Effects of Supplementation of Zinc, Manganese, or Copper and Different Phytase Levels in Serum and Bone Acid and Alkaline Phosphatases of Broiler Chicks

The knowledge of biological trace minerals and phytase requirements for modern broiler genotypes is not established and the pressure to reduce their usage in animal feeding due to environmental issues is increasing. Here, the alkaline phosphatase (ALP) and tartarate-resistant acid phosphatase (TRAP) of the tibia and serum of broilers fed with diets containing various levels of phytase and reduced levels of zinc, manganese, and copper was evaluated. The experiment was performed using 1,200 male Cobb broilers raised according to standard commercial husbandry techniques. Data were analyzed as a 4×3 factorial arrangement with four concentrations of zinc (0.34, 0.49, 0.64, and 0.79 ppm), manganese (0.18, 0.43, 0.68, and 0.93 ppm), or copper (0.09, 0.12, 0.15, and 0.18 ppm) and three concentrations of phytase (0, 500, and 1,000 FTU/kg) for age periods of 1-21 and 36-42 days. While the dietary supplementation with copper did not induce a significant effect in bone tissue biochemical markers, serum TRAP activity of 42-day old broilers increased with higher copper levels. Increasing dietary zinc levels linearly increased ALP activity in tibia growth, suggesting that zinc is essential for longitudinal bone growth. Phytase significantly promoted the increase of TRAP and ALP activities, suggesting that manganese increased growth plate activity, accelerated calcification, and remodeled the newly formed tissue into trabecular bone. Although not every enzymatic activity was affected by the treatments, the phytase use, along with trace minerals, improved the animal response to the rapid growth required nowadays and provided the nutrients for adequate bone metabolism.(AU)

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT-Raman spectroscopy / Cultivo de células-tronco mesenquimais de rato sobre arcabouços de PHBV-PCL: análise do meio de cultura condicionado por espectroscopia de FT-Raman

Abstract Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37oC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

Resumo As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a análise. Não houve variações nas leituras entre as amostras estudadas, concluindo-se que a FT-Raman não atendeu às expectativas nas condições estudadas.

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT Raman spectroscopy / Cultivo de células-tronco mesenquimais de rato sobre arcabouços de PHBV-PCL: análise do meio de cultura condicionado por espectroscopia de FT-Raman

Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37ºC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a [...].

Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT Raman spectroscopy / Cultivo de células-tronco mesenquimais de rato sobre arcabouços de PHBV-PCL: análise do meio de cultura condicionado por espectroscopia de FT-Raman

Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37ºC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.(AU)

As células-tronco mesenquimais (MSCs) possuem grande potencial para aplicação em procedimentos terapêuticos ligados a terapia celular e engenharia de tecidos, considerando-se a plasticidade e capacidade de formação em diferentes tipos celulares por elas. Dada a abrangência no emprego das MSCs, há necessidade de se compreender melhor algumas propriedades relacionadas à diferenciação osteogênica, particularmente liga à biomateriais usados em engenharia de tecidos. Este projeto objetiva o desenvolvimento de uma metodologia de análise empregando-se a FT-Raman para identificação e quantificação de componentes bioquímicos presentes em meios de cultura condicionados por MSCs, com ou sem indução à diferenciação osteogênica. Todos os experimentos foram realizados entre as passagens 3 e 5. Para essas análises, as MSCs foram cultivadas sobre arcabouços de polímeros biorreabsorvíveis de poli (hidroxibutirato-co-hidroxivalerato) (PHBV) e o poli (ε-caprolactona) (PCL). As MSCs (GIBCO®) foram inoculadas nos polímeros puros e na mistura 75:25 de PHBV / PCL (amostras densas e porosas). As células foram mantidas em DMEM (com baixa glicose) contendo GlutaMAX® e 10% de SFB a 37oC com 5% de CO2 por 21 dias. A própria placa foi usada como controle. Os meios de cultura condicionados foram coletados e analisadas em FT-Raman para sondagem de grupos funcionais, bem como possíveis variações moleculares associadas com a diferenciação e metabolismo celular. Foi possível discernir grupos funcionais de moléculas específicas no meio condicionado, como colesterol, fosfatidilinositol, triglicerídeos, forma Beta de polipeptídeos, regiões de amida e ligações de hidrogênio de proteínas, além da expressão de DNA. Na presente avaliação, a FT-Raman apresentou como uma técnica de resolução limitada, uma vez que modos vibracionais de estiramento próximos ou mesmo iguais podem ser expressos por moléculas diferente, dificultando a [...].(AU)

Effect of strength training versus raloxifene on bone weight, blood glucose, lipid and antioxidant profile in ovariectomized rats

We compared the effect of the treatment with strength training (ST) and raloxifene (RALOX) on bone weight, blood glucose, lipid, and antioxidant profile in ovariectomized rats. Twenty-four Wistar rats were distributed into four groups: ovariectomy + VEHICLE (control); ovariectomy + RALOX; ovariectomy + ST; ovariectomy + RALOX + ST. Thirty days after ovariectomy, the animals underwent the treatment with RALOX (750 µcg day-1) and/or ST (three sessions week-1). Thirty days after, all groups were scarified, tibia and femur were weighed, and the blood was collected for analysis of the lipid profile, glucose, and antioxidants catalase (CAT) and glutathione (GSH). The ST group showed greater femur weight (0.82 ± 0.18 g) and RALOX + ST had greater tibia weight (0.61± 0.17 g) than CONTROL with femur weight of 0.65 ± 0.08 g and tibia of 0.49 ± 0.08 g with no differences between treatments (p > 0.05). ST group showed significantly higher catalase (181.7 ± 15.4 µM g-1) compared to the other groups. In contrast, the GSH value was lower in ST group (89.2 ± 8.1 µM g-1) compared to RALOX (175.9 ± 17.1 µM g-1) and RALOX + ST (162.8 ± 12.1 µM g-1), but the values of these two groups did not differ from CONTROL(115.3 ± 21.1 µM g-1). Total cholesterol did not differ between groups (p > 0.05), but exercise alone(54.3 ± 2.5 mg dL-1) or with RALOX (53.0 ± 1.5 mg dL-1) resulted in higher HDL cholesterol than CONTROL (45.5 ± 2.5 mg dL-1). Only RALOX+ST presented lower glucose (140.3 ± 9.7 mg dL-1) values than CONTROL (201.7 ± 30.6 mg dL-1). In conclusion, ST promotes similar benefits on bone and metabolic parameters compared to pharmacological treatment in ovariectomized rats.(AU)

Different Calcium Levels and Two Limestone Granulometries in the Diet of Laying Hens: Performance and Bone Characteristics

The effect of different calcium levels and two limestone granulometries on performance, egg quality, biometry of digestive organs and bone characteristics of light laying hens were evaluated. A total of 270 laying hens were used during 112 days, distributed in a completely randomized design in a 3 x 2 factorial scheme, with 3 levels of calcium (3.8, 4.0 and 4.2%) and 2 limestone granulometries (0.222 and 1.922 mm), totaling 6 treatments with 5 repetitions each. The parameters evaluated were: feed intake, egg production, egg weight and mass, feed conversion per mass and per dozen eggs, albumen, yolk and shell percentage, specific gravity of eggs and shell thickness. In addition, the relative weight of the heart, proventricle, gizzard, liver, pancreas, intestine weight and length, and bone characteristics were also evaluated. A significant interaction was observed for tibia mineral matter. Egg production was influenced by the limestone granulometry, presenting greater value in the treatments which contained fine granulometry limestone. Higher calcium levels of 4.0 and 4.2% provide greater specific density of eggs. Biometric variables and bone characteristics were not influenced by the treatments. In conclusion, it is recommended to use fine-grained limestone (0.222 mm) and a 4% calcium level in diets for light commercial hens, as they improve performance and quality of eggs, without interfering in biometrics of digestive organs and bone characteristics.(AU)

The mechanism study of inhibition effect of prepared Radix Rehmanniainon combined with Radix Astragali osteoporosis through PI3K-AKT signaling pathway

Purpose: To observe the mechanism of prepared Radix Rehmanniainon combined with Radix Astragali in treating osteoporosis. Methods: Osteoporosis rat model was established by bilateral ovariectomy combined with low-calcium diet feeding. Bone mineral density was measured by bone densitometer. Bone metabolism markers in serum were detected by enzyme linked immunosorbent assay (ELISA), bone tissue structure was observed by hematoxylin-eosin staining, and the effect of prepared Radix Rehmanniainon combined with Radix Astragali on PI3K-AKT signaling pathway was investigated by immunohistochemistry and reverse transcription polymerase chain reaction. Results: Compared with the model group, the bone tissue structure and imbalance of bone metabolism were improved, and the bone mineral density was significantly increased in the prepared Radix Rehmanniainon combined with Radix Astragali groups. After intervention with prepared Radix Rehmanniainon combined with Radix Astragali, the positive expression of PIK3CA and Akt1 in rat bone tissue was enhanced, and the expression levels of Akt1 mRNA were significantly increased. Conclusions: Prepared Radix Rehmanniainon combined with Radix Astragali may treat osteoporosis by activating PI3K/AKT pathway.

Calcium Requirement and Vitamin D Supplementation in Meat-Type Quail at First Stage of Growth

Currently, feed formulations for quail are based on foreign data or data from other species; this undermines productivity with the use of excessive or deficient levels of some nutrients. Therefore, this study aimed to determine the nutritional requirement levels of calcium (Ca) and supplementation vitamin D in meat-type quail from 1 to 14 d of age. The experiment was a 4 × 4 factorial arrangement of treatments (levels of Ca: 0.42, 0.58, 0.74 and 0.90% and levels of vitamin D: 1,000, 2,000, 3,000 and 4,000 IU) with 3 replicate pens and 43 quails per pen. Weight gain increased quadratically with increased levels of Ca and vitamin D; this variable yielded estimated levels of 0.73% Ca and 2,883 IU vitamin D. There was no Ca x vitamin D interaction effect on bone variables. Bone Ca, bone phosphorus, bone ash, femoral bone strength, tibial bone strength and the femoral Seedor index increased quadratically with an increase in Ca levels. These variables yielded estimated values of 0.78, 0.71, 0.78, 0.77, 0.83, and 0.71% Ca, respectively. It is concluded that the requirements of Ca and vitamin D for the maximum weight gain of meat-type quails, in the period from one to 14 days of age, were 0.73% Ca (p=0.001) and 2,883 IU vitamin D (p=0.02), respectively.(AU)

Calcium Requirement and Vitamin D Supplementation in Meat-Type Quail at First Stage of Growth

Currently, feed formulations for quail are based on foreign data or data from other species; this undermines productivity with the use of excessive or deficient levels of some nutrients. Therefore, this study aimed to determine the nutritional requirement levels of calcium (Ca) and supplementation vitamin D in meat-type quail from 1 to 14 d of age. The experiment was a 4 × 4 factorial arrangement of treatments (levels of Ca: 0.42, 0.58, 0.74 and 0.90% and levels of vitamin D: 1,000, 2,000, 3,000 and 4,000 IU) with 3 replicate pens and 43 quails per pen. Weight gain increased quadratically with increased levels of Ca and vitamin D; this variable yielded estimated levels of 0.73% Ca and 2,883 IU vitamin D. There was no Ca x vitamin D interaction effect on bone variables. Bone Ca, bone phosphorus, bone ash, femoral bone strength, tibial bone strength and the femoral Seedor index increased quadratically with an increase in Ca levels. These variables yielded estimated values of 0.78, 0.71, 0.78, 0.77, 0.83, and 0.71% Ca, respectively. It is concluded that the requirements of Ca and vitamin D for the maximum weight gain of meat-type quails, in the period from one to 14 days of age, were 0.73% Ca (p=0.001) and 2,883 IU vitamin D (p=0.02), respectively.

Standardized ileal phosphorus digestibility of meat and bone meal and poultry byproduct meal for broilers

Two experiments were conducted using direct method to determine the standardized ileal phosphorus (P) digestibility coefficient (SIPDC) of meat and bone meal (MBM) and poultry byproduct meal (PBM) for broilers. In the first experiment, three MBM from different batches were evaluated, with four treatments (MBMI, MBMII, MBMIII, and a P-free diet) and nine replicates of seven birds each, totaling 252 birds distributed across a completely randomized design. The P-free diet was used to determine endogenous P losses. Titanium dioxide (5 g kg−1) was used as an indigestible indicator. In the second experiment, three PBM from different batches were evaluated, with 189 birds distributed over three treatments (PBMI, PBMII, and PBMIII) and nine replicates of seven birds each. These two experiments were conducted when birds were 19-22 days old. The birds were euthanized to collect the ileal content. Using the P-free diet, endogenous P losses were determined at 94.9 mg kg−1 dry matter intake. Standardized ileal P digestibility of MBMI (0.65) and MBMIII (0.69) was higher compared with MBMII (0.49). The SIPDC determined for three PBM sources were similar and 0.96, 0.96, and 0.93 for PBMI, PBMII, and PBMIII, respectively. The present study demonstrates that P in MBM is not as available as it is believed and that the variability between batches should be considered by nutritionists, while PBM can be considered an excellent P source to broiler diets.

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin

ABSTRACT Purpose Herein we evaluated the effects of platelet concentrate (PC) and platelet-poor plasma (PPP) on bone repair using noncritical defects in the calvaria of rabbits and compared them to the presence of TGF-β1 and osteocalcin on reparative sites. Methods Five noncritical defects of 8.7 mm in diameter were created on the calvaria of 15 animals. Each defect was treated differently, using autograft (ABG), ABG associated with PC (ABG + PC), ABG with PPP (ABG + PPP), isolated PPP, and blood clot (control). The animals were submitted to euthanasia on the second, fourth and sixth week post-surgery. Results The defects that received ABG+PC or PPP demonstrated lower bone formation when compared to specimens that received ABG in the same period. These results coincided to significant higher immunopositivity for TGF-β1 for specimens that received PC, and lower presence of cytokine in the group PPP. However, either higher or lower presence of TGF-β1 were also correlated to lower presence of osteocalcin. Likewise, these results were similar to findings in specimens treated only with PPP when compared to control. Conclusions PC and PPP were not effective when applied in association with ABG. Similarly, isolated use of PPP was not beneficial in optimizing the bone repair.

Superdosing Phytases in the Diets of Light Laying Hens: Productive Performance and Bone Quality

Two experiments were carried out with the objective of evaluating the effect of superdosing of two phytases on performance, egg quality, digestive organ biometry and bone quality of light hens in the first (58 weeks) and second (87 weeks) productive cycle. In the first cycle, 270 light hens were used, in which a completely randomized design was adopted in a 2 x 2 + 1 factorial scheme (bacterial phytase or fungal phytase x 450 FTUs or 900 FTUs + control diet). In the second cycle, 270 hens were used, following the same design as the previous experiment. The performance and quality of the eggs were evaluated in both cycles, and the biometry of the digestive organs and bone characteristics were also evaluated in the first experiment. There was no effect in the comparison between the means for the performance and egg quality (in both experiments), or for the biometric variables and bone characteristics (first experiment). There was no significant interaction between the factors for any of the variables in the two experiments. In the first experiment, egg production was higher with bacterial phytase and egg weight with fungal phytase. The mineral matter showed greater value with 450 FTUs. It is recommended to use bacterial phytase produced from Escherichia coli as it improves the performance of light laying hens. The dosage of 450 FTUs improves the mineral content of light laying hens and the use of phytase in the laying hen diet implies a lower feed cost.

Effect of IL-33 on pyroptosis of macrophages in mice with sepsis via NF-κB/p38 MAPK signaling pathway

ABSTRACT Purpose To demonstrate the effect of IL-33 on the macrophage pyroptosis in mice with sepsis through the NF-kB/p38 MAPK signal pathway. Methods In total, 24 C57BL/6 mice were divided into the sham operation group (sham) and the cecal ligation and puncture group (CLP). After CLP, 24 IL-33-/- mice were divided into the IL-33-/- group and the IL-33-/- intervention group. The latter group was intraperitoneally injected with IL-33. Mouse mortality was observed after CLP. Macrophage apoptosis in peritoneal lavage fluid was detected by flow cytometry. Serum inflammatory factor level was detected by ELISA. Apoptotic protein expression and NF-κB/p38 MAKP signaling pathway protein expression were detected by qRT-PCR and Western blot. Results Knocking out IL-33 significantly reduced the mortality of CLP mice, as well as the mRNA expression of IL-33 and the levels of serum inflammatory factors, including IL-33, IL-1β, and IL-18. It also reduced the rate of macrophage apoptosis and the expression of the apoptotic protein caspase-1 p10; increased the expression of IκBα; and reduced the protein expression of NF-κB and p38 MAPK. These effects were reversed after exogenous injection of IL-33. Conclusions IL-33 can increase the level of macrophage pyroptosis in mice with sepsis (by activating the NF-kB/p38MAPK signal pathway) and the mortality of these mice.

Superdosing Phytases in the Diets of Light Laying Hens: Productive Performance and Bone Quality

Two experiments were carried out with the objective of evaluating the effect of superdosing of two phytases on performance, egg quality, digestive organ biometry and bone quality of light hens in the first (58 weeks) and second (87 weeks) productive cycle. In the first cycle, 270 light hens were used, in which a completely randomized design was adopted in a 2 x 2 + 1 factorial scheme (bacterial phytase or fungal phytase x 450 FTUs or 900 FTUs + control diet). In the second cycle, 270 hens were used, following the same design as the previous experiment. The performance and quality of the eggs were evaluated in both cycles, and the biometry of the digestive organs and bone characteristics were also evaluated in the first experiment. There was no effect in the comparison between the means for the performance and egg quality (in both experiments), or for the biometric variables and bone characteristics (first experiment). There was no significant interaction between the factors for any of the variables in the two experiments. In the first experiment, egg production was higher with bacterial phytase and egg weight with fungal phytase. The mineral matter showed greater value with 450 FTUs. It is recommended to use bacterial phytase produced from Escherichia coli as it improves the performance of light laying hens. The dosage of 450 FTUs improves the mineral content of light laying hens and the use of phytase in the laying hen diet implies a lower feed cost.(AU)

Effect of crushed sugarcane on performance, bone characteristics and intestinal morphology of free-range chicken lines / Efeito da cana-de-açúcar triturada sobre desempenho, características ósseas e morfologia intestinal de linhagens de frangos caipiras

The objective of this study was to evaluate the effect of different levels of crushed sugarcane in the diet on the performance, bone characteristics and intestinal morphology of slow-growing broiler strains. A total of 448 chickens at 35 days of age were allotted to two randomized blocks in a 4×2 factorial arrangement (levels of feed replacement with crushed sugarcane, on a fresh-matter basis: 0, 15, 30, and 45%) and two strains (Pesadão and Label Rouge (LR)). Two replicates were used per block over time (containing the treatments and the strains), and 14 individuals were used per experimental unit. Weight gain and feed intake decreased linearly in response to the increasing sugarcane levels. Feed intake was higher in the Pesadão strain than in LR. Chicken foot weight was lower in Pesadão than in LR. There were no effects of sugarcane levels or strains on the tarsometatarsal bone weight, breaking strength, maximum load and deformation. The drymatter (DM) content of the tarsometatarsal bone was lower in LR than in Pesadão. The replacement of feed with sugarcane induced a linear increase in bone ash content and a decrease in bone phosphorus and calcium contents. Label Rouge chickens showed a greater crypt depth and lower values of villus/crypt ratio and mucosal layer thickness. The performance and intestinal morphology of broiler chickens are negatively affected by increasing sugarcane levels in the diet. In addition, the increasing intake of sugarcane changesthe bone mineral composition without reducing mechanical resistance, in both the Pesadão and Label Rouge strains.

O presente estudo foi realizado objetivando avaliar o desempenho, as características ósseas e a morfologia intestinal de duas linhagens de frangos tipo caipira alimentados com dietas contendo cana-de-açúcar triturada. Foram utilizados 448 frangos aos 35 dias de idade, distribuídos em dois blocos casualizados, em esquema fatorial 4x2, sendo quatro níveis de substituição (0; 15; 30 e 45%) da ração por cana-de-açúcar triturada e duas linhagens (Pesadão e Label Rouge (LR)), em duas repetições por bloco e 14 aves por unidade experimental. Foi observado redução linear com o aumento dos níveis de cana-de-açúcar sobre o ganho em peso e consumo de ração. O consumo de ração foi maior na linhagem Pesadão em relação à linhagem LR. No entanto, o peso do pé das aves foi menor para a linhagem Pesadão em comparação com a linhagem LR. Não foi observado efeitos da substituição da ração por cana-de-açúcar e das linhagens sobre o peso do osso tarsometatarso, carga de ruptura, força máxima e deformação. O teor de matéria seca do osso tarsometatarso foi menor para os animais LR em comparação com a linhagem Pesadão. A substituição da ração por cana-de-açúcar aumentou linearmente o teor de cinzas dos ossos e diminuiu o teor de fósforo e o de cálcio. A linhagem LR apresentou maior profundidade de criptas, menor relação vilosidade/cripta e espessura da camada da mucosa. O desempenho e a morfologia intestinal de frangos caipiras são afetados negativamente com o aumento dos níveis de cana-de-açúcar na dieta. Além disso, o aumento da ingestão de cana-de-açúcar altera a composição mineral óssea, sem prejudicar a resistência mecânica das linhagens Pesadão e LR.

Effect of crushed sugarcane on performance, bone characteristics and intestinal morphology of free-range chicken lines / Efeito da cana-de-açúcar triturada sobre desempenho, características ósseas e morfologia intestinal de linhagens de frangos caipiras

The objective of this study was to evaluate the effect of different levels of crushed sugarcane in the diet on the performance, bone characteristics and intestinal morphology of slow-growing broiler strains. A total of 448 chickens at 35 days of age were allotted to two randomized blocks in a 4×2 factorial arrangement (levels of feed replacement with crushed sugarcane, on a fresh-matter basis: 0, 15, 30, and 45%) and two strains (Pesadão and Label Rouge (LR)). Two replicates were used per block over time (containing the treatments and the strains), and 14 individuals were used per experimental unit. Weight gain and feed intake decreased linearly in response to the increasing sugarcane levels. Feed intake was higher in the Pesadão strain than in LR. Chicken foot weight was lower in Pesadão than in LR. There were no effects of sugarcane levels or strains on the tarsometatarsal bone weight, breaking strength, maximum load and deformation. The drymatter (DM) content of the tarsometatarsal bone was lower in LR than in Pesadão. The replacement of feed with sugarcane induced a linear increase in bone ash content and a decrease in bone phosphorus and calcium contents. Label Rouge chickens showed a greater crypt depth and lower values of villus/crypt ratio and mucosal layer thickness. The performance and intestinal morphology of broiler chickens are negatively affected by increasing sugarcane levels in the diet. In addition, the increasing intake of sugarcane changesthe bone mineral composition without reducing mechanical resistance, in both the Pesadão and Label Rouge strains.(AU)

O presente estudo foi realizado objetivando avaliar o desempenho, as características ósseas e a morfologia intestinal de duas linhagens de frangos tipo caipira alimentados com dietas contendo cana-de-açúcar triturada. Foram utilizados 448 frangos aos 35 dias de idade, distribuídos em dois blocos casualizados, em esquema fatorial 4x2, sendo quatro níveis de substituição (0; 15; 30 e 45%) da ração por cana-de-açúcar triturada e duas linhagens (Pesadão e Label Rouge (LR)), em duas repetições por bloco e 14 aves por unidade experimental. Foi observado redução linear com o aumento dos níveis de cana-de-açúcar sobre o ganho em peso e consumo de ração. O consumo de ração foi maior na linhagem Pesadão em relação à linhagem LR. No entanto, o peso do pé das aves foi menor para a linhagem Pesadão em comparação com a linhagem LR. Não foi observado efeitos da substituição da ração por cana-de-açúcar e das linhagens sobre o peso do osso tarsometatarso, carga de ruptura, força máxima e deformação. O teor de matéria seca do osso tarsometatarso foi menor para os animais LR em comparação com a linhagem Pesadão. A substituição da ração por cana-de-açúcar aumentou linearmente o teor de cinzas dos ossos e diminuiu o teor de fósforo e o de cálcio. A linhagem LR apresentou maior profundidade de criptas, menor relação vilosidade/cripta e espessura da camada da mucosa. O desempenho e a morfologia intestinal de frangos caipiras são afetados negativamente com o aumento dos níveis de cana-de-açúcar na dieta. Além disso, o aumento da ingestão de cana-de-açúcar altera a composição mineral óssea, sem prejudicar a resistência mecânica das linhagens Pesadão e LR. (AU)

Hipocalcemia pós-parto em vacas de leite

A hipocalcemia se refere a um quadro causado por decúbito prolongado e persistente durante o puerpério, acometendo vacas de alta produção de leite. Para minimizar a redução da concentração de cálcio, o organismo tenta suprir essa perda através da absorção intestinal e reabsorção óssea. Os sinais clínicos variam conforme o estágio da doença e o tratamento deve ser imediato assim que observados os primeiros sinais. A prevenção é a melhor estratégia e a mais viável de manter a qualidade de vida do rebanho. O objetivo desse trabalho foi realizar uma revisão de literatura sobre hipocalcemia pós-parto em vacas de leite.

Hypocalcemia refers to a condition caused by prolonged and persistent recumbency during the puerperium, affecting cows with high milk production. To minimize the reduction in calcium concentration, the body tries to compensate for this loss through intestinal absorption and bone resorption. Clinical signs vary according to the stage of the disease and treatment should be immediate as soon as the first signs are observed. Prevention is the best and most viable strategy to maintain the herd's quality of life. The objective of this work was to carry out a literature review on postpartum hypocalcemia in dairy cows.

A utilização do laser de baixa intensidade na cicatrização de ferida em cão / The use of low intensity laser in wound healing in dog

Background: Low-intensity laser is effective in cellular metabolism, analgesia, and tissue repair. The bioelectric, bioenergetic, and biochemical effects of laser therapy stimulate local circulation, collagen formation, and epithelization. The objective of this study was to report the use of gallium-aluminum arsenide (GaAlAs) laser (830 nm) in healing two woundscaused by necrosis in a dog. Case: An 8-year-old bitch, a rescued victim of animal abuse, had tibiotarsal dislocation in the left posterior limb. Orthopedic surgery was performed with the placement of an external fixation device with six pins in the distal third of the left posterior limb. A radiographic examination performed 42 days after the surgery revealed the loss of the tibiotarsal ratio, decreased radiopacity of the carpal bones, edema, inflammation, muscle atrophy, and the rotation of the anatomical axis of the metatarsals and phalanges. It was then decided to perform a second orthopedic surgery, in which a bone graft and arthrodesis with an 8-screw titanium plate were performed. Four of the plate screws were placed proximally to the tibia and four distally to the tarsus in addition to a screw and a pin transfixed through the distal metaphyseal region of the tibia and calcaneus. Traction was felt during suturing in the dermis and epidermis of the limb, suggesting the possibility of dehiscence of the stitches. After 3 h of surgery, the limb was cold and edematous. After 5 days, the tissue of the medial region of the tarsus developed necrosis, exposing a wound 6 cm long, 1.5 cm wide, and 2 mm deep (wound 1). Another necrotic lesion was observed in the dorsal region of the tarsos - 6.5 cm long, 2 cm wide, 3 mm deep, and exposing 5 cm of the titanium plate (wound 2). Therapy with GaAlAs laser was then performed over the entire length of the wounds. In each therapeutic session, the laser was first used in punctual mode at 10 J/cm², 830 nm, 20 s at each point of the length of the lesion and then in scan mode at 10 J/cm², 830 nm, 1000 Hz, and 40 s continuously. Laser therapy sessions were conducted at 2-4 day intervals, with a 12-day interval between the eighth and ninth sessions. During the entire treatment, cleansing and debridement of the wounds were performed every 48 h with saline and chlorhexidine digluconate, using a compression bandage, as described earlier. Wound 1 healed completely after two laser therapy sessions. In wound 2, tissue repair stagnated after the ninth and last laser session, leaving 4 cm of the titanium plate still exposed. The limb could not support the body weight of the patient, and radiographic examination revealed that the anatomical axis of the metatarsals and the phalanges was rotated and bone conformation was poor. Given the anatomical conditions associated with the patient's clinical picture, it was decided to amputate the limb between the femur and tibia. Discussion: Therapeutic lasers act on mitochondrial respiration, increasing respiratory metabolism and stimulating DNA synthesis and tissue proliferation. In the present case, there was a decrease in inflammatory cells, edema, and the size of the wounds. The pathological conditions of the affected site, influence of trauma, and degree of tissue damage affected the results of the laser therapy. Despite the size difference between wounds 1 and 2, the exposed titanium plate was a determining factor for the partial healing of wound 2.

Apamin-induced alterations in J774 1. 6 macrophage metabolism / [Alterações induzidas por apamina no metabolismo de macrófagos J774 1. 6]

Among the immune system cells, macrophages have an important role. Apamin, a bee venom constituent, is important in the defense of these insects. Thus, we aimed to evaluate the metabolism of J774 1.6 macrophage cell line when exposed to isolated and purified apamin, using cytotoxicity tests by MTT reduction and analysis by flow cytometry (apoptosis / necrosis, production of reactive oxygen species (ROS), membranous lipoperoxidation (LPO), electrical potential of the mitochondrial membrane (mMP) and DNA fragmentation). None of the tested concentrations (10 to 100µg/mL) were cytotoxic according to MTT reductions. Apoptosis rates decreased at concentrations of 2.5, 5.0, and 10.0µg/mL (P<0.05), while necrosis rates increased (P<0.05). However, rates of healthy cells at the highest tested concentration (10µg/mL) did not differ from control (P>0.05). Apamin did not alter ROS, LPO, or DNA fragmentation. Therefore, all analyzed concentrations (1.25 to 10µg/mL) decreased mMP. Such decrease in apoptosis might be due to a suppression of mitochondrial pro-apoptotic messengers, as this peptide causes no oxidative stress, lipid peroxidation, and DNA damage. Highly sensitive techniques are majorly important for proper interpretation of cellular toxicity mechanisms, combined with routine laboratory methods.(AU)

Das células do sistema imunológico, macrófagos desempenham um papel fundamental. Apamina, constituinte do veneno de abelhas, é importante na defesa destas. Objetivou-se avaliar o metabolismo da linhagem de macrófagos J774 1.6 expostos à apamina isolada e purificada, avaliando-se citotoxicidade por redução de MTT e análise por citometria de fluxo (apoptose / necrose, produção de espécies reativas de oxigênio (EROs), lipoperoxidação membranosa (LPO), potencial elétrico da membrana mitocondrial (MMP) e fragmentação do DNA). Nenhuma concentração testada (10 a 100µg / mL) foi citotóxica. As taxas de apoptose diminuíram nas concentrações 2,5, 5,0 e 10,0µg / mL (P<0,05), enquanto as de necrose aumentaram (P<0,05). Entretanto, as taxas de células saudáveis na maior concentração testada (10µg / mL) não diferiram do controle (P>0,05). A apamina não alterou as ERO, a LPO nem a fragmentação do DNA. Portanto, todas as concentrações analisadas (1,25 a 10µg / mL) diminuíram a mMP. Tal diminuição na apoptose pode ser por uma supressão de mensageiros pró-apoptóticos mitocondriais, já que este peptídeo não causa estresse oxidativo, peroxidação lipídica nem dano ao DNA. Técnicas altamente sensíveis são importantes para adequada interpretação dos mecanismos de citotoxicidade.(AU)

Energia metabolizável e aminoácidos digestíveis de coprodutos de trigo e de farinhas de origem animal em dietas para frangos de corte / [Metabolizable energy and digestible amino acids of wheat co-products and animal origin meals in broiler diets]

The objective of this study was to determine the metabolizable energy, standardized ileal amino acid digestibility (SIAAD) and digestible amino acid values of wheat meal (WM), wheat gluten meal (WGM), meat bone meal (MBM) and poultry-by-product meal (PPM) in diets for broilers. In the first experiment, the total excreta collection method was used, with 180 chicks with 14 days, distributed in a completely randomized design, with 4 feeds + reference diet with 6 replicates and 6 broilers each. In the second experiment, 180 broilers from 24 to 28 days old were used, distributed in a completely randomized design, with 4 feeds + one protein free diet with 6 replicates and 6 broilers each. The AMEn values of WM, WGM, MBM and PPM were 3052, 3174, 1556 and 3200 (kcal/kg), respectively. The average SIAAD of the essential and non-essential amino acids of WM, WGM, MBM and PPM were 74.21, 97.50, 69.28 and 83.08; 76.32, 96.42, 63.93 and 77.66, respectively. The values of the main digestible limiting amino acids were 0.18, 1.42, 0.41 and 0.94 for methionine; 0.20, 1.48, 1.73 and 2.73 for lysine; 0.23, 1.67, 0.72 and 1.89 for threonine; and 0.35, 3.22, 1.38 and 2.47 for valine, respectively for the feedstuffs in study.(AU)