Which detergent is most suitable for the generation of an acellular pancreas bioscaffold?

Pancreatic bioengineering is a potential therapeutic alternative for type 1 diabetes (T1D) in which the pancreas is decellularized, generating an acellular extracellular matrix (ECM) scaffold, which may be reconstituted by recellularization with several cell types to generate a bioartificial pancreas. No consensus for an ideal pancreatic decellularization protocol exists. Therefore, we aimed to determine the best-suited detergent by comparing sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), and Triton X-100 at different concentrations. Murine (n=12) and human pancreatic tissue from adult brain-dead donors (n=06) was harvested in accordance with Institutional Ethical Committee of the University of São Paulo Medical School (CEP-FMUSP) and decellularized under different detergent conditions. DNA content, histological analysis, and transmission and scanning electron microscopy were assessed. The most adequate condition for pancreatic decellularization was found to be 4% SDC, displaying: a) effective cell removal; b) maintenance of extracellular matrix architecture; c) proteoglycans, glycosaminoglycans (GAGs), and collagen fibers preservation. This protocol was extrapolated and successfully applied to human pancreas decellularization. The acellular ECM scaffold generated was recelullarized using human pancreatic islets primary clusters. 3D clusters were generated using 0.5×104 cells and then placed on top of acellular pancreatic slices (25 and 50 µm thickness). These clusters tended to connect to the acellular matrix, with visible cells located in the periphery of the clusters interacting with the ECM network of the bioscaffold slices and continued to produce insulin. This study provided evidence on how to improve and accelerate the pancreas decellularization process, while maintaining its architecture and extracellular structure, aiming at pancreatic bioengineering.

Shotgun proteomics of detergent-solubilized proteins from Trypanosoma evansi.

Trypanosoma evansi, the causative agent of surra, is the most prevalent pathogenic salivarian trypanosome and affects the majority of domesticated and wild animals in endemic regions. This work aimed to analyze detergent-solubilized T. evansi proteins and identify potential diagnostic biomarkers for surra. Triton X-114-extracted membrane-enriched proteins (MEP) of T. evansi bloodstream forms were analyzed using a gel-free technique (LC-ESI-MS/MS). 247 proteins were identified following the MS analysis of three biological and technical replicates. Two of these proteins were predicted to have a GPI-anchor, 100 (40%) were predicted to have transmembrane domains, and 166 (67%) were predicted to be membrane-bound based on at least one of six features: location (WolfPSORT, DeepLoc-2.0, Protcomp-9.0), transmembrane, GPI, and gene ontology. It was predicted that 76 (30%) of proteins had membrane evidence. Typical membrane proteins for each organelle were identified, among them ISG families (64, 65, and 75 kDa), flagellar calcium-binding protein, 24 kDa calflagin, syntaxins and oligosaccharyltransferase some of which had previously been studied in other trypanosomatids. T. evansi lacks singletons and exclusive orthologous groups, whereas three distinct epitopes have been identified. Data are available via ProteomeXchange with identifier PXD040594. SIGNIFICANCE: Trypanosoma evansi is a highly prevalent parasite that induces a pathological condition known as "surra" in various species of ungulates across five continents. The infection gives rise to symptoms that are not pathognomonic, thereby posing challenges in its diagnosis and leading to substantial economic losses in the livestock industry. A significant challenge arises from the absence of a diagnostic test capable of distinguishing between Trypanosoma equiperdum and T. evansi, both of which are implicated in equine diseases. Therefore, there is a pressing need to conduct research on the biochemistry of the parasite in order to identify proteins that could potentially serve as targets for differential diagnosis or therapeutic interventions.

Biofilm formation by Listeria monocytogenes from the meat processing industry environment and the use of different combinations of detergents, sanitizers, and UV-A radiation to control this microorganism in planktonic and sessile forms.

This study aimed to evaluate the ability of biofilm formation by L. monocytogenes from the meat processing industry environment, as well as the use of different combinations of detergents, sanitizers, and UV-A radiation in the control of this microorganism in the planktonic and sessile forms. Four L. monocytogenes isolates were evaluated and showed moderate ability to form biofilm, as well as carried genes related to biofilm production (agrB, agrD, prfA, actA, cheA, cheY, flaA, sigB), and genes related to tolerance to sanitizers (lde and qacH). The biofilm-forming isolates of L. monocytogenes were susceptible to quaternary ammonium compound (QAC) and peracetic acid (PA) in planktonic form, with minimum inhibitory concentrations of 125 and 75 ppm, respectively, for contact times of 10 and 5 min. These concentrations are lower than those recommended by the manufacturers, which are at least 200 and 300 ppm for QAC and PA, respectively. Biofilms of L. monocytogenes formed from a pool of isolates on stainless steel and polyurethane coupons were subjected to 14 treatments involving acid and enzymatic detergents, QAC and PA sanitizers, and UV-A radiation at varying concentrations and contact times. All treatments reduced L. monocytogenes counts in the biofilm, indicating that the tested detergents, sanitizers, and UV-A radiation exhibited antimicrobial activity against biofilms on both surface types. Notably, the biofilm formed on polyurethane showed greater tolerance to the evaluated compounds than the biofilm on stainless steel, likely due to the material's surface facilitating faster microbial colonization and the development of a more complex structure, as observed by scanning electron microscopy. Listeria monocytogenes isolates from the meat processing industry carry genes associated with biofilm production and can form biofilms on both stainless steel and polyurethane surfaces, which may contribute to their persistence within meat processing lines. Despite carrying sanitizer tolerance genes, QAC and PA effectively controlled these microorganisms in their planktonic form. However, combinations of detergent (AC and ENZ) with sanitizers (QAC and PA) at minimum concentrations of 125 ppm and 300 ppm, respectively, were the most effective.

Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep diets.

Chia (Salvia hispanica L.) seed (CS) and Pumpkin (Cucurbita moschata) seed (PS) are used in ruminant diets as energy sources. The current experiment studied the impact of dietary inclusion of CS and PS on nutrient intake and digestibility, milk yield, and milk composition of dairy sheep. Twelve primiparous Texel × Suffolk ewes [70 ± 5 days in milk (DIM); 0.320 ± 0.029 kg milk yield] were distributed in a 4 × 3 Latin square design and fed either a butter-based control diet [CON; 13 g/kg dry matter] or two diets with 61 g/kg DM of either CS or PS. Dietary inclusion of CS and PS did not alter live weight (p >0.1) and DM intake (p >0.1). However, compared to the CON, dietary inclusion of both CS and PS increased the digestibility of neutral detergent fiber (p <0.001) and acid detergent lignin (p < 0.001). Milk production (p = 0.001), fat-corrected milk (p < 0.001), and feed efficiency (p < 0.001) were enhanced with PS, while the highest milk protein yield (p < 0.05) and lactose yield (p < 0.001) were for CS-fed ewes. Compared to the CON diet, the ingestion of either CS and/or PS decreased (p < 0.001) the C16:0 in milk. Moreover, both CS and PS tended to enhance the content of C18:3n6 (p > 0.05) and C18:3n3 (p > 0.05). Overall short-term feeding of CS and/or PS (up to 6.1% DM of diet) not only maintains the production performance and digestibility of nutrients but also positively modifies the milk FA composition.

Evaluation of internal markers in digestibility estimation for sheep and goats.

The objective of this study was to compare the indirect methods of obtaining digestibility with the direct method of total fecal collection to estimate the apparent digestibility of nutrients in sheep and goats supplemented with non-protein nitrogen. Five goats and five sheep with no defined racial pattern were used, distributed in two 5 × 5 Latin squares, with split plots, considering the diets as plots and the apparent digestibility determination methodologies as subplots. The diets were composed of buffelgrass hay and the addition, via ruminal infusion, of increasing amounts of nitrogen supplementation in order to gradually raise the CP level of the basal diet in intervals of 2% points, that is, + 2, +4, + 6 and + 8%. Samples of the feeds offered, and the leftovers were collected daily during the five days of collection to determine the nutrient intake, as well as the total collection of feces to determine the apparent digestibility of the nutrients. The amount of fecal dry matter excreted was estimated by the concentration of Indigestible Acid Detergent Fiber (ADFi), Indigestible Neutral Detergent Fiber (NDFi), Indigestible Dry Matter at 244 h (DMi 244 h) and Indigestible Dry Matter at 264 h (DMi 264 h). Among the evaluated markers, DMi 264 h had the lowest accuracy in estimating fecal excretion and nutrient digestibility. For the goat species, the markers ADFi and DMi 244 h proved to be able to adequately predict fecal excretion and digestibility indices, while NDFi stood out for both species. Among the evaluated markers, NDFi is the one that most accurately estimates the nutrient digestibility of the diet for goats and sheep.

Assessment of the risk of contamination of enteral nutrition bottles based on the simulation of home use conditions and hygiene procedures.

BACKGROUND: Home-prepared enteral formulations are supplied to patients through enteral nutrition bottles, via a gravity bag or other container, which may be inadequately sanitized and reused more times than recommended by the manufacturer. Such procedures increase the risk of contamination and can compromise the patient's clinical outcome. In light of this, the present study aimed to assess the risk of contamination of enteral nutrition bottles by simulating home use conditions and hygiene procedures. METHODS: A simulation of bottle usage was conducted across the three categories of enteral nutrition (homemade enteral preparations, blended enteral preparations, and commercial enteral formulas) for 3 days, using three hygiene procedures reported by caregivers: use of detergent (DET); use of detergent and boiling water (DET+BW); and use of detergent and bleach (DET+BL). The microbiological contamination was determined by the analysis of aerobic mesophilic microorganisms. RESULTS: The bottles that were used for 3 days, regardless of the enteral nutrition category, were within the acceptable limit for aerobic mesophilic microorganisms (between <4 and 8.0 colony-forming units [CFU]/cm2) when sanitized using the DET+BW and DET+BL procedures. The enteral nutrition bottles, when cleaned using the DET procedure during the 3 days of usage, showed low microbial contamination (between <4 and 3.0 CFU/cm2) in blended preparation and commercial formula only. CONCLUSION: Thus, regardless of the enteral nutrition category, we found that the bottles can be used for 3 days, as long as the DET+BW or DET+BL hygiene procedure is applied and safe food handling measures are adopted.

A proof-of-principle for decontamination of transplantation kidney through UV-C exposition of the perfusate solution.

Kidney transplantation is a common yet highly demanding medical procedure worldwide, enhancing the quality of life for patients with chronic kidney disease. Despite its prevalence, the procedure faces a shortage of available organs, partly due to contamination by microorganisms, leading to significant organ disposal. This study proposes utilizing photonic techniques associated with organ support machines to prevent patient contamination during kidney transplantation. We implemented a decontamination system using ultraviolet-C (UV-C) irradiation on the preservation solution circulating through pigs' kidneys between harvest and implant. UV-C irradiation, alone or combined with ultrasound (US) and Ps80 detergent during ex-vivo swine organ perfusion in a Lifeport® Kidney Transporter machine, aimed to reduce microbiological load in both fluid and organ. Results show rapid fluid decontamination compared to microorganism release from the organ, with notable retention. By including Ps80 detergent at 0.5% during UV-C irradiation 3 log10 (CFU mL-1) of Staphylococcus aureus bacteria previously retained in the organ were successfully removed, indicating the technique's feasibility and effectiveness.

Sodium dodecylbenzene sulphonate (SDBS) present in detergents: action on the gills, skin, and blood of D. rerio fish.

1. Sodium dodecylbenzene sulphonate (SDBS) is one of the surfactants used worldwide in detergents which, due to high residual discharges, has great potential to cause ecotoxicological impacts. Therefore, the sublethal effects of SDBS on the gills and skin of male Danio rerio fish were investigated.2. The fish were distributed into three groups: GC (control), GT1 (0.25 mg/L of SDBS), and GT2 (0.5 mg/L of SDBS) and exposed for 21 days. After the experiment, histopathological analyses of the gills, histochemical analyses (counting of mucous cells), and biochemical analyses (antioxidant defense enzyme analysis, SOD, and CAT) were conducted.3. A significant increase (p < 0.05) in the incidence of circulatory disorders, progressive, and regressive alterations occurred in the GT1 and GT2 groups. Due to these changes, the total histopathological index of the gills was higher in these groups. Mucous cells in the gills and skin increased. There was an increase in SOD activity and a reduction in CAT activity in these groups. Haematology revealed neutrophilia and lymphocytosis in the blood of GT1 and GT2.4. The results clearly demonstrate that a 21-day exposure to SDBS causes severe morphophysiological damage to the gills, skin, and blood of D. rerio fish.

Impact of different Meloidogyne species on the development of sugarcane plants.

This study aimed to evaluate the impact of Meloidogyne javanica, Meloidogyne incognita, and Meloidogyne arenaria on different aspects of the development of sugarcane plants under greenhouse conditions. For this purpose, seedlings of the RB867515 genotype were individually inoculated with 5,000 eggs + second-stage juveniles of their respective nematodes/plant, and non-inoculated plants were used as control. After 330 days of inoculation, the plants were removed from the pots, and the following characteristics were evaluated: fresh mass of the aerial part and root system; leaf area; leaf chlorophyll index; culm diameter; fresh mass of culms; broth volume; contents of neutral and acid detergent fiber, cellulose, hemicellulose, lignin, apparent sucrose in broth, and reducing sugars in broth; total soluble solids concentration. Subsequently, the final population of nematodes in the root system of inoculated plants was determined to calculate the reproduction factor of nematodes. The results showed that all tested Meloidogyne species negatively affected plant development and the composition of some analyzed fractions, in comparison to the non-inoculated control. However, the presence of the root-knot nematode in sugarcane plants increased the contents of neutral and acid detergent fiber, cellulose, hemicellulose, lignin, and reducing sugars, regardless of the Meloidogyne species.

Nutrient intake, productive and metabolic parameters of Nellore bulls feed a forage free diet using cottonseed cake as a fiber source.

We hypothesized that cottonseed cake in confined Nellore young bulls' diet as fiber source, could maintain or improve the nutrient intake, and productive and metabolic parameters. It was evaluated the total replacement of whole plant corn silage (WPCS) by cottonseed cake (CSC) as a source of fiber in the diet. A completely randomized design with two treatments and 12 replications was used. The treatments were two experimental diets containing 300 g/kg of WPCS or CSC as roughage. All animals at 0, 15, 30, 60 and 112 days were weighed to monitor productive performance and nutrient intake. Dry matter intakes, organic matter, neutral detergent fiber, physically effective neutral detergent fiber, crude protein and non-fiber carbohydrates of the WPCS treatment cattle were higher (p < 0.05), and the ethereal extract intake of cattle fed CSC diet was higher. The BW of cattle fed WPCS was higher in the two initial periods of confinement (p < 0.05). Animals from both treatments showed linear growth during the confinement period. There was a fiber source × period interaction for total weight gain (TWG), average daily gain (ADG) and feed efficiency (FE) (p < 0.05), with the lowest in the first 15 days and the highest at 30 days, reflecting a compensatory gain in animals fed with CSC. Bulls fed with CSC showed negative TWG and ADG in the first period (p < 0.05), but recovered in the following periods, surpassing that of the SC treatment in the last two periods. There was negative EF of bulls fed with CSC in the first period (P < 0.05), but in the following periods the CSC diet did not differ from the WPCS diet. The animals fed CSC diet showed a higher concentration of cholesterol and glucose in the blood plasma (p < 0.05). The forage-free diet containing cottonseed cake as a source of fiber replacing corn silage promoted better animal performance.

Effects of leaf and stem maturation on nutritional value in Megathyrsus maximus.

BACKGROUND: Megathyrsus maximus is a forage grass native to Africa but widely cultivated in tropical and subtropical regions of the world where it is part of the grazing food chain. This study aimed to evaluate five M. maximus genotypes for the effect of maturity on their morpho-agronomic traits, nutritional composition and digestibility, and to correlate their leaf blade and stem anatomy with their nutritional value. RESULTS: The proportion of sclerenchyma tissues increased as maturity was reached, while lignin accumulation was differentiated between genotypes. Gatton Panic, Green Panic and Mutale genotypes maintained their acid detergent lignin (ADL) values for leaf blades in the three cuts evaluated. In sacco ruminal dry matter disappearance was lower in Green Panic genotype at the vegetative stage for stems, but not for leaf blades. Significant positive correlations were found between dry matter disappearance and mesophyll tissues, and the latter were negatively correlated with neutral detergent fiber (NDF) and ADL. CONCLUSION: Our results strongly indicate that cutting age and genotype affected the nutritional value of M. maximus leaf blades and stems, with a more pronounced loss of quality in stems than in leaf blades. We recommend increasing the frequency of grazing at early stage or anticipating the stage of stem elongation in Green Panic to produce forage with better nutritional value. © 2023 Society of Chemical Industry.

Influence of different growing stages of whole-plant soybeans on their nutrient content and silage quality for cattle.

The objective of this study was to determine the effects of growing stage (GS) on morphological and chemical composition of whole-plant soybean (WPS), and fermentative profile and chemical composition of whole-plant soybean silage (SS). This study was divided into two trials conducted in a complete randomised block design. The first trial evaluated the effect of GS from R1 to R8 (59-135 d after sowing) on morphological and chemical composition of WPS and its botanical components. The second trial determined the effects of GS from R3 (71 d after sowing) to R7 (124 d after sowing) on dry matter (DM) losses, fermentative profile, chemical composition and aerobic stability of SS. The proportion of leaves in WPS was reduced, while stem and pod proportions were increased as the GS progressed. Ensiling WPS at R6 and R7 decreased the contents of acetic acid, lactic acid and branched-chain fatty acids, and ethanol, and increased the contents of propionic acid and NH3-N. However, silage butyric acid concentrations in R6 and R7 were relatively high (18.1 and 19.9 g/kg DM, respectively). Butyric acid and buffering capacity varied according to GS with the lowest values observed in silages derived from GS R3, R5 and R7, and the highest values observed in silages made from GS R5. Later GS resulted in greater contents of DM, crude protein and ether extract, and lower contents of acid detergent fibre and non-fibre carbohydrate in SS. The high fat of SS produced from later GS limits high inclusion levels in ruminant diets. Morphological components impacted chemical composition of SS, whereas the R7 stage improved fermentative profile and resulted in an SS with greater in situ degradability of DM and neutral detergent fibre.

Isolation and Characterization of Thermus thermophilus Strain ET-1: An Extremely Thermophilic Bacterium with Extracellular Thermostable Proteolytic Activity Isolated from El Tatio Geothermal Field, Antofagasta, Chile.

The present study describes the isolation of an extremely thermophilic bacterium from El Tatio, a geyser field in the high planes of Northern Chile. The thermophile bacterium named Thermus thermophilus strain ET-1 showed 99% identity with T. thermophilus SGO.5JP 17-16 (GenBank accession No. CP002777) by 16S rDNA gene analysis. Morphologically, the cells were non-sporeforming Gram-negative rods that formed colonies with yellow pigmentation. This strain is able to proliferate between 55 and 80 °C with a pH range of 6-10, presenting an optimum growth rate at 80 °C and pH 8. The bacterium produces an extracellular protease activity. Characterization of this activity in a concentrated enzyme preparation revealed that extracellular protease had an optimal enzymatic activity at 80 °C at pH 10, a high thermostability with a half-life at 80 °C of 10 h, indicating that this enzyme can be classified as an alkaline protease. The proteolytic enzyme exhibits great stability towards chelators, divalent ions, organic solvents, and detergents. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF), implying that it was a serine protease. The high thermal and pH stability and the resistance to chelators/detergents suggest that the protease activity from this T. thermophilus. strain could be of interest in biotechnological applications.

Detergent-free parasite transformation and replication assay for drug screening against intracellular Leishmania amastigotes.

Leishmaniasis is an infectious disease caused by protozoan species in the genera Leishmania and Endotrypanum. Current antileishmanial drugs are limited due to adverse effects, variable efficacy, the development of resistant parasites, high cost, parenteral administration and lack of availability in endemic areas. Therefore, active searching for new antileishmanial drugs has been done for years, mainly by academia. Drug screening techniques have been a challenge since the intracellular localization of Leishmania amastigotes implies that the host cell may interfere with the quantification of the parasites and the final estimation of the effect. One of the procedures to avoid host cell interference is based on its detergent-mediated lysis and subsequent transformation of viable amastigotes into promastigotes, their proliferation and eventual quantification as an axenic culture of promastigotes. However, the use of detergent involves additional handling of cultures and variability. In the present work, cultures of intracellular amastigotes were incubated for 72 h at 26 °C after exposure to the test compounds and the transformation and proliferation of parasites took place without need of adding any detergent. The assay demonstrated clear differentiation of negative and positive controls (average Z´ = 0.75) and 50% inhibitory concentrations of compounds tested by this method and by the gold standard enumeration of Giemsa-stained cultures were similar (p = 0.5002) and highly correlated (r = 0.9707). This simplified procedure is less labor intensive, the probability of contamination and the experimental error are reduced, and it is appropriate for the automated high throughput screening of compounds.

Bamboo forage in Peruvian Amazon: a potential feed for cattle.

During the dry and rainy seasons of the Northeastern Zone of Peru, a chemical characterization of five species of bamboo prevalent in the area (Guadua lynnclarkiae, G. takahashiae, Bambusa vulgaris, G. weberbaueri, and Dendrocalamus asper) was conducted. Then, the effect of supplementing bamboo leaves (0, 20, and 40% inclusion of D. asper) on the intake and live weight gain of 18 Gyr × Holstein heifers was evaluated for 28 days. Among the species evaluated, D. asper has the greatest crude protein (CP) concentration (158-166 g/kg Dry matter- DM), post-ruminal CP supply (127 g/kg DM), and in vitro organic matter digestibility (444-456 g/kg DM) but similar concentrations of crude ash (124 g/kg DM), calcium (2.4-2.8 mg/g), phosphorus (0.7-2.1 mg/g), protein fractions A, B1, B2, B3, C (45, 5, 35, 56, and 17g/kg DM, respectively), rumen-undegraded CP (31% CP), neutral detergent fiber (NDF, 685g/kg DM), and acid detergent fiber (ADF, 357 g/kg DM) than the other species evaluated. Dry matter intake was higher in the control treatment and in the 20% bamboo leaf inclusion treatment than in the 40% bamboo inclusion treatment. Intake of CP and NDF decreased with the increase in bamboo inclusion. Despite the differences in DM, CP, and NDF intake, the live weight gain remained similar across treatments. However, there was a greater feed conversion in the 20% bamboo leaf inclusion treatment. During the dry season, bamboo leaves can be used as an alternative supplement at a maximum inclusion of 20% without affecting the live weight gain.

Susceptibility of Cu(I) transport ATPases to sodium dodecyl sulfate. Relevance of the composition of the micellar phase.

Sodium dodecyl sulfate (SDS) is a well-known protein denaturing agent. A less known property of this detergent is that it can activate or inactivate some enzymes at sub-denaturing concentrations. In this work we explore the effect of SDS on the ATPase activity of a hyper-thermophilic and a mesophilic Cu(I) ATPases reconstituted in mixed micelles of phospholipids and a non-denaturing detergent. An iterative procedure was used to evaluate the partition of SDS between the aqueous and the micellar phases, allowing to determine the composition of micelles prepared from phospholipid/detergent mixtures. The incubation of enzymes with SDS in the presence of different amounts of phospholipids reveals that higher SDS concentrations are required to obtain the same degree of inactivation when the initial concentration of phospholipids is increased. Remarkably, we found that, if represented as a function of the mole fraction of SDS in the micelle, the degree of inactivation obtained at different amounts of amphiphiles converges to a single inactivation curve. To interpret this result, we propose a simple model involving active and inactive enzyme molecules in equilibrium. This model allowed us to estimate the Gibbs free energy change for the inactivation process and its derivative with respect to the mole fraction of SDS in the micellar phase, the latter being a measure of the susceptibility of the enzyme to SDS. Our results showed that the inactivation free energy changes are similar for both proteins. Conversely, susceptibility to SDS is significantly lower for the hyperthermophilic ATPase, suggesting an inverse relation between thermophilicity and susceptibility to SDS.

Metabolic responses and performance of Holstein × Gyr heifers grazing Brachiaria decumbens supplemented with varied crude protein levels.

We aimed to evaluate the effect of supplemental CP on the nutritional characteristics and performance of Holstein × Gyr crossbreed heifers grazing intensively-managed Brachiaria decumbens throughout the year. Thirty-eight heifers with average initial body weight of 172.5 ± 11.15 kg (mean ± SE) and 8.2 ± 0.54 mo of age were randomly assigned to four treatments: three protein supplements (SUP) composed of soybean meal and ground corn fed at 5g/kg of BW, plus a control group (CON). The supplements had 12, 24 and 36% of CP for treatments S12 (n = 9), S24 (n = 10), and S36 (n = 9), respectively. The experiment latest one year, subdivided into four seasons: rainy, dry, rainy-dry transition (RDT), and dry-rainy transition (DRT). Feces and pasture samples were collected for 4 days in each season, using chromium oxide, titanium dioxide, and indigestible neutral detergent fiber (NDF) to estimate fecal excretion, supplement, and pasture intake, respectively. The data were analyzed using PROC GLIMMIXED of the SAS with repeated measures. No effects of supplementation were detected on pasture and NDF intake. However, SUP animals had a greater intake of DM, metabolizable energy, and metabolizable protein. A positive linear response on metabolizable protein intake was observed among SUP animals. We observed an interaction between treatment and season for all digestibility variables, with a positive linear response in CP digestibility among SUP animals during all seasons. For neutral detergent fiber (NDF) digestibility, we observed a positive linear response in RDT and rainy seasons and a quadratic response during the dry season. Furthermore, SUP animals had greater average daily gain (ADG) than non-supplemented animals, and among SUP animals, there was a quadratic response to ADG, with the greatest gain observed in S24. We observed greater nitrogen retention coefficient in SUP animals than in non-supplemented animals and a positive linear effect among SUP animals. Supplemental CP did not affect microbial protein production and efficiency. We observed an interaction between treatment and season for blood glucose, with SUP animals having greater glucose concentration in all seasons than non-supplemented animals. Additionally, we observed a quadratic response among SUP animals only during RDT and dry season, with the greatest glucose concentration in S24. SUP animals had greater blood concentrations of urea and IGF-1. In conclusion, SUP animals had greater intake, digestibility, and performance than non-supplemented animals, with the 24% CP supplement demonstrating the best metabolic responses and performance.

Perfusion and Ultrasonication Produce a Decellularized Porcine Whole-Ovary Scaffold with a Preserved Microarchitecture.

The application of decellularized scaffolds for artificial tissue reconstruction has been an approach with great therapeutic potential in regenerative medicine. Recently, biomimetic ovarian tissue reconstruction was proposed to reestablish ovarian endocrine functions. Despite many decellularization methods proposed, there is no established protocol for whole ovaries by detergent perfusion that is able to preserve tissue macro and microstructure with higher efficiency. This generated biomaterial may have the potential to be applied for other purposes beyond reproduction and be translated to other areas in the tissue engineering field. Therefore, this study aimed to establish and standardize a protocol for porcine ovaries' decellularization based on detergent perfusion and ultrasonication to obtain functional whole-ovary scaffolds. For that, porcine ovaries (n = 5) were perfused with detergents (0.5% SDS and 1% Triton X-100) and submitted to an ultrasonication bath to produce acellular scaffolds. The decellularization efficiency was evaluated by DAPI staining and total genomic DNA quantification. ECM morphological evaluation was performed by histological, immunohistochemistry, and ultrastructural analyses. ECM physico-chemical composition was evaluated using FTIR and Raman spectroscopy. A cytocompatibility and cell adhesion assay using murine fibroblasts was performed. Results showed that the proposed method was able to remove cellular components efficiently. There was no significant ECM component loss in relation to native tissue, and the scaffolds were cytocompatible and allowed cell attachment. In conclusion, the proposed decellularization protocol produced whole-ovaries scaffolds with preserved ECM composition and great potential for application in tissue engineering.

Does macauba cake alter nutrient digestibility and microorganism population in the rumen of sheep?

Twenty Santa Inês male sheep were used in a randomized block design to evaluate the effects of different levels of macauba cake (MC) on nutrient digestibility and the population of microorganism in the rumen. The animals were divided into four groups according to MC levels (0, 10, 20, and 30% of DM) and initial body weight ranging from 32.75 to 52.17 kg. Diets were isonitrogenous and formulated to attend metabolizable energy requirements, and feed intake was regulated with 10% allowance for leftovers. Each experimental period lasted 20 days, with the final 5 days reserved for sample collection. Macauba cake inclusion did not affect the dry matter, organic matter, or crude protein intake but increased the ether extract, neutral detergent fiber, and acid detergent fiber intake, mainly because of changes in the concentrations of these components in diets with a higher level of MC. With MC inclusion, a linear decreasing effect was observed for the dry matter and organic matter digestibility, and a quadratic effect with a maximum point of 21.5% was observed for the acid detergent fiber digestibility. A relative reduction of 73% in anaerobic fungal populations was observed with the lowest level of MC inclusion, and a relative increase of 162% in methanogenic populations was observed with the highest level of MC inclusion. The increasing level of macauba cake up to 30% of the diet of lambs reduced the dry matter digestibility and reduced anaerobic fungi but increased methanogenic population.

Effect of physically effective neutral detergent fiber on nutrient intake and digestibility, ruminal and blood parameters, and ingestive behavior of confined beef cattle.

We aimed to evaluate the effect of levels of physically effective neutral detergent fiber from forage (NDFfor) on confined cattle's metabolism and ingestive behavior. Four crossbred steers (514.0 ± 4.54 kg BW) rumen-cannulated were used. The animals were randomly distributed in a 4×4 Latin square design, and the treatments were diets with 9.5%, 5.5%, 2.5%, and 0.0% NDF from whole plant corn silage. The trial was divided into 4 periods of 21 days. The dry matter, organic matter (OM), crude protein, neutral detergent fiber (NDF), physically effective NDF 8mm (peNDF8mm) and NDF1.18mm intakes, and the digestibility of OM and NDF showed a quadratic behavior. The rumen pH values showed a decreasing linear, and the time at pH below 5.8 increased linearly in the diets with lower NDFfor. The production of volatile fatty acids and the proportions of propionate and butyrate showed an increasing quadratic behavior. On the other hand, the proportion of acetate fitted a decreasing quadratic equation. With the lower participation of forage in the diets, the time spent on rumination activity decreased in a quadratic way, and the time spent in idleness increased in a quadratic way. The corn silage can be reduced to 135 g/kg DM in the diet to obtain a minimum of 5.5% of NDF from roughage.