Fraction B2 Research Articles

Platelet activation and inflammation in transgender women using hormone therapy

ObjectiveThe pathophysiological process behind the increased risk of cardiovascular disease (CVD) in transgender women remains to be elucidated. This exploratory study aimed to investigate whether changes in platelet activation and inflammation after the start of feminizing gender-affirming hormone therapy (GAHT) could be a contributing mechanism. DesignLongitudinal cohort study. MethodsVenous blood was collected from 17 transgender women at 0, 12 and 52 weeks after GAHT initiation, consisting of estradiol and testosterone suppression. Platelet activation markers plasma thromboxane B2, Closure Time, CD63, CD62p, platelet-leukocyte complexes and immature platelet fraction were measured. CRP and 11 cytokines were measured as inflammation markers. ResultsCD63, CD62p and platelet-leukocyte complexes tended to increase after 12 weeks of GAHT. After 52 weeks, all platelet activation markers showed anti-aggregatory changes. Eight out twelve inflammation markers exhibited a decreasing tendency at week 12. Equivalently, after 52 weeks, eight inflammation markers tended to decrease, seven of which had also exhibited a decrease at week 12. ConclusionsThe collective findings suggest that platelet activation fluctuates during feminizing GAHT, exhibiting an initial increase followed by a decrease. Additionally, inflammation markers tend to decrease. Within the scope of this study, we could not identify GAHT induced platelet activation as a definite contributing factor in the increased risk of CVD in transgender women. Studies with larger numbers of participants and longer follow-up duration are needed to further investigate the effect of feminizing gender-affirming hormone therapy on platelet activation and inflammation. Trial registrationEudraCT #2017-003072-31.

Nutritional Value Evaluation of Corn Silage from Different Mesoregions of Southern Brazil

Corn silage is widely used in livestock farming; however, its quality is easily altered, and one of the factors that has a high influence in this regard is the region of production. The objective was to evaluate the chemical–bromatological composition of 498 samples of corn silage from mesoregions in Southern Brazil during the 2022/2023 summer harvest. The following were studied in relation to our objective: nutritional composition, dry matter, mineral matter, ether extract, starch, crude protein, neutral detergent fiber, acid detergent fiber, acid detergent lignin, total digestible nutrients, total carbohydrates, and fractions of carbohydrates. The silages from Central South-PR had higher levels of starch and ether extract (30.68% ± 6.24% and 3.41% ± 0.92%, respectively), whereas in West-SC, the silages had higher levels in the A + B1 fraction of carbohydrates (49.59% ± 6.34%). Silages in North-PR had higher concentrations of neutral detergent fiber and acid detergent fiber (49.86% ± 5.92% and 29.70% ± 4.38%, respectively), while in Northwest-RS and West-PR, silages had higher levels of the B2 carbohydrate fractions (46.25% ± 1.98% and 44.55% ± 3.84%, respectively). The nutritional composition differences presented were due to the variables of each mesoregion, interfering in the scenario of formulating diets and animal nutrition.

Effectiveness of near-infrared spectroscopy to predict the chemical composition of feces and total-tract apparent nutrients digestibility estimated with undigestible neutral detergent fiber or acid-insoluble ash in lactating buffaloes' feces

Following a comparison of nutrient total-tract digestibility estimates in lactating buffaloes using single-point undigestible NDF (uNDF) or acid-insoluble ash (AIA) as internal markers, the potential of fecal near-infrared spectroscopy (NIRS) to provide calibration equations for the assessment of the chemical composition of feces and nutrient total-tract digestibility estimated with internal markers was explored. Chemical analyses were performed on 147 fecal samples from lactating buffaloes reared on 5 farms in central Italy (Naples). Each farm fed a silage-based TMR to the buffaloes, and the TMR was sampled in the 2 d before the fecal collection. The TMR and individual fecal samples were collected and analyzed for DM, OM, ash, AIA, ether extract (EE), starch, fiber fractions (amylase-treated NDF without residual ash [aNDFom], amylase-treated NDF inclusive of residual ash [aNDF], ADF without residual ash [ADFom], ADF, hemicellulose, cellulose, ADL, uNDF), N, CP and CP bound to aNDF (NDICP) and to ADF (ADICP). The uNDF content was determined through a 240-h in vitro fermentation and employed, together with AIA as markers, to estimate the total-tract apparent digestibility and total-tract digestibility of DM, OM, ash, N, CP, EE, aNDFom, aNDF, NDIP, ADFom, and ADF, ADIN, ADL, hemicellulose, cellulose, starch, NFC, and the B3 fraction of N (NB3). No correlation was found between DM and OM digestibility estimated with AIA and uNDF as internal markers. Weak correlations were detected for all the other nutients digestibilities, and strong correlations were observed for EE, ADFom, hemicellulose, NDIN, ADIN, NB3, NFC, and starch. The sample set (n = 147) was divided in a calibration set (n = 111) and a validation set (n = 36) to "train" and "validate" the fecal NIRS curve through an external validation process. An estimation usable for preliminary or initial evaluation was obtained for N, CP, and aNDF fecal content. An excellent prediction was obtained for total tract digestibility of ADIN (R2 = 0.90) when estimated with uNDF as the internal marker. The NIRS technology was not able to accurately predict all the other traits and the estimated nutrient digestibility of lactating buffalo diets from fecal spectra.

Chemical Properties, Ruminal Fermentation, Gas Production and Digestibility of Silages Composed of Spineless Cactus and Tropical Forage Plants for Sheep Feeding.

The aim was to evaluate the chemical composition, carbohydrates, protein fractionation and in vitro gas production of silages composed of spineless cactus and tropical forages and their effect on sheep performance. Treatments consisted of silages: corn silage (CS), spineless cactus silage (SCS), spineless cactus + gliricidia (SCG), spineless cactus + buffel grass silage (SCBG) and spineless cactus + pornunça (SCP). Silos were opened 60 days after ensiling, and analyses were carried out. The digestibility test lasted for 36 days, with eight animals per treatment. A completely randomized design was adopted. Considering carbohydrate fractionation, CS, SCS and SCBG silages had higher total carbohydrate content (p = 0.001). The SCS silage presented a higher A + B1 fraction (p = 0.001). The SCBG and SCG silages showed a higher B2 fraction (p < 0.0001) compared to the CS and SCS silages. The SCBG and SCP silages presented a higher C fraction (p = 0.001). For protein fractionation, the SCP and SCG silages showed higher crude protein contents (p = 0.001). The CS and SCS silages showed a higher A fraction (p = 0.001). The SCBG silage presented a higher B1 + B2 fraction (p = 0.001). The SCG silage showed a higher B3 fraction (p = 0.006) compared to SCBG silage. The SCS and SCP silages showed a higher C fraction (p = 0.001). Exclusive SCS silage showed higher in vitro dry matter digestibility (p = 0.001), dry matter degradability (p = 0.001) and total gas production (p = 0.001). The use of the SCBG, SCP and SCG silages to feed sheep increased the dry matter intake (p < 0.001). Sheep fed the SCG silage showed greater dry matter and crude protein digestibility compared to the sheep fed the CS, SCS and SCP silages (p = 0.002). There was a higher water intake (p < 0.001) with the use of the SCS and SCG silages to feed the sheep. The SCP and SCG silages provided a greater intake (p < 0.001) and excretion (p < 0.001) of nitrogen by the animals. Although there were no differences between the treatments for daily gains, lambs that received the spineless cactus-based silage associated with tropical forages showed higher gains (160-190 g/day) than lambs that received CS silage (130 g/day). Thus, the use of spineless cactus associated with buffelgrass, pornunça and gliricidia to prepare mixed silages (60:40) to feed sheep has potential use to feed sheep, with positive effects on nutrient degradation and increases in dry matter intake. Under experimental conditions, we recommend the exclusive use of spineless cactus silage associated with buffel grass, pornunça and gliricidia in feeding sheep in semi-arid regions, as it provides nutrients, water and greater daily gains compared to corn silage.

Effect of different alternative additives on nutritional value, in vitro digestibility, and carbohydrate fractionation of whole-plant sorghum silage

This study aimed to evaluate the impact of different additives on nutritional value, in vitro digestibility, and fractionation of carbohydrates of sorghum silage. The experimental design was completely randomized with four replications and seven treatments: 1) Control (C), whole-plant sorghum with no additives; 2) Microbial (M), whole-plant sorghum + microbial additive (0.2% of original matter); 3) Sugar (S), whole-plant sorghum + brown sugar (4.0% of original matter); 4) Microbial plus sugar additives (MS), whole-plant sorghum + microbial additive (0.2% of original matter) + brown sugar (4.0% of original matter); 5) Whey (W), whole-plant sorghum + whey (3% of original matter); 6) Microbial plus whey additives (MW), whole-plant sorghum + microbial additive (0.2% of original matter) + whey (3% of original matter); 7) Microbial plus sugar plus whey (MSW), whole-plant sorghum + microbial additive (0.2% of original matter) + brown sugar (4.0% of original matter) + whey (3% of original matter). The sorghum was cut 111 days after emergence, in the milky-floury stage, and ensiled in polyvinyl chloride silos, adapted with a Bunsen valve. After 35 days of storage, the silos were opened, and samples were collected for the respective analyzes. The additives only presented differences for dry matter, mineral matter, organic matter, crude protein, ether extract, neutral detergent fiber, acid detergent fiber, total carbohydrates, and A + B1 fraction of the carbohydrates. In the present study, microbial inoculants, brown sugar, whey, and their interactions were effective in improving the chemical profile as well as the soluble carbohydrate fraction of whole-plant sorghum silage.

Chemical constituents and antioxidant activity of Britton's wild petunia (Ruellia brittoniana) flower

Abstract. Sianturi GLR, Trisnawati EW, Koketsu M, Suryanti V. 2023. Chemical constituents and antioxidant activity of Britton's wild petunia (Ruellia brittoniana) flower. Biodiversitas 24: 3665-3672. Britton's wild petunia orkencana ungu (Ruellia brittoniana), which has purple flowers, is widely used as a decorative plant. Isolation and structure determination of the antioxidant compound of R. brittoniana flower were carried out. Extraction was carried out by maceration, and compound isolation was conducted by column chromatography. Structure elucidation was accomplished by FTIR, NMR, and MS Spectrometry. Apigenin (4',5,7-trihydroxyflavone) was isolated from the B1 fraction using chloroform: methanol (10: 1) eluent. Fraction A1, obtained from elution by chloroform: methanol (10: 0), consisted of 24 compounds. Two compounds were identified as 1-hexadecanol and 1-phenyl ethanone, which have antioxidant properties of fraction A1. Fraction B2 was eluted by chloroform: methanol (10:1), consisting of 20 compounds. A compound was identified as 2-methoxy phenol, contributingto the antioxidant properties of fraction B2. The total phenolic content of R. brittoniana flower extract was 1.033 mg GAE/g, and its total anthocyanin content was 16.97%. Fractions A1 and B2 possessed strong antioxidant activities due to their phenolic compounds. Apigenin has three -OH groups in the 4’, 5, and 7 positions; only the -OH group at the 4’ position contributes to its antioxidant activity. Therefore, apigenin has moderate antioxidant activity by DPPH assay. The modification structure of apigenin needs further study to enhance its antioxidant activity. This research shows that R. brittoniana flower can be a source of apigenin, also known to have anticancer properties.

Ensiling and thermic treatment effects on ruminal carbohydrate fermentation and post-ruminal crude protein concentration in partial-crop peas and faba beans

This study was conducted to examine effects of ensiling, toasting or ensiling plus toasting in partial-crop field peas and faba beans harvested each with 375 g dry matter/kg (Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH) scale 79 and 81, respectively) on gas production and post-ruminal crude protein (PRCP) concentration byin vitro incubation in ruminal fluid batch-cultures. The silages made from partial-crop field peas and faba beans (Rostock Model Silages) had a pH of 4.3 and 4.6, respectively, and were not typically lactic acid dominated. The silages remained stable after opening for 100 h (peas) and 168 h (faba beans). Toasting was simulated in a drying oven at 160 °C for 60 min. Post-incubation pH and gas accumulation profiles were little affected by treatment. Ensiling did not alter effective PRCP. Toasting and ensiling plus toasting increased effective PRCP up to 25 and 20%-points in peas and up to 35 and 11%-points in faba beans, respectively. Ensiling increased non-protein nitrogen and soluble protein concentration, whilst toasting decreased soluble protein. Significant correlations existed between protein fraction B3 (neutral detergent-insoluble protein) and effective PRCP (r≥0.84;P&lt;0.05) and fraction C (acid detergent-insoluble protein) and effective PRCP (r≥0.79;P&lt;0.05). Ensiling and toasting both decreased arginine and lysine levels. It was concluded that partial-crop peas and faba beans with BBCH 79 and 81, respectively, can provide readily available nutrients and high-quality fibre in the residual plant. However, preserving by ensiling required balance between the reduction of non-protein nitrogen and fermentability characteristics. Toasting reduced protein solubility and increased PRCP, but it was not clear if PRCP was usable for ruminants or was partially bound into Maillard polymers.

Mineral profile, carbohydrates fractionation, nitrogen compounds and in vitro gas production of elephant grass silages associated with cactus pear

BackgroundThe high amount of soluble carbohydrates and the reduced dry matter content in cactus pear can cause excessive fermentation, resulting in nutrient losses, when it is preserved in the silage form. Thus, the association of cactus pear with elephant grass in the production of mixed silages may reduce nutritional losses during the ensiling process. Thus, the aim was to evaluate the mineral profile, carbohydrates fractionation, nitrogen compounds, and in vitro gas production of elephant grass silages associated with a cactus pear levels (0, 150, 300, and 450 g/kg on dry matter basis). The study was carried out in a completely randomized design, with 4 treatments and 5 replications, totaling 20 experimental silos.ResultsThe increase in cactus pear levels in elephant grass silages composition provided an increase in the contents of K (P = 0.013), Ca (P < 0.001), Mg (P < 0.001), Na (P = 0.001), B (P = 0.044) and Zn (P = 0.016), reduced P (P = 0.039) and promoted a quadratic effect for Fe (P = 0.045) content. The addition of cactus pear levels in elephant grass silages increased the total carbohydrates (P < 0.001) and A + B1 fraction (P = 0.002) and promoted a quadratic effect for B2 fraction (P = 0.032). For nitrogen compounds, the increase in cactus pear levels in elephant grass silages composition reduced the B1 + B2 fraction (P = 0.002) and increased the C fraction (P = 0.007). There was no effect of cactus pear levels on the in vitro gas production of elephant grass silages (P > 0.05).ConclusionsUnder the experimental conditions, the addition of cactus pear in elephant grass silage at levels up to 450 g/kg dry matter does not affect the in vitro gas production, however, it improves the mineral profile, contributes to the increase in the total carbohydrate content and the A + B1 fraction in the silages. Besides this, cactus pear addition reduces the B1 + B2 protein fraction, increasing the fraction C content in the silages, suggesting the necessity to supply an additional soluble nitrogen source for good ruminal functioning.

Fermentation profile, aerobic stability, chemical and mineral composition of silages of mango combined with cocoa pod husk meal

The objective was to evaluate the fermentation profile, aerobic stability, chemical composition, and mineral content of silages of mango combined with cocoa pod husk meal. A completely randomized design was adopted, including four levels (65, 70, 75, and 80%) and five repetitions, totaling 20 experimental silos that were opened after 90 days of sealing. Increasing mango levels in the silages increased the dry matter recovery, total carbohydrates, and fraction B2 of carbohydrates, and reduced gas losses, dry matter, and mineral matter. The quadratic effect was found for pH, buffering capacity, potassium, boron, iron, and nitrogen fractions A and B1 + B2. Using unconventional products such as mango combined with cocoa pod husk meal for silage making can reduce the cost of food supplementation for ruminants, and the environmental contamination.

Relation of phase fraction to superplastic behavior of multi-principal element alloy with a multi-phase structure

An Al0.3CoCrNi medium-entropy alloy was annealed in two conditions to differentiate the phase fraction of nano-scaled B2 and sigma precipitates in FCC microstructure, processed by high-pressure torsion, and subjected to a superplasticity test. The Al0.3CoCrNi with different microstructure exhibited distinct superplastic performance, with maximum elongation of 1900% and 1735% from tensile deformation at 1073 K at the strain rate at 5 × 10−3 s−1 and 10−2 s−1, respectively. The microstructural analysis revealed that the superplasticity is dominantly governed by the grain boundary sliding mechanism with the accommodation of intragranular dislocations. The Al0.3CoCrNi alloy, after superplastic deformation, consists of the mixture of FCC, B2, and sigma phases with different phase fractions. The comparison of this multi-phase structured alloy at two conditions and the sister alloy, Al0.5CoCrFeMnNi high-entropy alloy, suggests that increment of B2 and sigma phase fraction is beneficial to promote high-strain rate superplasticity by limiting the grain growth.

ISOLASI DAN KARAKTERISASI SENYAWA FENOLIK DARI DAUN PUTAT (Planchonia valida Blume) (ISOLATION AND CHARACTERIZATION OF PHENOLIC COMPOUND FROM PUTAT LEAVES (Planchonia valida Blume))

Gallic acid have been isolated from the leaves of the Putat plant (Planchonia valida Blume). This study aimed to isolate and characterize isolates from Putat leaves by phytochemical and FTIR spectrophotometry. The research procedures included extraction by maceration, partitioning by fractionation, vacuum liquid chromatography (KCV), gravity column chromatography (KKG), thin layer chromatography (TLC), and FTIR analysis. A total of 1400 grams of putat leaf powder was macerated to produce 159.243 grams (11.38%) of methanol extract. A total of 101 grams of methanol extract was dechlorophyllated to obtain 21.430 grams (21.21%) of dechlorophyllated extract. The dechlorophyllated extract was partitioned gradually with n-hexane and ethyl acetate to produce n-hexane fractions of 0.341 grams (1.591%), and ethyl acetate of 10.143 grams (47.33%), and methanol of 9.170 grams (42.79%). A phytochemical test showed that in the dechlorophyllated extract, the methanol and ethyl acetate fractions contained alkaloids, phenolics, flavonoids, and terpenoids while the n-hexane fraction contained alkaloids and flavonoids. A total of 7 grams of ethyl acetate fraction was fractionated by vacuum liquid chromatography to produce fractions B1 to B8. The B4 fraction was continued to the separation step by gravity column chromatography so that the fractions S1 to S9 were obtained. The S4 fraction was continued to the separation stage using preparative thin layer chromatography (TLC) to produce isolates S4.1 to S4.5. Isolate S4.1 was tested for purity by two-dimensional TLC showed that the isolate was not pure. The results of the phytochemical test with 5% FeCl3 spray reagent indicated a positive phenolic compound which was strengthened by FTIR characterization of the presence of –OH, -CH aromatic, -CH aliphatic, C=C, C=O and C-O goups. Based on these results, it was concluded that the isolate from the dechlorophyllated extract of Putat leaves belonged to phenolic compounds.

Fractionation analysis and risk assessment of potential toxic elements in reservoir sediments in central China.

The pollution of potential toxic elements (PTEs) in the environment is a persistent issue and draws public attention constantly. However, there is no study comprehensively assessing the PTE pollution of water supply reservoirs in central China. This study determined the total contents and fractions of seven PTEs in 10 water supply reservoirs of central China, and evaluated PTE pollution risks by adopting various models. The results indicated that PTE pollution in water supply reservoirs was lower comparing that in estuary or river sediments, and Mn (manganese) was the predominant PTE in central China reservoirs. Fractionation analysis revealed that for B1 fraction (exchangeable/acid-soluble fraction) of Mn, Zn (zinc) and B2 fraction (reducible fraction) of Pb (lead) was high, while the effective state portion of Cr (chromium) was small. The monomial potential ecological risk indices and Igeo values were low among most samples, indicating overall modest PTE pollution. However, the results of the individual contamination factor (ICF) suggested that Pb might lead to potential health risks to human beings.

Estimation of Nutritive Value and Protein Degradability of Trifolium Repens and Poa Pratensis as the Dominant Pasture Species, Under Simulated Rotational Grazing

Summary A cutting trial was conducted to determine the nutritional value, crude protein (CP) fractions distribution and CP ruminal degradability of white clover (Trifolium repens L.) and Kentucky bluegrass (Poa pratensis L.) as the dominant species of permanent grassland. The legume and grass herbage was obtained in the field experiment carried out on pasture throughout a spring growth season where rotational grazing was simulated using three consecutive cuts, which were compared. The fractionation of CP, according to Cornell Net Carbohydrate and Protein System (CNCPS v6.5), was used to predict protein degradability of the legume and grass forage. A significant (p &lt;0.05) lowering of the CP content in the legume and grass herbage was found, while NDF (neutral detergent fiber) and ADF (acid detergent fiber) significantly increased (p &lt;0.05) as the spring growth season progressed. A significant decrease (p &lt;0.05) of fractions A1 and A2 was noted in the legume and grass herbage, with a significant (p &lt;0.05) increase of CP fractions B2 and C as the vegetation season progressed. A higher rumen degradable protein (RDP) content (p &lt;0.05) of white clover and Kentucky bluegrass herbage was obtained in the first harvest, compared with the second and third one. Significantly larger values (p &lt;0.05) of net energy were observed in the white clover and Kentucky bluegrass forage obtained from the first cut, compared with later regrowths, and white clover was characterized by a higher energy content of 12.4–33.1% (dry matter basis), relative to Kentucky bluegrass.

Distribution pattern and ecological risk assessment of heavy metals in Henan section of the Yellow River

Abstract Heavy metals (HMs) pollution in the environment is a persistent issue and draws public attention constantly. However, there is little research assessing the pollution level of HMs in the Henan section of the Yellow River although the river is one of the most important water supply rivers in Henan province. In this research, the fraction contents of six HMs in the Henan section of the Yellow River were investigated by adopting the European Community Bureau of Reference (BCR) sequential extraction procedure. Moreover, the potential health risk caused by HMs was evaluated by adopting both the geo-accumulation index (Igeo) and ecological hazard index method. The results indicate the presence of a light HM pollution level in the Yellow River generally. However, the health risk caused by Cd cannot be neglected. Fraction analysis indicates that the content of the B2 fraction is high, which needs more attention. Overall, branch afflux might be one of the important pathways through which anthropogenic activities influence the Yellow River. The results of both the Igeo and ecological hazard index indicate that the potential health risk caused by HMs is low in the Yellow River, and the Yellow River protection policies implemented by Henan province may be the underlying reason.

In vitro ruminal fermentation kinetics of diets with crambe cake protein replacing soybean meal protein by gas production technique

The objective of this study was to evaluate ingredients and diets containing increasing levels of crambe cake protein replacing soybean meal protein, with in vitro ruminal fermentation parameters using a gas production technique. Diets were formulated for feedlot lambs and contained different levels of crambe cake protein (0, 250, 500, 750, and 1000 g kg-1) replacing soybean meal protein. Corn silage was used as roughage. Carbohydrate digestion rates were estimated using the in vitro gas production technique and the cumulative gas production kinetics were analyzed using the bicompartmental logistic model. The parameters values of ruminal degradation kinetics were generated using the R statistical program with the Gauss-Newton algorithm and then subjected to analysis of variance and regression (when necessary) according to a completely randomized experimental design with five treatments and five replications. Upon carbohydrate fractionation of ingredients and experimental diets, it was observed that corn grain and corn silage presented the highest levels of total carbohydrates (TC), with values of 128.3 and 464.8 g kg-1 dry matter (DM) in fraction B2, respectively. Lower TC content was found for soybean meal and crambe cake (CC). There was a predominance of fractions A + B1 in the ingredients and experimental diets. The B2 fraction decreased in the diets with the inclusion of the CC protein, and CC presented the highest C fraction. Protein fractionation (g kg-1 DM and g kg-1 crude protein - CP), the ingredients and diets showed a higher proportion of fractions A and B1 + B2. In in vitro degradation, the diet without CC (0 g kg-1 DM) showed the highest final cumulative gas production (365.04 mL g-1 of incubated DM), while the CC presented the lowest volume (166.68 mL g-1 of incubated DM). The gas volume of non-fibrous carbohydrate fermentation and fibrous carbohydrate degradation rate exhibited a quadratic effect according increasing levels of CC (Pmax = 265.8 g kg-1 DM and Pmin = 376.3 g kg-1 DM, respectively). The lag time and final gas volume showed a decreasing linear effect with increasing levels of CC protein. The degradation rate of non-fibrous carbohydrates and the final volume of fibrous carbohydrates did not differ. Replacing soybean meal protein with CC protein at the level of 250 g kg-1 of dry matter in diets formulated for feedlot lambs leads to good profiles of ruminal fermentation kinetics with respect to the degradation of fibrous and non-fibrous carbohydrates.

Effects of nitrogen fertilization on protein and carbohydrate fractions of Marandu palisadegrass

The effects of nitrogen (N) fertilization levels on protein and carbohydrate fractions in Marandu palisadegrass pasture [Urochloa brizantha (Hochst. ex A. Rich.) R.D. Webster] were investigated in a pasture over five years. The experimental design was completely randomized with four levels of N (0, 90, 180, and 270 kg N ha-1, as urea) for five years, and with three replicates. The study was conducted in a continuously stocked pasture during the forage growing season (December to April) in a tropical region. The effects of N fertilization were similar across the five years. With increasing N fertilization, the concentrations of crude protein (CP) increased from 103 to 173 g kg−1 (P < 0.001), soluble fractions (Fraction A + B1) increased from 363 to 434 g kg−1 of total CP (P = 0.006); neutral detergent fiber (NDF) decreased from 609 to 556 g kg−1 (P = 0.037); indigestible NDF (P = 0.046), potentially degradable neutral detergent fiber (P = 0.037), and acid detergent fiber decreased (P = 0.05), and total digestible nutrient (TDN) increased (P < 0.001). Increasing N fertilization decreased the concentrations of Fraction C (P = 0.014) and total carbohydrates (P < 0.0001), and increased CP:organic matter digestibility (P < 0.01). Concentrations of neutral detergent fiber free of ash and protein (P = 0.003), indigestible neutral detergent fiber (P < 0.001), neutral detergent fiber potentially degradable (P = 0.11), CP (P < 0.001), Fraction A + B1 (P < 0.001), Fraction B2 (P < 0.001), Fraction B3 (P < 0.01), and non-structural carbohydrates differed (P < 0.001) across years. Therefore, N fertilization can be used to increase CP, soluble protein, and TDN.

Lopinavir and Nelfinavir Induce the Accumulation of Crystalloid Lipid Inclusions within the Reservosomes of Trypanosoma cruzi and Inhibit Both Aspartyl-Type Peptidase and Cruzipain Activities Detected in These Crucial Organelles.

Several research groups have explored the repositioning of human immunodeficiency virus aspartyl peptidase inhibitors (HIV-PIs) on opportunistic infections caused by bacteria, fungi and protozoa. In Trypanosoma cruzi, HIV-PIs have a high impact on parasite viability, and one of the main alterations promoted by this treatment is the imbalance in the parasite’s lipid metabolism. However, the reasons behind this phenomenon are unknown. In the present work, we observed by transmission electron microscopy (TEM) that the treatment of T. cruzi epimastigotes with the HIV-PIs lopinavir and nelfinavir induced a huge accumulation of crystalloid-shaped lipids within the reservosomes, most of them deforming these key organelles. As previously reported, those structures are characteristic of lipid inclusions formed mostly of cholesterol and cholesterol-esters. The fractionation of nontreated epimastigotes generated two distinct fractions enriched in reservosomes: one mostly composed of lipid inclusion-containing reservosomes (Fraction B1) and one where lipid inclusions were much less abundant (Fraction B2). Interestingly, the extract of Fraction B2 presented enzymatic activity related to aspartyl-type peptidases 3.5 times higher than that found in the extract obtained from Fraction B1. The cleavage of cathepsin D substrate by this class of peptidases was strongly impaired by pepstatin A, a prototypical aspartyl PI, and the HIV-PIs lopinavir and nelfinavir. In addition, both HIV-PIs also inhibited (to a lesser extent) the cruzipain activity present in reservosomes. Finally, our work provides new evidence concerning the presence and supposed participation of aspartyl peptidases in T. cruzi, even as it adds new information about the mechanisms behind the alterations promoted by lopinavir and nelfinavir in the protozoan.

Replacing Alfalfa with Paper Mulberry in Total Mixed Ration Silages: Effects on Ensiling Characteristics, Protein Degradation, and In Vitro Digestibility.

Simple SummaryThe usage of alfalfa (Medicago sativa L.) as a dietary protein source for ruminants in China is limited by forage quality and planting scale. Paper mulberry (Broussonetia papyrifera L., RY) has emerged as a new and representative high-protein woody forage resource for ruminants. However, information is less available regarding how substituting RY for alfalfa affect the fermentation and protein quality in total mixed ration (TMR) silages. This study evaluated ensiling characteristics, protein quality, and in vitro digestibility in TMR silages by mixing RY with alfalfa at different rations. The TMR were made with alfalfa and RY mixtures (36.0%), maize meal (35.0%), oat grass (10.0%), soybean meal (7.5%), brewers’ grain (5.0%), wheat bran (5.0%), premix (1.0%), and salt (0.5%) on a dry matter basis, respectively. The alfalfa and RY mixtures were made in the following ratios of dry matter: 36:0 (RY0), 27:9 (RY9), 18:18 (RY18), 9:27 (RY27), and 0:36 (RY36). The results showed that RY substitution had no adverse effect on fermentation quality and nutritional composition, but inhibited true protein degradation, while decreasing in vitro dry matter and crude protein digestibility. Therefore, RY and alfalfa mixtures at a ratio of 18:18 is suitable for silage-based TMR.To develop an alternative high-protein forage resource to alleviate ruminant feed shortages, we investigated the effects of replacing alfalfa (Medicago sativa L.) with different ratios of paper mulberry (Broussonetia papyrifera L., RY) on fermentation quality, protein degradation, and in vitro digestibility of total mixed ration (TMR) silage. The TMR were made with alfalfa and RY mixtures (36.0%), maize meal (35.0%), oat grass (10.0%), soybean meal (7.5%), brewers’ grain (5.0%), wheat bran (5.0%), premix (1.0%), and salt (0.5%) on a dry matter basis, respectively. The alfalfa and RY mixtures were made in the following ratios of dry matter: 36:0 (RY0), 27:9 (RY9), 18:18 (RY18), 9:27 (RY27), and 0:36 (RY36). After ensiling for 7, 14, 28, and 56 days, fermentation quality, protein degradation, and microbial counts were examined, and chemical composition and in vitro digestibility were analyzed after 56 days of ensiling. All TMR silages, irrespective of the substitution level of RY, were well preserved with low pH and ammonia nitrogen content, high lactic acid content, and undetectable butyric acid. After ensiling, the condensed tannin content for RY18 silages was higher than the control, but non-protein nitrogen, peptide nitrogen, and free amino acid nitrogen contents was lower, while the fraction B1 (buffer-soluble protein) was not different among all the silages. Dry matter and crude protein digestibility for RY27 and RY36 silages was lower than the control, but there was no difference between control and RY18 silages. This study suggested that ensiling RY with alfalfa inhibited true protein degradation, but decreased in vitro dry matter and crude protein digestibility of TMR silages, and that 18:18 is the optimal ratio.

Protein and Carbohydrate Fractions in Warm-Season Pastures: Effects of Nitrogen Management Strategies

Nitrogen (N) management affects herbage production and chemical composition; however, information on the impact of tropical herbage on N and carbohydrate fractions is scarce. A two-year study was conducted to investigate the potential use of pintoi peanut (Arachis pintoi) compared with N fertilization of palisade grass (Brachiariabrizantha cv. Marandu) by evaluating the herbage chemical composition (fractionation of protein and carbohydrate), herbage mass and accumulation rate, herbage disappearance rate, and stocking rate of pastures. The experiment was conducted in a completely randomized design with three treatments, and four replications (paddocks) were used with twenty-one non-lactating crossbred dairy heifers. Treatments consisted of pastures of palisade grass without a N source (control), fertilized with urea (150 kg/ha/year; fertilized), or mixed with pintoi peanut (mixed). Inclusion of the legume increased concentrations of fractions A (p = 0.009), which is the soluble N compound, and B3 (p &lt; 0.001), which is slowly degraded true protein, compared with pastures fertilized with N and non-fertilized pastures. Nitrogen fertilization increased fraction B1 + B2 (p = 0.046), mainly true proteins, and decreased fraction C (p = 0.0007), indigestible protein, and neutral detergent fiber concentrations (p = 0.0003), contributing to increasing the nutritive value of the herbage. Additionally, N fertilization increased herbage mass (p = 0.004) and herbage allowance (p = 0.0001). Both N fertilization and biologically fixed N increased herbage allowance (p = 0.02) and accumulation rate (p = 0.02), as well as the crude protein content of herbage (p &lt; 0.0001) compared with non-fertilized pastures. Nitrogen fertilization increased true protein and decreased indigestible protein of herbage and promoted a greater herbage mass production, while the inclusion of legumes increased soluble protein and decreased the slowly degraded true protein of herbage. Both N management strategies increased herbage allowance and accumulation rate.

Effects of Harvest and Fertilization Frequency on Protein Yield and Extractability From Flood-Tolerant Perennial Grasses Cultivated on a fen Peatland

Paludiculture, and in particular the cultivation of perennial grasses as biomass feedstock for green biorefineries, may be an economic and environmentally sustainable option for agricultural peatlands in temperate regions. However, the optimal biomass quality for protein extraction from flood-tolerant grasses is largely unknown. The aim of this study was to define the combined effect of harvest and fertilization frequency, with one to five annual cuts, on protein yield and extractability for the grasses tall fescue (TF) and reed canary grass (RCG), cultivated on an agricultural fen peatland in Denmark.The content of protein fractions was determined according to the Cornell Net Carbohydrate and Protein System (CNCPS). We assessed protein extractability by lab-scale biorefinery techniques using a screw-press followed by acid precipitation of true protein. The two methods were compared to correlate potential extractable protein yields with actual biorefinery outputs. We found the highest annual biomass and crude protein (CP) yields in the two cut treatments, with 13.4 and 15.6 t dry matter (DM) ha−1 year−1, containing 2.9–3.4 t CP ha−1 year−1for TF and RCG, respectively. The highest neutral-extractable (fractions B1and B2) true protein yields of 1.1 and 1.5 t ha−1 year−1were found in the two cut treatments, representing 39% (TF) - 45% (RCG) of total CP. Using biorefining techniques, we were able to precipitate up to 2.2 t DM ha−1 year−1of protein concentrate, containing up to 39% CP. Significant correlations between methods were found, with a distinct relationship between CNCPS fractions B1+ B2and CP yield of the protein concentrate, indicating the suitability of the CNCPS as an indicator for extractable protein yields. Biomass and CP yields were not significantly improved beyond two annual cuts. However, timing and harvest frequencies significantly affected plant maturity and consequently extractable CP contents and protein concentrate yields. We conclude that TF and RCG are promising feedstocks for green biorefineries due to high biomass, extractable CP, and protein concentrate yields, and highlight the potential of flood-tolerant grasses, cultivated on wet agricultural peatlands, for an enhanced valorisation beyond the common utilisation for bioenergy.