Down-regulation of complement genes in lipopolysaccharidechallenged zebrafish (Danio rerio) larvae exposed to Indonesian propolis / Regulação negativa dos genes do complemento em larvas de peixe-zebra (Danio rerio) desafiadas com lipopolissacarídeo expostas à própolis indonésia

Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 μg/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 μg/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 μg/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.

Embora a própolis tenha sido relatada por ter atividade anti-inflamatória, seus efeitos no sistema complemento, uma parte do sistema imunológico inato, não foram muito estudados. Esta pesquisa foi conduzida para descobrir os efeitos da própolis da Indonésia nos níveis de expressão de C3, C1r/s, Bf, MBL e C6 em larvas de peixe-zebra induzidas por lipopolissacarídeo (LPS). Foram realizadas contagens de macrófagos que migram para o saco vitelino e histologia do fígado. As larvas foram divididas em quatro grupos: CON (cultivadas apenas em meio E3), LPS (cultivadas em meio contendo 0,5 μg/L de LPS), LPSIBU (cultivadas em meio contendo LPS e, em seguida, tratadas com 100 μg/L de ibuprofeno por 24 horas) e LPSPRO (cultivado em meio contendo LPS, e então imerso em própolis 14,000 μg/L por 24 horas). Os resultados mostraram que a expressão do gene do complemento em larvas dos grupos LPSIBU e LPSPRO foi geralmente menor que em larvas do grupo LPS. O número de migrações de macrófagos para a gema no grupo LPSPRO também foi menor que no grupo LPS. A estrutura histológica do fígado em todos os grupos foi considerada normal. Este estudo mostra que a própolis indonésia tem potencial para ser utilizada como alternativa na substituição dos AINEs (anti-inflamatórios não esteroides).

Down-regulation of complement genes in lipopolysaccharidechallenged zebrafish (Danio rerio) larvae exposed to Indonesian propolis / Regulação negativa dos genes do complemento em larvas de peixe-zebra (Danio rerio) desafiadas com lipopolissacarídeo expostas à própolis indonésia

Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 μg/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 μg/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 μg/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.(AU)

Embora a própolis tenha sido relatada por ter atividade anti-inflamatória, seus efeitos no sistema complemento, uma parte do sistema imunológico inato, não foram muito estudados. Esta pesquisa foi conduzida para descobrir os efeitos da própolis da Indonésia nos níveis de expressão de C3, C1r/s, Bf, MBL e C6 em larvas de peixe-zebra induzidas por lipopolissacarídeo (LPS). Foram realizadas contagens de macrófagos que migram para o saco vitelino e histologia do fígado. As larvas foram divididas em quatro grupos: CON (cultivadas apenas em meio E3), LPS (cultivadas em meio contendo 0,5 μg/L de LPS), LPSIBU (cultivadas em meio contendo LPS e, em seguida, tratadas com 100 μg/L de ibuprofeno por 24 horas) e LPSPRO (cultivado em meio contendo LPS, e então imerso em própolis 14,000 μg/L por 24 horas). Os resultados mostraram que a expressão do gene do complemento em larvas dos grupos LPSIBU e LPSPRO foi geralmente menor que em larvas do grupo LPS. O número de migrações de macrófagos para a gema no grupo LPSPRO também foi menor que no grupo LPS. A estrutura histológica do fígado em todos os grupos foi considerada normal. Este estudo mostra que a própolis indonésia tem potencial para ser utilizada como alternativa na substituição dos AINEs (anti-inflamatórios não esteroides).(AU)

Down-regulation of complement genes in lipopolysaccharide-challenged zebrafish (Danio rerio) larvae exposed to Indonesian propolis

Abstract Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 g/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 g/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 g/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.

Resumo Embora a própolis tenha sido relatada por ter atividade anti-inflamatória, seus efeitos no sistema complemento, uma parte do sistema imunológico inato, não foram muito estudados. Esta pesquisa foi conduzida para descobrir os efeitos da própolis da Indonésia nos níveis de expressão de C3, C1r/s, Bf, MBL e C6 em larvas de peixe-zebra induzidas por lipopolissacarídeo (LPS). Foram realizadas contagens de macrófagos que migram para o saco vitelino e histologia do fígado. As larvas foram divididas em quatro grupos: CON (cultivadas apenas em meio E3), LPS (cultivadas em meio contendo 0,5 g/L de LPS), LPSIBU (cultivadas em meio contendo LPS e, em seguida, tratadas com 100 g/L de ibuprofeno por 24 horas) e LPSPRO (cultivado em meio contendo LPS, e então imerso em própolis 14,000 g/L por 24 horas). Os resultados mostraram que a expressão do gene do complemento em larvas dos grupos LPSIBU e LPSPRO foi geralmente menor que em larvas do grupo LPS. O número de migrações de macrófagos para a gema no grupo LPSPRO também foi menor que no grupo LPS. A estrutura histológica do fígado em todos os grupos foi considerada normal. Este estudo mostra que a própolis indonésia tem potencial para ser utilizada como alternativa na substituição dos AINEs (anti-inflamatórios não esteroides).

Early ontogeny of tetra Markiana nigripinnis (Characiformes: Characidae)

The early development of Markiana nigripinnis is described by morphological characters, pigmentation, and morphometry. Larvae were obtained through semi-natural breeding, collected, fixed, and identified according to their development. Eighty individuals with standard lengths ranging from 3.1 to 24.3 mm were analyzed. Larvae are poorly developed at hatching, with a relatively large yolk sac and finfold. At the preflexion stage, the eyes are pigmented, the mouth and anus are functional, the yolk is completely absorbed, and the pectoral fin bud emerges. At flexion, the first rays of the caudal, anal, and dorsal fins become evident. The pelvic fin bud emerges only at the postflexion stage, in addition to the complete absorption of the finfold. Pigmentation is distributed throughout the body, with a greater concentration on the top of the head, around the mouth, and at the base of the caudal fin. The myomere total number ranged from 34 to 49 (16-23 preanal, and 18-27 postanal). Juveniles show morphological characteristics like adults. The fins ray number are pectoral: 11-13, pelvic: 5-7, dorsal: 8-11, caudal: 16-27, and anal 30-47. The morphometric relationships reveal variations in growth along the early ontogeny of the species.

O desenvolvimento inicial de Markiana nigripinnis foi descrito considerando os caracteres morfológicos, a pigmentação e a morfometria. Indivíduos foram obtidos por meio de reprodução seminatural, coletados, acondicionados, fixados e identificados conforme seu período e estágio de desenvolvimento. Foram analisados 80 indivíduos com comprimento padrão variando de 3,1 a 24,3 mm. As larvas são pouco desenvolvidas à eclosão, com saco vitelino relativamente grande e presença de membrana embrionária. Em pré-flexão, os olhos estão pigmentados, a boca e o ânus são funcionais, o vitelo é completamente absorvido e surge o botão da nadadeira peitoral. Em flexão, os primeiros raios das nadadeiras caudal, anal e dorsal tornam-se evidentes. O botão da nadadeira pélvica aparece somente em pós-flexão, além da completa absorção da nadadeira embrionária. A pigmentação se distribui pelo corpo todo, com maior concentração no topo da cabeça, ao redor da boca e na base da nadadeira caudal. O número total de miômeros variou de 34 a 49 (16-23 pré e 18-27 pós-anal). Os juvenis apresentaram características morfológicas semelhantes ao adulto. O número de raios das nadadeiras é peitoral: 11-13, pélvica: 5-7, dorsal: 8-11, caudal: 16-27 e anal 30-47. As relações morfométricas revelam variações no crescimento ao longo da ontogenia inicial da espécie.

Down-regulation of complement genes in lipopolysaccharide-challenged zebrafish (Danio rerio) larvae exposed to Indonesian propolis / Regulação negativa dos genes do complemento em larvas de peixe-zebra (Danio rerio) desafiadas com lipopolissacarídeo expostas à própolis indonésia

Abstract Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 μg/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 μg/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 μg/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.

Resumo Embora a própolis tenha sido relatada por ter atividade anti-inflamatória, seus efeitos no sistema complemento, uma parte do sistema imunológico inato, não foram muito estudados. Esta pesquisa foi conduzida para descobrir os efeitos da própolis da Indonésia nos níveis de expressão de C3, C1r/s, Bf, MBL e C6 em larvas de peixe-zebra induzidas por lipopolissacarídeo (LPS). Foram realizadas contagens de macrófagos que migram para o saco vitelino e histologia do fígado. As larvas foram divididas em quatro grupos: CON (cultivadas apenas em meio E3), LPS (cultivadas em meio contendo 0,5 μg/L de LPS), LPSIBU (cultivadas em meio contendo LPS e, em seguida, tratadas com 100 μg/L de ibuprofeno por 24 horas) e LPSPRO (cultivado em meio contendo LPS, e então imerso em própolis 14,000 μg/L por 24 horas). Os resultados mostraram que a expressão do gene do complemento em larvas dos grupos LPSIBU e LPSPRO foi geralmente menor que em larvas do grupo LPS. O número de migrações de macrófagos para a gema no grupo LPSPRO também foi menor que no grupo LPS. A estrutura histológica do fígado em todos os grupos foi considerada normal. Este estudo mostra que a própolis indonésia tem potencial para ser utilizada como alternativa na substituição dos AINEs (anti-inflamatórios não esteroides).

Cysteine and Aspartic Proteases Underlie the Digestion of Egg Yolk Proteins during the Development of Columba livia domestica Embryo

Yolk proteins undergo digestion either inside the egg yolk or in the surrounding yolk sac membrane (YSM) before being consumed by the developing avian embryo. However, the mechanisms underlying the digestion of yolk proteins during embryogenesis are largely unexplored in the pigeon Columba livia domestica. To better understand these mechanisms, the present study examined the classes of activated proteases in the egg yolk and the gene expression patterns of cathepsin B (CTSB) and cathepsin D (CTSD), which encode for lysosomal cysteine and aspartic proteases, respectively, in the YSM. We investigated the activated proteases by applying different types of protease inhibitors to yolk samples taken from incubation day 16. Then, we detected the mRNA levels of CTSB and CTSD in the YSM at incubation days 6, 8, 10, and 12-17. Both cysteine and aspartic proteases appeared to be activated in the egg yolk. Moreover, CTSB expression increased progressively and reached the maximum value on day 13; however, it decreased significantly on days 14 and 15 and further reduced toward hatching (day 17). In contrast, CTSD expression was weak and fluctuated insignificantly during development. Our results suggest that the degradation of yolk proteins at late developmental stages largely occurs in the egg yolk itself, probably by the activated cysteine and aspartic proteases. Furthermore, cathepsin B in the YSM seems to have a primary role in protein digestion, but this role decreases toward hatching.(AU)

The main channel and river confluences as spawning sites for migratory fishes in the middle Uruguay River

Evidence indicates that migratory fish reproduce in the middle Uruguay River, but the location of spawning sites remains unknown. To identify spawning sites in the basin, fish eggs and larvae were sampled monthly between October 2016 and January 2017. The sampling was carried out in three sections along the middle Uruguay River, covering two environments: the main channel and the mouth of tributaries (Comandaí, Ijuí, and Piratinim rivers). A total of 11,519 eggs and 3,211 larvae were captured, belonged to ten migratory species. Eggs and larvae of migratory fishes, were widely distributed, with predominance of segmented eggs, and larvae in yolk-sac and pre-flexion stages, with higher densities near the confluence with the Piratinim River. Larvae assemblages showed spatial variation, indicating that spawning sites differ among migratory species. This study provides new information about fish reproduction in the middle Uruguay River, revealing that migratory species spawn in different localities along the main channel and tributaries. This region may function as a critical site for fish reproduction, although it is currently threatened by the risk of hydropower expansion, emphasizing the need for measures that preserve their environmental conditions, hydrological connectivity and ecological functions.(AU)

Evidências indicam que os peixes migradores se reproduzem no médio rio Uruguai, mas a localização dos locais de desova ainda permanece desconhecida. Para identificar estes locais, foram realizadas mensalmente entre outubro de 2016 e janeiro de 2017, amostragens de ovos e larvas de peixes em três seções ao longo do médio rio Uruguai, cobrindo dois ambientes: o canal principal e a foz dos afluentes (rios Comandaí, Ijuí e Piratinim). Foram capturados 11.519 ovos e 3.211 larvas, pertencentes a dez espécies migradoras. Ovos e larvas de peixes migradores, foram amplamente distribuídos, com predomínio de ovos segmentados e larvas em estágios de larval-vitelino e pré-flexão, com maiores densidades próximo à confluência com o rio Piratinim. As assembleias de larvas mostraram variação espacial, indicando que os locais de desova diferem entre as espécies migradoras. Este estudo fornece novas informações sobre a reprodução de peixes no médio rio Uruguai, revelando que espécies migradoras desovam em diferentes localidades ao longo do canal principal e afluentes. Esta região pode funcionar como um local crítico para a reprodução de peixes, embora atualmente esteja ameaçada pelo risco de expansão da energia hidrelétrica, enfatizando a necessidade de medidas que preservem suas condições ambientais, conectividade hidrológica e funções ecológicas.(AU)

Egg transport and larval behavior of curimba, Prochilodus lineatus (Valenciennes, 1836; Characiformes, Prochilodontidae) in a drift simulator channel

Landscape changes and hydrologic alterations associated with dam construction have greatly hindered reproductive success and the recruitment of migratory species. However, little is known about the flow conditions that are necessary for the reproductive success of migrators. We aimed to analyze one of the important migrants of the Paraná River, the curimba, Prochilodus lineatus(Valenciennes, 1836), to determine the minimum velocities necessary for egg drift and analyze the behavior of larvae at different flow velocities. Thus, we experimentally evaluated the behavior of eggs and larvae released in a drift simulator channel (DSC) under four flow velocities. Additionally, we investigate the swimming ability of larvae in relation to certainmorphometric measures of their bodies. We also performed morphometry and calculations of larval body and fin proportions to determine which structures influence swimming ability. The results indicated that environments with flow velocities greater than 0.081 m s-1were favorable for egg drift and increased the chances of successful transportof eggs to downstream areas. We also observed that soon after hatching, the larvae positioned themselves against the flow, indicating that they had a perception of position and orientation. The larvae were able to maintain and disperse against the flow within the channel 32h after hatching at all velocities, which was mainly due to the development of pectoral and caudal fins that were actively involved in the swimming process, as well as the reduction in the dimensions of the yolk sac, which reduced drag.(AU)

Effects of broiler breeder age on immune system development of progeny

The objective of this study was to determine the effects that breeder age has on digestive and immune system development; the transfer of immunoglobulins to egg yolk, yolk sac, and neonate chicks; and the immune response of chicks up to 35 days old. Three ages (32, 42, and 52 weeks) of Hubbard breeders were studied with ages as treatments. A total of 425 eggs were weighed for each of the three treatments and incubated. After hatching, a total of 300 1-day-old chicks were used in each treatment. We studied the development of the gastrointestinal tract and immune system of progeny and IgY transfer from breeder to progeny. Chicks from 52-week-old breeders had greater gastrointestinal tract growth up to seven days of life and greater body weight at 14 days. Older breeders (52 weeks) had higher amounts of IgY in serum and egg yolk. Chicks from the youngest breeders (32-weeks-old) had a better immune response at two weeks post-vaccination. It can be concluded that the older breeders have a greater capacity to immunize progeny up to 14 days. Strategies can be developed to increase IgY in the serum of young breeders and, consequently, increase the innate immunity of the newly-hatched chicks.(AU)

Characterization of hematopoietic stem cells from the canine yolk sac

The characterization of hematopoietic stem cells (HSC) from the canine yolk sac (cYS) can contribute to future gene therapies because it is possible to obtain information about the beginning of the development of the circulatory system through the characterization. The cYS is a likely source of HSC, which is a source of blood cell development in mammals. Studies in this field have been conducted for decades; however, interest in cellular therapy is currently at its peak with greater visibility, and these cells are a promising therapeutic tool for the treatment of diseases related to animals and humans. The aim of this study was to isolate and characterize HSC from the cYS embryos at 30 to 45 days of gestational age. Our results showed that the cYS was macroscopically located in the ventral region with a central portion and extremities. The cells in culture presented a circular morphology and cell clusters. The average cell viability was 22.55% dead cells out of 6.5 × 104 total cells. The cells were also able to form colonies on methylcellulose. Flow cytometry analysis revealed the expression of CD34, CD117, and CD45. Our results suggest that the cYS can be used as a source of hematopoietic cells, and this study is very important to understand the mechanism and development of the hematopoietic system in dogs.(AU)

Characterization of hematopoietic stem cells from the canine yolk sac

Abstract The characterization of hematopoietic stem cells (HSC) from the canine yolk sac (cYS) can contribute to future gene therapies because it is possible to obtain information about the beginning of the development of the circulatory system through the characterization. The cYS is a likely source of HSC, which is a source of blood cell development in mammals. Studies in this field have been conducted for decades; however, interest in cellular therapy is currently at its peak with greater visibility, and these cells are a promising therapeutic tool for the treatment of diseases related to animals and humans. The aim of this study was to isolate and characterize HSC from the cYS embryos at 30 to 45 days of gestational age. Our results showed that the cYS was macroscopically located in the ventral region with a central portion and extremities. The cells in culture presented a circular morphology and cell clusters. The average cell viability was 22.55% dead cells out of 6.5 × 104 total cells. The cells were also able to form colonies on methylcellulose. Flow cytometry analysis revealed the expression of CD34, CD117, and CD45. Our results suggest that the cYS can be used as a source of hematopoietic cells, and this study is very important to understand the mechanism and development of the hematopoietic system in dogs.

Sodium butyrate in post-hatch feed for broiler chicks / [Uso de butirato de sódio em rações pós-eclosão para pintos de corte]

This study was designed to determine adequate levels for sodium butyrate inclusion in pre-hatching and pre-starter feed in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, each comprising five replicates of ten birds each. Five dietetic levels of sodium butyrate (control, 0.050, 0.075, 0.100 and 0.200%) were used in the pre-starter feed offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated after 24 hours of feed access and at 7 days of age. A metabolic trial was performed when the chicks were seven to ten days of age. Supplementation of 0.1% sodium butyrate increased the development of broiler chicks' intestinal villi at seven days of age but was not able to improve performance in the pre-starter phase. Supplementation with sodium butyrate in a diet offered in the transportation box does not improve broiler performance in the pre-starter phase. Supplementation of up to 0.16% is recommended, in order to improve the metabolizability of the ether extract for broilers at seven days of age.(AU)

Objetivou-se, com este estudo, encontrar níveis adequados de inclusão de butirato de sódio em ração pós-eclosão e pré-inicial, buscando minimizar os efeitos negativos do jejum de pintos de corte. Os animais foram distribuídos em delineamento inteiramente ao acaso, com cinco tratamentos e cinco repetições de 10 aves cada. Foram utilizadas cinco suplementações de butirato de sódio (controle; 0,05; 0,075; 0,100 e 0,200%) na ração pré-inicial, fornecidas aos pintos na caixa de transporte e durante a fase pré-inicial. Após 24 horas de acesso ao alimento e aos sete dias de idade, foram avaliados desempenho, retração do saco vitelino, concentração de glicose plasmática, peso e histomorfometria do intestino delgado. De sete a 10 dias de idade, realizou-se ensaio de metabolizabilidade. A suplementação de 0,10% de butirato de sódio aumentou o desenvolvimento das vilosidades intestinais dos pintos de corte aos sete dias de idade, mas não foi capaz de melhorar o desempenho na fase pré-inicial. A suplementação com butirato de sódio em dieta oferecida na caixa de transporte não melhora o desempenho dos frangos na fase pré-inicial. Recomenda-se suplementação de até 0,16%, a fim de melhorar a metabolizabilidade do extrato etéreo para frangos de corte aos sete dias de idade.(AU)

Use of Glutamine in Post-Hatch Feed for Broiler Chicks

Post-hatch delayed placement damages, physical and physiological development of broiler chicks. The objective of this study was to ascertain adequate levels of glutamine inclusion in post-hatch and pre-starter feed, in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. The birds were distributed in a completely randomized design, with four treatments and five replicates of ten birds each. Four levels of glutamine supplementation (0, 1, 2 and 3%) were used in the pre-starter feed, which was given to the chicks in the transportation box and during the pre-starter phase. After 24 hours of access to this feed, and at 7 days of age, the chicks performance, yolk sac retraction, plasma glucose concentration, weight and small-intestine histomorphometry were evaluated. From seven to ten days of age, a metabolizability assay was performed. Glutamine supplementation tended to increase the use of the yolk sac, the concentration of plasma glucose (p 0.10) and the depth of the crypt in the ileum after 24 hours of life (p 0.05). There was no statistical difference between the glutamine supplementation levels, in relation to glucose, histomorphometry or metabolizability in the pre-starter phase. Glutamine concentrations did not differ in relation to performance, histomorphometry of the small intestine or metabolizability of nutrients in the pre-starter phase.(AU)

Use of Glutamine in Post-Hatch Feed for Broiler Chicks

Post-hatch delayed placement damages, physical and physiological development of broiler chicks. The objective of this study was to ascertain adequate levels of glutamine inclusion in post-hatch and pre-starter feed, in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. The birds were distributed in a completely randomized design, with four treatments and five replicates of ten birds each. Four levels of glutamine supplementation (0, 1, 2 and 3%) were used in the pre-starter feed, which was given to the chicks in the transportation box and during the pre-starter phase. After 24 hours of access to this feed, and at 7 days of age, the chicks performance, yolk sac retraction, plasma glucose concentration, weight and small-intestine histomorphometry were evaluated. From seven to ten days of age, a metabolizability assay was performed. Glutamine supplementation tended to increase the use of the yolk sac, the concentration of plasma glucose (p 0.10) and the depth of the crypt in the ileum after 24 hours of life (p 0.05). There was no statistical difference between the glutamine supplementation levels, in relation to glucose, histomorphometry or metabolizability in the pre-starter phase. Glutamine concentrations did not differ in relation to performance, histomorphometry of the small intestine or metabolizability of nutrients in the pre-starter phase.

Effects of breeder age and short-term temperature stimulation during incubation on performance, organ weight, and carcass yield in male and female broilers / Efeitos da idade da matriz e de alterações curtas de temperatura de incubação no desempenho, peso dos órgãos e rendimento de carcaça em frangos de corte machos e fêmeas

Birds develop thermoregulatory control during the last days of incubation. Different temperature conditioning programs have been proposed to prepare broiler chicks for post-hatch life. This study aimed to investigate the effects of short-term temperature (STT) stimulation and breeder age on hatching performance, embryo development, yolk absorption, and post-hatch performance of male and female broilers. A 2 × 2 factorial completely randomized design was used, with two breeder ages (30 and 60 wk), two temperature incubation programs (control and STT), and 2,520 eggs per treatment. Eggs were distributed in two large-scale commercial incubators with a capacity of 120,960 eggs. The control group was subjected to a standard single-stage incubation program (37.2-37.4 °C), whereas the STT group was subjected to a temperature increment of 1 °C for 4 h on embryonic days 16, 17, 18, and 19. Embryos were analyzed at 16 and 19 days of incubation, and chicks at hatch. At 19 days of incubation, STT conditioning resulted in lower yolk sac weight in embryos from 60-wk-old breeders and higher relative weight of the gastrointestinal tract in embryos from 30-wk-old breeders. At hatch, males had lower residual yolk weight, females had greater length, and chicks from 60-wk-old breeders subjected to STT had higher body weight and relative weight of the gastrointestinal tract. Eggs from 30-wk-old breeders showed higher fertility and hatchability and lower infertility, mortality, and second-grade chick percentages. Regarding performance, it was found that males had higher body weight from 7 to 42 days of age and higher whole leg yields. Females, on the other hand, had higher fat yields. Breeder age and sex influenced gastrointestinal tract development, carcass yield, and performance. A 1 °C increase in incubation temperature for 4 h from days 16 to 19 of incubation affected yolk absorption and digestive tract development in chicks from 60-wk-old breeders without, however, influencing performance results.(AU)

As aves desenvolvem o controle do sistema termorregulatório durante os últimos dias de incubação, dessa forma, diferentes programas temperatura de incubação têm sido descritos para preparar os pintos de corte para a vida pós-eclosão. Objetivou-se nesse estudo, estabelecer os efeitos de alterações curtas na temperatura de incubação e da idade da matriz no desempenho de eclosão, desenvolvimento embrionário, absorção da gema e desempenho pós-eclosão de frangos de corte machos e fêmeas. O delineamento experimental foi inteiramente casualizado em fatorial 2 x 2 com 2 idades de matriz (30 e 60 semanas) e 2 programas de incubação de temperatura, controle (CT) e grupo de treinamento de temperatura de curto prazo (STT) e 2.520 ovos em cada tratamento. Os ovos foram distribuídos em 2 incubadoras comerciais de grande escala com capacidade para 120.960 ovos. O grupo CT seguiu um programa de incubação de estágio único padrão (37,2-37,4°C) e o STT teve um incremento em + 1 ° C por 4 hs nos dias 16, 17, 18 e 19 de incubação. Os embriões foram analisados aos 16 e 19 dias e pintinhos na eclosão. Aos 19 dias de incubação, embriões de matrizes com 60 semanas apresentaram menor peso do saco vitelino e embriões de matrizes com 30 semanas apresentaram maior porcentagem de trato gastrointestinal quando em STT. Na eclosão, os machos apresentaram menor vitelo residual, e as fêmeas apresentaram maior comprimento, pintinhos mais pesados e maior trato gastrointestinal quando originárias de matrizes de 60 semanas de idade e expostos ao STT. Na eclosão, os ovos com 30 semanas de idade apresentaram maior porcentagem de fertilidade e eclodibilidade e menor infertilidade, mortalidade e pintos de secunda categoria. No desempenho, os machos apresentaram maior peso corporal do 7° ao 42° dia e maiores valores de porcentagem de pernas e as fêmeas maiores valores de porcentagem de gordura. A idade das matrizes e o sexo influenciaram o desenvolvimento visceral, o rendimento de carcaça e o desempenho dos frangos de corte. O aumento de 1°C durante 4 horas entre os dias 16 e 19 de incubação afetou a absorção da gema e o trato digestivo em pintos de matrizes com 60 semanas de idade, sem refletir nos resultados de desempenho.(AU)

BPA toxicity during development of zebrafish embryo / Toxicidade do BPA durante o desenvolvimento de embriões de zebrafish

Bisphenol A (BPA) is a monomer used in the production of polycarbonate, a polymer commonly found in plastics, epoxy resins and thermal papers. The presence of BPA in food, water, air and dust has been of great concern in recent years not only due to environmental and ecological issues but also because of its supposed risk to public health related to its mutagenic and carcinogenic potential. In this study we evaluated the toxicity of bisphenol A in zebrafish embryos (Danio rerio) and determined the 50% lethal concentration (LC50) of this chemical. BPA was used at concentrations ranging from 1 M to 100 M in E3 medium/0.5% dimethylsulfoxide (DMSO) from previously prepared stock solutions in 100% DMSO. Controls included embryos exposed only to E3 medium or supplemented with 0.5% DMSO. Camptothecin (CPT), a known inhibitor of cell proliferation was used as positive control at a concentration of 0.001 M in E3 medium/0.5% DMSO. Adults zebrafish were placed for breeding a day before the experimental set up, then, viable embryos were collected and selected for use. Experiments were carried out in triplicates, according to specifications from Organization for Economic Cooperation and Development (OECD). One embryo/well (25 embryos per concentration) was distributed in 96 well microplates in presence or absence of the chemicals. The plates were kept in BOD incubators with a controlled temperature of 28.5 ºC and with photoperiod of 14 h light:10 h dark. After 24h, 48h, 72h and 96h exposure, the exposed embryos were evaluated according to the following parameters: mortality, coagulation, rate of heartbeat, hatching and presence of morphological abnormalities. Photography was obtained by photomicroscopy. Apoptosis was evaluated by DNA ladder assay. DNA was extracted by phenol:chloroform method and analyzed by 2% agarose gel electrophoresis. DNA fragments were visualized after ethidium bromide staining in ultraviolet transilluminator. The LC50 determined for BPA was 70 M after 24 hours, 72 M after 48 hours, 47 M after 72 hours and 31 M after 96 hours exposure. BPA induced morphological and physiological alterations such as yolk sac and pericardial edema, hatching delay or inhibition, spine deformation, decreasing in heartbeat rate and mortality. In conclusion, this study demonstrated that BPA induced marked malformations in zebrafish embryos at concentrations above 25 M corroborating the current concerns related to the widespread presence of BPA in the air, food and water used by humans as well as in the bodily fluids and tissues.(AU)

Bisfenol A (BPA) é um monômero utilizado na produção de policarbonato, um polímero comumente encontrado em plásticos, resinas epóxi e papéis térmicos. A presença de BPA em alimentos, água, ar e poeira tem sido motivo de grande preocupação nos últimos anos, não só devido a questões ambientais e ecológicas, mas também ao suposto risco para a saúde pública relacionado ao seu potencial mutagênico e carcinogênico. Neste estudo avaliamos a toxicidade do bisfenol A em embriões de peixe-zebra (Danio rerio) e determinamos a concentração letal 50% (LC50) deste composto químico. O BPA foi usado na faixa de concentração entre 1 M e 100M em meio E3/0,5% de dimetilsulfóxido (DMSO), preparado a partir de soluções estoques em 100% DMSO. Os controles negativos incluíram embriões expostos apenas ao meio E3 ou suplementado com 0,5% DMSO. Camptotecina (CPT), um conhecido inibidor da proliferação celular, foi usado como controle positivo a uma concentração de 0,001 M em meio E3/0,5% DMSO. Peixes-zebra adultos foram colocados para reprodução um dia antes da montagem experimental, em seguida, embriões viáveis foram coletados e selecionados para uso. Os experimentos foram realizados em triplicata, de acordo com as especificações da Organização para Cooperação e Desenvolvimento Econômico (OCDE). Um embrião/ poço (25 embriões por concentração) foi distribuído em microplacas de 96 poços na presença ou ausência dos compostos químicos. As placas foram mantidas em incubadoras BOD com temperatura controlada de 28,5 ºC e com fotoperíodo de 14h claro:10h escuro. Após 24h, 48h, 72h e 96h, os embriões expostos foram avaliados de acordo com os seguintes parâmetros: mortalidade, presença de coagulação, taxa do batimento cardíaco, eclosão e presença de anormalidades morfológicas. Fotografias foram obtidas por fotomicroscopia. A apoptose foi avaliada pelo ensaio de DNA ladder. O DNA foi extraído pelo método fenol:clorofórmio e analisado por eletroforese em gel de agarose a 2%. Fragmentos de DNA foram visualizadas após coloração com brometo de etídio em um transiluminador ultravioleta. A LC50 determinada para o BPA foi 70 M após 24 horas, 72 M após 48 horas, 47 M após 72 horas e 31 M após exposição por 96 horas. O BPA induziu alterações morfológicas e fisiológicas como edema de saco vitelino e edema pericárdico, atraso no tempo ou inibição da eclosão, deformação da coluna vertebral, diminuição da taxa de batimentos cardíacos e mortalidade. Em conclusão, este estudo demonstrou que o BPA induziu grande número de malformações em embriões de peixe-zebra em concentrações acima de 25 M, corroborando as preocupações atuais relacionadas a presença generalizada do BPA no ar, alimento e água usados pelos seres humanos bem como nos fluidos e tecidos corporais.(AU)

BPA toxicity during development of zebrafish embryo / Toxicidade do BPA durante o desenvolvimento de embriões de Zebrafish

Bisphenol A (BPA) is a monomer used in the production of polycarbonate, a polymer commonly found in plastics, epoxy resins and thermal papers. The presence of BPA in food, water, air and dust has been of great concern in recent years not only due to environmental and ecological issues but also because of its supposed risk to public health related to its mutagenic and carcinogenic potential. In this study we evaluated the toxicity of bisphenol A in zebrafish embryos (Danio rerio) and determined the 50% lethal concentration (LC50) of this chemical. BPA was used at concentrations ranging from 1 µM to 100 µM in E3 medium/0.5% dimethylsulfoxide (DMSO) from previously prepared stock solutions in 100% DMSO. Controls included embryos exposed only to E3 medium or supplemented with 0.5% DMSO. Camptothecin (CPT), a known inhibitor of cell proliferation was used as positive control at a concentration of 0.001 µM in E3 medium/0.5% DMSO. Adults zebrafish were placed for breeding a day before the experimental set up, then, viable embryos were collected and selected for use. Experiments were carried out in triplicates, according to specifications from Organization for Economic Cooperation and Development (OECD). One embryo/well (25 embryos per concentration) was distributed in 96 well microplates in presence or absence of the chemicals. The plates were kept in BOD incubators with a controlled temperature of 28.5 ºC and with photoperiod of 14 h light:10 h dark. After 24h, 48h, 72h and 96h exposure, the exposed embryos were evaluated according to the following parameters: mortality, coagulation, rate of heartbeat, hatching and presence of morphological abnormalities. Photography was obtained by photomicroscopy. Apoptosis was evaluated by DNA ladder assay. DNA was extracted by phenol:chloroform method and analyzed by 2% agarose gel electrophoresis. DNA fragments were visualized after ethidium bromide staining in ultraviolet transilluminator. The LC50 determined for BPA was 70 µM after 24 hours, 72 µM after 48 hours, 47 µM after 72 hours and 31 µM after 96 hours exposure. BPA induced morphological and physiological alterations such as yolk sac and pericardial edema, hatching delay or inhibition, spine deformation, decreasing in heartbeat rate and mortality. In conclusion, this study demonstrated that BPA induced marked malformations in zebrafish embryos at concentrations above 25 µM corroborating the current concerns related to the widespread presence of BPA in the air, food and water used by humans as well as in the bodily fluids and tissues.

Bisfenol A (BPA) é um monômero utilizado na produção de policarbonato, um polímero comumente encontrado em plásticos, resinas epóxi e papéis térmicos. A presença de BPA em alimentos, água, ar e poeira tem sido motivo de grande preocupação nos últimos anos, não só devido a questões ambientais e ecológicas, mas também ao suposto risco para a saúde pública relacionado ao seu potencial mutagênico e carcinogênico. Neste estudo avaliamos a toxicidade do bisfenol A em embriões de peixe-zebra (Danio rerio) e determinamos a concentração letal 50% (LC50) deste composto químico. O BPA foi usado na faixa de concentração entre 1 µM e 100µM em meio E3/0,5% de dimetilsulfóxido (DMSO), preparado a partir de soluções estoques em 100% DMSO. Os controles negativos incluíram embriões expostos apenas ao meio E3 ou suplementado com 0,5% DMSO. Camptotecina (CPT), um conhecido inibidor da proliferação celular, foi usado como controle positivo a uma concentração de 0,001 µM em meio E3/0,5% DMSO. Peixes-zebra adultos foram colocados para reprodução um dia antes da montagem experimental, em seguida, embriões viáveis foram coletados e selecionados para uso. Os experimentos foram realizados em triplicata, de acordo com as especificações da Organização para Cooperação e Desenvolvimento Econômico (OCDE). Um embrião/ poço (25 embriões por concentração) foi distribuído em microplacas de 96 poços na presença ou ausência dos compostos químicos. As placas foram mantidas em incubadoras BOD com temperatura controlada de 28,5 ºC e com fotoperíodo de 14h claro:10h escuro. Após 24h, 48h, 72h e 96h, os embriões expostos foram avaliados de acordo com os seguintes parâmetros: mortalidade, presença de coagulação, taxa do batimento cardíaco, eclosão e presença de anormalidades morfológicas. Fotografias foram obtidas por fotomicroscopia. A apoptose foi avaliada pelo ensaio de DNA ladder. O DNA foi extraído pelo método fenol:clorofórmio e analisado por eletroforese em gel de agarose a 2%. Fragmentos de DNA foram visualizadas após coloração com brometo de etídio em um transiluminador ultravioleta. A LC50 determinada para o BPA foi 70 µM após 24 horas, 72 µM após 48 horas, 47 µM após 72 horas e 31 µM após exposição por 96 horas. O BPA induziu alterações morfológicas e fisiológicas como edema de saco vitelino e edema pericárdico, atraso no tempo ou inibição da eclosão, deformação da coluna vertebral, diminuição da taxa de batimentos cardíacos e mortalidade. Em conclusão, este estudo demonstrou que o BPA induziu grande número de malformações em embriões de peixe-zebra em concentrações acima de 25 µM, corroborando as preocupações atuais relacionadas a presença generalizada do BPA no ar, alimento e água usados pelos seres humanos bem como nos fluidos e tecidos corporais.

Effects of placement time on performance and gastrointestinal tract growth of male broiler chickens

The main objective of this study was to evaluate the effect of placement time on the performance of broiler chickens and the development of their gastrointestinal tract. Two methodologies for measuring broiler performance were compared, one considering day of pulling as the first day, the other considering day of placement as the first day. A total of 1,056 one-day old male Cobb® 500 broiler chicks were subjected to treatments of different placement times after pulling from the hatchery: 3, 24, 48, and 72 h. The studied traits were: feed intake, body weight, feed conversion, viability, and gastrointestinal tract development. When day of pulling was considered the first day, feed intake and body weight at 39 days decreased as placement time increased. However, when day of placement was considered the first day, fasting up to 72 h did not have any negative effect on broiler performance at 39 days post-placement. Placement time did not affect yolk sac utilization or liver weight. At nine days post-placement, weights of gizzard + proventriculus, pancreas, and small intestine increased with increasing placement time. At seven days of age, there was no effect of placement time on villus height or crypt depth. It is possible to place broiler chicks up to 72 h post-hatching with no negative effects when day of placement is considered the first day for evaluating broiler performance.(AU)

Use of Betaine in Post-Hatch Feed for Broiler Chicks

Post-hatch delayed placement damages the physical and physiological development of broiler chicks. This study was designed to find adequate levels of betaine inclusion in pre-hatching and pre-starter feed, in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, five replicates of ten birds each. Five dietetic levels of betaine (control, 0.070, 0.130, 0.200 and 0.260%) were used in the pre-starter ration, offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated, after 24 hours of feed access and at 7 days of age. A metabolic trial was performed from seven to ten days of age. Betaine supplementation linearly influenced the chicks ileum crypt depth after 24 hours in the transportation box. There was a quadratic effect, with an increase in feed intake up to 0.152% betaine supplementation. There was an improvement in the quality of the jejunum with betaine supplementation above 0.1%. The metabolizable coefficient of the ether extract was improved until reaching 0.163% of betaine supplementation. Betaine supplementation around 0.150% increases feed intake, the use of ether extract and interferes with the intestinal villi of chicks at seven days of age.(AU)

BPA toxicity during development of zebrafish embryo

Abstract Bisphenol A (BPA) is a monomer used in the production of polycarbonate, a polymer commonly found in plastics, epoxy resins and thermal papers. The presence of BPA in food, water, air and dust has been of great concern in recent years not only due to environmental and ecological issues but also because of its supposed risk to public health related to its mutagenic and carcinogenic potential. In this study we evaluated the toxicity of bisphenol A in zebrafish embryos (Danio rerio) and determined the 50% lethal concentration (LC50) of this chemical. BPA was used at concentrations ranging from 1 M to 100 M in E3 medium/0.5% dimethylsulfoxide (DMSO) from previously prepared stock solutions in 100% DMSO. Controls included embryos exposed only to E3 medium or supplemented with 0.5% DMSO. Camptothecin (CPT), a known inhibitor of cell proliferation was used as positive control at a concentration of 0.001 M in E3 medium/0.5% DMSO. Adults zebrafish were placed for breeding a day before the experimental set up, then, viable embryos were collected and selected for use. Experiments were carried out in triplicates, according to specifications from Organization for Economic Cooperation and Development (OECD). One embryo/well (25 embryos per concentration) was distributed in 96 well microplates in presence or absence of the chemicals. The plates were kept in BOD incubators with a controlled temperature of 28.5 ºC and with photoperiod of 14 h light:10 h dark. After 24h, 48h, 72h and 96h exposure, the exposed embryos were evaluated according to the following parameters: mortality, coagulation, rate of heartbeat, hatching and presence of morphological abnormalities. Photography was obtained by photomicroscopy. Apoptosis was evaluated by DNA ladder assay. DNA was extracted by phenol:chloroform method and analyzed by 2% agarose gel electrophoresis. DNA fragments were visualized after ethidium bromide staining in ultraviolet transilluminator. The LC50 determined for BPA was 70 M after 24 hours, 72 M after 48 hours, 47 M after 72 hours and 31 M after 96 hours exposure. BPA induced morphological and physiological alterations such as yolk sac and pericardial edema, hatching delay or inhibition, spine deformation, decreasing in heartbeat rate and mortality. In conclusion, this study demonstrated that BPA induced marked malformations in zebrafish embryos at concentrations above 25 M corroborating the current concerns related to the widespread presence of BPA in the air, food and water used by humans as well as in the bodily fluids and tissues.

Resumo Bisfenol A (BPA) é um monômero utilizado na produção de policarbonato, um polímero comumente encontrado em plásticos, resinas epóxi e papéis térmicos. A presença de BPA em alimentos, água, ar e poeira tem sido motivo de grande preocupação nos últimos anos, não só devido a questões ambientais e ecológicas, mas também ao suposto risco para a saúde pública relacionado ao seu potencial mutagênico e carcinogênico. Neste estudo avaliamos a toxicidade do bisfenol A em embriões de peixe-zebra (Danio rerio) e determinamos a concentração letal 50% (LC50) deste composto químico. O BPA foi usado na faixa de concentração entre 1 M e 100M em meio E3/0,5% de dimetilsulfóxido (DMSO), preparado a partir de soluções estoques em 100% DMSO. Os controles negativos incluíram embriões expostos apenas ao meio E3 ou suplementado com 0,5% DMSO. Camptotecina (CPT), um conhecido inibidor da proliferação celular, foi usado como controle positivo a uma concentração de 0,001 M em meio E3/0,5% DMSO. Peixes-zebra adultos foram colocados para reprodução um dia antes da montagem experimental, em seguida, embriões viáveis foram coletados e selecionados para uso. Os experimentos foram realizados em triplicata, de acordo com as especificações da Organização para Cooperação e Desenvolvimento Econômico (OCDE). Um embrião/ poço (25 embriões por concentração) foi distribuído em microplacas de 96 poços na presença ou ausência dos compostos químicos. As placas foram mantidas em incubadoras BOD com temperatura controlada de 28,5 ºC e com fotoperíodo de 14h claro:10h escuro. Após 24h, 48h, 72h e 96h, os embriões expostos foram avaliados de acordo com os seguintes parâmetros: mortalidade, presença de coagulação, taxa do batimento cardíaco, eclosão e presença de anormalidades morfológicas. Fotografias foram obtidas por fotomicroscopia. A apoptose foi avaliada pelo ensaio de DNA ladder. O DNA foi extraído pelo método fenol:clorofórmio e analisado por eletroforese em gel de agarose a 2%. Fragmentos de DNA foram visualizadas após coloração com brometo de etídio em um transiluminador ultravioleta. A LC50 determinada para o BPA foi 70 M após 24 horas, 72 M após 48 horas, 47 M após 72 horas e 31 M após exposição por 96 horas. O BPA induziu alterações morfológicas e fisiológicas como edema de saco vitelino e edema pericárdico, atraso no tempo ou inibição da eclosão, deformação da coluna vertebral, diminuição da taxa de batimentos cardíacos e mortalidade. Em conclusão, este estudo demonstrou que o BPA induziu grande número de malformações em embriões de peixe-zebra em concentrações acima de 25 M, corroborando as preocupações atuais relacionadas a presença generalizada do BPA no ar, alimento e água usados pelos seres humanos bem como nos fluidos e tecidos corporais.