This study aimed to examine the effects of re-ensiling time and Lactobacillus buchneri on the fermentation profile, chemical composition and aerobic stability of sugarcane silages. The experiment was set up as a repeated measure design consisting of four air-exposure periods (EP)(0, 6, 12, and 24 h) microbial additive (A) (L. buchneri; or lack of there), with five replicates. Sugarcane was ground through a stationary forage chopper and ensiled in four plastic drums of 200-L capacity. After 210 days of storage, the drums were opened and half of the silage mass was treated with L. buchneri at the concentration of 105 cfu/g of forage. Subsequently, the silages were divided into stacks. The re-ensiling process was started immediately, at 0, 6, 12 and 24-hour intervals, by transferring the material to PVC mini-silos. Silos were opened after 120 days of re-ensiling. The use of L. buchneri reduced butyrate concentration but did not change ethanol or acetic acid concentrations and aerobic stability. An interaction effect between L. buchneri and re-ensiling time was observed for dry matter (DM) losses and composition. Lactobacillus buchneri is not effective in improving aerobic stability in re-ensiled sugarcane silages. However, less DM is lost in silages treated with L. buchneri and exposed to air for 24 h. Re-ensiling sugar cane in up to 24 h of exposure to air does not change final product quality.

A growing number of infectious pathogens are spreading among geographic regions. Some pathogens that were previously not considered to pose a general threat to human health have emerged at regional and global scales, such as Zika and Ebola Virus Disease. Other pathogens, such as yellow fever virus, were previously thought to be under control but have recently re-emerged, causing new challenges to public health organisations. A wide array of new modelling techniques, aided by increased computing capabilities, novel diagnostic tools, and the increased speed and availability of genomic sequencing allow researchers to identify new pathogens more rapidly, assess the likelihood of geographic spread, and quantify the speed of human-to-human transmission. Despite some initial successes in predicting the spread of acute viral infections, the practicalities and sustainability of such approaches will need to be evaluated in the context of public health responses.

Mud volcanoes are singular seafloor structures classified as ‘sensitive habitats’. Here we report on the sponge fauna from a field of eight mud volcanoes located in the Spanish margin of the northern Gulf of Cadiz (North-eastern Atlantic), at depths ranging from 380 to 1146 m. Thirty-eight beam-trawl samplings were conducted (covering over 61,000 m2) from 2010 to 2012, in the frame of a EC-LIFE + INDEMARES grant. A total of 1659 specimens were retrieved, belonging to 82 species, from which 79 were in the Class Demospongiae and three in Hexactinellida. Two species were new to science (Jaspis sinuoxea sp. nov.; Myrmekioderma indemaresi sp. nov.) and three others recorded for the first time in the Atlantic Ocean (Geodia anceps, Coelosphaera cryosi and Petrosia raphida). Five additional species were ‘Atlantic oddities’, since this study provides their second record in the Atlantic Ocean (Lanuginella cf. pupa, Geodia cf. spherastrella, Cladocroce spathiformis, Cladocroce fibrosa and Haliclona pedunculata). Basic numerical analyses indicated a significant linear relationship between the species richness per m2 and the number of sponge individuals per m2, meaning that in most volcanoes many species occur in equivalent, moderate abundance. Likewise, sponge species richness increased with depth, while the abundance of hard substrata resulting from carbonate precipitation and the fishing activities around the volcanoes had no detectable effect on the sponge fauna. However, in the latter case, a negative trend – lacking statistical support – underlaid the analyses, suggesting that a more extensive sampling would be necessary to derive more definitive conclusions in this regard.

The current study aimed to test whether organic matter intake by free-ranging ruminants could be estimated from the amount of nitrogen (N) excreted in faeces and to compare this approach to conventional techniques. An equation describing the relationship between excreted N and nutrient intake was developed in indoor digestibility trials conducted with male sheep (n = 36) and cattle (n = 24) housed in metabolism cages and solely fed hay harvested from a local rangeland. Faecal N excretion was linearly related to organic matter (OM) intake without a significant animal species effect. To evaluate the linear equation, data from free-ranging trials conducted with sheep and cattle were used. The faecal N approach was compared with either in situ digestibility plus external marker (n = 123) or n-alkanes (n = 272) to estimate OM intake and digestible OM intake. Estimates obtained through the faecal N approach did not closely fit those obtained with either conventional technique for any variable. Averaging all individual values, the supply of metabolizable energy (ME) estimated through faecal N was similar to the required level, whereas both the in situ and n-alkanes techniques overestimated ME supply. In conclusion, OM intake by free-ranging sheep and cattle can be estimated based on the amount of N excreted in faeces with some advantages over conventional techniques: knowledge about herbage attributes is not required and it accounts for individual variability on selectivity and digestion processes.

American tegumentary leishmaniasis is caused by different species of Leishmania. This protozoan employs several mechanisms to subvert the microbicidal activity of macrophages and, given the limited efficacy of current therapies, the development of alternative treatments is essential. Animal venoms are known to exhibit a variety of pharmacological activities, including antiparasitic effects. Crotoxin (CTX) is the main component of Crotalus durissus terrificus venom, and it has several biological effects. Nevertheless, there is no report of CTX activity during macrophage – Leishmania interactions. Thus, the main objective of this study was to evaluate whether CTX has a role in macrophage M1 polarization during Leishmania infection murine macrophages, Leishmania amazonensis promastigotes and L. amazonensis-infected macrophages were challenged with CTX. MTT [3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide] toxicity assays were performed on murine macrophages, and no damage was observed in these cells. Promastigotes, however, were affected by treatment with CTX (IC50 = 22·86 µg mL−1) as were intracellular amastigotes. Macrophages treated with CTX also demonstrated increased reactive oxygen species production. After they were infected with Leishmania, macrophages exhibited an increase in nitric oxide production that converged into an M1 activation profile, as suggested by their elevated production of the cytokines interleukin-6 and tumour necrosis factor-α and changes in their morphology. CTX was able to reverse the L. amazonensis-mediated inhibition of macrophage immune responses and is capable of polarizing macrophages to the M1 profile, which is associated with a better prognosis for cutaneous leishmaniasis treatment.

In this work we explore the radiocarbon (14C) signal as an independent tool to assess the year of formation of individual tree rings of tropical species in northern Brazil. Three different species were analyzed in this work: Dipteryx magnifica, Enterolobium maximum, and Hymenolobium petraeum. The studied samples are from the stem of only one individual of each species, all cut down in 2008 in Porto Trombetas, Pará, Brazil. Individual tree rings were identified based on wood anatomy and they were counted from bark to pith. Several rings were selected for 14C analysis in order to cover the overall shape of the 14C atmospheric bomb peak (after 1955). The 14C content was measured at Laboratory of Radiocarbon, Universidade Federal Fluminense (LAC-UFF). Results are compared with the Southern Hemisphere 14C atmospheric calibration curves. For E. maximum and H. petraeum the 14C signal exhibits an overall good match with the SH zone 3 and suggests annual seasonality in the growth-ring formation. These species offer suitable characteristics for dendrochronology. The D. magnifica shows mismatches in the 14C measurements that are likely a result of difficulties in identifying ring boundaries with certainty. Additional techniques may be helpful to disentangle the origin of this discrepancy.

Parasites of the genera Plasmodium and Haemoproteus (Apicomplexa: Haemosporida) are a diverse group of pathogens that infect birds nearly worldwide. Despite their ubiquity, the ecological and evolutionary factors that shape the diversity and distribution of these protozoan parasites among avian communities and geographic regions are poorly understood. Based on a survey throughout the Neotropics of the haemosporidian parasites infecting manakins (Pipridae), a family of Passerine birds endemic to this region, we asked whether host relatedness, ecological similarity and geographic proximity structure parasite turnover between manakin species and local manakin assemblages. We used molecular methods to screen 1343 individuals of 30 manakin species for the presence of parasites. We found no significant correlations between manakin parasite lineage turnover and both manakin species turnover and geographic distance. Climate differences, species turnover in the larger bird community and parasite lineage turnover in non-manakin hosts did not correlate with manakin parasite lineage turnover. We also found no evidence that manakin parasite lineage turnover among host species correlates with range overlap and genetic divergence among hosts. Our analyses indicate that host switching (turnover among host species) and dispersal (turnover among locations) of haemosporidian parasites in manakins are not constrained at this scale.

A micro heteregenous reprocessed fuel spiked with thorium in a PWR fuel element considering (TRU-Th) cycle was simulated using three different configurations and different fissile materials that varied from 6.0% to 7.0%. The reprocessed fuels were obtained using the ORIGEN 2.1 code from a burned PWR standard fuel (33,000 MWd/tHM burned), with 3.1% of initial enrichment, which was remained in the cooling pool for five years and then reprocessed using UREX+ technique. The keff and plutonium generation during the burnup were evaluated and compared with the standard fuel. This study was performed using the SCALE 6.0.

Brazil is one of the most important soybean producers in the world. Soybean is a very important crop for the country as it is used for several purposes, from food to biodiesel production. The levels of soybean yield in the different growing regions of the country vary substantially, which results in yield gaps of considerable magnitude. The present study aimed to investigate the soybean yield gaps in Brazil, their magnitude and causes, as well as possible solutions for a more sustainable production. The concepts of yield gaps were reviewed and their values for the soybean crop determined in 15 locations across Brazil. Yield gaps were determined using potential and attainable yields, estimated by a crop simulation model for the main maturity groups of each region, as well as the average actual famers’ yield, obtained from national surveys provided by the Brazilian Government for a period of 32 years (1980–2011). The results showed that the main part of the yield gap was caused by water deficit, followed by sub-optimal crop management. The highest yield gaps caused by water deficit were observed mainly in the south of Brazil, with gaps higher than 1600 kg/ha, whereas the lowest were observed in Tapurah, Jataí, Santana do Araguaia and Uberaba, between 500 and 1050 kg/ha. The yield gaps caused by crop management were mainly concentrated in South-central Brazil. In the soybean locations in the mid-west, north and north-east regions, the yield gap caused by crop management was <500 kg/ha. When evaluating the integrated effects of water deficit and crop management on soybean yield gaps, special attention should be given to Southern Brazil, which has total yield gaps >2000 kg/ha. For reducing the present soybean yield gaps observed in Brazil, several solutions should be adopted by growers, which can be summarized as irrigation, crop rotation and precision agriculture. Improved dissemination of agricultural knowledge and the use of crop simulation models as a tool for improving crop management could further contribute to reduce the Brazilian soybean yield gap.

The insertion of reprocessed fuel spiked with thorium in a typical PWR fuel element considering (TRU-Th) cycle was simulated using different fissile materials that varied from 5.5% to 7.0%. The reprocessed fuels were obtained using the ORIGEN 2.1 code from a burned PWR standard fuel (33,000 MWd/tHM burned), with 3.1% of initial enrichment, which was remained in the cooling pool for five years and then reprocessed using UREX+ technique. The kinf, hardening spectrum and the fuel evolution during the burnup were evaluated. This study was performed using the SCALE 6.0

Leptospirosis is a zoonosis of worldwide distribution, caused by infection with pathogenic spirochaetes of the genus Leptospira. The wild boar (Sus scrofa), an important hunting species in Europe, seems to play a significant role in the epidemiological cycle of leptospirosis. A total of 101 serum samples from wild boar hunted in Northern Portugal were analysed for leptospiral antibodies detection by microscopic agglutination test. Sera were collected during hunting seasons (2011–2013) and tested with 17 different pathogenic serovars of Leptospira. Antibodies against nine serovars were detected in 66 (65·4%) of these sera. Serovars Tarassovi and Altodouro exhibited the highest seroreactivity rates (23·8% and 16·8%, respectively), followed by Autumnalis (7·9%) and Bratislava (6·9%). Age and district of origin were found to be risk factors for the presence of leptospiral antibodies in contrast to gender. From a One Health perspective, this study revealed that wild boar should be considered as a potential source of leptospirosis dissemination for humans and animal species (domestic and wild) in shared environments, particularly in the Trás-os-Montes region.

Next-generation 454 sequencing techniques were used to re-examine diversity of mitochondrial cytochrome b lineages of avian malaria (Plasmodium relictum) in Hawaii. We document a minimum of 23 variant lineages of the parasite based on single nucleotide transitional changes, in addition to the previously reported single lineage (GRW4). A new, publicly available portal (Integroomer) was developed for initial parsing of 454 datasets. Mean variant prevalence and frequency was higher in low elevation Hawaii Amakihi (Hemignathus virens) with Avipoxvirus-like lesions (P = 0·001), suggesting that the variants may be biologically distinct. By contrast, variant prevalence and frequency did not differ significantly among mid-elevation Apapane (Himatione sanguinea) with or without lesions (P = 0·691). The low frequency and the lack of detection of variants independent of GRW4 suggest that multiple independent introductions of P. relictum to Hawaii are unlikely. Multiple variants may have been introduced in heteroplasmy with GRW4 or exist within the tandem repeat structure of the mitochondrial genome. The discovery of multiple mitochondrial lineages of P. relictum in Hawaii provides a measure of genetic diversity within a geographically isolated population of this parasite and suggests the origins and evolution of parasite diversity may be more complicated than previously recognized.

The objective of this study was to evaluate the addition of IGF-I to pig insemination doses stored at 15°C, in conjunction with the addition of different amounts of vitamin E (α-tocopherol). Semen samples (n = 12) from four boars were treated by the addition of different concentrations of vitamin E, ranging up to 400 μg/ml. Immediately after processing and after the doses had been stored at 15°C for 24 or 72 h, samples were warmed at 37°C and 30 ng/ml of IGF-I was added. The assessments were made after 10 and 120 min of IGF-I addition. There was a minor effect of the vitamin E added before cooling and IGF-I added after storage on sperm quality. The addition of 400 μg/ml of vitamin E to diluted semen reduced (P < 0.01) the malondialdehyde (MDA) production in boar semen stored at 15°C for 72 h, regardless of the addition of IGF-I as additive during a 120 min incubation period at 37°C. In these conditions, IGF-I also reduced (P < 0.05) the MDA production in semen samples without addition of vitamin E. IGF-I in the presence of vitamin E reduced (P = 0.03) the glucose intake in freshly diluted boar semen samples before cooling. It was concluded that the addition of 400 μg/ml of vitamin E reduces the MDA production in boar semen stored at 15°C for 72 h, regardless of the presence of IGF-I additive. The addition of IGF-I in doses stored for 72 h with vitamin E ensures higher sperm motility after 120 min of incubation at 37°C.

Tributyrin (TBT) is a TAG composed of three butyric acids that has beneficial effects on ulcerative colitis due to its trophic, anti-inflammatory, pro-apoptotic and anti-carcinogenic properties. The goal of the present study was to evaluate the efficacy and mechanisms of action of TBT supplementation in the prevention of mucosal damage in experimental colitis. Mice received either a control diet or a TBT-supplemented diet for 15 d. Colitis was induced by dextran sodium sulphate administration during the last 7 d. Mucosal damage and the activation of immune cells and cytokines were determined by histological score, flow cytometry and ELISA. Leucocyte rolling and adhesion were assessed by intravital microscopy. Oxidative stress was determined by monitoring hydroperoxide concentration and evaluating superoxide dismutase (SOD) and catalase activities. Intestinal permeability was analysed using diethylenetriaminepentaacetate acid (99mTcDTPA). Compared with the colitis group, the animals in the colitis+TBT group had reduced mucosal damage and neutrophil and eosinophil mucosal infiltration, which were associated with a higher percentage of regulatory T cells (Treg) and higher levels of transforming growth factor β and IL-10 in the lamina propria. The level of in vivo leucocyte adhesion in the colon microvasculature was reduced after TBT supplementation. A lower level of hydroperoxide and higher levels of SOD and catalase activities were associated with TBT supplementation. TBT-supplemented mice showed reduced intestinal permeability to the levels intermediate between the control and colitis groups. In conclusion, the present results show that TBT has positive effects on colonic restructuring in experimental colitis. Additionally, TBT supplementation changes the immune response by controlling inflammation and regulating the expression of anti-inflammatory cytokines and Treg.

An ATP diphosphohydrolase (EC 3.6.1.5) activity was identified in a Leishmania (Viannia) braziliensis promastigotes preparation (Lb). Ultrastructural cytochemical microscopy showed this protein on the parasite surface and also stained a possible similar protein at the mitochondrial membrane. Isolation of an active ATP diphosphohydrolase isoform from Lb was obtained by cross-immunoreactivity with polyclonal anti-potato apyrase antibodies. These antibodies, immobilized on Protein A-Sepharose, immunoprecipitated a polypeptide of approximately 48 kDa and, in lower amount, a polypeptide of approximately 43 kDa, and depleted 83% ATPase and 87% of the ADPase activities from detergent-homogenized Lb. Potato apyrase was recognized in Western blots by IgG antibody from American cutaneous leishmaniasis (ACL) patients, suggesting that the parasite and vegetable proteins share antigenic conserved epitopes. Significant IgG seropositivity in serum samples diluted 1:50 from ACL patients (n=20) for Lb (65%) and potato apyrase (90%) was observed by ELISA technique. Significant IgG antibody reactivity was also observed against synthetic peptides belonging to a conserved domain from L. braziliensis NDPase (80% seropositivity) and its potato apyrase counterpart (50% seropositivity), in accordance with the existence of shared antigenic epitopes and demonstrating that in leishmaniasis infection the domain r82-103 from L. braziliensis NDPase is a target for the human immune response.

Evolutionary and closer structural relationships are demonstrated by phylogenetic analysis, peptide prediction and molecular modelling between Solanum tuberosum apyrase, Schistosoma mansoni SmATPase 2 and Leishmania braziliensis NDPase. Specific protein domains are suggested to be potentially involved in the immune response, and also seem to be conserved during host and parasite co-evolution. Significant IgG antibody reactivity was observed in sera from patients with American cutaneous leishmaniasis (ACL) and schistosomiasis using potato apyrase as antigen in ELISA. S. mansoni adult worm or egg, L. braziliensis promastigote (Lb) and Trypanosoma cruzi epimastigote (EPI) have ATP diphosphohydrolases, and antigenic preparations of them were evaluated. In ACL patients, IgG seropositivity was about 43% and 90% for Lb and potato apyrase, respectively, while IgM was lower (<19%) for both. In schistosomiasis patients IgM (>40%) or IgG (100%) seropositivity for both soluble egg (SEA) and adult worm (SWAP) antigens was higher than that found for potato apyrase (IgM=10%; IgG=39%). In Chagas disease, IgG seropositivity for EPI and potato apyrase was 97% and 17%, respectively, while the IgM was low (3%) for both antigens. The study of the conserved domains from both parasite proteins and potato apyrase could lead to the development of new drug targets or molecular markers.