Effects of sunflower oil infusions of Asparagopsis taxiformis on in vitro ruminal methane production and biohydrogenation of polyunsaturated fatty acids.

Asparagopsis taxiformis inhibits ruminal methane (CH4) production due to its bromoform (CHBr3) content. The immersion of A. taxiformis in edible vegetable oils allows the extraction and stabilization of the highly volatile CHBr3 in the oil phase. The objectives of this study were to explore the effects of adding sunflower oils with increasing concentrations of CHBr3 on in vitro ruminal methanogenesis and biohydrogenation. Five batches of 48-h in vitro incubations were performed in 14 fermentation bottles, using rumen inocula collected shortly after the slaughter of young crossbred bulls and 1 g of dry matter (DM) from a total diet of mixed feed without added oil (control) or with 60 µL of sunflower oil per gram of DM as the substrate. The treatments were the CHBr3 content in the oil added: 0 µg (B0), 25 µg (B25), 50 µg (B50), 75 µg (B75), 100 µg (B100), and 150 µg (B150) of CHBr3 per gram of substrate DM. Organic matter (OM) degradability, total gas, CH4, volatile fatty acids (VFA), long-chain fatty acids, and dimethyl acetals (DMA) were analyzed at the end of each incubation. Data were analyzed with a model considering the treatments as the fixed effect and the run as a random block and using orthogonal contrasts. Degradability of OM was higher in the control group and was unaffected by CHBr3 concentration. Total gas production per gram of degraded OM was unaffected by treatments and averaged 205 ± 29.8 mL/g. Methane (mL) production decreased linearly with increasing CHBr3 concentrations, with 33%, 47%, and 87% reductions for B75, B100, and B150, respectively. Total VFA concentration was unaffected by oil inclusion but was reduced by 20% in CHBr3-containing treatments, although without any dose-response pattern. The molar percentage of acetate decreased linearly, whereas propionate and butyrate increased linearly with the increasing CHBr3 dosage. Including oil in the diet decreased the branched-chain fatty acids and DMA content. Increasing CHBr3 concentrations did not affect branched-chain fatty acids, but linearly increased most of the identified DMA. Adding oil to the control diet increased the 18:2n-6, whereas increasing the concentration of CHBr3 had no effect on 18:2n-6 but decreased linearly the 18:0 and increased the trans-18:1 isomers. The results obtained provide evidence that oil immersions of A. taxiformis can successfully inhibit ruminal production of CH4 in vitro at doses of 100 and 150 µg/g DM, and simultaneously modulate biohydrogenation.

Statistical properties of the entropy from ordinal patterns.

The ultimate purpose of the statistical analysis of ordinal patterns is to characterize the distribution of the features they induce. In particular, knowing the joint distribution of the pair entropy-statistical complexity for a large class of time series models would allow statistical tests that are unavailable to date. Working in this direction, we characterize the asymptotic distribution of the empirical Shannon's entropy for any model under which the true normalized entropy is neither zero nor one. We obtain the asymptotic distribution from the central limit theorem (assuming large time series), the multivariate delta method, and a third-order correction of its mean value. We discuss the applicability of other results (exact, first-, and second-order corrections) regarding their accuracy and numerical stability. Within a general framework for building test statistics about Shannon's entropy, we present a bilateral test that verifies if there is enough evidence to reject the hypothesis that two signals produce ordinal patterns with the same Shannon's entropy. We applied this bilateral test to the daily maximum temperature time series from three cities (Dublin, Edinburgh, and Miami) and obtained sensible results.

Insectivorous bats as biomonitor of metal exposure in the megalopolis of Mexico and rural environments in Central Mexico.

The Megalopolis of Mexico is one of the largest cities in the world and presents substantial problems of metal pollution. Insectivorous bats that inhabit this city are potentially exposed to metals and could therefore serve as a good biomonitor. We collected 70 adult male individuals of Tadarida brasiliensis (Chiroptera: Molossidae) from two areas inside the Megalopolis (Cuautitlán and Xochimilco) and two rural environments in Central Mexico (Tequixquiac and Tlalcozotitlán). We analyzed livers to determine the total concentrations of ten metals by the ICP-MS technique, compared concentrations among study sites to provide evidence of metal exposure, and explored the associations between metals and their accumulation patterns in bats. The hepatic metal concentrations we recorded were generally consistent with those of similar studies in insectivorous bats. Higher concentrations of Cu and Zn in Cuautitlán and Xochimilco bats were associated with vehicular traffic. Higher concentrations of V, Cr, and Co in Tequixquiac bats and Cd in Tlalcozotitlán bats were linked with industrial, agricultural, or sewage sources. Variations in Fe and Mn concentrations were related to geogenic sources or local conditions. Similar Ni and Pb concentrations were linked with strong homeostatic controls or historical pollution. Accumulation patterns showed that all urban bats belonged to a single population with similar degrees of metal exposure, while rural bats belonged to two different populations exposed to different metals. Our results highlight the need to monitor the emissions generated by particular sources in each study site.

Extending the biosynthetic repertoires of cyanobacteria and chloroplasts.

Chloroplasts in plants and algae and photosynthetic microorganisms such as cyanobacteria are emerging hosts for sustainable production of valuable biochemicals, using only inorganic nutrients, water, CO2 and light as inputs. In the past decade, many bioengineering efforts have focused on metabolic engineering and synthetic biology in the chloroplast or in cyanobacteria for the production of fuels, chemicals and complex, high-value bioactive molecules. Biosynthesis of all these compounds can be performed in photosynthetic organelles/organisms by heterologous expression of the appropriate pathways, but this requires optimization of carbon flux and reducing power, and a thorough understanding of regulatory pathways. Secretion or storage of the compounds produced can be exploited for the isolation or confinement of the desired compounds. In this review, we explore the use of chloroplasts and cyanobacteria as biosynthetic compartments and hosts, and we estimate the levels of production to be expected from photosynthetic hosts in light of the fraction of electrons and carbon that can potentially be diverted from photosynthesis. The supply of reducing power, in the form of electrons derived from the photosynthetic light reactions, appears to be non-limiting, but redirection of the fixed carbon via precursor molecules presents a challenge. We also discuss the available synthetic biology tools and the need to expand the molecular toolbox to facilitate cellular reprogramming for increased production yields in both cyanobacteria and chloroplasts.

Elucidating constraints for differentiation of major human Klebsiella pneumoniae clones using MALDI-TOF MS.

The establishment of matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) in routine microbial identification boosted many developments towards high-throughput applications, including bacterial typing. However, results are still controversial for different bacterial species. We aim to evaluate the suitability of MALDI-TOF MS for typing clinically relevant multidrug resistant (MDR) Klebsiella pneumoniae subsp. pneumoniae clones using routine conditions and a previously validated chemometric analysis workflow. Mass spectra of 83 K. pneumoniae clinical isolates representing major human MDR clones [11 sequence types (STs), 22 PFGE-types] recovered in Portugal and Spain during outbreaks and non-outbreak situations (2003-2012) were obtained from cell extracts (CE) and intact cells (IC), and analysed with different chemometric tools. We observed a highly consistent peak pattern among isolates from different clones either with CE or IC, suggesting a high degree of conservation of biomolecules analysed (a large part corresponding to ribosomal proteins). Moreover, the low degree of agreement between MALDI-TOF MS and other methods (from 34.9 % to 43.4 % of correct assignments for CE and from 40.8 % to 70.1 % for IC) corroborates the low discriminatory potential of the technique at infraspecies level. Our results suggest a low discriminatory power of MALDI-TOF MS for clinically relevant MDR K. pneumoniae clones and highlight the need of developing tools for high-resolution typing in this species.

Inference of genetic diversity in popcorn S3 progenies.

Molecular markers are a useful tool for identification of complementary heterotic groups in breeding programs aimed at the production of superior hybrids, particularly for crops such as popcorn in which heterotic groups are not well-defined. The objective of the present study was to analyze the genetic diversity of 47 genotypes of tropical popcorn to identify possible heterotic groups for the development of superior hybrids. Four genotypes of high genetic value were studied: hybrid IAC 125, strain P2, and varieties UENF 14 and BRS Angela. In addition, 43 endogamous S3 progenies obtained from variety UENF 14 were used. Twenty-five polymorphic SSR-EST markers were analyzed. A genetic distance matrix was obtained and the following molecular diversity parameters were estimated: number of alleles, number of effective alleles, polymorphism information content (PIC), observed and expected heterozygosities, Shannon diversity index, and coefficient of inbreeding. We found a moderate PIC and high diversity index, indicating that the studied population presents both good discriminatory ability and high informativeness for the utilized markers. The dendrogram built based on the dissimilarity matrix indicated six distinct groups. Our findings demonstrate the genetic diversity among the evaluated genotypes and provide evidence for heterotic groups in popcorn. Furthermore, the functional genetic diversity indicates that there are informative genetic markers for popcorn.

Effects of Combining Feed Grade Urea and a Slow-release Urea Product on Performance, Dietary Energetics and Carcass Characteristics of Feedlot Lambs Fed Finishing Diets with Different Starch to Acid Detergent Fiber Ratios.

Recent findings have shown that microbial nitrogen flow and digestible energy of diets are increased when urea is combined with a slow-release urea (SRU) in diets with a starch to acid detergent fibre ratio (S:F) 4:1. This affect is attributable to enhanced synchrony between ruminal N availability for microbial growth and carbohydrate degradation. To verify the magnitude of this effects on lamb performance, an experiment was conducted to evaluate the effects of combining urea and a SRU in diets containing S:F ratios of 3:1, 4:1, or 5:1 on performance, dietary energetics and carcass characteristics of finishing lambs. For that, 40 Pelibuey×Katahdin lambs (36.65±3 kg) were assigned to one of five weight groupings in 20 pens (5 repetition/treatments). The S:F ratio in the diet was manipulated by partially replacing the corn grain and dried distiller's grain with solubles by forage (wheat straw) and soybean meal to reach S:F ratios of 3:1, 4:1 or 5:1. An additional treatment of 4:1 S:F ratio with 0.8% urea as the sole source of non-protein nitrogen was used as a reference for comparing the effect of urea combination vs. conventional urea at the same S:F ratio. There were no treatment effects on dry matter intake (DMI). Compared the urea combination vs urea at the same S:F ratio, urea combination increased (p<0.01) average daily gain (ADG, 18.3%), gain for feed (G:F, 9.5%), and apparent energy retention per unit DMI (8.2%). Irrespective of the S:F ratio, the urea combination improved the observed-to-expected dietary ratio and apparent retention per unit DMI was maximal (quadratic effect, p≤0.03) at an S:F ratio of 4:1, while the conventional urea treatment did not modify the observed-to-expected net energy ratio nor the apparent retention per unit DMI at 4:1 S:F ratio. Urea combination group tended (3.8%, p = 0.08) to have heavier carcasses with no effects on the rest of carcass characteristics. As S:F ratio increased, ADG, G:F, dietary net energy, carcass weight, dressing percentage and longissimus thoracis (LM) area increased linearly (p≤0.02). Combining urea and a slow-release urea product results in positive effects on growth performance and dietary energetics, but the best responses are apparently observed when there is a certain proportion (S:F ratio = 4:1) of starch to acid detergent fibre in the diet.

Functional molecular markers (EST-SSR) in the full-sib reciprocal recurrent selection program of maize (Zea mays L.).

This study aimed to improve grain yield in the full-sib reciprocal recurrent selection program of maize from the North Fluminense State University. In the current phase of the program, the goal is to maintain, or even increase, the genetic variability within and among populations, in order to increase heterosis of the 13th cycle of reciprocal recurrent selection. Microsatellite expressed sequence tags (EST-SSRs) were used as a tool to assist the maximization step of genetic variability, targeting the functional genome. Eighty S1 progenies of the 13th recur-rent selection cycle, 40 from each population (CIMMYT and Piranão), were analyzed using 20 EST-SSR loci. Genetic diversity, observed heterozygosity, information content of polymorphism, and inbreeding co-efficient were estimated. Subsequently, analysis of genetic dissimilarity, molecular variance, and a graphical dispersion of genotypes were conducted. The number of alleles in the CIMMYT population ranged from 1 to 6, while in the Piranão population the range was from 2 to 8, with a mean of 3.65 and 4.35, respectively. As evidenced by the number of alleles, the Shannon index showed greater diversity for the Piranão population (1.04) in relation to the CIMMYT population (0.89). The genic SSR markers were effective in clustering genotypes into their respective populations before selection and an increase in the variation between populations after selection was observed. The results indicate that the study populations have expressive genetic diversity, which cor-responds to the functional genome, indicating that this strategy may contribute to genetic gain, especially in association with the grain yield of future hybrids.

Genetic diversity of popcorn genotypes using molecular analysis.

In this study, we analyzed dominant molecular markers to estimate the genetic divergence of 26 popcorn genotypes and evaluate whether using various dissimilarity coefficients with these dominant markers influences the results of cluster analysis. Fifteen random amplification of polymorphic DNA primers produced 157 amplified fragments, of which 65 were monomorphic and 92 were polymorphic. To calculate the genetic distances among the 26 genotypes, the complements of the Jaccard, Dice, and Rogers and Tanimoto similarity coefficients were used. A matrix of Dij values (dissimilarity matrix) was constructed, from which the genetic distances among genotypes were represented in a more simplified manner as a dendrogram generated using the unweighted pair-group method with arithmetic average. Clusters determined by molecular analysis generally did not group material from the same parental origin together. The largest genetic distance was between varieties 17 (UNB-2) and 18 (PA-091). In the identification of genotypes with the smallest genetic distance, the 3 coefficients showed no agreement. The 3 dissimilarity coefficients showed no major differences among their grouping patterns because agreement in determining the genotypes with large, medium, and small genetic distances was high. The largest genetic distances were observed for the Rogers and Tanimoto dissimilarity coefficient (0.74), followed by the Jaccard coefficient (0.65) and the Dice coefficient (0.48). The 3 coefficients showed similar estimations for the cophenetic correlation coefficient. Correlations among the matrices generated using the 3 coefficients were positive and had high magnitudes, reflecting strong agreement among the results obtained using the 3 evaluated dissimilarity coefficients.

Construction of an integrated genetic map for Capsicum baccatum L.

Capsicum baccatum L. is one of the five Capsicum domesticated species and has multiple uses in the food, pharmaceutical and cosmetic industries. This species is also a valuable source of genes for chili pepper breeding, especially genes for disease resistance and fruit quality. However, knowledge of the genetic structure of C. baccatum is limited. A reference map for C. baccatum (2n = 2x = 24) based on 42 microsatellite, 85 inter-simple sequence repeat, and 56 random amplified polymorphic DNA markers was constructed using an F2 population consisting of 203 individuals. The map was generated using the JoinMap software (version 4.0) and the linkage groups were formed and ordered using a LOD score of 3.0 and maximum of 40% recombination. The genetic map consisted of 12 major and four minor linkage groups covering a total genome distance of 2547.5 cM with an average distance of 14.25 cM between markers. Of the 152 pairs of microsatellite markers available for Capsicum annuum, 62 were successfully transferred to C. baccatum, generating polymorphism. Forty-two of these markers were mapped, allowing the introduction of C. baccatum in synteny studies with other species of the genus Capsicum.

MALDI-TOF mass spectrometry as a tool for the discrimination of high-risk Escherichia coli clones from phylogenetic groups B2 (ST131) and D (ST69, ST405, ST393).

Reliable, quick and low-cost methods are needed for the early detection of multidrug-resistant and highly virulent high-risk B2 and D Escherichia coli clones or clonal complexes (HiRCC). Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) seems to have a good discriminatory potential at different subspecies levels, but it was never evaluated for the discrimination of E. coli clones. We assessed the potential of MALDI-TOF MS coupled to multivariate data analysis to discriminate representative E. coli B2 and D HiRCC. Seventy-three E. coli isolates from B2 (including ST131 and B2 non-ST131 clones) and D (ST69, ST393, ST405) with variable pulsed-field gel electrophoresis (PFGE) patterns, origins and dates (1980-2010) were tested. MS spectra were acquired from independent extracts obtained from different plate cultures in two different Microflex LT MALDI-TOF devices (Bruker) after a standard extraction procedure. MALDI-TOF MS fingerprinting analysis revealed a good discriminatory ability between the four HiRCC analysed (ST131, ST69, ST405, ST393) and between B2 ST131 and other B2 non-ST131 isolates. Clusters defined by MALDI-TOF MS were consistent with the clonal complexes assigned by multilocus sequence typing (MLST), although differences were detected regarding the composition of clusters obtained by the comparison of PFGE profiles. We demonstrate, for the first time, that characteristic mass fingerprints of different E. coli HiRCC are sufficiently discriminatory and robust to enable their differentiation by MALDI-TOF MS, which might represent a promising tool for the optimisation of infection control, individual patient management and large-scale epidemiological studies of public health relevance. The good correlation between phenotypic and genotypic features further corroborates phylogenetic relationships delineated by MLST.

Genetic-molecular characterization of backcross generations for sexual conversion in papaya (Carica papaya L.).

The low number of improved cultivars limits the expansion of the papaya crop, particularly because of the time required for the development of new varieties using classical procedures. Molecular techniques associated with conventional procedures accelerate this process and allow targeted improvements. Thus, we used microsatellite markers to perform genetic-molecular characterization of papaya genotypes obtained from 3 backcross generations to monitor the inbreeding level and parental genome proportion in the evaluated genotypes. Based on the analysis of 20 microsatellite loci, 77 genotypes were evaluated, 25 of each generation of the backcross program as well as the parental genotypes. The markers analyzed were identified in 11 of the 12 linkage groups established for papaya, ranging from 1 to 4 per linkage group. The average values for the inbreeding coefficient were 0.88 (BC1S4), 0.47 (BC2S3), and 0.63 (BC3S2). Genomic analysis revealed average values of the recurrent parent genome of 82.7% in BC3S2, 64.4% in BC1S4, and 63.9% in BC2S3. Neither the inbreeding level nor the genomic proportions completely followed the expected average values. This demonstrates the significance of molecular analysis when examining different genotype values, given the importance of such information for selection processes in breeding programs.

Use of elephant grass for energy production in Campos dos Goytacazes-RJ, Brazil.

Elephant grass is a tropical forage plant widely distributed throughout Brazil. It was first exclusively used in the livestock sector as cattle feed. The grass is characterized by its high productivity and photosynthetic capacity and is considered as an alternative source of renewable energy. Here, we estimated the general combining ability of the parents and specific combining ability of the hybrids based on morpho-agronomic biomass-quality traits. The experiment was conducted in a randomized block design with 3 replicates. The diallel was composed of 16 hybrids and 2 groups of genitors. In the diallel analysis of variance, we observed a significant difference among treatments. A significant difference was observed among genitors for dry matter production (DMP). For the general combining ability of group 1, the traits leaf blade width, DMP, height, percentage of neutral detergent fiber, percentage of hemicellulose, percentage of lignin, percentage of acid detergent fiber, and percentage of cellulose were significant. For the estimates of general combining ability of DMP, parents Porto Rico 534-B, Vruckwona, Taiwan A-146, and Mercker S. E. A. were 0.4748, 3.2819, 1.1659, and 0.4317. The parents of Mercker S. E. A. and Porto Rico 534-B produced the highest percentage of detergent fiber and percentage of lignin with values of 0.1482 and 0.0856. Thus, parents Vruckwona, Porto Rico 534-B, and Taiwan A-146 are promising for integration into breeding programs. The best hybrid combinations for DMP were 1 x 5, 1 x 8, 2 x 6, 3 x 7, and 4 x 5.

Influence of protein and energy level in finishing diets for feedlot hair lambs: growth performance, dietary energetics and carcass characteristics.

Forty-eight Pelibuey×Katahdin male intact lambs (23.87±2.84 kg) were used in an 84-d feeding trial, with six pens per treatment in a 2×2 factorial design arrangement. The aim of the study was to evaluate the interaction of two dietary energy levels (3.05 and 2.83 Mcal/kg ME) and two dietary protein levels (17.5% and 14.5%) on growth performance, dietary energetics and carcass traits. The dietary treatments used were: i) High protein-high energy (HP-HE); ii) High protein-low energy (HP-LE); iii) Low protein-high energy (LP-HE), and iv) Low protein-low energy (LP-LE). With a high-energy level, dry matter intake (DMI) values were 6.1% lower in the low-protein diets, while with low-energy, the DMI values did not differ between the dietary protein levels. Energy levels did not influence the final weight and average daily gain (ADG), but resulted in lower DMI values and higher gain efficiencies. No effects of protein level were detected on growth performance. The observed dietary net energy (NE) ratio and observed DMI were closer than expected in all treatments and were not affected by the different treatments. There was an interaction (p<0.03) between energy and protein level for kidney-pelvic and heart fat (KPH), KPH was higher in lambs fed high energy and high protein diet but not in high energy and low protein diet. The KPH was increased (20.2%, p = 0.01) in high-energy diets, while fat thickness was increased (21.7%, p = 0.02) in high-protein diets. Therefore, it is concluded that dietary energy levels play a more important role in feed efficiency than protein levels in finishing lambs with a high-energy diet (>2.80 Mcal/kg ME). Providing a level of protein above 14.5% does not improves growth-performance, dietary energetics or carcass dressing percentage.

Effects of Combining Feed Grade Urea and a Slow-release Urea Product on Characteristics of Digestion, Microbial Protein Synthesis and Digestible Energy in Steers Fed Diets with Different Starch:ADF Ratios.

As a result of the cost of grains, the replacement of grains by co-products (i.e. DDGS) in feedlot diets is a common practice. This change produces diets that contain a lower amount of starch and greater amount of fibre. Hypothetically, combining feed grade urea (U) with slow release urea (Optigen) in this type of diet should elicit a better synchrony between starch (high-rate of digestion) and fibre (low-rate of digestion) promoting a better microbial protein synthesis and ruminal digestion with increasing the digestible energy of the diet. Four cannulated Holstein steers (213±4 kg) were used in a 4×4 Latin square design to examine the combination of Optigen and U in a finishing diet containing different starch:acid detergent fibre ratios (S:F) on the characteristics of digestive function. Three S:F ratios (3.0, 4.5, and 6.0) were tested using a combination of U (0.80%) and Optigen (1.0%). Additionally, a treatment of 4.5 S:F ratio with urea (0.80% in ration) as the sole source of non-protein nitrogen was used to compare the effect of urea combination at same S:F ratio. The S:F ratio of the diet was manipulated by replacing the corn grain by dried distillers grain with solubles and roughage. Urea combination did not affect ruminal pH. The S:F ratio did not affect ruminal pH at 0 and 2 h post-feeding but, at 4 and 6 h, the ruminal pH decreased as the S:F ratio increased (linear, p<0.05). Ruminal digestion of OM, starch and feed N were not affected by urea combination or S:F ratio. The urea combination did not affect ADF ruminal digestion. ADF ruminal digestion decreased linearly (p = 0.02) as the S:F ratio increased. Compared to the urea treatment (p<0.05) and within the urea combination treatment (quadratic, p<0.01), the flow of microbial nitrogen (MN) to the small intestine and ruminal microbial efficiency were greater for the urea combination at a S:F ratio of 4.5. Irrespective of the S:F ratio, the urea combination improved (2.8%, p = 0.02) postruminal N digestion. As S:F ratio increased, OM digestion increased, but ADF total tract digestion decreased. The combination of urea at 4.5 S:F improved (2%, p = 0.04) the digestible energy (DE) more than expected. Combining urea and Optigen resulted in positive effects on the MN flow and DE of the diet, but apparently these advantages are observed only when there is a certain proportion of starch:ADF in the diet.

Hibernacula of bats in Mexico, the southernmost records of hibernation in North America.

Although Mexico holds the southernmost hibernating bats in North America, information on winter behavior and hibernacula microclimate use of temperate Mexican bats is limited. We studied hibernating bats at high altitudes (>1,000 m a.s.l.) in northern and central Mexico during 5 consecutive winters. Our aims were to document and describe the hibernacula, winter behavior (such as abundance and roost pattern), and microclimates (estimated as adjacent substrate temperature) of cave-hibernating bats in Mexico. We found 78 hibernacula and 6,089 torpid bats of 10 vespertilionid species, increasing by over 50% the number of cave-hibernating bat species and quadrupling the number of hibernacula for Mexico. Hibernacula were at altitudes between 1,049 and 3,633 m a.s.l., located in 3 mountain ranges, mainly in oak and conifer forests. Myotis velifer was the most common species, followed by Corynorhinus townsendii and C. mexicanus. We recorded the adjacent substrate temperatures from 9 species totaling 1,106 torpid bats and found differences in microclimate use among the 3 most common species. In general, abundance of torpid bats in our region of study was similar to those in the western United States, with aggregations of tens to a few hundred individuals per cave, and was lower than in the eastern United States where a cave may hold thousands of individuals. Knowledge of bat hibernation is crucial for developing conservation and management strategies on current conditions while accommodating environmental changes and other threats such as emerging diseases.

Aunque México tiene los murciélagos hibernantes más sureños en Norteamérica, la información sobre el comportamiento invernal y el uso de microclimas en los refugios de hibernación de los murciélagos templados mexicanos es limitada. Estudiamos a los murciélagos hibernantes en altitudes altas (>1000 msnm) en el norte y centro de México durante cinco inviernos consecutivos. Nuestros objetivos fueron documentar y describir las cuevas de hibernación, el comportamiento invernal (como la abundancia y patrón de percha), y el uso de microclimas (estimado como la temperatura del sustrato adyacente), de los murciélagos que hibernan en cuevas en México. Encontramos 78 cuevas de hibernación con 6089 murciélagos en torpor de 10 especies de vespertiliónidos, incrementando en más del 50% el número de especies de murciélagos que hibernan en cuevas y cuadriplicando el número de cuevas de hibernación para México. Las cuevas de hibernación estuvieron en elevaciones entre 1049 y 3633 msnm, localizadas en tres cadenas montañosas, principalmente en bosques de encinos y coníferas. Myotis velifer fue la especie más común, seguida por Corynorhinus townsendii y C. mexicanus. Reportamos las temperaturas del sustrato adyacente de 1106 murciélagos en torpor de nueve especies y encontramos diferencias en el uso de microclimas entre las tres especies más comunes. Aquí proveemos información relevante para especies de murciélagos templados en la ocurrencia más sureña de hibernación de murciélagos en Norteamérica. En general, la abundancia de murciélagos en torpor que encontramos fue similar a las del oeste de Estados Unidos, con agregaciones de decenas y algunos cientos de individuos por cueva; y fue menor que las del este de Estados Unidos, donde las cuevas pueden albergar miles de murciélagos. El conocimiento de la hibernación de murciélagos es crucial para el desarrollo de estrategias de conservación y manejo adecuadas en la actualidad y mientras se adaptan a los cambios ambientales y a otras amenazas tales como las enfermedades emergentes.

In vitro activity of cefiderocol and other newly approved antimicrobials against multi-drug resistant Gram-negative pathogens recovered in intensive care units in Spain and Portugal.

OBJECTIVE: The antimicrobial resistance is a significant public health threat, particularly for healthcare-associated infections caused by carbapenem-resistant Gram-negative pathogens which are increasingly reported worldwide. The aim of this study was to provide data on the in vitro antimicrobial activity of cefiderocol and that of commercially available comparator antibiotics against a defined collection of recent clinical multi-drug resistant (MDR) microorganisms, including carbapenem resistant Gram-negative bacteria collected from different regions in Spain and Portugal. METHODS: A total of 477 clinical isolates of Enterobacterales, Pseudomonas aeruginosa, Acinetobacter baumannii and Stenotrophomonas maltophilia were prospectively (n=265) and retrospectively (n=212) included (2016-2019). Susceptibility testing was performed using standard broad microdilution and results were interpreted using CLSI-2021 and EUCAST-2021 criteria. RESULTS: Overall, cefiderocol showed a good activity against Enterobacterales isolates, being 99.5% susceptible by CLSI and 94.5% by EUCAST criteria. It also demonstrated excellent activity against P. aeruginosa and S. maltophilia isolates, all being susceptible to this compound considering CLSI breakpoints. Regarding A. baumannii (n=64), only one isolate was resistant to cefiderocol. CONCLUSIONS: Our results are in agreement with other studies performed outside Spain and Portugal highlighting its excellent activity against MDR gram-negative bacteria. Cefiderocol is a therapeutic alternative to those available for the treatment of infections caused by these MDR bacteria.

Effect of the dietary supplementation with sunflower oil-enriched bromoform from Asparagopsis taxiformis on lambs' growth, health, and ruminal methane production.

The red seaweed Asparagopsis taxiformis has a potent antimethanogenic effect, which has been proven both in vitro and in vivo. Vegetable oil immersions of this seaweed (hereafter Bromoil) help stabilise the bromoform (CHBr3) responsible for its antimethanogenic effect. We evaluate the effects of increasing the levels of CHBr3 in lamb diets on growth performance, methane (CH4) production, animal health and meat quality. Twenty-four Merino Branco ram lambs were fed a ground complete compound feed, supplemented with 50 mL/kg DM of sunflower oil with different CHBr3 content. The treatments were defined by the CHBr3 doses in the oil: 0 mg (control - B0), 15 mg (B15), 30 mg (B30) and 45 mg (B45) of CHBr3 per kg of feed DM. The feed was prepared daily by mixing Bromoil with the compound feed. At the end of the experiment, the lambs were sacrificed, the ruminal content was collected for in vitro fermentation to evaluate CH4 production and organic matter (OM) degradability, and the rumen mucosa was sampled for histological examination. Meat samples were collected for chemical composition and CHBr3 analysis. The half-life of CHBr3 in the air-exposed feed was 3.98 h making it very difficult to establish the practiced level of CHBr3 supplementation. Lambs-fed treatments B30 and B45 decreased DM intake by up to 28%. Average daily gain was also reduced due to CHBr3 supplementation, with B45 showing results 40% lower than B0. DM feed conversion ratio was similar for all treatments. The degradability of OM, the volume of total gas and of gas without CH4 were unaffected by the experimental treatments, evaluated by the in vitro method. However, the volume of CH4 decreased by up to 75% for treatments above 30 mg/kg DM, while the yield of CH4/g OM degraded was reduced by up to 78% with treatments above 30 mg/kg DM. Meat chemical composition was not affected by Bromoil supplementation and no traces of CHBr3 were found in meat samples. During this experiment, the animals presented normal health and behaviour. However, postslaughter examination of the rumen showed distinct lesions on the ventral region of the rumen mucosa of animals supplemented with Bromoil. These lesions were more severe in the animals receiving treatments B30 and B45. This research determined that although concentrations of CHBr3 in the diet above 30 mg/kg DM helped to reduce CH4 emissions, it negatively affected the performance and rumen wall.

PAHs in franciscana dolphins from the Southwestern Atlantic Ocean: Concentration and maternal transfer assessments.

Polycyclic aromatic hydrocarbons (PAHs) are organic compounds ubiquitous in the environment and known for their toxic, mutagenic, and carcinogenic effects. These compounds can bioaccumulate in the biota and be transferred through trophic webs. The franciscana dolphin (Pontoporia blainvillei), as top predators, can be an environmental sentinels. Thus, this study aimed to provide data about PAHs concentration in their hepatic tissue collected on the coast of Espírito Santo (Franciscana Management Area, FMA Ia), Rio de Janeiro (FMA IIa), and São Paulo states (FMA IIb), in Southeastern Brazil. PAHs were detected in 86 % of franciscana dolphins (n = 50). The highest ∑PAHsTotal median concentration was reported in FMA Ia followed by FMA IIb and FMA IIa (1055.6; 523.9, and 72.1 ng.g-1 lipid weight, respectively). Phenanthrene was detected in one fetus and two neonates, showing maternal transfer of PAHs in these dolphins. Evaluating PAHs with potential toxic effects is of utmost importance for the conservation of a threatened species.

Rapid detection of urinary tract infections caused by Proteus spp. using PNA-FISH.

We developed a fluorescence in situ hybridization (FISH) method for the rapid detection of Proteus spp. in urine, using a novel peptide nucleic acid (PNA) probe. Testing on 137 urine samples from patients with urinary tract infections has shown specificity and sensitivity values of 98 % (95 % CI, 93.2-99.7) and 100 % (95 % CI, 80,8-100), respectively, when compared with CHROMagar Orientation medium. Results indicate that PNA-FISH is a reliable alternative to traditional culture methods and can reduce the diagnosis time to approximately 2 h.