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The objectives of this study were to describe alterations that age and dietary inclusion of direct-fed microbial (DFM) Bacillus subtilis (BS) and a specific essential oil (EO) blend (carvacrol, cinnamaldehyde, cineol, and pepper extract) causes in the activity of digestive enzymes (maltase: MALT; aminopeptidase-N: APN; intestinal alkaline phosphate: IAP) and expression patterns of genes related to transport (oligopeptide transporter gene: SLC15A1; Na+-dependent glucose and galactose transporter gene: SLC5A1; Na+-independent glucose, galactose, and fructose transporter gene: SLC2A2; ATPase, Na+/K+ transporting gene: ATP1A1) and digestion (aminopeptidase-N gene: ANPEP; maltase-glucoamylase gene: MGAM; Sucrase-isomaltase gene: SI) of carbohydrates and proteins in the small intestine of broilers. Also, the objective was to analyze if growth performance of broilers is affected by supplementation (BS and EO blend). Day-old male broiler chicks (n = 1,320) were assigned to 5 treatments. Diets included a basal diet (BD) as a negative control (CON); experimental diets were BD + BS; BD + BS + EO; BD + EO; BD + antibiotic growth promoter (AGP) avilamycin was the positive control. Performance was evaluated between 1 to 42 d. Transcript abundance of transport-related genes and digestion-related genes were assayed by RT-qPCR and determined at d 7, 21, and 42. MALT-, APN-, and IAP-specific activities were determined at d 7, 21, and 42. Broilers fed BS had greater SLC15A1 mRNA abundance compared to CON, while EO and AGP were related to higher activities of IAP and APN. Analysis over time revealed higher abundance of MGAM, SLC2A2, SLC15A1, SLC5A1 and SI mRNA at d 42 when compared to d 7. Activity of IAP decreased after d 7 and activity of MALT increased with age. The current study suggests that age had effect over carbohydrate and protein transport and carbohydrate digestion. The supplementation of BS DFM hade evident effect over protein transport and that the use of EO in the diet enhanced the activities of carbohydrate and protein digestion, reflecting improvement in digestive and transport physiology of birds. Changes performed by BS DFM and EO did not favor performance.

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The effect of replacing corn with low-tannin sorghum on broiler performance, carcass yield, integrity of mucosa of small intestine segments, and activity of membrane enzymes of the jejunum is investigated. A total of 594 male Cobb-500 broiler chicks were randomly assigned to 3 dietary treatments: 100% corn (control), 50% corn replacement with low-tannin sorghum (low sorghum), and 100% corn replacement with low-tannin sorghum (high sorghum). Body weight gain, feed consumption, feed conversion, and carcass yield were determined at 7, 21, and 42 d, and segments of the small intestine were collected. Feed conversion and weight gain were impaired at d 42 in broilers fed the high-sorghum diet, but no differences were observed for carcass yield among the treatments (P > 0.05). Crypt cell mitotic index of the jejunum and ileum at d 21 and 42 was lower in broilers fed the control diet than in those fed low- and high-sorghum diets (P < 0.05). Aminopeptidase activity was higher in broilers fed the control diet than in those fed low- and high-sorghum diets irrespective of age (P < 0.05). Conversely, intestinal alkaline phosphatase activity in the small intestine did not differ among the dietary treatments (P > 0.05). Our results indicate that 50% corn replacement with low-tannin sorghum is suitable for broiler diets, whereas 100% corn replacement with low-tannin sorghum had negative effects on the intestinal mucosa and performance of broilers at 42 d.

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Experimental studies have shown that hatching rate depends, among other factors, on the main physical characteristics of the eggs. The physical parameters used in our work were egg weight, eggshell thickness, egg sphericity, and yolk per albumen ratio. The relationships of these parameters in the incubation process were modeled by Fuzzy logic. The rules of the Fuzzy modeling were based on the analysis of the physical characteristics of the hatching eggs and the respective hatching rate using a commercial hatchery by applying a trapezoidal membership function into the modeling process. The implementations were performed in software. Aiming to compare the Fuzzy with a statistical modeling, the same data obtained in the commercial hatchery were analyzed using multiple linear regression. The estimated parameters of multiple linear regressions were based on a backward selection procedure. The results showed that the determination coefficient and the mean square error were higher using the Fuzzy method when compared with the statistical modeling. Furthermore, the predicted hatchability rates by Fuzzy Logic agreed with hatching rates obtained in the commercial hatchery.

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Two experiments were carried out to study the effect of breeder age on incubation parameters (hatchability, eggshell thickness, egg surface temperature and chick weight). In Exp. 1, fertile eggs (30- and 60-wk-old breeders) were incubated at three different temperatures (36.8, 37.8 and 38.8 ºC). Eggshell surface temperature was measured by attaching a thermocouple to the shell and data were collected in a datalogger every ten minutes. This study was conducted according to a 3 x 2 factorial design (three temperatures and two breeder ages). Data revealed that eggshell surface temperature changed according to incubation temperature, with the main increase occurring between 10 and 13 days of incubation, and that the maximum increase in eggshell surface temperature was not higher than +0.6 ºC, irrespective of incubator temperature. The incubator temperature affected total incubation period and hatchability (%) at 38.8 ºC, independent of breeder age. Heavier eggs resulted in heavier chicks, irrespective of incubator temperature. In Exp 2, the eggs (30- and 60-wk-old breeders) were incubated at 37.8 ºC and eggs characteristics (weight, specific gravity, total hatchability and chicks weight) were evaluated according to a randomized experimental design. The data showed that breeder age affected eggshell thickness and chick weight (heavier eggs resulted in heavier chicks), but not specific gravity, eggshell surface temperature or hatchability. The findings of this study revealed that hatchability can be influenced by incubation temperature, but not by the breeder age. Breeder age can affect eggshell thickness, egg weight and eggshell surface temperature, but not specific gravity.(AU)

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This study was carried out to verify if Saccharomyces cerevisiae cell wall (SCCW) dietary supplementation (0.2%) was capable of protecting the intestinal mucosa of broiler chickens vaccinated against coccidiosis. Body weight gain, feed intake, feed conversion and intestinal mucosa morphometric parameters and epithelial loss were evaluated. In the experiment,400 day-old male chicks were distributed according to a completely randomized design in a 2x2 factorial arrangement. The following treatments were applied: T1 - no vaccination/ no SCCW supplementation; T2 - no vaccination/SCCW supplementation; T3 - vaccination/no SCCW supplementation; and T4 - vaccination/SCCW supplementation to four replicates of 25 birds each. Birds were vaccinated on the first day of age using a spray vaccine (Coccivac B®, Coopers), containing E. acervulina, E. maxima, E. mivati and E. tenella. S. cerevisiae cell wall was supplied from the first day of age. Live performance, intestinal morphometric parameters and epithelial loss were evaluated at 14, 21 and 28 days of age. Performance was affected by vaccination only at 21-days of age, when body weight gain was reduced in the vaccinated birds, but no body weight difference was observed on day 28. Vaccine also increased the crypt depth (p<0.05) in the duodenum and jejunum, suggesting a high cell activity in the crypt:villus transition area to maintain the epithelial cell turnover. Villi number/area (103,269 µm²) was not affected (p>0.05) by vaccine or cell wall supplementation, and epithelial loss was more pronounced in the duodenum and jejunum. In conclusion, the findings of this study suggest that S. cerevisiae cell wall supplementation may be an useful management tool to maintain the intestinal integrity of broilers vaccinated against coccidiosis.(AU)

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ETHNOPHARMACOLOGICAL RELEVANCE: Polygonum ferrugineum Wedd. (Polygonaceae) is used to heal infected wounds and as antiseptic, antibiotic or antifungal in the traditional Argentinean medicine. The present investigation was carried out to evaluate the antifungal properties of different extracts of aerial parts of Polygonum ferrugineum, in order to give support to its ethnopharmacological use and to isolate the compounds responsible for the antifungal properties. The most active compounds were tested for their capacity of producing hyphae malformations, similar to those previously observed for crude extracts. MATERIALS AND METHODS: Agar Dilution Method (ADM) and Agar Overlay Bioautography (AOB) were used for bioassay-guided fractionation of the aerial part extracts against a panel of human opportunistic pathogenic fungi. The Neurospora crassa assay, followed by Optical Microscopy and Scanning Electron Microscopy observation, was used for studies of mechanisms of action. RESULTS: MeOH extract and DCM and Hex sub-extracts, but not Aq, EtOAc or BuOH ones possess antifungal activity. Of the seven isolated compounds, cardamonin 2 showed a selective inhibition of Epidermophyton floccosum with a very low MIC (=6.2 µg/mL) and pashanone 1 possessed moderate antifungal activity (MICs=25-50 µg/mL) but a broader spectrum of action. Chalcone 2, but not 1, induced swelling and shortening of the Neurospora crassa hyphae, similar as those caused by the crude DCM extract. CONCLUSIONS: The bioassay-guided fractionation of Polygonum ferrugineum DCM extract allowed the isolation of five active compounds. Among them, cardamonin 2 showed the highest antifungal activity and selectivity towards Epidermophyton floccosum; in addition, it induced Neurospora crassa malformations that are similar than those produced by the crude DCM extract. These results give additional support to the ethnopharmacological use of Polygonum ferrugineum as antifungal agent.

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Nitric oxide (NO) plays a key role in body temperature (Tb) regulation of mammals, acting on the brain to stimulate heat loss. Regarding birds, the putative participation of NO in the maintenance of Tb in thermoneutrality or during heat stress and the site of its action (periphery or brain) is unknown. Thus, we tested if NO participates in the maintenance of chicks' Tb in those conditions. We investigated the effect of intramuscular (im; 25, 50, 100mg/kg) or intracerebroventricular (icv; 22.5, 45, 90, 180 microg/animal) injections of the non selective NO synthase inhibitor L-NAME on Tb of 5-day-old chicks at thermoneutral zone (TNZ; 31-32 degrees C) and under heat stress (37 degrees C for 5-6h). We also verified plasma and diencephalic nitrite/nitrate levels in non-injected chicks under both conditions. At TNZ, 100mg/kg (im) or 45, 90, 180 microg (icv) of L-NAME decreased Tb. A significant correlation between Tb and diencephalic, but not plasma, nitrite/nitrate levels was observed. Heat stress-induced hyperthermia was inhibited by all tested doses of L-NAME (im and icv). Tb was correlated neither with plasma nor with diencephalic nitrite/nitrate levels during heat stress. These results indicate the involvement of brain NO in the maintenance of Tb of chicks, an opposite action of that observed in mammals, and may modulate hyperthermia.

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The effect of breeder age on long bone development was studied in chicken embryos from 12 days of incubation until hatching. Fertile eggs were incubated and randomly assigned in a 2 x 6 factorial arrangement (two breeder ages - 38 and 60 weeks and six incubation days - 12, 14, 16, 18, 20, and 21). Enzymatic activity of acid and alkaline phosphatases in tibial epiphyses and weights as well as length and width in tibias and femurs of the embryos were determined. Tartrate-resistant acid and alkaline phosphatases activity in epiphyses was not affected by breeder age. Absolute weight and width of femur and tibia were larger in 60-week-old embryos compared to 38-week-old. Enzymatic activity and morphometric measurements increased with incubation day, independently of breeder age. The results showed that the process of endochondral ossification during the last two thirds of embryo development was not influenced by the age of the breeders. Although, in terms of absolute weight, the long bones of embryos from older breeders were heavier, which was associated with the larger width of the bones, but and not with their length.

O efeito da idade da matriz sobre o desenvolvimento dos ossos longos foi estudado em embriões de frango de 12 dias de incubação até o nascimento. Ovos férteis foram incubados e distribuídos em delineamento inteiramente casualizado em arranjo fatorial 2 x 6 (duas idades de matriz - 38 e 60 semanas e seis dias de incubação - 12, 14, 16, 18, 20 e 21 dias). Determinou-se a atividade enzimática das fosfatase alcalina e ácida-resistente ao tartrato no peso e nas epífises da tíbia, no comprimento e na largura da tíbia e do fêmur. A atividade das fosfatases não foi afetada pela idade da matriz. O peso absoluto e a largura de fêmur e tíbia foram maiores nos embriões das matrizes com 60 semanas de idade. Atividade enzimática e medidas morfométricas aumentaram com o dia de incubação independentemente da idade da matriz. Concluiu-se que o processo de ossificação endocondral durante os dois últimos terços de desenvolvimento embrionário não foi influenciado pela idade das matrizes. No entanto, em termos de peso absoluto, os ossos longos de embriões provenientes de matrizes velhas foram mais pesados o que foi associado à maior largura e não ao maior comprimento dos ossos.

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The effect of breeder age on long bone development was studied in chicken embryos from 12 days of incubation until hatching. Fertile eggs were incubated and randomly assigned in a 2 x 6 factorial arrangement (two breeder ages - 38 and 60 weeks and six incubation days - 12, 14, 16, 18, 20, and 21). Enzymatic activity of acid and alkaline phosphatases in tibial epiphyses and weights as well as length and width in tibias and femurs of the embryos were determined. Tartrate-resistant acid and alkaline phosphatases activity in epiphyses was not affected by breeder age. Absolute weight and width of femur and tibia were larger in 60-week-old embryos compared to 38-week-old. Enzymatic activity and morphometric measurements increased with incubation day, independently of breeder age. The results showed that the process of endochondral ossification during the last two thirds of embryo development was not influenced by the age of the breeders. Although, in terms of absolute weight, the long bones of embryos from older breeders were heavier, which was associated with the larger width of the bones, but and not with their length.(AU)

O efeito da idade da matriz sobre o desenvolvimento dos ossos longos foi estudado em embriões de frango de 12 dias de incubação até o nascimento. Ovos férteis foram incubados e distribuídos em delineamento inteiramente casualizado em arranjo fatorial 2 x 6 (duas idades de matriz - 38 e 60 semanas e seis dias de incubação - 12, 14, 16, 18, 20 e 21 dias). Determinou-se a atividade enzimática das fosfatase alcalina e ácida-resistente ao tartrato no peso e nas epífises da tíbia, no comprimento e na largura da tíbia e do fêmur. A atividade das fosfatases não foi afetada pela idade da matriz. O peso absoluto e a largura de fêmur e tíbia foram maiores nos embriões das matrizes com 60 semanas de idade. Atividade enzimática e medidas morfométricas aumentaram com o dia de incubação independentemente da idade da matriz. Concluiu-se que o processo de ossificação endocondral durante os dois últimos terços de desenvolvimento embrionário não foi influenciado pela idade das matrizes. No entanto, em termos de peso absoluto, os ossos longos de embriões provenientes de matrizes velhas foram mais pesados o que foi associado à maior largura e não ao maior comprimento dos ossos.(AU)

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