more synapses per neuron within the restricted range of 6,500-9,500 across cortical sites. Further, local variations in the glial/neuron ratio are not correlated with local variations in the number of synapses per neuron or local synaptic densities. These findings suggest that it is not that larger neurons, neurons with more synapses, or even sites with more synapses demand more energy, but simply that larger neurons (in low density sites) have more energy available per cell and for the totality of its synapses than smaller neurons (in high density sites) due to competition for limited resources supplied by a capillary bed of fairly homogeneous density throughout the brain.

Avoiding the effects of translocation on the estimates of the metabolic rates across an elevational gradient.

Body maintenance costs are often considered a proxy for performance in fitness traits. Maintenance energy requirements are measured as minimal metabolic rate of inactive, postabsorptive individuals in the laboratory. For mountain-dwelling species, translocation to the laboratory often means that they are also moved to another elevation. Due to physiological adaptations to local oxygen pressure, rapid elevational change can alter metabolic rate and translocation may result in erroneous estimates of body maintenance costs. In this study, we measured resting metabolic rate (RMR) of three populations of the Mesquite lizard (Sceloporus grammicus, Wiegmann 1828) at their native elevations (i.e., 2600, 3200 and 4100 m). Our results showed that at native elevations, mass specific RMR of lizards from the high elevation population (4100 m) did not differ from the RMR of the other populations (i.e., 2600 and 3200 m), whereas the lizards from the low elevation (2600 m) had lower RMR than those from the intermediate population. These results differ from a previous study in which the RMR of lizards from the same populations were reported to increase with native elevation when translocated and measured at an intermediate elevation. Hence, our results show that translocation in elevation can affect metabolic measures. We caution researchers that changes in elevation may preclude accurate measures of RMR in some animals and may therefore incorrectly predict performance of fitness-related traits.

Disruption of neuromedin B receptor improves mitochondrial oxidative phosphorylation capacity in gastrocnemius muscle of female mice.

Neuromedin B (NB), a bombesin-like peptide, exerts its specific actions by binding to the neuromedin B receptor (NBR), a G protein-coupled receptor. Female NBR-knockout (NBR-KO) mice exhibit resistance to diet-induced obesity, without hyperphagia, suggesting possible increase in energy expenditure. Skeletal muscle (SM) is crucial for whole body energy homeostasis, however, the presence of NB-NBR signaling and its effects in SM are unknown. Here, we show that male and female wild type express Nmbr and Nmb mRNA in SM, with higher levels in females. Female NBR-KO gastrocnemius showed increased Myh7 mRNA level, which characterizes type I fibers (oxidative profile). Their permeabilized gastrocnemius fibers, studied by high-resolution respirometry, exhibited higher consumption of O2 coupled to ATP synthesis and unaltered uncoupled respiration. NBR-KO gastrocnemius had higher protein levels of ATP-synthase and Nduf9 mRNA, corresponding to mitochondrial complex I subunit. NBR-KO gastrocnemius exhibited slight increase in mitochondria number, increased thickness of Z line at electron microscopy, and unaltered mitochondrial dynamics markers. Therefore, in the females' gastrocnemius, a predominantly glycolytic SM, the NBR absence promotes changes that favor mitochondrial oxidative phosphorylation capacity. In addition, in L6 myocytes, NB treatment (5 µg/mL/16 h) promoted lower O2 consumption coupled to ATP synthesis, suggesting direct action at SM cells. Altogether, the study reinforces the hypothesis that inhibition of NB-NBR signaling enhances the capacity for oxidative phosphorylation of white SM, encouraging future studies to elucidate their contribution on other types of SM and whole body energy expenditure, which may lead to a new target to drug development for obesity treatment.NEW & NOTEWORTHY This study describes neuromedin B (NB) and NB receptor as new regulators of skeletal muscle mitochondrial function. The white skeletal muscle mitochondrial oxidative phosphorylation capacity was increased by NB receptor genetic disruption in female mice. These findings may contribute to the resistance to diet-induced obesity, previously found in these mice, which requires future studies. Thus, investigations are necessary to clarify if blockade of NB receptor may be an approach to develop drugs to combat obesity.

Phenotypic plasticity in the energy metabolism of a small Andean rodent: Effect of short-term thermal acclimation and developmental conditions.

The study of phenotypic variation within species in response to different environments is a central issue in evolutionary and ecological physiology. Particularly, ambient temperature is one of the most important factors modulating interactions between animals and their environment. Phyllotis xanthopygus, a small Andean rodent, exhibits intraspecific differences along an altitudinal gradient in traits relevant to energy balance that persist after acclimation to common experimental temperatures. Therefore, we aim to explore geographic variations in energetic traits of P. xanthopygus and to assess the contribution of phenotypic plasticity to population differences. We compared metabolic rate and thermal conductance in response to different acclimation temperatures in animals collected at distinct altitudes (F0 generation) and in their offspring, born and raised under common-garden conditions (F1 generation). We found intraspecific differences in resting metabolic rate (RMR) of animals collected at different altitudes that were no longer evident in the F1 generation. Furthermore, although both generations showed the same pattern of RMR flexibility in response to acclimation temperature, its magnitude was lower for the F1 individuals. This suggests that developmental conditions affect the short-term acclimation capacity of this trait during adulthood. On the other hand, thermal conductance (C) showed irreversible plasticity, as animals raised in the laboratory at stable warm conditions had a relatively higher C than the animals from the field, showing no adjustments to thermal acclimation during adulthood in either group. In sum, our results support the hypothesis that the developmental environment shapes energetic traits, emphasizing the relevance of incorporating ontogeny in physiological studies.

Energetic and structural effects of the Tanford transition on ligand recognition of bovine ß-lactoglobulin.

Bovine ß-lactoglobulin, an abundant protein in whey, is a promising nanocarrier for peroral administration of drug-like hydrophobic molecules, a process that involves transit through the different acidic conditions of the human digestive tract. Among the several pH-induced conformational rearrangements that this lipocalin undergoes, the Tanford transition is particularly relevant. This transition, which occurs with a midpoint around neutral pH, involves a conformational change of the E-F loop that regulates accessibility to the primary binding site. The effect of this transition on the ligand binding properties of this protein has scarcely been explored. In this study, we carried out an energetic and structural characterization of ß-lactoglobulin molecular recognition at pH values above and below the zone in which the Tanford transition occurs. The combined analysis of crystallographic, calorimetric, and molecular dynamics data sheds new light on the interplay between self-association, ligand binding, and the Tanford pre- and post-transition conformational states, revealing novel aspects underlying the molecular recognition mechanism of this enigmatic lipocalin.

Fur Color and Nutritional Status Predict Hair Cortisol Concentrations of Dogs in Nicaragua.

This study examined the relationships between hair cortisol concentrations (HCC) and sex, age, nutritional status (as determined by body condition scores, or BCS), and body mass (geometric mean calculated from morphometric measurements), as well as the potential influence of hair pigmentation (light, dark, or agouti/mixed) on HCC in dogs of the Bosawas Biosphere Reserve, Nicaragua. The dogs examined in this study live in a marginal environment where disease, malnutrition, and mortality rates are high. For fur color, HCC was significantly higher in light fur than in than dark and mixed fur (p < 0.001). In addition, BCS scores were found to have a negative effect on HCC (p < 0.001). Measures of sex and body size exhibited inconclusive effects on HCC, and when compared to adult dogs, juvenile dogs did not exhibit significantly different HCC. Repeated measures of dogs over time reveal a moderate intra-class correlation, suggesting that there are unmeasured sources of individual-level heterogeneity. These findings imply a need to account for fur color in studies of HCC in dogs, and the study suggests an overlooked relationship between cortisol and body condition scores in undernourished dogs in diverse settings.

Mechanical and energetic determinants of impaired gait following stroke: segmental work and pendular energy transduction during treadmill walking.

Systems biology postulates the balance between energy production and conservation in optimizing locomotion. Here, we analyzed how mechanical energy production and conservation influenced metabolic energy expenditure in stroke survivors during treadmill walking at different speeds. We used the body center of mass (BCoM) and segmental center of mass to calculate mechanical energy production: external and each segment's mechanical work (Wseg). We also estimated energy conservation by applying the pendular transduction framework (i.e. energy transduction within the step; Rint). Energy conservation was likely optimized by the paretic lower-limb acting as a rigid shaft while the non-paretic limb pushed the BCoM forward at the slower walking speed. Wseg production was characterized by greater movements between the limbs and body, a compensatory strategy used mainly by the non-paretic limbs. Overall, Wseg production following a stroke was characterized by non-paretic upper-limb compensation, but also by an exaggerated lift of the paretic leg. This study also highlights how post-stroke subjects may perform a more economic gait while walking on a treadmill at preferred walking speeds. Complex neural adaptations optimize energy production and conservation at the systems level, and may fundament new insights onto post-stroke neurorehabilitation.This article has and associated First Person interview with the first author of the paper.

Convergent changes in muscle metabolism depend on duration of high-altitude ancestry across Andean waterfowl.

High-altitude environments require that animals meet the metabolic O2 demands for locomotion and thermogenesis in O2-thin air, but the degree to which convergent metabolic changes have arisen across independent high-altitude lineages or the speed at which such changes arise is unclear. We examined seven high-altitude waterfowl that have inhabited the Andes (3812-4806 m elevation) over varying evolutionary time scales, to elucidate changes in biochemical pathways of energy metabolism in flight muscle relative to low-altitude sister taxa. Convergent changes across high-altitude taxa included increased hydroxyacyl-coA dehydrogenase and succinate dehydrogenase activities, decreased lactate dehydrogenase, pyruvate kinase, creatine kinase, and cytochrome c oxidase activities, and increased myoglobin content. ATP synthase activity increased in only the longest established high-altitude taxa, whereas hexokinase activity increased in only newly established taxa. Therefore, changes in pathways of lipid oxidation, glycolysis, and mitochondrial oxidative phosphorylation are common strategies to cope with high-altitude hypoxia, but some changes require longer evolutionary time to arise.

Resting metabolic rates increase with elevation in a mountain-dwelling lizard.

Individuals that inhabit broad elevational ranges may experience unique environmental challenges. Because temperature decreases with increased elevation, the ectotherms living at high elevations have to manage limited activity time and high thermoregulatory effort. The resting metabolic rate (RMR) of a postabsorptive animal is related to its total energy requirements as well as many other fitness traits. Mesquite lizards (Sceloporus grammicus) living on La Malinche Volcano, Mexico, inhabit a wide elevational range with some populations apparently thriving above the tree line. We measured the RMR of lizards from different elevations (i.e., 2,600, 3,200, and 4,100 m) at four ecologically relevant temperatures (i.e., 15, 25, 30, and 35 °C) and found that RMR of mesquite lizards increased with temperature and body mass. More importantly, lizards from the high-elevation population had mass specific RMR that was higher at all temperatures. While the higher RMRs of high-elevation populations imply higher metabolic costs at a given temperature these lizards were also smaller. Both of these traits may allow these high elevation populations to thrive in the face of the thermal challenges imposed by their environment.

Effects of temperature and meal size on the postprandial metabolic response of Leptodactylus latrans (Anura, Leptodactylidae).

The postprandial increment of metabolism, often referred to as specific dynamic action (SDA), encompasses the summed costs of meal ingestion, digestion, absorption, and assimilation. Different SDA parameters, such as its magnitude, duration, and relative cost, are affected by a diverse set of environmental and physiological determinants, including meal size and body temperature. While the influence of these variables has been thoroughly examined in most ectothermic vertebrate groups, few studies have focused on the determinants and consequences of the SDA response in anuran amphibians. Thus, we examined the effects of meal size and body temperature on the SDA response of a Neotropical frog, Leptodactylus latrans, by measuring the rates of oxygen consumption of frogs while fasting and after being fed meals of different sizes at different temperatures. SDA lasted from 3 to 5 days and increased with meal size for frogs fed meals equivalent to 5-15% of their body mass. SDA cost was not affected by meal size and averaged 14.6% of the caloric content of the meal. Temperature increment was accompanied by proportional increases in metabolic rate and shorter SDA duration. The relative SDA cost averaged 14.8% of the meal caloric content at 20°C and 25°C, but increased to 23.5% at 30°C. Our results indicate that meal size imposes no physiological or energetic constraint to L. latrans feeding. Digestion at temperatures near the thermal preference of the species seems to optimize energetic return, whereas the shortening of SDA duration at higher temperatures may provide significant ecological advantages.

A biophysical and structural study of two chitinases from Agave tequilana and their potential role as defense proteins.

Plant chitinases are enzymes that have several functions, including providing protection against pathogens. Agave tequilana is an economically important plant that is poorly studied. Here, we identified a chitinase from short reads of the A. tequilana transcriptome (AtChi1). A second chitinase, differing by only six residues from the first, was isolated from total RNA of plants infected with Fusarium oxysporum (AtChi2). Both enzymes were overexpressed in Escherichia coli and analysis of their sequences indicated that they belong to the class I glycoside hydrolase family19, whose members exhibit two domains: a carbohydrate-binding module and a catalytic domain, connected by a flexible linker. Activity assays and thermal shift experiments demonstrated that the recombinant Agave enzymes are highly thermostable acidic endochitinases with Tm values of 75 °C and 71 °C. Both exhibit a molecular mass close to 32 kDa, as determined by MALDI-TOF, and experimental pIs of 3.7 and 3.9. Coupling small-angle x-ray scattering information with homology modeling and docking simulations allowed us to structurally characterize both chitinases, which notably show different interactions in the binding groove. Even when the six different amino acids are all exposed to solvent in the loops located near the linker and opposite to the binding site, they confer distinct kinetic parameters against colloidal chitin and similar affinity for (GlnNAc)6, as shown by isothermal titration calorimetry. Interestingly, binding is more enthalpy-driven for AtChi2. Whereas the physiological role of these chitinases remains unknown, we demonstrate that they exhibit important antifungal activity against chitin-rich fungi such as Aspergillus sp. DATABASE: SAXS structural data are available in the SASBDB database with accession numbers SASDDE7 and SASDDA6. ENZYMES: Chitinases (EC3.2.1.14).

Altered Movement Biomechanics in Chronic Ankle Instability, Coper, and Control Groups: Energy Absorption and Distribution Implications.

CONTEXT: Patients with chronic ankle instability (CAI) exhibit deficits in neuromuscular control, resulting in altered movement strategies. However, no researchers have examined neuromuscular adaptations to dynamic movement strategies during multiplanar landing and cutting among patients with CAI, individuals who are ankle-sprain copers, and control participants. OBJECTIVE: To investigate lower extremity joint power, stiffness, and ground reaction force (GRF) during a jump-landing and cutting task among CAI, coper, and control groups. DESIGN: Cross-sectional study. SETTING: Laboratory. PATIENTS OR OTHER PARTICIPANTS: A total of 22 patients with CAI (age = 22.7 ± 2.0 years, height = 174.6 ± 10.4 cm, mass = 73.4 ± 12.1 kg), 22 ankle-sprain copers (age = 22.1 ± 2.1 years, height = 173.8 ± 8.2 cm, mass = 72.6 ± 12.3 kg), and 22 healthy control participants (age = 22.5 ± 3.3 years, height = 172.4 ± 13.3 cm, mass = 72.6 ± 18.7 kg). INTERVENTION(S): Participants performed 5 successful trials of a jump-landing and cutting task. MAIN OUTCOME MEASURE(S): Using motion-capture cameras and a force plate, we collected lower extremity ankle-, knee-, and hip-joint power and stiffness and GRFs during the jump-landing and cutting task. Functional analyses of variance were used to evaluate between-groups differences in these dependent variables throughout the contact phase of the task. RESULTS: Compared with the coper and control groups, the CAI group displayed (1) up to 7% of body weight more posterior and 52% of body weight more vertical GRF during initial landing followed by decreased GRF during the remaining stance and 22% of body weight less medial GRF across most of stance; (2) 8.8 W/kg less eccentric and 3.2 W/kg less concentric ankle power, 6.4 W/kg more eccentric knee and 4.8 W/kg more eccentric hip power during initial landing, and 5.0 W/kg less eccentric knee and 3.9 W/kg less eccentric hip power; and (3) less ankle- and knee-joint stiffness during the landing phase. Concentric power patterns were similar to eccentric power patterns. CONCLUSIONS: The CAI group demonstrated altered neuromechanics, redistributing energy absorption from the distal (ankle) to the proximal (knee and hip) joints, which coincided with decreased ankle and knee stiffness during landing. Our data suggested that although the coper and control groups showed similar landing and cutting strategies, the CAI group used altered strategies to modulate impact forces during the task.

Ocean acidification exacerbates the effects of paralytic shellfish toxins on the fitness of the edible mussel Mytilus chilensis.

High latitudes are considered particularly vulnerable to ocean acidification, since they are naturally low in carbonate ions. The edible mussel Mytilus chilensis is a common calcifier inhabiting marine ecosystems of the southern Chile, where culturing of this species is concentrated and where algal blooms produced by the toxic dinoflagellate A. catenella are becoming more frequent. Juvenile Mytilus chilensis were exposed to experimental conditions simulating two environmental phenomena: pCO2 increase and the presence of paralytic shellfish toxins (PST) produced by the dinoflagellate Alexandrium catenella. Individuals were exposed to two levels of pCO2: 380 µatm (control condition) and 1000 µatm (future conditions) over a period of 39 days (acclimation), followed by another period of 40 days exposure to a combination of pCO2 and PST. Both factors significantly affected most of the physiological variables measured (feeding, metabolism and scope for growth). However, these effects greatly varied over time, which can be explained by the high individual variability described for mussels exposed to different environmental conditions. Absorption efficiency was not affected by the independent effect of the toxic diet; however, the diet and pCO2 interaction affected it significantly. The inhibition of the physiological processes related with energy acquisition by diets containing PST, may negatively impact mussel fitness, which could have important consequences for both wild and cultured mussel populations, and thus, for socioeconomic development in southern Chile.

The Role of Feeding Specialization on Post-Prandial Metabolic Rate in Snakes of the Genus Bothrops.

Feeding specialization is a recurrent issue in the evolution of snakes and is sometimes associated to morphological and/or behavioral adaptations that improve snake performance to exploit a particular food type. Despite its importance for animal fitness, the role of physiological traits has been much less studied than morphological and behavioral traits in the evolution of feeding specialization in snakes. In this context, the energetic cost of post-prandial period is an important physiological factor due to the remarkable effect on the snake energy budget. We collected data on post-prandial metabolic rate (SDA) in five species of pit vipers from the genus Bothrops with different degrees of mammal feeding specialization to test the hypothesis that feeding specialist species have lower energy costs during the digestion of their regular food item when compared to species with a more generalist diet. Our results support this hypothesis and suggest that ontogenetic changes in diet can be accompanied by changes in energy cost of the digestion process.