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Métodos de conservação e determinação de triptofano e serotonina em sangue por HPLC-FLD

2026-03-19T22:10:01Z

Title: Métodos de conservação e determinação de triptofano e serotonina em sangue por HPLC-FLD Autor: Markmann, Ryana Cristina Primeiro orientador: Valente, Ériton Egidio Lisboa Abstract: Tryptophan follows three metabolic pathways, including the serotonin pathway (5-HT), which acts in intestinal, immunological, endocrine, and metabolic processes, especially in ruminants. The detection of 5-HT in biological fluids is performed by chromatographic techniques, with HPLC-FLD being promising but still poorly established for blood fractions. As 5-HT is unstable and easily degraded, it requires antioxidants and controlled storage for reliable analysis. This study was conducted with the aim of developing a rapid and accurate methodology for storing and quantifying 5-HT together with tryptophan (TRP) using highperformance liquid chromatography with fluorescence detection (HPLC-FLD) and evaluating temporal variations and variations in blood fractions. The analytical method was isocratic with fluorescence detection. The mobile phase was evaluated with 50 mM potassium phosphate buffer at different pH values (3.5, 4.0, 4.5) and methanol:buffer ratios (25:75, 20:80, 15:85). The stability of analytes in blood serum samples with different preservatives was evaluated for tryptophan and serotonin stability in blood serum samples. To analyze the temporal dynamics of serotonin in blood fractions, an experiment was conducted with six male Holstein cattle (436.8 ± 36.4 kg). Blood samples were collected at time points 0 (before feeding), 6, 12, and 18 hours after the first feeding. The mobile phase composed of 80% KH₂PO₄ buffer (pH 3.5) and 20% methanol showed the best results with excellent correlation for TRP (R² = 1.0000) and 5-HT (R² = 0.9997). The limits of quantification (LOQ) and detection (LOD) found were 14.54 pmol/mL and 4.80 pmol/mL for TRP and 12.27 pmol/mL and 4.058 pmol/mL for 5-HT. The recoveries obtained were 105% for TRP and 103% for 5-HT. Treatment of serum samples with HClO₄ and methanol (40%) resulted in better preservation of TRP and 5-HT stored at -20°C and -80°C for 56 days. Serum 5-HT levels remained stable throughout the day, while tryptophan levels fluctuated. The results demonstrate that the method developed is sensitive and reliable for the quantification of serotonin and tryptophan in biological matrices. The joint addition of HClO4 and methanol allows TRP and 5-HT stored at -80°C for at least 56 days. Publisher: Universidade Estadual do Oeste do Paraná Tipo do documento: Dissertação

Simbiótico na alimentação de matrizes suínas em lactação

2026-02-25T22:29:32Z

Title: Simbiótico na alimentação de matrizes suínas em lactação Autor: Bickel, Amanda Gabriela Primeiro orientador: Carvalho, Paulo Levi de Oliveira Abstract: The supplementation of synbiotics in swine production has become increasingly relevant due to their positive effects on intestinal microbiota modulation and animal health, in addition to their use as performance enhancers. In this context, a study was conducted to evaluate the effect of synbiotic supplementation in diets of lactating sows on productive efficiency, hematological, biochemical and immunological profiles, nutrient digestibility, intestinal health of sows, and the productive performance of their offspring. The experiment involved 84 TN70 sows (Topigs Norsvin®), with an initial body weight of 236.0 ± 22.9 kg and a final body weight of 206.5 ± 27.6 kg, allocated in a randomized complete block design (RCBD), with 21 replicates and four treatments, where the pen containing the sow and her litter was considered the experimental unit. Treatments consisted of: (1) control diet (CD); (2) CD + 0.5 kg synbiotic/ton of feed (CD05); (3) CD + 1.0 kg synbiotic/ton of feed (CD10); and (4) CD + 1.5 kg synbiotic/ton of feed (CD15). The synbiotic consisted of Bifidobacterium bifidum, Bifidobacterium lactis, Enterococcus faecium, Lactobacillus casei, Saccharomyces boulardii, Saccharomyces cerevisiae, galacto-oligosaccharides (GOS), and mannanoligosaccharides (MOS). Sows from the CD15 group showed a higher body condition score at the end of lactation (P = 0.048) compared with those from the CD group, and a lower body condition score loss (P = 0.009). Piglets from sows receiving synbiotic supplementation had greater weaning body weight (P = 0.003) compared with piglets from the CD group. Furthermore, piglets from the CD group showed a higher incidence of diarrhea (P = 0.013) than piglets from the synbiotic groups. Sows from the CD05 group presented lower values of the enzyme aspartate aminotransferase (AST) (P = 0.059) compared with those from the CD group. Females receiving synbiotic supplementation (CD05, CD10, and CD15) exhibited higher values of digestible energy (DE) (P < 0.0001) than those from the CD group. As synbiotic inclusion levels increased, there was an improvement in digestible protein (DP) values of sows (P < 0.0001). In addition, sows from the CD05 and CD15 groups showed a high abundance of bacteria belonging to the Methanobacteriaceae family and the genus Methanobrevibacter, suggesting a potential intensification of methanogenic activity, associated with a more anaerobic intestinal environment and enhanced fermentation of fibrous substrates. In conclusion, synbiotic supplementation in diets of lactating sows improved sow and litter performance, reduced the incidence of diarrhea in piglets, enhanced DE and DP values in sows, and modulated the intestinal microbiota, without negatively affecting other variables. Under the experimental conditions, synbiotics can be used as a performance-enhancing additive during lactation. Publisher: Universidade Estadual do Oeste do Paraná Tipo do documento: Dissertação

Potencial forrageiro de gliricídia (Gliricidia sepium (Jacq.) Steud.) sob diferentes intervalos de corte e sua inclusão em dietas para frangos de corte

2026-01-06T14:13:44Z

Title: Potencial forrageiro de gliricídia (Gliricidia sepium (Jacq.) Steud.) sob diferentes intervalos de corte e sua inclusão em dietas para frangos de corte Autor: Silva, Haroldo Wilson da Primeiro orientador: Neres, Marcela Abbado Abstract: This research was conducted in two complementary trials. In the first trial, the objective was to evaluate the productivity and bromatological composition of gliricidia as a function of different cutting intervals, as well as to characterize its amino acid profile. Initially, the soil was previously prepared with a leveling harrow to uniform the terrain, and samples from the 0-20 cm depth layer were collected for chemical analysis before and after the crop cycle. The gliricidia crop was established from seedlings produced in a greenhouse, which, after sixty days under regular irrigation and subsequent acclimation to sunlight, were transplanted to the field. Planting was carried out in a dense regime, with a spacing of 1.0 m between rows and 0.5 m between plants, totaling 20,000 plants per hectare. At 150 days after planting, a uniformity cut was performed at 50 cm above the ground to standardize the stand, from which subsequent cuts, maintaining the same height, were carried out according to the intervals defined for each treatment. The research was conducted in a Completely Randomized Design (CRD). The treatments consisted of four cutting intervals (45, 60, 75, and 90 days), with seven replications each, totaling 28 experimental units. The experimental unit was the plot, composed of three crop rows with 135 plants each. For the analyses, data from the three rows were summarized by the mean to represent the value of each plot, ensuring data independence for statistical analysis. The data were subjected to analysis of variance (ANOVA), and when significant differences were detected (p<0.05), the treatment means were compared by Tukey's test. Polynomial regression analysis was also performed to evaluate the behavior of the variables as a function of the cutting intervals, selecting the best-fit model based on the coefficient of determination and the significance of the parameters. The trial on the impact of different cutting intervals (45, 60, 75, and 90 days) on the yield and nutritional quality of gliricidia revealed differences. In annual dry matter production, the 60-day interval recorded 233.34 kg, a value 14.6% higher than the 45-day interval (203.62 kg) and 18.3% higher than the 75-day interval (197.25 kg). Compared to the 60-day interval, the 90-day interval (184.40 kg) showed a reduction of 21.0%. Regarding the bromatological composition, the crude protein content decreased with increasing cutting interval: the 32.45% at 45 days reduced to 31.41% (-3.2%) at 60 days, 30.37% (-3.3% compared to the previous) at 75 days, and 28.60% (-5.8%) at 90 days, totaling an 11.9% reduction between the extremes. The total dry matter/crude protein ratio (kg/ha) at the 60-day interval (122.18 kg/ha) was 27.5% higher than the 45-day interval (95.85 kg/ha) and 28.1% higher than the 90-day interval (87.88 kg/ha). The analysis of total crude protein production per hectare (CP/ha), which synthesizes dry matter productivity and its protein content, confirms the superiority of the 60-day cutting interval. It is concluded, therefore, that the 60-day cutting interval was the most balanced, providing the highest dry matter yield and the best dry matter/total protein ratio per hectare. Although the crude protein content is maximized at shorter intervals (45 days), the sharp drop in overall productivity makes management at 60 days more advantageous when the goal is to optimize the joint production of matter and nutrients. In the second trial, the objective was to evaluate the effects of including gliricidia leaf meal in broiler diets on zootechnical performance, carcass characteristics, jejunal morphometry, short-chain fatty acids, and biochemical parameters. The gliricidia meal and flour were produced from leaves and tender stems harvested at four different cutting intervals (45, 60, 75, and 90 days), originating from the first trial of this thesis. For this trial, the cutting intervals did not influence the production of the two derivatives, since all material, after harvest and weighing, was homogenized before processing both products. The material was dried in a forced ventilation oven at 55°C for 72 hours and, in order to increase the protein content, the petioles were manually separated from the leaf blades during the production of the meal, since a previous analysis had proven that the petioles have a lower protein content compared to the leaf blades. Then, the meal was ground in a Willey-type mill with a 2 mm mesh sieve to produce the flour, intended for bird feed and to be subjected to chemical-bromatological and amino acid analysis. The research was conducted with 480 male broiler chickens, Ross lineage, in a completely randomized design, consisting of five levels of gliricidia meal inclusion (0%, 0.75%, 1.5%, 3%, and 4.5%), six replications, and 16 birds per experimental unit. Zootechnical performance parameters (weight gain, feed intake, and feed conversion ratio) were evaluated at 21 and 42 days of age. At the end of the experimental period (42 days), biological samples were collected for specific analyses: jejunal fragments for intestinal histomorphometry (villus height, crypt depth, and absorption area); cecal content for quantification of short-chain fatty acids; and blood for serum biochemical profile (metabolites and liver enzymes), analyzed by automatic spectrophotometry. At 42 days, carcass and cut yields were evaluated, as well as meat quality parameters pH (at 15 minutes and 24 hours post mortem), color (at 15 minutes and 24 hours post mortem), water holding capacity, cooking loss, shear force, and lipid oxidation by thiobarbituric acid reactive substances (0, 30, and 90 days). The collected data were subjected to analysis of variance, with mean comparisons by Dunnett's test and polynomial regression analysis, adopting a significance level of 5%. The inclusion of gliricidia meal in broiler feed presents a dual dose-dependent effect, acting as a pharma-nutrient agent. In the initial phase (1 to 21 days), a reduction in feed intake was observed in treatments with 1.5%, 3%, and 4.5% inclusion, with a minimum value of 863.04 g at the 4.5% level compared to 1182.80 g for the control. The 0.75% level (1142.17 g) did not differ from the control. Regarding weight gain, all inclusion levels resulted in values lower than the control (909.55 g), with the lowest value recorded at the 4.5% level (623.00 g). Feed conversion was worse at the 3% (1.376) and 4.5% (1.386) levels, while the other treatments remained equivalent to the control (1.301). In the total phase (1 to 42 days), a difference in feed intake was found at all experimental levels, including 0.75% (4383 g versus 4614 g for the control). Weight gain was lower in the group with 4.5% inclusion (2466 g versus 3127 g). Feed conversion was worse at the 1.5% level (1.501 versus 1.476 for the control). The productive efficiency index was lower in treatments with 1.5%, 3%, and 4.5% (479, 436, and 404, respectively) compared to the control (524). Carcass yield was lower at the 3% (68.91%) and 4.5% (68.17%) levels, with no differences in the other evaluated parameters. No changes were observed in the values of thiobarbituric acid reactive substances, cooking loss, shear force, water holding capacity, pH, meat color, or cecal short-chain fatty acid concentration. A reduction in serum concentrations of uric acid (up to 3.11 mg/dL) and cholesterol (up to 100.83 mg/dL) was recorded in treatments with 3% and 4.5% inclusion, with no differences at the other levels compared to the control. At doses above 0.75%, its bioactive compounds impair zootechnical performance, although they promote metabolic benefits at higher levels (3% to 4.5%), without affecting meat quality or intestinal integrity. However, for commercial poultry farming, the production losses outweigh any metabolic advantages, making its use unfeasible at the tested levels. Publisher: Universidade Estadual do Oeste do Paraná Tipo do documento: Tese

Blend de proteases na alimentação de leitões em fase de creche

2026-01-06T12:39:18Z

Title: Blend de proteases na alimentação de leitões em fase de creche Autor: Tillmann, Aline Carolina Primeiro orientador: Carvalho, Paulo Levi de Oliveira Abstract: The objective of the study was to evaluate the effects of including a blend of proteases in the diet of piglets during the nursery phase on zootechnical performance parameters, incidence of diarrhea, apparent digestibility of nutrients and energy, hematological and biochemical blood profile, intestinal morphometry, pH and weight of gastrointestinal tract organs, and cecal microbiology. Ninety-six male piglets with high genetic potential were used, weaned at 28 days of age, with an initial body weight of 7.82 ± 0.55 kg. The experiment lasted 35 days, until the animals reached an average body weight of 23.18 ± 3.07 kg. The animals were distributed in a completely randomized design (CRD) with four treatments: PC – Positive control; NC – Negative control, with a 5% reduction in crude protein requirements and 2% reduction in amino acid profile and no protease addition; NC100 – Negative control + 100 g of protease/ton of feed and NC150 – Negative control + 150 g of protease/ton, with eight replicates per treatment, totaling 32 experimental units with three animals each. The protease tested consisted of a blend of fungal and bacterial protease, formulated from Aspergillus niger, Bacillus subtilis, and Bacillus licheniformis. The results showed that animals receiving the NC100 treatment had higher daily weight gain (P = 0.046) when compared to animals consuming NC. Piglets receiving the NC150 treatment had a lower incidence of diarrhea (P = 0.020) when compared to the others. There was a difference in the variables of apparent dry matter digestibility (P = 0.0013) and organic matter digestibility (P = 0.0045), in which animals in the PC treatment had better nutrient digestibility when compared to those in the NC treatment, while animals that consumed the NC100 diet had higher digestible protein (DP) (P <.0001) than the PC and NC treatments. Animals in the NC100 treatment also showed better energy efficiency (P <.0001). Animals in the NC treatment had higher plasma alanine aminotransferase concentrations (P = 0.036). Animals in the NC100 treatment had lower concentrations of glucose (P = 0.033) and triglycerides (P = 0.031) in the bloodstream, while animals in the NC treatment had higher total protein content (P = 0.006) in the blood. The use of protease reduced the incidence of diarrhea, in addition to improving the apparent digestibility of nutrients and blood parameters, and under the experimental conditions, it may be a viable alternative for reducing crude protein in diets without compromising performance. Publisher: Universidade Estadual do Oeste do Paraná Tipo do documento: Dissertação