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Dietary protein adjustments can reduce environmental impact and economic losses in production systems. However, we lack information regarding nitrogen (N) metabolism and protein requirements for maintenance of crossbred animals such as Red Norte breed, precluding a precise dietary management. The objective was to evaluate the effect of increasing dietary CP levels (9%, 11%, 13%, 15% and 17%) on intake, digestibility and N balance, as well as to estimate the metabolizable protein requirements for maintenance (MPm) of growing Red Norte bulls. Thirty five animals averaging 280 ± 4.0 kg BW were fed during 45 days in a 60 : 40 forage : concentrate ratio diet in which the last 5 days were used for the digestibility trial. Intakes of CP and non-fibrous carbohydrates (NFCs) and feed efficiency linearly increased (P < 0.05) as CP levels increased, while DM, NDF, nitrogen efficiency use and ether extract were not influenced by CP levels (P > 0.05). Digestibilities of DM, organic matter, ether extract, NFC and CP as well as metabolizable energy intake linearly increased (P < 0.05), and true digestibility of CP was not affected (P > 0.05) by treatments. Urinary N and retained N linearly increased (P < 0.05) with the increase in dietary N. The MPm were estimated as 4.46 g/BW0.75 and the efficiency of use of MPm was 0.673. In conclusion, obtained MPm requirements of growing Red Norte bulls are greater than the values reported in literature for Zebu cattle and dietary CP levels of 15% and 17% exhibited great responses for growing Red Norte cattle. However, a cost-benefit evaluation should be done before its use.
The rocky shores of the north-east Atlantic have been long studied. Our focus is from Gibraltar to Norway plus the Azores and Iceland. Phylogeographic processes shape biogeographic patterns of biodiversity. Long-term and broadscale studies have shown the responses of biota to past climate fluctuations and more recent anthropogenic climate change. Inter- and intra-specific species interactions along sharp local environmental gradients shape distributions and community structure and hence ecosystem functioning. Shifts in domination by fucoids in shelter to barnacles/mussels in exposure are mediated by grazing by patellid limpets. Further south fucoids become increasingly rare, with species disappearing or restricted to estuarine refuges, caused by greater desiccation and grazing pressure. Mesoscale processes influence bottom-up nutrient forcing and larval supply, hence affecting species abundance and distribution, and can be proximate factors setting range edges (e.g., the English Channel, the Iberian Peninsula). Impacts of invasive non-native species are reviewed. Knowledge gaps such as the work on rockpools and host–parasite dynamics are also outlined.
The search for life in the Universe is a fundamental problem of astrobiology and modern science. The current progress in the detection of terrestrial-type exoplanets has opened a new avenue in the characterization of exoplanetary atmospheres and in the search for biosignatures of life with the upcoming ground-based and space missions. To specify the conditions favourable for the origin, development and sustainment of life as we know it in other worlds, we need to understand the nature of global (astrospheric), and local (atmospheric and surface) environments of exoplanets in the habitable zones (HZs) around G-K-M dwarf stars including our young Sun. Global environment is formed by propagated disturbances from the planet-hosting stars in the form of stellar flares, coronal mass ejections, energetic particles and winds collectively known as astrospheric space weather. Its characterization will help in understanding how an exoplanetary ecosystem interacts with its host star, as well as in the specification of the physical, chemical and biochemical conditions that can create favourable and/or detrimental conditions for planetary climate and habitability along with evolution of planetary internal dynamics over geological timescales. A key linkage of (astro)physical, chemical and geological processes can only be understood in the framework of interdisciplinary studies with the incorporation of progress in heliophysics, astrophysics, planetary and Earth sciences. The assessment of the impacts of host stars on the climate and habitability of terrestrial (exo)planets will significantly expand the current definition of the HZ to the biogenic zone and provide new observational strategies for searching for signatures of life. The major goal of this paper is to describe and discuss the current status and recent progress in this interdisciplinary field in light of presentations and discussions during the NASA Nexus for Exoplanetary System Science funded workshop ‘Exoplanetary Space Weather, Climate and Habitability’ and to provide a new roadmap for the future development of the emerging field of exoplanetary science and astrobiology.
The enrichment of meat with selenium is important to improve the intake of selenium by humans. The effects of supranutritional doses of sodium selenite or selenium-enriched yeast on performance, carcass characteristics and meat quality were evaluated using 63 Nellore cattle in a completely randomized design with two sources (sodium selenite and selenium-enriched yeast), three levels (0.3, 0.9 and 2.7 mg Se/kg DM) and control treatment (without addition of selenium). Final body weight (BW), average daily gain, dry matter intake and gain to feed ratio (G : F) at the end of 84 days of supplementation were not influenced by treatments (P>0.05). Values of pH, ribeye area, back fat thickness and marbling score were also not influenced by treatments (
P>0.05). Dressing percentage was greater (P=0.02) in Nellore cattle supplemented with organic Se (58.70%) compared to animals supplemented with inorganic Se (57.94%). Hot carcass weight increased (
P=0.05) with the increasing of Se levels in the diet. Colour, shear force (SF), cooking and drip loss remained unchanged (
P>0.05); however thiobarbituric acid reactive substances was 15.51% higher with inorganic Se compared with organic Se. The selenium concentration in the meat of animals receiving organic selenium was higher (
P<0.001) than that of animals receiving sodium selenite, at all levels (0.3; 0.9 and 2.7 mg/kg DM). The meat of animals receiving 2.7 mg of organic Se/kg of DM presented concentration of 372.7 μg Se/kg in the L.dorsi muscle, and the intake of 150 g of this meat by humans provides approximately 100% of the recommended Se intake (55 μg Se/day for adults). Therefore, the use of supranutritional doses of 2.7 mg Se/kg of DM, regardless of source, is a way of naturally producing selenium-enriched meat without compromising performance, carcass characteristics and quality of Nellore bovine meat.
Intake in sugar-rich diets can be limited either via rumen fill or excessive rumen fermentation and source of non-fibre carbohydrate (NFC) in the diet can affect both factors. The aim of the current study was to quantify the effect of partially replacing ground maize (GM) with steam-rolled maize (SRM) or pelleted citrus pulp (PCP) at two concentrate levels in sugarcane-based diets on digestibility, rumen ecosystem and metabolism of Nellore steers. Six rumen-cannulated steers were assigned to a 6 × 6 Latin square, replicated in time, in a 2 × 3 factorial arrangement of treatments with two levels of concentrate (600 or 800 g concentrate/kg dry matter [DM]) and three NFC sources. Each steer within a period was considered an experimental unit. Feeding more concentrate increased total tract digestibility of organic matter and decreased fibre intake and passage rate. It also reduced rumen populations of Fibrobacter succinogenes and Streptococcus bovis and increased Ruminococcus flavefaciens. Substituting PCP for GM increased rumen pH, acetic acid and organic matter digestibility. Feeding PCP also reduced R. flavefaciens and R. amylophilus rumen populations. Substituting SRM for GM increased starch digestibility and rumen propionic acid, but decreased rumen ammonia concentration. Feeding SRM increased rumen populations of Megasphaera elsdenii with the high-concentrate diet but reduced Ruminococcus albus populations at both concentrate levels. In conclusion, partial replacement of GM by PCP decreased intake in sugar-rich diets, while increasing total tract neutral detergent fibre digestibility. Replacement of GM with SRM increases rumen fermentation and total tract digestibility of starch.
The use of tail chalk and estrus/heat expression scores (HEATSC) evaluation is instrumental in identifying cows with greater estrus expression and greater artificial insemination pregnancy rates (P/AI) in cows submitted to timed artificial insemination (TAI), and cows with low or no estrus expression present lower P/AI. It was intended in this study to improve the pregnancy rates in TAI for Bos indicus beef cows, and gonadotrophin-releasing hormone (GnRH) injection was hypothesized to increase pregnancy rates in a TAI program for cows submitted to progesterone–estradiol-based protocols with low or no estrus expression, evaluated by HEATSC. Cows (n= 2284) received a progesterone device and 2 mg estradiol benzoate, after 8 days the device was removed and 1 mg estradiol cypionate, 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotropin was administered. All cows were marked with chalk and HEATSC evaluated (scales 1 to 3) at TAI performed on day 10. Animals with HEATSC1 and HEATSC2 (n= 937) received 100 μg de gonadorelin (GNRH group; n= 470), or 1 ml saline (Control group; n= 467), and cows with HEATSC3 (named HEAT group; n= 1347) received no additional treatment. The larger dominant follicle, evaluated on day 8and at TAI (day 10), was greater in HEAT group (P= 0.0145 and P <0.001, respectively). Corpus luteum (CL) area and progesterone concentration was evaluated on day 17, and CL area was larger in HEAT group, intermediary in Control and lower in GnRH group (Control= 2.68 cm2, GnRH= 2.37 cm2, HEAT group= 3.07 cm2, P <0.001). Greater progesterone concentrations were found in HEAT group than in Control and GnRH groups (Control= 4.74 ng/ml, GnRH= 4.29 ng/ml, HEAT group= 6.08 ng/ml, P<0.001). There was a difference in ovulation rate, greater in HEAT group than GnRH and Control groups (Control= 72.5%; GnRH= 81.25%; HEAT group= 90.71%; P= 0.0024). Artificial insemination pregnancy rates was greater in HEAT group (57.09% (769/1347) than in Control and GNRH groups, with positive effect of GnRH injection at the time of TAI in P/AI (Control= 36.18% (169/467), GnRH= 45.95% (216/470); P<0.0001). In conclusion, GnRH application in cows with low HEATSC (1 and 2) is a simple strategy, requiring no changes in TAI management to increase pregnancy rates in postpartum beef cows submitted to progesterone–estradiol-based TAI protocols, without reaching, however, the pregnancy rates of cows that demonstrate high estrus expression at the TAI.
This paper describes a model of electron energization and cyclotron-maser emission applicable to astrophysical magnetized collisionless shocks. It is motivated by the work of Begelman, Ergun and Rees [Astrophys. J. 625, 51 (2005)] who argued that the cyclotron-maser instability occurs in localized magnetized collisionless shocks such as those expected in blazar jets. We report on recent research carried out to investigate electron acceleration at collisionless shocks and maser radiation associated with the accelerated electrons. We describe how electrons accelerated by lower-hybrid waves at collisionless shocks generate cyclotron-maser radiation when the accelerated electrons move into regions of stronger magnetic fields. The electrons are accelerated along the magnetic field and magnetically compressed leading to the formation of an electron velocity distribution having a horseshoe shape due to conservation of the electron magnetic moment. Under certain conditions the horseshoe electron velocity distribution function is unstable to the cyclotron-maser instability [Bingham and Cairns, Phys. Plasmas 7, 3089 (2000); Melrose, Rev. Mod. Plasma Phys. 1, 5 (2017)].
Chagas disease (CD) is a neglected parasitic condition endemic in the Americas caused by Trypanosoma cruzi. Patients present an acute phase that may or not be symptomatic, followed by lifelong chronic stage, mostly indeterminate, or with cardiac and/or digestive progressive lesions. Benznidazole (BZ) and nifurtimox are the only drugs approved for treatment but not effective in the late chronic phase and many strains of the parasite are naturally resistant. New alternative therapy is required to address this serious public health issue. Repositioning and combination represent faster, and cheaper trial strategies encouraged for neglected diseases. The effect of imatinib (IMB), a tyrosine kinase inhibitor designed for use in neoplasias, was assessed in vitro on T. cruzi and mammalian host cells. In comparison with BZ, IMB was moderately active against different strains and forms of the parasite. The combination IMB + BZ in fixed-ratio proportions was additive. Novel 14 derivatives of IMB were screened and a 3,2-difluoro-2-phenylacetamide (3e) was as potent as BZ on T. cruzi but had low selectivity index. The results demonstrate the importance of phenotypic assays, encourage the improvement of IMB derivatives to reach selectivity and testify to the use of repurposing and combination in drug screening for CD.
The mechanisms involved in kidney disturbances during development, induced by vitamin D3 deficiency in female rats, that persist into adulthood were evaluated in this study. Female offspring from mothers fed normal (control group, n=8) or vitamin D-deficient (Vit.D-, n=10) diets were used. Three-month-old rats had their systolic blood pressure (SBP) measured and their blood and urine sampled to quantify vitamin D3 (Vit.D3), creatinine, Na+, Ca+2 and angiotensin II (ANGII) levels. The kidneys were then removed for nitric oxide (NO) quantification and immunohistochemical studies. Vit.D- pups showed higher SBP and plasma ANGII levels in adulthood (P<0.05) as well as decreased urine osmolality associated with increases in urinary volume (P<0.05). Decreased expression of JG12 (renal cortex and glomeruli) and synaptopodin (glomeruli) as well as reduced renal NO was also observed (P<0.05). These findings showed that renal disturbances in development in pups from Vit.D- mothers observed in adulthood may be related to the development of angiogenesis, NO and ANGII alterations.
In this work, we report evidences of the improvement of X-ray attenuation efficiency by the addition of a very small amount of Graphene Oxide (GO) in polymer-based nanocomposite. Poly(vinylidene fluoride) (PVDF) homopolymer and barium sulfate (BaSO4) nanoparticles were mixed. PVDF/BaSO4 nanocomposite was found to attenuate 9.14% of a 20 kV X-ray beam. The addition of only 4.0 wt % of GO nanosheets to the nanocomposite improved this X-Ray attenuation efficiency to 24.56%. The respective linear attenuation coefficients (μ) were 39.9 cm-1 and 54.4 cm-1, respectively. The X-ray attenuation gradually decreases until 6.71% and 17.62%, respectively, for the X-ray beam with higher energy (100 kV). Fourier transform infrared data revealed that, due to the lack of the bending vibration modes of CF2 molecule at 656 cm-1, 688 cm-1, 723 cm-1, 776 cm-1and 796 cm-1, characteristics of the γ-crystalline phase of PVDF, the nanocomposites casted from solution are mostly in the β-ferroelectric phase of PVDF, besides the γ-paraelectric phase. SEM micrographs were used to evaluate the dispersion state of graphene sheets and the BaSO4 nanoparticles into the polymeric matrix. UV-Vis spectrometry and Differential Scanning Calorimetry (DSC) were also performed in order to complement the structural analysis. The results confirm that the addition of graphene sheets in PVDF polymer-based nanocomposites enhances the X-ray shielding efficiency. The phenomenon is discussed in terms of the reported anomalous negative thermal expansion coefficient of graphene sheets
In the current study, phage-exposed mimotopes as targets against tegumentary leishmaniasis (TL) were selected by means of bio-panning cycles employing sera of TL patients and healthy subjects, besides the immune stimulation of peripheral blood mononuclear cells (PBMCs) collected from untreated and treated TL patients and healthy subjects. The clones were evaluated regarding their specific interferon-γ (IFN-γ) and interleukin-4 (IL-4) production in the in vitro cultures, and selectivity and specificity values were calculated, and those presenting the best results were selected for the in vivo experiments. Two clones, namely A4 and A8, were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of IFN-γ, IL-2, IL-12, tumour necrosis factor-α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls. In conclusion, we presented a strategy to identify new mimotopes able to induce Th1 response in PBMCs from TL patients and healthy subjects, and that were successfully used to protect against L. amazonensis infection.
Keto analogues and amino acids (KAAA) supplementation can reduce blood ammonia concentrations in athletes undergoing high-intensity exercise under both ketogenic and thermoneutral conditions. This study evaluated the acute effects of KAAA supplementation on ammonia metabolism during extenuating endurance exercise in rats fed a ketogenic diet. In all, eighty male Fischer rats at 90 d of age were divided into eight groups, and some were trained using a swimming endurance protocol. A ketogenic diet supplemented with keto analogues was administered for 10 d. Administration of the ketogenic diet ended 3 d before the exhaustion test (extenuating endurance exercise). A ketogenic diet plus KAAA supplementation and extenuating endurance exercise (trained ketogenic diet supplemented with KAAA (TKKa)) increased blood ammonia concentrations by approximately 50 % compared with the control diet (trained control diet supplemented with KAAA (TCKa)) and similar training (effect size=1·33; statistical power=0·50). The KAAA supplementation reduced blood urea concentrations by 4 and 18 % in the control and ketogenic diet groups, respectively, compared with the groups fed the same diets without supplementation. The trained groups had 60 % lower blood urate concentrations after TCKa treatment than after TKKa treatment. Our results suggest that KAAA supplementation can reduce blood ammonia concentrations after extenuating endurance exercise in rats fed a balanced diet but not in rats fed a ketogenic diet.
Moringa oleifera seeds are currently being used as a livestock feed across tropical regions of the world due to its availability and palatability. However, limited knowledge exists on the effects of the raw seeds on ruminant metabolism. As such, the rumen stimulation technique was used to evaluate the effects of substituting increasing concentrations of ground Moringa seeds (0, 100, 200 and 400 g/kg concentrate dry matter (DM)) in the diet on rumen fermentation and methane production. Two identical, Rusitec apparatuses, each with eight fermenters were used with the first 8 days used for adaptation and days 9 to 16 used for measurements. Fermenters were fed a total mixed ration with Urochloa brizantha as the forage. Disappearance of DM, CP, NDF and ADF linearly decreased (P<0.01) with increasing concentrations of Moringa seeds in the diet. Total volatile fatty acid production and the acetate to propionate ratio were also linearly decreased (P<0.01). However, only the 400 g/kg (concentrate DM basis) treatment differed (P<0.01) from the control. Methane production (%), total microbial incorporation of 15N and total production of microbial N linearly decreased (P<0.01) as the inclusion of Moringa seeds increased. Though the inclusion of Moringa seeds in the diet decreased CH4 production, this arose from an unfavourable decrease in diet digestibility and rumen fermentation parameters.
The use of fast and non-destructive techniques for identifying sugarcane varieties enables the development of automatic sorting systems, contributing towards improving pre-processing steps in the alcohol and sugar industries. In this context, principal component analysis (PCA), factorial discriminant analysis (FDA), stepwise forward discriminant analysis (SFDA) and partial least-squares discriminant analysis (PLS-DA) were used to classify four Brazilian sugarcane varieties based on visible/near infrared (Vis/NIR) spectral reflectance measurements (450–1000 nm range) of stalks. All wavelengths contributed towards discriminating the sugarcane varieties, but the 600–750 nm range was most relevant. When evaluating PCA results considering the four sugarcane varieties, two of them overlapped and it was only possible to use classifiers of three varieties. Factorial discriminant analysis, PLS-DA and SFDA reached correct classifications of 0.81, 0.82 and 0.74, respectively, when considering the external validation data and the four sugarcane varieties evaluated. Results showed that Vis/NIR spectroscopy combined with discriminating methods is a promising tool for non-destructive and fast sugarcane variety classification, which can be used in the agro-food industry or directly in the field.
The aim of the current study was to evaluate the effect of different forms of soybean lipids on enteric methane emission, intake, performance, digestibility and meat quality of 40 young Nellore bulls (initial body weight (BW) 444 ± 10.2 kg and 24 ± 2.1 months). The dietary treatments were as follows: NF = no dietary additional fat (46.0 g ether extract (EE)/kg diet), SO = soybean oil (62.0 g EE/kg diet), SB = soy beans (without any processing; 62.0 g EE/kg diet) and RPF = rumen-protected fat based on soybean oil (62.0 g EE/kg diet). The intake of dry matter (DM), organic matter (OM), crude protein (CP) and neutral detergent fibre (NDF) was greater in the SO diet than those fed with NF. The SO diet decreased digestibility of NDF when compared with the NF and RPF diets. The diets did not affect digestibility of DM, OM, CP or emission of enteric methane. Animals fed with SO had greater average daily gain and feed efficiency in relation to the other diets tested. The SO diet increased hot and cold carcass weights and subcutaneous fat thickness of carcasses when compared with the NF diet. The proportions of saturated and unsaturated, mono and polyunsaturated, and n-3 and n-6 fatty acids were not affected significantly by treatments. The SO diets were demonstrated to be more beneficial for animal performance compared with diets without supplemental fat.
This experiment evaluated the impacts of supplementing a yeast-derived product (Celmanax; Church & Dwight Co., Inc., Princeton, NJ, USA) on productive and health responses of beef steers, and was divided into a preconditioning (days 4 to 30) and feedlot receiving phase (days 31 to 69). In all, 84 Angus × Hereford steers were weaned on day 0 (BW=245±2 kg; age=186±2 days), and maintained in a single group from days 0 to 3. On day 4, steers were allocated according to weaning BW and age to a 21-pen drylot (4 steers/pen). Pens were randomly assigned to (n=7 pens/treatment): (1) no Celmanax supplementation during the study, (2) Celmanax supplementation (14 g/steer daily; as-fed) from days 14 to 69 or (3) Celmanax supplementation (14 g/steer daily; as-fed) from days 31 to 69. Steers had free-choice access to grass-alfalfa hay, and were also offered a corn-based concentrate beginning on day 14. Celmanax was mixed daily with the concentrate. On day 30, steers were road-transported for 1500 km (24 h). On day 31, steers returned to their original pens for the 38-day feedlot receiving. Shrunk BW was recorded on days 4, 31 and 70. Feed intake was evaluated daily (days 14 to 69). Steers were observed daily (days 4 to 69) for bovine respiratory disease (BRD) signs. Blood samples were collected on days 14, 30, 31, 33, 35, 40, 45, 54 and 69, and analyzed for plasma cortisol, haptoglobin, IGF-I, and serum fatty acids. Preconditioning results were analyzed by comparing pens that received (CELM) or not (CONPC) Celmanax during the preconditioning phase. Feedlot receiving results were analyzed by comparing pens that received Celmanax from days 14 to 69 (CELPREC), days 31 to 69 (CELRECV) or no Celmanax supplementation (CON). During preconditioning, BRD incidence was less (P=0.03) in CELM v. CONPC. During feedlot receiving, average daily gain (ADG) (P=0.07) and feed efficiency (P=0.08) tended to be greater in CELPREC and CELRECV v. CON, whereas dry matter intake was similar (P⩾0.29) among treatments. No other treatment effects were detected (P⩾0.20). Collectively, Celmanax supplementation reduced BRD incidence during the 30-day preconditioning. Moreover, supplementing Celmanax tended to improve ADG and feed efficiency during the 38-day feedlot receiving, independently of whether supplementation began during preconditioning or after feedlot entry. These results suggest that Celmanax supplementation benefits preconditioning health and feedlot receiving performance in beef cattle.
Phosphorus (P) is an essential nutrient in livestock production, involved in most metabolic processes in the body, and with an impact on bone development and growth. Diet supplementation with P can improve productivity, but at excess levels the P utilization can be low, and the high P excretion will lead to environmental pollution (Tamminga, 2003). P supplementation also adds substantially to the cost of feed. Using the isotopic dilution technique (Vitti et al., 2000), we have studied the effect of different P levels in diet on P absorption and excretion in Brazilian sheep, and identified the level at which true P absorption saturates.
The physical processes driving the chemical evolution of galaxies in the last ~ 11Gyr cannot be understood without directly probing the dust-obscured phase of star-forming galaxies and active galactic nuclei. This phase, hidden to optical tracers, represents the bulk of the star formation and black hole accretion activity in galaxies at 1 < z < 3. Spectroscopic observations with a cryogenic infrared observatory like SPICA, will be sensitive enough to peer through the dust-obscured regions of galaxies and access the rest-frame mid- to far-infrared range in galaxies at high-z. This wavelength range contains a unique suite of spectral lines and dust features that serve as proxies for the abundances of heavy elements and the dust composition, providing tracers with a feeble response to both extinction and temperature. In this work, we investigate how SPICA observations could be exploited to understand key aspects in the chemical evolution of galaxies: the assembly of nearby galaxies based on the spatial distribution of heavy element abundances, the global content of metals in galaxies reaching the knee of the luminosity function up to z ~ 3, and the dust composition of galaxies at high-z. Possible synergies with facilities available in the late 2020s are also discussed.
The present study aimed to explore the molecular factors underlying differences in Calpain/Calpastatin proteolytic system in Nellore and Angus cattle. Longissimus muscle samples were collected in Nellore (n = 6; body weight (BW) = 373 ± 37·3 kg) and Angus (n = 6; BW = 383 ± 23·9 kg) cattle at slaughter for analysis of gene and protein expression, and Calpastatin enzyme activity. Additionally, the myofibrillar fragmentation index was used to quantify the extension of proteolysis in longissimus muscle samples. A greater myofibrillar fragmentation was observed in skeletal muscle of Angus compared with Nellore cattle. Conversely, no differences were found between breeds for mRNA expression of Calpain 1 (CAPN1) and Calpastatin (CAST). Similarly, no differences were observed for the abundance of Calpain and Calpastatin proteins between skeletal muscles of Nellore and Angus cattle. Despite the lack of differences in mRNA and protein abundance, a greater activity of Calpastatin was observed in skeletal muscle of Nellore compared with Angus cattle. These data indicate that the greater proteolysis in skeletal muscle of Angus compared with Nellore cattle is mainly driven by a greater Calpastatin activity rather than Calpain or Calpastatin mRNA and protein expression.
In this study, a Leishmania hypothetical protein, LiHyS, was evaluated regarding its antigenicity, immunogenicity and protective efficacy against visceral leishmaniasis (VL). Regarding antigenicity, immunoblottings and an enzyme-linked immunosorbent assay using human and canine sera showed high sensitivity and specificity values for the recombinant protein (rLiHyS) in the diagnosis of VL. When evaluating the immunogenicity of LiHyS, which is possibly located in the parasite's flagellar pocket, proliferative assays using peripheral blood mononuclear cells from healthy subjects or VL patients showed a high proliferative index in both individuals, when compared to the results obtained using rA2 or unstimulated cultures. Later, rLiHyS/saponin was inoculated in BALB/c mice, which were then challenged with Leishmania infantum promastigotes. The vaccine induced an interferon-γ, interleukin (IL)-12 and granulocyte-macrophage colony-stimulating factor production, which was maintained after infection and which was associated with high nitrite and IgG2a antibody levels, as well as low IL-4 and IL-10 production. Significant reductions in the parasite load in liver, spleen, bone marrow and draining lymph nodes were found in these animals. In this context, the present study shows that the rLiHyS has the capacity to be evaluated as a diagnostic marker or vaccine candidate against VL.