Ruminal microbiome changes across lactation in primiparous Holstein cows with varying methane intensity: Heritability assessment.

Methane is a potent greenhouse gas produced during the ruminal fermentation and is associated with a loss of feed energy. Therefore, efforts to reduce methane emissions have been ongoing in the last decades. Methane production is highly influenced by factors such as the ruminal microbiome and host genetics. Previous studies have proposed to use the ruminal microbiome to reduce long-term methane emissions, as ruminal microbiome composition is a moderately heritable trait and genetic improvement accumulates over time. Lactation stage is another important factor that might influence methane production but potential associations with the ruminal microbiome have not been evaluated previously. This study sought to examine the changes in ruminal microbiome over the lactation period of primiparous Holstein cows differing in methane intensity and estimate the heritability of the abundance of relevant microorganisms. Ruminal content samples from 349 primiparous Holstein cows with 14 - 378 d in milk were collected from May 2018 to June 2019. Methane intensity (MI) of each cow was calculated as methane concentration/milk yield. Up to 64 taxonomic features (TF) from 20 phyla had a significant differential abundance between cows with low and high MI early in lactation, 16 TF during mid lactation, and none late in lactation. Taxonomical features within the Firmicutes, Proteobacteria, Melainabacteria, Cyanobacteria, Bacteroidetes and Actinobacteria phyla were associated to low MI, whereas eukaryotic TF and those within the Euryarchaeota, Verrucomicrobia, Kiritimatiellaeota, Lentisphaerae phyla were associated to high MI. Out of the 60 TF that were found to be differentially abundant between early and late lactation in cows with low MI, 56 TF were also significant when cows with low and high MI were compared in the first third of the lactation. In general, microbes associated with low MI were more abundant early in lactation (e.g., Acidaminococcus, Aeromonas and Weimeria genera) and showed low to moderate heritabilities (0.03 to 0.33). These results suggest some potential to modulate the rumen microbiome composition through selective breeding for lower MI. Differences in the ruminal microbiome of cows with extreme MI levels likely result from variations in the ruminal physiology of these cows and were more noticeable early in lactation probably due to important interactions between the host phenotype and environmental factors associated to that period. Our results suggest that the ruminal microbiome evaluated early in lactation may be more precise for MI difference, and hence, this should be considered to optimize sampling periods to establish a reference population in genomic selection scenarios.

A new graph of soil Rn-gas transport: Radon-rose plot.

Soil radon gas movement depends on soil geology, environmental thermodynamic parameters and, micro-seismic telluric activity. Mapping radon time dependent concentration at the relaxation depth in a selected area, provide transport direction in a seismically high-risk region. Nuclear track methodology is employed to determine main gradient vector for radon transport. Applying the gradient definition, a "radon rose" graph is constructed from which prone area can be promptly identified. Results show that short time interval, Rn-transport direction may change unpredictably, however, the length of each "spoke" around the circle provides information on the soil Rn-gas probable shifts towards or from a direction per time interval. The new graph is a novelty and provide improved approach for environmental protection and radon dosimetry.

Mitigation of greenhouse gases in dairy cattle via genetic selection: 1. Genetic parameters of direct methane using noninvasive methods and proxies of methane.

Records of methane emissions from 1,501 cows on 14 commercial farms in 4 regions of Spain were collected from May 2018 to June 2019. Methane concentrations (MeC) were measured using a nondispersive infrared methane detector installed within the feed bin of the automatic milking system during 14- to 21-d periods. Rumination time (RT; min/d) was collected using collars with a tag that registered time (minutes) spent eating and ruminating. The means of MeC and methane production (MeP) were 1,254.28 ppm and 182.49 g/d, respectively; mean RT was 473.38 min/d. Variance components for MeC, MeP, and RT were estimated with REML using pedigree and genomic information in a single-step model. Heritabilities for MeC and MeP were 0.11 and 0.12, respectively. Rumination time showed a slightly larger heritability estimate (0.17). The genetic correlation between MeP and MeC was high (>0.95), suggesting that selection on either trait would lead to a positive correlated response on the other. Negative correlations were estimated between RT and MeC (-0.24 ± 0.38) and MeP (-0.43 ± 0.35). Methane concentration and MeP had slightly positive correlations with milk yield (0.17 ± 0.39 and 0.21 ± 0.36), protein percentage (0.08 ± 0.32 and 0.30 ± 0.45), protein yield (0.22 ± 0.41 and 0.31 ± 0.35), fat percentage (0.02 ± 0.40 and 0.27 ± 0.36), and fat yield (0.27 ± 0.28 and 0.29 ± 0.28) from bivariate analyses. Rumination time had positive correlations with milk yield (0.41 ± 0.75) and protein yield (0.26 ± 0.57) and negative correlations with fat yield (-0.45 ± 0.32), protein percentage (-0.15 ± 0.38), and fat percentage (-0.40 ± 0.47). A positive approximated genetic correlation was estimated between fertility and MeC (0.10 ± 0.05) and MeP (0.18 ± 0.05), resulting in slightly higher CH4 production when selecting for better fertility [days open estimated breeding values (EBV) are expressed with mean 100 and SD 10, inversely related to days from calving to conception; that is, greater days open EBV implies better fertility]. Positive correlations were also estimated for stature with MeC and MeP (0.30 ± 0.04 and 0.43 ± 0.04, respectively). Other type traits (chest width, udder depth, angularity, and capacity) were positively correlated with methane traits, possibly because of higher milk yield and higher feed intake from these animals. Rumination time showed positive EBV correlations with production traits and type traits, and negative correlations with somatic cell count and body condition score. Based on the genetic correlations and heritabilities estimated in this study, methane is measurable and heritable, and estimates of genetic correlations suggest no strong opposition to current breeding objectives in Spanish Holsteins.

Shielding for transporting an 241Am-Be source for industrial applications.

Paraffin, water and water-extended polyester (WEP) were used as main moderator to design a mobile shielding for a 666 GBq 241Am-Be source used in oil industry. The shielding performance was estimated using Monte Carlo methods where the γ-rays induced by the neutron interaction with the shielding materials were also included. The spectra of neutrons and γ-rays around the shielding were estimated, as well as the total neutron and g-ray total fluences per history. The neutron source strength was used to calculate ambient dose equivalent rates, aiming to satisfy the international recommendation (2 mSv/h at the shielding surface). Moderators modify the neutron spectrum of the source reducing the amount of fast neutrons (0.5-11 MeV) and producing epithermal and thermal neutrons. During neutron transport in the moderator neutron capture and inelastic scattering produce gamma-rays. Paraffin has the best shielding performance however it is not suitable because can be affected by extreme temperatures. WEP and water have similar shielding performance, however water do not satisfy the international recommendations because at one site the neutron and γ-ray doses rates are larger than 2 mSv/h. WEP is the best option because fulfill the international recommendations, it is stable and temperatures where paraffin fails. The WEP-based shielding weights 66 kg.

Brown bear attacks on humans: a worldwide perspective.

The increasing trend of large carnivore attacks on humans not only raises human safety concerns but may also undermine large carnivore conservation efforts. Although rare, attacks by brown bears Ursus arctos are also on the rise and, although several studies have addressed this issue at local scales, information is lacking on a worldwide scale. Here, we investigated brown bear attacks (n = 664) on humans between 2000 and 2015 across most of the range inhabited by the species: North America (n = 183), Europe (n = 291), and East (n = 190). When the attacks occurred, half of the people were engaged in leisure activities and the main scenario was an encounter with a female with cubs. Attacks have increased significantly over time and were more frequent at high bear and low human population densities. There was no significant difference in the number of attacks between continents or between countries with different hunting practices. Understanding global patterns of bear attacks can help reduce dangerous encounters and, consequently, is crucial for informing wildlife managers and the public about appropriate measures to reduce this kind of conflicts in bear country.

Human prostasomes from normozoospermic and non-normozoospermic men show a differential protein expression pattern.

Prostasomes are exosomes such as extracellular vesicles, produced in the prostatic epithelium and released into the seminal plasma, that play an important role enhancing male fertility. Although some studies have demonstrated that prostasomes have a rich proteomic content, it is still unclear if that proteomic content varies depending on the male fertility status. Prostasomes from 12 normozoospermic and 14 non-normozoospermic seminal samples were isolated by differential ultracentrifugation. Protein content was studied by quantitative mass spectrometry and compared between both cohorts. We identified 1282 proteins with 745 of them (57.8%) being present in all seven prostasome pools. Forty-seven of those commonly present proteins showed differential expression levels in both cohorts. Specifically, prostasomes from non-normozoospermic samples showed a pattern of protein underexpression for a group of proteins including several proteins from the spermatozoa's energy production pathways as well as some proteins directly implicated in sperm activity. Variations in prostasomal protein content levels may have a relevant correlation with male fertility and thus could be of great utility as a biomarker of fertility status.

Feeding broilers with dry whey powder and whey protein concentrate affected productive performance, ileal digestibility of nutrients and cecal microbiota community.

Dietary interventions are a common practice in the poultry industry to promote optimal performance and health of animals. Here, we aim at assessing the influence of supplementing broiler diets with dry whey powder (DWP) and whey protein concentrate (WPC) on nutrient coefficient of apparent ileal digestibility (CAID) and productive performance. Cecal microbiota composition was also determined using Illumina amplicon sequencing. Dietary treatments were control diet (no supplementation of DWP or WPC), 60-DWP (60 g/kg of DWP), and 80-WPC (80 g/kg of WPC). One-day-old male broilers were randomly assigned to one of three treatments, and housed in floor pens. In Trial 1, 90 1-day-old chicks were allocated to three pens/treatment, with 10 birds/pen, during 21 days for CAID evaluation. Diet 60-DWP increased Ca CAID (P=0.041), while diet 80-WPC improved Ca and P CAID (P<0.001 and 0.002, respectively) when compared with control diet. In Trial 2, 810 one-day-old chicks were allocated to nine pens/treatment, with 30 birds/pen, during 42 days. Feeding chickens with 60-DWP and 80-WPC increased their BW, average daily gain (ADG) and feed intake (FI) during the starter (P<0.001 for all variables) and grower-finisher periods (P<0.001 for BW and FI, and P=0.048 for ADG), and during the entire feeding period (P<0.05), when compared with control diet. Diets 60-DWP and 80-WPC reduced the feed conversion ratio of chickens during the starter period (P<0.001 and 0.003, respectively), while 60-DWP reduced this parameter during the entire feeding period (P=0.048), when compared to control diet. At day 42, cecal microbial communities of chickens that were fed with 60-DWP and 80-WPC differed from those fed with control diet (R=0.776, P=0.008; and R=0.740, P=0.008, respectively). The abundance of Bacteroides fragilis, Bacteroides spp., Escherichia coli/Shigella flexneri and Megamonas furniformis increased when 60-DWP and 80-WPC diets were offered, while the presence of Helicobacter pullorum decreased. Lactobacillus salivarius consistently increased in chickens with better feed conversion ratio, which were those fed with 60-DWP. The results obtained in the present study indicate that growth of chickens is improved by DWP and WPC supplementation because of a higher mineral digestibility, increased feed intake and modulation of cecal microbiota communities.

Short communication: Signs of host genetic regulation in the microbiome composition in 2 dairy breeds: Holstein and Brown Swiss.

This study aimed to evaluate whether the host genotype exerts any genetic control on the microbiome composition of the rumen in cattle. Microbial DNA was extracted from 18 samples of ruminal content from 2 breeds (Holstein and Brown Swiss). Reads were processed using mothur (https://www.mothur.org/) in 16S and 18S rRNA gene-based analyses. Then, reads were classified at the genus clade, resulting in 3,579 operational taxonomic units (OTU) aligned against the 16S database and 184 OTU aligned against the 18S database. After filtering on relative abundance (>0.1%) and penetrance (95%), 25 OTU were selected for the analyses (17 bacteria, 1 archaea, and 7 ciliates). Association with the genetic background of the host animal based on the principal components of a genomic relationship matrix based on single nucleotide polymorphism markers was analyzed using Bayesian methods. Fifty percent of the bacteria and archaea genera were associated with the host genetic background, including Butyrivibrio, Prevotella, Paraprevotella, and Methanobrevibacter as main genera. Forty-three percent of the ciliates analyzed were also associated with the genetic background of the host. In total, 48% of microbes were associated with the host genetic background. The results in this study support the hypothesis and provide some evidence that there exists a host genetic component in cattle that can partially regulate the composition of the microbiome.

Evaluation of antimicrobial persistent activity of alcohol-based hand antiseptics against bacterial contamination.

The purpose of this study was to evaluate persistent activity of three alcohol-based antiseptics widely used in the clinical routine containing chlorhexidine, triclosan or mecetronium after hand disinfection. Four tests were used to evaluate the antimicrobial activity of antiseptics on: (i) resident microbiota; (ii) nutrient agar plates (NAP) subsequently inoculated with a test organism; and transient microbiota acquired by contact with dry hands (iii), or (iv) a wet inert surface that had been artificially contaminated. Four reference strains (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis) and an Acinetobacter baumannii strain responsible for a hospital outbreak were used as transient microbiota. Antimicrobial activity was calculated according to the CFUs reduction by reference to non-disinfected control hands. The antiseptics were applied according to European Standard EN1500. Solutions containing chlorhexidine or triclosan showed some persistent effects on transient microbiota on inert humid surfaces and NAP, but not on contaminated dry hands. Solutions containing mecetronium showed no persistent effect on transient flora in any of the trials. All alcohol-based antiseptics tested were more effective against resident flora than soap. No persistent activity was observed for A. baumannii in any of the trials. Chlorhexidine and triclosan are preferred when an antiseptic with persistent activity is desired, but a moist environment appears to be necessary for that antibacterial activity. Accordingly, relevant conclusions regarding the persistent activity of antiseptics for clinical practice and protection against bacterial contamination cannot be derived from this study for the alcoholic solutions tested.

Use of chitosans to modulate ruminal fermentation of a 50:50 forage-to-concentrate diet in sheep.

The objective of this study was to investigate the effects of chitosan (CHI) on ruminal and cecal fermentation and on apparent digestibility in sheep. In a first trial, 4 ruminally fistulated sheep fed an alfalfa hay and concentrate diet formulated to meet 1.2 times their maintenance energy requirements were assigned to 2 treatments [no-additive control (CTR), or 136 mg/kg of BW of CHI] for 19 d in a 2 x 2 crossover design. Samples of ruminal fluid and feces were taken for VFA and NH(3)-N concentrations. At the end of each period of the crossover, ruminal fluid was used as inoculum for batch cultures (24 and 96 h), and the effects on rate and extent of gas production and on fermentation variables for starch, cellulose, and the same diet fed to the animals were calculated. In a second trial, 4 sheep fed the alfalfa hay and concentrate diet were assigned to 2 treatments (CTR and CHI), as for the first trial, but this time for 14 d in a 2 x 2 crossover design. Apparent digestibility was studied. In the first trial, in terms of ruminal fermentation, CHI did not affect (P = 0.12) total VFA concentration or acetate proportion, but increased propionate proportion (P = 0.004) and propionate-to-acetate ratio (C3:C2; P = 0.007) and decreased branched-chain VFA (BCVFA) proportion (P = 0.003) and NH(3)-N concentration (P = 0.045). In feces, CHI decreased total VFA concentration (P = 0.01) and acetate (P = 0.045) and butyrate (P = 0.025) proportions, and increased propionate (P = 0.04), valerate (P = 0.01), and BCVFA (P = 0.032) proportions and C3:C2 ratio (P = 0.045). In batch cultures, when starch was used as substrate, CHI decreased acetate proportion (P = 0.016) and methane production (P = 0.03) and increased propionate and valerate proportions and C3:C2 ratio (P < 0.001). When cellulose was used as substrate, CHI decreased in vitro OM digestibility (P = 0.033), total VFA concentration, propionate proportion (P < 0.001), C3:C2 ratio (P = 0.006), methane production (P < 0.001), NH(3)-N concentration (P = 0.019), and gas production rate (P = 0.002), and increased butyrate (P = 0.029) and BCVFA (P = 0.004) proportions and pH (P < 0.001). When the alfalfa hay and concentrate diet was used as substrate, CHI decreased butyrate (P < 0.001) and BCVFA (P = 0.008) proportions, methane (P = 0.005) and asymptotic gas production (P = 0.02), and gas production rate (P = 0.05). In the second trial, CHI decreased NDF apparent digestibility (P = 0.09). In conclusion, CHI shifted ruminal fermentation toward energetically more efficient routes, without reducing OM apparent digestibility.

[NUTRISOL: a computer programme for communitary and hospital nutritional evaluation of free access]. / NUTRISOL: un programa informático para la evaluación nutricional comunitaria y hospitalaria de acceso libre.

We have developed the computer programme NUTRISOL, a nutritional programme destined to analysis of dietary intake by means of the food transformation to nutrient. It has been performed under Windows operative system, using Visual Basic 6.0. It is presented in a CD-Rom. We have used the Spanish CSIC Food Composition Table and domestic food measures commonly used in Spain which could be modified and updated. Diverse kind of diets and reference anthropometric data are also presented. The results may be treated using various statistical programmes. The programme contains three modules: 1) Nutritional epidemiology, which allows to create or open a data base, sample management, analyse food intake, consultation of nutrient content and exportation of data to statistical programmes. 2) Analyses of diets and recipes, creation or modification of new ones. 3) To ask different diets for prevalent pathologies. Independent tools for modifying the original tables, calculate energetic needs, recommend nutrient intake and anthropometric indexes are also offered. In conclusion, NUTRISOL Programme is an application which runs in PC computers with minimal equipment in a friendly interface, of easy use, freeware, which may be adapted to each country, and has demonstrated its usefulness and reliability in different epidemiologic studies. Furthermore, it may become an efficient instrument for clinical nutrition and health promotion.

18F-FDG metabolism in a rat model of chronic infarction: a 17-sector semiquantitative analysis.

UNLABELLED: Strategies to establish the functional benefit of cell therapy in cardiac regeneration and the potential mechanism are needed. AIMS: Development of a semi-quantitative method for non invasive assessment of cardiac viability and function in a rat model of myocardial infarction (MI) based on the use of microPET. ANIMALS, METHODS: Ten rats were subjected to myocardial imaging 2, 7, 14, 30, 60 and 90 days after left coronary artery ligation. Intravenous 18F-fluoro-2-deoxy-2-D-glucose (18F-FDG) was administered and regional 18F activity concentrations per unit area were measured in 17 regions of interest (ROIs) drawn on cardiac polar maps. By comparing the differences in 18F uptake between baseline and each of the follow up time points, parametric polar maps of statistical significance (PPMSS) were calculated. Left ventricular ejection fraction (LVEF) was blindly assessed echocardiographically. All animals were sacrificed for histopathological analysis after 90 days. RESULTS: The diagnostic quality of 18F-FDG microPET images was excellent. PPMSS demonstrated a statistically significant decrease in 18F concentrations as early as 48 hours after MI in 4 of the 17 ROIs (segments 7, 13, 16 and 17; p < 0.05) that persisted throughout the study. Semiquantitative analysis of 18F-FDG uptake correlated with echocardiographic decrease in LVEF (p < 0.001). CONCLUSION: The use of PPMSS based on 18F-FDG-microPET provides valuable semi-quantitative information of heart glucose metabolism allowing for non-invasive follow up thus representing a useful strategy for assessment of novel therapies in cardiac regeneration.

Study of lithium ion exchange by two synthetic zeolites: Kinetics and equilibrium.

We examined the exchange of univalent cations (Na+ and H+) retained on two commercially available synthetic zeolites with Li+ ions present in aqueous solutions in contact with the solids with a view to preparing effective controlled-release pharmaceutical forms. The studied zeolites were manufactured by Merck and featured channel diameters of 0.5 (Zeolite 5A, Ref. 1.05705.250, designated Z-05 in this work) and 1.0 nm (Zeolite 13X, Ref. 1.05703.250, designated Z-10 here). The XRD technique revealed that Z-05 possesses an LTA structure derived from that of sodalite and Z-10 a faujasite-type structure. Their exchange capacities were found to be 2.72 and 3.54 meq/g. The Z-Na + Li(+) / Z-Li + Na(+) and Z-H + Li(+) / Z-Li + H(+) ion-exchange processes were found to be reversible and their kinetic laws to obey the equation (-dC/dt) = k(a) x C(n) x (1-theta) - (k(d) x theta), with n = 1 for Z-10 and n = 2 for Z-05. Based on the equilibrium results, the overall processes involve one (with Z-05) or two single ion-exchange processes (with Z-10). In both cases, the equations that govern equilibrium are direct results of the kinetic laws. Thus, the first process-the one with only Z-05-involves the retention of Li+ cations at anionic sites on the outer surface of the solid and their access to the larger pores; the second process-which occurs with Z-10 only-involves the retention of lithium(I) cations within the zeolite channels. In both systems, the exchange with Li+ (from the aqueous solution) is easier than that with H+; this is consistent with our kinetic, equilibrium, and thermodynamic results.

Assessing the residual antibacterial activity of clinical materials disinfected with glutaraldehyde, o-phthalaldehyde, hydrogen peroxide or 2-bromo-2-nitro-1,3-propanediol by means of a bacterial toxicity assay.

This study investigated the use of a rapid bacterial toxicity test for detecting disinfectant residues released by disinfected materials. The test substances included an environmental disinfectant used in hospitals in high-risk areas, such as critical care units or emergency services, and three disinfectants used on clinical devices when a high level of disinfection is required. The test materials were polyurethane, polypropylene, glass, latex and cotton from different instruments and utensils used in hospitals. Of the four test disinfectants, o-phthalaldehyde (OPA) and 2-bromo-2-nitro-1,3-propanediol (BNP) showed the greatest inhibitory activity (as much as 300-fold greater than hydrogen peroxide in the case of OPA) according to the toxicity text. However, with the exception of hydrogen peroxide on latex, it was the most porous test materials, namely latex and cotton, that accumulated the least residue. BNP was the disinfectant that left the least residue on the five test materials, while the greatest residual concentration was left by hydrogen peroxide on latex (as much as 5 microg/cm2). The biotest used in this study permitted the detection of disinfectant residues released by different types of previously disinfected clinical materials, and can be adapted to simulate elution conditions similar to those existing in routine hospital practice.

Lithium adsorption by acid and sodium amberlite.

In this paper we used a previously reported model for examining the adsorption of nonelectrolytes in solution by solid adsorbents to study the adsorption of lithium(I) cations by acid and sodium amberlites, which is an ion-exchange process. Based on the results, both are equilibrium processes and obey a kinetic law with a unity partial order in the Li+ concentration. The kinetic results were used to calculate the specific rate constants and thermodynamic activation functions involved. Also, equilibrium isotherms were used to determine the corresponding ion-exchange capacities, the individual equilibrium constants, and the thermodynamic functions for the overall process.