Freezing and thawing in Antarctica: characterization of antifreeze protein (AFP) producing microorganisms isolated from King George Island, Antarctica.

Antarctic temperature variations and long periods of freezing shaped the evolution of microorganisms with unique survival mechanisms. These resilient organisms exhibit several adaptations for life in extreme cold. In such ecosystems, microorganisms endure the absence of liquid water and exhibit resistance to freezing by producing water-binding molecules such as antifreeze proteins (AFP). AFPs modify the ice structure, lower the freezing point, and inhibit recrystallization. The objective of this study was to select and identify microorganisms isolated from different Antarctic ecosystems based on their resistance to temperatures below 0 °C. Furthermore, the study sought to characterize these microorganisms regarding their potential antifreeze adaptive mechanisms. Samples of soil, moss, permafrost, and marine sediment were collected on King George Island, located in the South Shetland archipelago, Antarctica. Bacteria and yeasts were isolated and subjected to freezing-resistance and ice recrystallization inhibition (IR) tests. A total of 215 microorganisms were isolated, out of which 118 were molecularly identified through molecular analysis using the 16S rRNA and ITS regions. Furthermore, our study identified 24 freezing-resistant isolates, including two yeasts and 22 bacteria. A total of 131 protein extracts were subjected to the IR test, revealing 14 isolates positive for AFP production. Finally, four isolates showed both freeze-resistance and IR activity (Arthrobacter sp. BGS04, Pseudomonas sp. BGS05, Cryobacterium sp. P64, and Acinetobacter sp. M1_25C). This study emphasizes the diversity of Antarctic microorganisms with the ability to tolerate freezing conditions. These microorganisms warrant further investigation to conduct a comprehensive analysis of their antifreeze capabilities, with the goal of exploring their potential for future biotechnological applications.

Plant growth regulators and mobilization of reserves in imbibition phases of yellow passion fruit.

The production of seedlings of the passion fruit tree, usually, is sexual, and the seeds are not uniform in the seedling emergence, and soaking treatments of seeds can provide faster and more uniform germination. It was aimed to study the action of plant growth regulators and the mobilization of reserves in the stages of soaking of yellow passion fruit seeds. The seeds were soaked for five hours in solutions containing plant growth regulators, in a completely randomized design, in a factorial 8 x 4, with four replications. The first factor corresponds to eight plant growth regulators: T1 - distilled water (control); T2 - 6-benzylaminepurine ​​500 mg L-1; T3 - 4-(3-indolyl) butyric acid 500 mg L-1; T4 - gibberellic acid 500 mg L-1; T5 - spermine 250 mg L-1; T6 - spermine 750 mg L-1; T7 - spermidine 750 mg L-1; T8 - spermidine 1250 mg L-1; and the second factor, to the four soaking times: zero, four, 72 and 120 hours, corresponding, respectively, to the dry seed, and to phases I, II, and III of the imbibition curve. It was evaluated the biochemical composition of seeds (lipids, soluble sugars and starch). The seeds showed accumulation of lipids in phase III; the content of soluble sugars increased in phase I and decreased in phase II. The starch content increased until the phase II and decreased in phase III. Starch is the main reserve in the seeds and the main source of energy used in phase III; soaking the seeds in polyamines generates an accumulation of lipids in the seeds and soaking in plant growth regulators increases the burning of starch.

Potential of coffee straw biochal as a substrate conditioner in seed lettuce and sorghum germination and vigority.

The use of residues from coffee production to obtain biochar is a sustainable approach, which aims to minimize the environmental impact of these materials. In this study, the effect of adding coffee straw biochar on the physiological quality of lettuce and sorghum seeds was investigated. Thus, the objective of this work was to study the effect of adding different concentrations of coffee biochar in the substrate composition on the physiological quality of lettuce (Lactuca sativa) and sorghum (Sorghum bicolor) seeds. The experimental design used was completely randomized, with five concentrations of biochar (0; 7.5; 15; 30 and 60%), conducted with four replications of 25 seeds. The use of biochar in the concentrations studied does not provide an increase in the average germination percentage and vigor of lettuce and sorghum seeds. The increase in the concentration of biochar caused less seed vigor, suggesting a toxic effect. For seed germination, there was no significant difference between lettuce and sorghum species, regardless of treatment. For the germination speed index, sorghum seeds have higher means, except for the treatment with the addition of 15% coffee straw biochar. Lettuce seeds have higher shoot length averages, except for treatment with 100% commercial substrate. The sorghum seeds have higher mean root length and dry mass than lettuce, regardless of the treatment.

Maturation and harvest time of Ateleia glazioveana Baill. seeds.

Ateleia glazioveana Baill. is a pioneer, rustic and can be used for forest recovery. This work aimed to study the process of physiological maturation of this species. The research was carried out in the city of Alegre - ES, the trees were identified in the floral anthesis and accompanied during the filling of the fruits and development of the seeds until the complete maturation. The fruits were harvested at the following stages 7, 14, 21, 28, 35 and 42 days after anthesis, and characterized according to: morphometry, moisture, fresh and dry mass of fruits and seeds, germination, germination speed index, shoot and root length and dry mass of seedlings. The regression equations were adjusted for the main characteristics analyzed as a function of the harvest period. The point of physiological maturity of timbó occurred at 42 days after anthesis.

Multivariate analysis of agronomic, physicochemical, and physiological characters of passion fruit hybrids cultivated at different environments.

Simultaneous analysis studies of several agronomic traits in cultivated plants make it possible to identify phenotypic and genotypic differences due to environmental variations, such as altitude. Therefore, the objective was to evaluate, through multivariate analysis of agronomic, physicochemical and physiological characters, passion fruit hybrids cultivated in different environments. The hybrids used were Gigante Amarelo, Rubi do Cerrado and Sol do Cerrado, cultivated in the southern region of Espírito Santo in four municipalities/environments: Marataízes (41 m), Jerônimo Monteiro (104 m), Alegre (711 m), and Ibitirama (1016 m). The agronomic characters of the plants, the physical-chemical characteristics of the fruits and the physical, biochemical and physiological qualities of the seeds were analyzed. The Singh method was used to determine the most important differentiating characters between hybrids growing in different environments. Based on these characters, a dissimilarity matrix was generated and a principal coordinate analysis was performed. It was observed that the pulp yield was influenced by altitude. The three hybrids showed greater performance in terms of agronomic characters at altitude (41 m) than at altitude (104 m). The Sol do Cerrado hybrid showed high performance in the physical-chemical characteristics of the fruits at altitude (104 m).

Phytoscreening of Vochysiaceae species: Molecular identification by HPLC-ESI-MS/MS and evaluating of their antioxidant activity and inhibitory potential against human α-amylase and protein glycation.

Scientific research based on medicinal plants has been highlighted as a complementary treatment to T2DM, stand out the Vochysiaceae family, which have been widely used in folk medicine by traditional South American communities to treat some diseases. Our study aimed to investigate the antioxidant and antiglycation activities of ethanol extracts of leaves (LF) and stem barks (SB) of Vochysiaceae species, evaluated their capacities to inhibit glycoside and lipid hydrolases related to T2DM and molecular identification by HPLC-ESI-MS/MS. Our main findings indicate that the ethanolic extract of four of eight analyzed plants such as LF and SB of Q. grandiflora, Q. parviflora, V. elliptica and Calisthene major exhibited, respectively, potential of α-amylase inhibition (IC50 of LF: 5.7 ±â€¯0.6, 4.1 ±â€¯0.5, 5.8 ±â€¯0.5, 3.2 ±â€¯0.6 and IC50 of SB: 3.3 ±â€¯0.7, 6.2 ±â€¯2.0, 121.0 ±â€¯8.6 and 11.2 ±â€¯2.8 µg/mL), capacities of antioxidant (ORAC of LF: 516.2 ±â€¯0.1, 547.6 ±â€¯4.9, 544.3 ±â€¯6.1, 442.6 ±â€¯2.4 and ORAC of SB: 593.6 ±â€¯22.3, 497.7 ±â€¯0.8, 578 ±â€¯12.3, 593.6 ±â€¯19.5 µmol trolox eq/g; FRAP of LF: 796.1 ±â€¯0.9, 427.7 ±â€¯22.0, 81.0 ±â€¯1.9, 685 ±â€¯37.9 and FRAP of SB: 947.4 ±â€¯24.9, 738.6 ±â€¯24.3, 98.8 ±â€¯7.9, 970.8 ±â€¯13.9 µmol trolox eq/g; DPPH IC50 of LF: 14.2 ±â€¯1.8, 36.3 ±â€¯6.9, 11.8 ±â€¯1.9, 13.3 ±â€¯1.2 and DPPH IC50 of SB: 16.0 ±â€¯3.0, 15.5 ±â€¯1.9, 126.1 ±â€¯23. 6, 5.3 ±â€¯0.3 µg/mL, respectively) and antiglycation (BSA/Frutose IC50 of LF: 43.1 ±â€¯3.4, 52.1 ±â€¯6.0, 175.5 ±â€¯32, 8, 111.8 ±â€¯14.7 and BSA/Frutose IC50 of SB:, 40.1 ±â€¯11.9, 51.2 ±â€¯16. 7, 46.6 ±â€¯5.7, 53.5 ±â€¯13.6 µg/mL) and presence of polyphenols, such as flavonoids and condensed tannins. The extracts presented low ability to inhibit α-glycosidase and lipase enzymes in the initial assays, with values below 40% of inhibition. In BSA/methylglyoxal, only Q. grandiflora SB, V. eliptica LF and V. tucanorum LF showed activity (IC50: 655.5 ±â€¯208.5, 401.9 ±â€¯135.2 and 617.1 ±â€¯80.6 µg/mL, respectively) and only C. major LF and SB, in Arg/methylglyoxal (IC50: 485.1 ±â€¯130.8 and 468.0 ±â€¯150.5 µg/ml, respectively). This study presented new findings about the biological and pharmacological potential of some species of Vochysiaceae family, contributing to the understanding of the action and efficacy in use of these plants, in their management of postprandial hyperglycemia and in glycation and oxidative processes that contribute to managing diabetes mellitus.

Preparation of nano-hydroxyapatite/chitosan aqueous dispersions: From lab scale to continuous production using an innovative static mixer.

Chitosan is widely used in the preparation of organic-inorganic composite materials, such as n-HAp/CS composites, which find application for bone regeneration. The methods for their preparation are various, and usually based on the preparation of intermediate n-HAp/CS dispersions, which can greatly influence the final properties of the resulting composites since it is expected that homogenous and stable dispersions lead to composite materials with improved final properties. This work hypothesizes that, additionally to process parameters such as pH, n-HAp/CS weight ratio, mixing conditions and the presence of salts, chitosan itself has a high impact on dispersions stability. Thus, the importance of properly control the preparation of the n-HAp/CS intermediate dispersions is highlighted by doing a systematic study where relevant processing parameters were studied at lab scale using ultrasonication, alone or in the presence of chitosan, namely on particle size and zeta potential. Furthermore, and based on the best laboratorial conditions, the production of n-HAp/CS nanocomposite dispersions in continuous mode was attempted through NETmix® technology, an innovative static mixer and reactor developed at the Associate Laboratory LSRE-LCM of the Faculty of Engineering of the University of Porto (FEUP).

Supercritical CO2 assisted process for the production of high-purity and sterile nano-hydroxyapatite/chitosan hybrid scaffolds.

Hybrid scaffolds composed of hydroxyapatite (HAp), in particular in its nanometric form (n-HAp), and chitosan (CS) are promising materials for non-load-bearing bone graft applications. The main constraints of their production concern the successful implementation of the final purification/neutralization and sterilization steps. Often, the used purification strategies can compromise scaffold structural features, and conventional sterilization techniques can result in material's thermal degradation and/or contamination with toxic residues. In this context, this work presents a process to produce n-HAp/CS scaffolds mimicking bone composition and structure, where an innovative single step based on supercritical CO2 extraction was used for both purification and sterilization. A removal of 80% of the residual acetic acid was obtained (T = 75°C, p = 8.0 MPa, 2 extraction cycles of 2 h) giving rise to scaffolds exhibiting adequate interconnected porous structure, fast swelling and storage modulus compatible with non-load-bearing applications. Moreover, the obtained scaffolds showed cytocompatibility and osteoconductivity without further need of disinfection/sterilization procedures. Among the main advantages, the proposed process comprises only three steps (n-HAp/CS dispersion preparation; freeze-drying; and supercritical CO2 extraction), and the supercritical CO2 extraction show clear advantages over currently used procedures based on neutralization steps. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 965-975, 2018.

Phylogenomics and evolution of floral traits in the Neotropical tribe Malmeeae (Annonaceae).

Androdioecy is the rarest sexual system among plants. The majority of androdioecious species are herbaceous plants that have evolved from dioecious ancestors. Nevertheless, some woody and androdioecious plants have hermaphrodite ancestors, as in the Annonaceae, where androdioecious genera have arisen several times in different lineages. The majority of androdioecious species of Annonaceae belong to the Neotropical tribe Malmeeae. In addition to these species, Pseudoxandra spiritus-sancti was recently confirmed to be androdioecious. Here, we describe the morphology of male and bisexual flowers of Pseudoxandra spiritus-sancti, and investigate the evolution of androdioecy in Malmeeae. The phylogeny of tribe Malmeeae was reconstructed using Bayesian inference, maximum parsimony and maximum likelihood of 32 taxa, using DNA sequences of 66 molecular markers of the chloroplast genome, sequenced by next generation sequencing. The reconstruction of ancestral states was performed for characters associated with sexual systems and floral morphology. The phylogenetic analyses reconstructed three main groups in Malmeeae, (Malmea (Cremastosperma, Pseudoxandra)) sister to the rest of the tribe, and (Unonopsis (Bocageopsis, Onychopetalum)) sister to (Mosannona, Ephedranthus, Klarobelia, Oxandra, Pseudephedranthus fragrans, Pseudomalmea, Ruizodendron ovale). Hermaphroditism is plesiomorphic in the tribe, with four independent evolutions of androdieocy, which represents a synapomorphy of two groups, one that includes three genera and 14 species, the other with a single genus of seven species. Male flowers are unisexual from inception and bisexual flowers possess staminodes and functional stamens. Pseudoxandra spiritus-sancti is structurally androdioecious.

Short communication: Variability in milk urea nitrogen and dairy total mixed ration composition in the northeastern United States.

The main objective of this survey was to examine variability in milk urea nitrogen (MUN) for Dairy Herd Improvement Association (DHIA) herds in the northeastern United States (the Northeast), examine trends in dairy cow diet composition, and determine potential relationships for MUN and diet composition. Trends in milk fat and protein concentrations, milk yield, days in milk on test day, and lactation number of the cows were also evaluated. The data set for the survey included 10,839,461 DHIA dairy cow records from 2004 to 2015 for 13 states (CT, DE, MA, MD, ME, NH, NJ, NY, PA, RI, VA, VT, and WV) and was retrieved from Dairy Records Management Systems (Raleigh, NC). Average (across states and years) milk yield, milk fat, and milk protein were 31.6 ± 0.24 kg/d, 3.85 ± 0.021%, and 3.13 ± 0.013%, respectively. No obvious trends were observed for milk fat or protein content, but milk yield steadily increased during the survey period. Milk urea N concentration averaged 13.3 ± 0.13 mg/dL, with no obvious or consistent trends. Examination of variability in dairy feed cost and all milk price for the Northeast indicated that high MUN generally coincided with high feed cost and high milk price. For the diet composition survey, 9,707 records of total mixed ration (TMR) analyses, unrelated to the milk composition data set, from the Cumberland Valley Analytical Service (Maugansville, MD) database were examined. Concentration of TMR crude protein (CP) decreased from 17.1% in 2007 to 16.4% in 2015, but there was not an obvious trend in soluble protein concentration. Concentration of TMR neutral detergent fiber (NDF) and 24-h in vitro NDF degradability declined steadily during the survey period and was accompanied by a steady increase in TMR starch concentration. Examination of these unrelated data sets revealed lack of correlation between MUN and diet chemical composition. Thus, we conclude that individual cow MUN in Northeast dairy herds fluctuated between 2004 and 2015. It appeared that MUN followed variability in feed cost; however, ration feed ingredient data were not available to better define the reasons for the variations in MUN.

Using brown midrib 6 dwarf forage sorghum silage and fall-grown oat silage in lactating dairy cow rations.

Double cropping and increasing crop diversity could improve dairy farm economic and environmental sustainability. In this experiment, corn silage was partially replaced with 2 alternative forages, brown midrib-6 brachytic dwarf forage sorghum (Sorghum bicolor) or fall-grown oat (Avena sativa) silage, in the diet of lactating dairy cows. We investigated the effect on dry matter (DM) intake, milk yield (MY), milk components and fatty acid profile, apparent total-tract nutrient digestibility, N utilization, enteric methane emissions, and income over feed cost. We analyzed the in situ DM and neutral detergent fiber disappearance of the alternative forages versus corn silage and alfalfa haylage. Sorghum was grown in the summer and harvested in the milk stage. Oats were grown in the fall and harvested in the boot stage. Compared with corn silage, neutral detergent fiber and acid detergent fiber concentrations were higher in the alternative forages. Lignin content was highest for sorghum silage and similar for corn silage and oat silage. The alternative forages had less than 1% starch compared with the approximately 35% starch in the corn silage. Ruminal in situ DM effective degradability was similar, although statistically different, for corn silage and oat silage, but lower for sorghum silage. Diets with the alternative forages were fed in a replicated 3 × 3 Latin square design experiment with three 28-d periods and 12 Holstein cows. The control diet contained 44% (DM basis) corn silage. In the other 2 diets, sorghum or oat silages were included at 10% of dietary DM, replacing corn silage. Sorghum silage inclusion decreased DM intake, MY, and milk protein content but increased milk fat and maintained energy-corrected MY similar to the control. Oat silage had no effect on DM intake, MY, or milk components compared to the control. The oat silage diet increased apparent total-tract digestibility of dietary nutrients, except starch, whereas the sorghum diet slightly decreased DM, organic matter, crude protein, and starch digestibility. Cows consuming the oat silage diet had higher milk urea N and urinary urea N concentrations. Milk N efficiency was decreased by the sorghum diet. Diet did not affect enteric methane or carbon dioxide emissions. This study shows that oat silage can partially replace corn silage at 10% of the diet DM with no effect on MY. Brown midrib sorghum silage harvested at the milk stage with <1% starch may decrease DM intake and MY in dairy cows.

Effect of high-oleic-acid soybeans on production performance, milk fatty acid composition, and enteric methane emission in dairy cows.

The objective of this study was to investigate the effect of 3 soybean sources differing in fatty acid profile and processing method on productivity, milk composition, digestibility, rumen fermentation, and enteric methane emission in lactating dairy cows. The soybean sources were conventional, high-linoleic-acid variety extruded soybean meal (ESBM; 8.7% ether extract with 15% oleic and 54% linoleic acids); extruded Plenish (DuPont Pioneer, Johnston, IA), high-oleic-acid variety soybean meal (EPSBM; 8.4% ether extract with 73% oleic and 8% linoleic acids); and whole, heated Plenish soybeans (WPSB; 20.2% ether extract). The study involved 15 Holstein cows in a replicated 3 × 3 Latin square design experiment with three 28-d periods. The inclusion rate of the soybean sources in the diet was (dry matter basis) 17.1, 17.1, and 7.4% for ESBM, EPSBM, and WPSB, respectively, which resulted in ether extract concentration of the diets of 3.99, 3.94, and 4.18%, respectively. Compared with ESBM, the Plenish diets tended to increase dry matter intake and decreased feed efficiency (but had no effect on energy-corrected milk feed efficiency). The Plenish diets increased milk fat concentration on average by 5.6% and tended to increase milk fat yield, compared with ESBM. The WPSB diet tended to increased milk true protein compared with the extruded soybean meal diets. Treatments had no effect on rumen fermentation and enteric methane or carbon dioxide emissions, except pH was higher for WPSB versus EPSBM. The Plenish diets decreased the prevalence of Ruminococcus and increased that of Eubacterium and Treponema in whole ruminal contents. Total-tract apparent digestibility of organic matter and crude protein were decreased by WPSB compared with ESBM and EPSBM. Compared with the other treatments, urinary N excretion was increased by EPSBM and fecal N excretion was greater for WPSB. Treatments had marked effects on milk fatty acid profile. Generally, the Plenish diets increased mono-unsaturated (mostly cis-9 18:1) and decreased polyunsaturated, total trans-, and conjugated linoleic fatty acids concentrations in milk fat. In this study, compared with conventional, high-linoleic-acid variety extruded soybean meal, the Plenish soybean diets increased milk fat concentration and tended to increase fat yield, decreased feed efficiency, and modified milk fatty acid profile in a manner expected from the greater concentration of oleic acid in Plenish soybean oil.

Short communication: Preference for flavored concentrate premixes by dairy cows.

Flavor preferences may be used to stimulate feed intake in dairy cows, which may improve use of robotic milking systems and increase feed intake of sick cows. A cafeteria-design experiment was used to determine if dairy cows have flavor preferences. Sixteen lactating Holstein cows averaging 197±32d in milk, 1.9±0.8 lactations, 27.8±4.2kg/d of dry matter intake, and 41.5±7.4kg/d of milk yield were involved in the experiment. Cows were offered 7 flavored concentrate premixes (FCP) and 1 control premix. The FCP flavors were anise, fenugreek, honey, orange, thyme, molasses, and vanilla; the absence of flavor, neutral, acted as a control. The inclusion rate of the flavors in FCP was 250 to 300g/t on an as-is basis. Cows were not adapted to the flavors before the experiment. Cows were housed in a tiestall barn and offered, on each day, 4 different FCP (1kg each) in plastic bins placed in front of each cow. The experiment lasted 6 consecutive days. Each FCP was presented to each cow once every 2d, 2h after the morning feeding. Flavors and position of the bins in front of the cows were randomized. As a result, each flavor was presented to each cow 3 times during the experiment, at 3 different bin locations. Each cow had access to the FCP for 5min from the time they started eating. Eating time and amount eaten were recorded. The vanilla and fenugreek FCP were consumed the most, at 408 and 371g/5-min offering, respectively, whereas the orange and anise FCP were consumed the least, at 264 and 239g/5-min offering, respectively. Similarly, cows spent the most time eating the vanilla and fenugreek FCP at 99 and 75 s/offering, respectively, and the least amount of time eating the orange and anise FCP at 49 and 50 s/offering, respectively. We detected an effect of bin position: the 2 center FCP were consumed more than the outer 2 FCP. Flavor had no effect on consumption rate. In conclusion, relative to the control, concentrate intake was not affected by flavor, but dairy cows may prefer vanilla or fenugreek flavors when offered a novel choice.

Effects of rumen-protected methionine, lysine, and histidine on lactation performance of dairy cows.

The objective of this study was to evaluate the effects of supplementing a metabolizable protein (MP)-deficient diet with rumen-protected (RP) Met, Lys, and His, individually or combined, on the performance of lactating dairy cows. The experiment was a 9-wk randomized complete block design with 72 Holstein cows. Following a 2-wk covariate period, cows were blocked by days in milk, milk yield, and parity, and randomly assigned to 1 of the following 6 treatments: (1) MP-adequate diet [MPA; +243g/d MP balance, according to the National Research Council (2001) requirements]; (2) MP-deficient diet (MPD; -54g/d MP balance); (3) MPD supplemented with RPMet (MPDM); (4) MPD supplemented with RPLys (MPDL); (5) MPD supplemented with RPHis (MPDH); and (6) MPD supplemented with RPMet, RPLys, and RPHis (MPDMLH). Dry matter intake (DMI), yields of milk and milk components (fat, protein, lactose) and energy-corrected milk (ECM), feed and ECM feed efficiencies, and milk and plasma urea N were decreased by MPD, compared with MPA. Supplementation of the MPD diet with RPLys increased milk protein content and plasma glucose concentration and tended to increase milk urea N. Addition of RPHis tended to increase DMI, increased milk protein concentration, and numerically increased yields of milk fat, protein, and ECM. In addition to the trends for increased DMI and milk fat content, and higher milk protein concentration, supplementation of the 3 RP AA also increased yields of milk fat, protein, and ECM and ECM feed efficiency. Relative to MPA, milk N efficiency tended to be increased by MPD. Concentrations of plasma essential AA (except Met and Thr) were decreased by MPD compared with MPA. Supplementation of RPMet, RPLys, and RPHis increased plasma Met (except for MPDM), Lys, and His concentrations, respectively. Cows fed MPD had lower blood hemoglobin concentration and numerically higher plasma ghrelin than cows fed MPA. Concentration of total saturated fatty acids in milk fat were or tended to be higher for MPD compared with MPA and MPDMLH, respectively. Concentration of total polyunsaturated and yield of milk odd- and branched-chain fatty acids were or tended to be decreased by MPD compared with MPA. Overall, the results of this study confirm our previous data and suggest that His stimulates DMI and the combination of the 3 RP AA (Met, Lys, and His) has the potential to improve milk and milk component yields in dairy cows fed MP-deficient diets.

Effect of 3-nitrooxypropanol on methane and hydrogen emissions, methane isotopic signature, and ruminal fermentation in dairy cows.

The objective of this crossover experiment was to investigate the effect of a methane inhibitor, 3-nitrooxypropanol (3NOP), on enteric methane emission, methane isotopic composition, and rumen fermentation and microbial profile in lactating dairy cows. The experiment involved 6 ruminally cannulated late-lactation Holstein cows assigned to 2 treatments: control and 3NOP (60 mg/kg of feed dry matter). Compared with the control, 3NOP decreased methane emission by 31% and increased hydrogen emission from undetectable to 1.33 g/d. Methane emissions per kilogram of dry matter intake and milk yield were also decreased 34% by 3NOP. Milk production and composition were not affected by 3NOP, except milk fat concentration was increased compared with the control. Concentrations of total VFA and propionate in ruminal fluid were not affected by treatment, but acetate concentration tended to be lower and acetate-to-propionate ratio was lower for 3NOP compared with the control. The 3NOP decreased the molar proportion of acetate and increase those of propionate, butyrate, valerate, and isovalerate. Deuterium-to-hydrogen ratios of methane and the abundance of (13)CH3D were similar between treatments. Compared with the control, minor (4‰) depletion in the (13)C/(12)C ratio was observed for 3NOP. Genus composition of methanogenic archaea (Methanobrevibacter, Methanosphaera, and Methanomicrobium) was not affected by 3NOP, but the proportion of methanogens in the total cell counts tended to be decreased by 3NOP. Prevotella spp., the predominant bacterial genus in ruminal contents in this experiment, was also not affected by 3NOP. Compared with the control, Ruminococcus and Clostridium spp. were decreased and Butyrivibrio spp. was increased by 3NOP. This experiment demonstrated that a substantial inhibition of enteric methane emission by 3NOP in dairy cows was accompanied with increased hydrogen emission and decreased acetate-to-propionate ratio; however, neither an effect on rumen archaeal community composition nor a significant change in the isotope composition of methane was observed.

Lack of mutagenic effect by multi-walled functionalized carbon nanotubes in the somatic cells of Drosophila melanogaster.

Carbon nanotubes (CNTs) are formed by rolling up a single graphite sheet into a tube. Among the different types of CNTs, the multi-walled carbon nanotubes (MWCNTs) comprise a set of concentric nanotubes with perfect structures. Several uses for MWCNTs have been suggested to be included in biological applications such as manufacturing of biosensors, carriers of drugs. However, before these materials can be put on the market, it is necessary to know their genotoxic effects. Thus, this study aims to evaluate the mutagenicity of multi-walled carbon nanotubes (MWCNTs) functionalized in somatic cells of Drosophila melanogaster, using the somatic mutation and recombination test (SMART). This assay detects the loss of heterozygosity of marker genes expressed phenotypically on the wings of the fly. Larvae of three days were used, resulting from ST cross, with basal levels of the cytochrome P450 and larvae of high metabolic bioactivity capacity (HB cross). They were treated with different concentrations of MWCNTs functionalized. The MH descendants, analyzed in both ST and HB crosses, had no significant effects on the frequency of mutant. Based on the results and on the experimental conditions mentioned in this study, it was concluded that MWCNTs were not mutagenic in D. melanogaster.

EPR detection of paramagnetic chromium in liver of fish (Anguilla anguilla) treated with dichromate(VI) and associated oxidative stress responses-contribution to elucidation of toxicity mechanisms.

The impact of chromium (Cr) on fish health has been the subject of numerous investigations, establishing a wide spectrum of toxicity, attributed particularly to the hexavalent form [Cr(VI)]. However, reports on the simultaneous assessment of Cr toxicity in fish and its toxico-kinetics, namely involving metal speciation, are scarce. Therefore, keeping in view the understanding of the mechanisms of Cr(VI) toxicity, this work intended to detect the formation of paramagnetic Cr species in liver of Anguilla anguilla following short-term dichromate(VI) intraperitoneal treatment (up to 180 min), assessing simultaneously the pro-oxidant properties. The formation of Cr(V) and Cr(III) was examined by electron paramagnetic resonance (EPR), as an innovative approach in the context of fish toxicology, and related with the levels of total Cr. Cr(V) was successfully detected and quantified by EPR spectrometry, showing a transient occurrence, mostly between 15 and 90 min post-injection, with a peak at 30 min. The limitations of EPR methodology towards the detection and quantification of Cr(III) were confirmed. Although Cr(VI) exposure induced the antioxidant system in the eel's liver, the oxidative deterioration of lipids was not prevented. Overall, the results suggested that Cr(V), as a short-lived species, did not appear to be directly and primarily responsible for the cellular damaging effects observed, since stress responses persisted up to the end of exposure regardless Cr(V) drastic decay. Though further research is needed, ROS mediated pathways (suggested by superoxide dismutase and catalase activity induction) and formation of Cr(III) complexes emerged as the most plausible mechanisms involved in Cr(VI) toxicity.

Effect of feeding a reduced-starch diet with or without amylase addition on lactation performance in dairy cows.

The objective of this study was to determine lactation performance responses of high-producing dairy cows to a reduced-starch diet compared with a normal-starch diet and to the addition of exogenous amylase to the reduced-starch diet. Thirty-six multiparous Holstein cows (51+/-22 DIM and 643+/-49kg of body weight at trial initiation) were randomly assigned to 1 of 3 treatments in a completely randomized design: a 3-wk covariate adjustment period during which the cows were fed the normal-starch diet, followed by a 12-wk treatment period during which the cows were fed their assigned treatment diets. The normal-starch TMR did not contain exogenous amylase (NS-). The reduced-starch diets, formulated by partially replacing corn grain with soy hulls, were fed without (RS-) and with (RS+) exogenous amylase added to the TMR. Starch and NDF concentrations averaged 27.1 and 30.6%, 21.8 and 36.6%, and 20.7 and 36.6% (dry matter basis) for the NS-, RS-, and RS+ diets, respectively. Dry matter intake for cows fed the RS- diet was 2.4 and 3.2kg/d greater than for cows fed the NS- and RS+ diets, respectively. Intake of NDF ranged from 1.19 to 1.52% of body weight among the treatments, with the RS- diet being 28% greater than the NS- diet and 13% greater than the RS+ diet. Milk yield averaged 50.4kg/d and was unaffected by treatment. Fat-corrected milk yield was 2.9kg/d greater for cows fed the RS- diet than for cows fed the NS- diet. Body weight and body condition score measurements were unaffected by treatment. Fat-, solids-, and energy-corrected milk feed conversions (kilograms/kilogram of DMI) were 12 to 13% greater for cows fed the RS+ diet than for cows fed the RS- diet. Dry matter and nutrient digestibilities were lowest for cows fed the NS- diet and greatest for cows fed the RS+ diet, and were greater for cows fed the RS+ diet than for cows fed the RS- diet, with the exception of starch digestibility, which was similar. Greater conversion of feed to milk for dairy cows fed reduced-starch diets that include exogenous amylase may offer potential for improving economic performance.

Type of corn endosperm influences nutrient digestibility in lactating dairy cows.

An experiment was conducted to evaluate the effect of type of corn endosperm on nutrient digestibility in lactating dairy cows. Near-isogenic variants of an Oh43 x W64A normal dent endosperm hybrid carrying floury-2 or opaque-2 alleles were grown in spatial isolation in field plots and harvested as dry shelled corn. Six ruminally cannulated, multiparous Holstein cows (67 +/- 9 d in milk at trial initiation) were randomly assigned to a replicated 3 x 3 Latin square design with 14-d periods; the first 11 d of each period were for diet adaptation followed by 3 d of sampling and data collection. Treatment diets that contained dry rolled vitreous-, floury-, or opaque-endosperm corn [33% of dry matter (DM)], alfalfa silage (55% of DM) and protein-mineral-vitamin supplement (12% of DM) were fed as a total mixed ration. The percentage vitreous endosperm was zero for floury and opaque endosperm corns and 64 +/- 7% for the vitreous corn. Prolamin protein content of floury and opaque endosperm corns was 30% of the content found in vitreous corn. Degree of starch access and in vitro ruminal starch digestibility measurements were 32 and 42% greater on average, respectively, for floury and opaque endosperm corns than for vitreous corn. Dry matter and starch disappearances after 8-h ruminal in situ incubations were, on average, 24 and 32 percentage units greater, respectively, for floury and opaque endosperm corns than for vitreous corn. Ruminal pH and acetate molar percentage were lower, propionate molar percentage was greater, and acetate:propionate ratio was lower for cows fed diets containing floury and opaque endosperm corns than for cows fed vitreous corn. In agreement with laboratory and in situ measurements, total-tract starch digestibility was 6.3 percentage units greater, on average, for cows fed diets containing floury and opaque endosperm corns than vitreous corn. Conversely, apparent total-tract neutral detergent fiber (NDF) digestibility was lower for cows fed diets containing floury and opaque endosperm corns compared with vitreous corn. The type of endosperm in corn fed to dairy cows can have a marked effect on digestion of starch and NDF. Feeding less vitreous corn increased starch digestion but decreased NDF digestion.

On the characterization of energy networks of proteins.

The construction of a realistic theoretical model of proteins is determinant for improving the computational simulations of their structural and functional aspects. Modeling proteins as a network of non-covalent connections between the atoms of amino acid residues has shown valuable insights into these macromolecules. The energy-related properties of protein structures are known to be very important in molecular dynamics. However, these same properties have been neglected when the protein structures are modeled as networks of atoms and amino acid residues. A new approach for the construction of protein models based on a network of atoms is presented. This method, based on interatomic interaction, takes into account the energy and geometric aspects of the protein structures that were not employed before, such as atomic occlusion inside the protein, the use of solvation, protein modeling and analysis, and the use of energy potentials to estimate the energies of interatomic non-covalent contacts. As a result, we achieved a more realistic network model of proteins. This model has the virtue of being more robust in face of different unknown variables that usually are arbitrarily estimated. We were able to determine the most connected residues of all the proteins studied, so that we are now in a better condition to study their structural role.