In ovo injection of ascorbic acid and higher incubation temperature modulate blood parameters in response to heat exposure in broilers.

1. This study analysed whether in ovo injection of ascorbic acid before incubation and at high incubation temperature influenced blood characteristics and performance in broilers reared in different temperature conditions. 2. A total of 3,000 fertile eggs from broiler breeders (Cobb®) were randomly divided into three incubation treatments: no ascorbic acid injection and egg incubation at 37.5°C (control); no ascorbic acid injection and egg incubation at 39°C; in ovo ascorbic acid injection prior to incubation (6 µg AA/100 µl water) and egg incubation at 39°C. 3. Male chicks hatched from the three incubation treatments were submitted to three distinct rearing temperatures (control, cold and hot) from the third week of age onwards (540 chicks were divided into 6 treatments with 5 replicates per treatment). 4. Measurements at 42 d showed that, after egg incubation at 39°C, the haematocrit, haemoglobin values, ionised calcium and glucose concentrations were increased and base excess values were reduced. However, in ovo injection of ascorbic acid normalised all these parameters. 5. Partial CO2 and O2 pressure were higher with increased rearing temperature. Blood pH was lower when eggs were incubated at 39°C and injected with ascorbic acid. In ovo injection of ascorbic acid induced leucocytosis due to lymphocytosis and heterophilia, restored basophils rate and led to monocytopoenia. Leucocytosis was triggered by hot rearing temperature due to lymphocytosis, eosinophilia and heterophilia. 6. The results obtained in this study showed that in ovo injection of ascorbic acid before incubation may serve as a long-term stimulator and modulator of the broiler immune system, and that high incubation temperatures induce adaptations in the electrolytic balance, minimising or avoiding the occurrence of respiratory alkalosis under hot rearing temperature.

Incubation temperature alters thermal preference and response to heat stress of broiler chickens along the rearing phase.

The current study aimed to investigate whether embryonic temperature manipulation may alter thermal preference throughout the rearing phase of broiler chickens and how this manipulation may affect response to thermal challenge, metabolism, growth rate and feed intake rate. Eggs were exposed to a constant incubation temperature [machine temperatures: 36°C (Low), 37.5°C (Control), and 39°C (High); eggshell temperature of 37.4 ± 0.08°C, 37.8 ± 0.15°C, and 38.8 ± 0.33°C, respectively] from d 13 till hatching. Low treatment chickens showed lower plasma T3 and GH levels at d 1 of age and lower T3 level at d 42 of age compared to the Control treatment. Preferred ambient, rectal temperature, T4 level, growth rate, food intake rate, and response to thermal challenge were not altered in these chickens. On the other hand, High-treatment chickens exhibited high preferred ambient temperature and rectal temperature during the first 2 wk post-hatch, lower plasma T3 level at d 21 and 42 and a delayed increase in respiratory movement in response to thermal challenge compared to the Control treatment. However, chickens subjected to the Control and High treatments did not differ in T4 and GH level and performance. We conclude that exposure to high temperature during late embryonic development has long-lasting effects on the thermoregulatory system of broiler chickens by affecting the heat tolerance of these chickens. Moreover, the preferred ambient temperature of the chickens from heat-treated eggs correspond to those recommended for the strain under study, whereas for the cold-treated and control-chickens it was 1°C below, indicating that incubation temperature might have consequences on the ambient temperature chickens require during the rearing phase.

Incubation temperature manipulation during fetal development reduces adiposity of broiler hatchlings.

Broilers are known as an efficient source of lean meat. Genetic selection resulted in broiler strains with large body size and fast growth, but a concomitant increase in fat deposition also occurred. Other than reducing nutrient intake, there is a lack of alternative methods to control body fat composition of broilers. The present study assessed whether incubation temperature (machine temperatures: 36ºC, 37.5ºC, and 39ºC; eggshell temperatures: 37.4 ± 0.08°C, 37.8 ± 0.15ºC, and 38.8 ± 0.33°C, respectively.) from d 13 affects broiler hatchling fat deposition. We analyzed adipocyte hypertrophy and proliferation in 3 body regions; weight and chemical composition of yolk-free chicks and yolk sacs; and serum lipid profile. Increased incubation temperature reduced abdominal and cervical adipocyte size. Independently of temperature, cervical adipocytes were smaller and showed higher proliferation than adipocytes in the abdominal and thigh regions. Smaller cervical adipocytes were observed in birds from eggs incubated at 36ºC and 39ºC. With regard to weight and composition of chicks, ash content as a percentage of dry matter was the only variable affected by temperature; it was higher in chicks from eggs incubated at 36ºC than at 39ºC and showed no significant difference between chicks incubated at 39ºC and 37.5ºC. Absolute and relative weights of yolk sacs were higher from eggs incubated at 39ºC than at 36ºC, and these two treatments did not differ from the 37.5ºC control. Absolute measures of yolk sac lipids, moisture, dry matter, and crude protein content were lower in chicks from eggs incubated at 36ºC, and no significant differences were found for these variables between chicks from eggs incubated at 37.5ºC and 39ºC. Hatchlings from eggs incubated at 36°C had significantly higher cholesterol levels than chicks incubated at the other 2 temperatures, but no additional effects on blood lipids were detected. Incubation temperature manipulation during fetal development altered cervical and abdominal adipocyte size in broiler hatchlings and could become a tool in hatcheries to manipulate chick quality, although further studies are needed to evaluate its long-term effects.

Vitamin C prevents the effects of high rearing temperatures on the quality of broiler thigh meat1.

We investigated the effects of incubation temperatures and vitamin C injections into eggs (treatments: 37.5ºC, 39ºC, 39ºC+vitamin C) on resultant chick pectoralis major and sartorius muscle fiber hypertrophy, as well as their effects on the quality of breast and over-thigh meat of broilers reared under cold, control, or hot temperatures. Incubation at 39ºC increased the shear force and reduced meat redness in breast meat (P < 0.05). Vitamin C prevented these high temperature incubation effects [shear force (kgf cm(-2)): 37.5ºC = 2.34, 39ºC = 2.79, 39ºC+vitamin C = 2.44; redness: 37.5ºC = 2.64, 39ºC = 1.90, 39ºC+vitamin C = 2.30], but reduced water content (37.5ºC = 74.81%, 39ºC = 74.53%, 39ºC+vitamin C = 69.39%) (P < 0.05). Cold rearing temperatures increased breast meat redness (a*: cold = 2.78, control = 2.12, hot = 1.98), while hot rearing temperatures reduced the muscle fiber area (cold = 5.413 µm(2), control = 5.612 µm(2), hot = 4.448 µm(2)) (P < 0.05) without altering meat quality (P > 0.05). Hot rearing temperatures increased the cooking loss (cold = 30.10%, control = 33.66%, hot = 37.01%), shear force (cold = 3.05 kgf cm(-2), control = 3.43 kgf cm(-2), hot = 4.29 kgf cm(-2)) and redness (a*: cold = 4.63, control = 3.55, hot = 3.20) in the over-thigh meat of broilers from eggs incubated at 37.5ºC, increasing the area of muscle fibers, while cold rearing temperatures diminished cooking loss and shear force, reducing the muscle fiber area (P < 0.05). Incubation at 39ºC and 39ºC+vitamin C prevented the effects of hot and cold rearing temperatures, by diminishing and increasing the muscle fiber area, respectively.

Efeitos do espaço de alojamento e da idade sobre desenvolvimento, integridade e proteção da mucosa intestinal de frangos de corte / Effects of enclosure size and age on development, integrity and protection of the intestinal mucosa in broiler chickens

Foi avaliado se o espaço de alojamento influencia o desenvolvimento da mucosa do intestino delgado de frangos, sendo usado um esquema fatorial 4 x 2 (14, 28, 42, 55 dias de idade e alojamento pequeno: 0,11m²/ave e grande: 0,525m²/ave). Foram analisadas: altura, perímetro e número dos vilos, profundidade das criptas, número de células caliciformes e número de vilos sem e com perda de epitélio e exposição do conjuntivo. Em ambos os espaços de alojamento, o número de vilos diminuiu e a profundidade das criptas aumentou com a idade nas três regiões intestinais. A altura e o perímetro dos vilos aumentaram de 14 para 28 dias no jejuno e íleo, e o número de células caliciformes foi maior no duodeno e íleo a partir dos 42 dias. A perda de epitélio pelos vilos do íleo aumentou com a idade. Nas aves do alojamento grande, ocorreu aumento na altura e no perímetro dos vilos duodenais com a idade, possibilitado pelo aumento na profundidade das criptas, que resultou em vilos maiores nessas aves do que nas do alojamento pequeno aos 55 dias de idade. O número de células caliciformes diminuiu nas aves do alojamento pequeno aos 42 dias de idade. Os dados mostram que o tamanho do alojamento interferiu no modelo de desenvolvimento temporal da mucosa intestinal e que frangos apresentaram maior tamanho de vilo duodenal quando alojados em espaço maior.

This study evaluated if enclosure size and age influence the intestinal mucosa development of broiler chickens, using an experimental design in an 4x2 factorial arrangement (14, 18, 42 and 55 days of age and two enclosure spaces: small= 0.11m² /bird and large= 0.525m²/bird). Villous height, perimeter and number, and crypt depth, goblet cells number, and number of villous without and with epithelium loss and conjunctive tissue exposition were analyzed in duodenum, jejunum and ileum. Regardless of enclosure size, the villous number diminished and the crypt depth increased with the age in the three intestinal regions. From 14 to 28 days of age, villous height and perimeter increased in the jejunum and ileum. The number of goblet cells was greater from 42 days in duodenum and ileum, and epithelium loss increased until 42 days in the ileum. Broilers housed in small and large enclosures differed in the intestinal characteristics from 42 days. In broilers housed in large enclosure, duodenal villous height and perimeter increased with the age, with an increase in the crypt depth, reaching greater size at 55 days of age than duodenal villous of the broilers housed in a small enclosure. These later presented at 42 days a smaller number of goblet cells than the former. According these results, enclosure size influenced the temporal pattern of the intestinal mucosa development, and broilers presented major duodenal villous growth when housed in large enclosures.

Desempenho e rendimento de carcaça de frangos de corte alimentados com pectina na ração / Performance and carcass yield of broiler chickens fed with pectin in the ration

Avaliou-se o efeito da ingestão contínua de pectina em 720 frangos de corte sobre o desempenho e rendimento de carcaça e o consumo de água. O delineamento foi inteiramente ao acaso, em esquema de parcela subdividida, com níveis de pectina 0, 1, 3 e 5%, e idade primeira, segunda, terceira, quarta, quinta e sexta semanas, com seis repetições por tratamento. A viabilidade criatória não foi influenciada pela pectina na ração. O peso vivo, o consumo de ração e o ganho de peso diminuíram, e a conversão alimentar aumentou com a ingestão de pectina aos 35 e 42 dias. Não houve efeito da pectina sobre o consumo de ração (CR) e o consumo de água (CAg) na terceira semana. Na sexta semana, o CAg e a relação CAg:CR aumentaram com a ingestão de pectina. Aos 35e 42 dias, todos os parâmetros avaliados para rendimento de carcaça e corte diminuíram com o aumento da ingestão da fibra, sendo que apenas o rendimento de coxa+sobrecoxa não sofreu influência aos 42 dias. Os dados mostram que a ingestão contínua de até 1% de pectina na ração mantém o desempenho máximo das aves e que o consumo acima desse nível prejudica os parâmetros zootécnicos e aumenta o consumo de água na fase de maior crescimento.

The effect of the continuous intake of pectin was evaluated for 720 broilers regarding performance, carcass yield and water consumption. The design was completely randomized in a split plot design, with pectin levels of 0, 1, 3 and 5% and age 1st, 2nd, 3rd, 4th, 5th and 6th week, with six replicates. The viability was not influenced by dietary pectin. Body weight, feed intake and weight gain decreased and feed intake increased with pectin at 35 and 42 days. No effect of pectin on feed intake (FI) and water consumption (W) in the third week. On the sixth week regarding W and FI: W increased with the intake of pectin. At 35 and 42 days all parameters for carcass and cut decreased with increasing intake of fiber, and only the thigh + drumstick yield was not affected at 42 days. The data show that the continuous intake of up to 1% pectin in the diet maintains the best performance in birds and consumption above that level affects the performance parameters and increases the consumption of water in the higher growth phase.

Composição de fibras musculares esqueléticas de eqüinos jovens da raça Brasileiro de Hipismo / Composition of skeletal muscle fibers of young Brasileiro de Hipismo horse breed

The aim of this study was to typify the skeletal striated fibers of the gluteus medius muscle of young Brasileiro de Hipismo (BH) horses by means of histochemical analysis with m-ATPase and NADH-TR according to the sex and the biopsy depth. It was observed that the frequency (F; percent) and the relative cross sectional area (RCSA; percent) of the fibers type IIX were greater than the fibers type IIA, which F and RCSA were greater than the fibers type I. The comparison between sex and muscles depht, showed no significant difference in F and RCSA in the three types of fibers. The results of morphometry showed that the gluteus medius muscle has greater glycolitic metabolism and anaerobic capacity because of the presence of large proportion of type IIX fibers. This may be justified by the genetic influence of Thoroughbred in the formation of Brasileiro de Hipismo breed.

Morfometria de testículos escrotais, abdominais e inguinais de eqüinos criptórquios unilaterais / Morphometry of scrotal, abdominal and inguinal testes of unilateral cryptorchid horses

Foi realizado um estudo morfométrico de testículos escrotais e retidos de 10 eqüinos criptórquios unilaterais, sendo cinco abdominais e cinco inguinais. As características avaliadas foram espessura da albugínea testicular, área dos túbulos seminíferos e número de células de Leydig. A espessura da albugínea testicular mostrou-se reduzida (P< 0,05) nas gônadas criptórquias abdominais quando comparadas às escrotais contralaterais. Também foram observadas diferenças (P< 0,01) nas áreas dos túbulos seminíferos dos testículos abdominais e inguinais, que se apresentaram diminuídas quando comparadas às dos escrotais contralaterais. A diminuição foi de, aproximadamente, 45 por cento nos testículos abdominais e de 31 por cento nos inguinais. Não foram verificadas diferenças (P> 0,05) na contagem de células de Leydig das gônadas criptórquias comparadas às escrotais contralaterais. Nos eqüinos, o criptorquismo afetou com maior intensidade a área dos túbulos seminíferos dos testículos abdominais e inguinais e a espessura da albugínea daqueles retidos no abdome.

Aerobic training, but not creatine supplementation, alters the gluteus medius muscle.

The aim of the present study was to investigate the effect of oral supplementation of creatine on the muscular responses to aerobic training. Twelve purebred Arabian horses were submitted to aerobic training for 90 d, with and without creatine supplementation, and evaluated with respect to BW and BCS and to the area and frequency of the different types of muscle fibers in the gluteus medius. Supplementation consisted of the daily administration of 75 g of creatine monohydrate mixed into the ration for the 90 d of training. Physical conditioning was conducted on a high-performance treadmill, and training intensity was stipulated by calculating the velocity at which blood lactate reaches 4 mmol/L, determined monthly for each animal. The individual intensity of physical force at 80% of aerobic threshold was established. Morphometry of gluteus medius muscle fibers was performed on frozen sections processed for histochemical analysis of myosin adenosine triphosphatase and immunohistochemistry of slow-contracting myosin. The results demonstrated that the animals maintained a moderate BCS without alteration of BW during the course of training, providing evidence of equilibrium between food intake and caloric expenditure during the study period. The present study demonstrated that aerobic training for 90 d caused hypertrophy of fiber types I (P = 0.04), IIA (P = 0.04), and IIX (P = 0.01), as well as an increase in the relative area occupied by type I fibers (P = 0.02) at the expense of type IIX fibers (P = 0.03), resulting in modifications of the contractile and metabolic characteristics of the gluteus medius muscle. It was not possible to show any beneficial effect from creatine on the skeletal muscle characteristics examined.

Hemocyte types and total and differential counts in unparasitized and parasitized Anastrepha obliqua (Diptera, Tephritidae) larvae

The hemocyte types, in addition to total and differential hemocyte counts were studied in parasitized and unparasitized Anastrepha obliqua larvae at the beginning and at the end of the third instar. In both developmental phases, in parasitized and unparasitized larvae, prohemocytes, plasmatocytes, granulocytes, adipohemocytes, spherulocytes and oenocytoids cells were observed. Mitotic figures indicate prohemocytes as stem cells. Prohemocytes, plasmatocytes and granulocytes are the most numerous cells in the hemolymph of A. obliqua. Difference in the total number of hemocytes was observed between unparasitized and parasitized larvae at the end of the third instar, but not at the beginning

Hemocyte types and total and differential counts in unparasitized and parasitized Anastrepha obliqua (Diptera, Tephritidae) larvae.

The hemocyte types, in addition to total and differential hemocyte counts were studied in parasitized and unparasitized Anastrepha obliqua larvae at the beginning and at the end of the third instar. In both developmental phases, in parasitized and unparasitized larvae, prohemocytes, plasmatocytes, granulocytes, adipohemocytes, spherulocytes and oenocytoids cells were observed. Mitotic figures indicate prohemocytes as stem cells. Prohemocytes, plasmatocytes and granulocytes are the most numerous cells in the hemolymph of A. obliqua. Difference in the total number of hemocytes was observed between unparasitized and parasitized larvae at the end of the third instar, but not at the beginning.

Mispatterning in the ommatidia of Apis mellifera pupae treated with a juvenile hormone analogue.

To further understand the function of morphogenetic hormones in honeybee eye differentiation, the alterations in ommatidial patterning induced by pyriproxyfen, a juvenile hormone (JH) analogue, were studied by scanning and transmission electron microscopy. Prepupae of prospective honeybee workers were treated with pyriproxyfen and the effects on ommatidial differentiation were described at the end of the pupal development. The results show that the entire ommatidia, i.e., the dioptric as well as the receptor systems, were affected by the JH analogue. The wave of ommatidial differentiation, which progresses from the posterior to the anterior region of the pupal eyes, was arrested. In treated pupae, the rhabdomeres only differentiated at the apical axis of the retinula, the secondary and tertiary pigment cells did not develop their cytoplasm protrusions, and the cone cell quartet did not pattern correctly. Simultaneously, an intense vacuolization was observed in cells forming ommatidia. In a previous study we showed that pyriproxyfen exerts an inhibition on pupal ecdysteroid secretion. In this sense, the arrested ommatidial differentiation in pyriproxyfen-treated pupae could be due to a secondary effect resulting from an alteration in pupal ecdysteroid titers.

Influência da suplementaçäo de glutamina sobre o desempenho e o desenvolvimento de vilos e criptas do intestino delgado de frangos / Influence of glutamine supplementation on performance and intestinal villous and crypt development in broiler chickens

Investigou-se o efeito da suplementaçäo de glutamina na dieta sobre o consumo de raçäo, ganho de peso e conversäo alimentar e sobre a estrutura da mucosa intestinal de frangos. Foram utilizados 320 pintos de corte machos distribuídos em um delineamento inteiramente ao acaso com dois tratamentos e quatro repetiçöes, sendo T1 suplementado com 1 por cento de L-glutamina na dieta e T2 controle. Os índices de desempenho foram analisados aos 7, 21 e 49 dias de idade das aves. Aos 7 e 14 dias de idade oito aves foram sacrificadas para colheita de fragmentos de cada porçäo do intestino delgado para avaliaçäo da morfometria intestinal em microscopia de luz em sistema analisador de imagens "Video Plan". As variáveis estudadas foram altura dos vilos, profundidade de cripta e relaçäo vilo: cripta. A adiçäo de 1 por cento de glutamina à dieta de frangos näo influenciou (P>0,05) o seu desempenho zootécnico. Entretanto, 1 por cento de glutamina na raçäo foi capaz de alterar (P<0,01) a altura do vilo, a profundidade de cripta e a relaçäo vilo:cripta no duodeno, bem como a altura de vilo do íleo de frangos no sétimo dia de idade

Regression of the lateral oviducts during the larval-adult transformation of the reproductive system of Melipona quadrifasciata and Frieseomelitta varia.

Toward the end of the larval phase (prepupa), the reproductive systems of Melipona quadrifasciata and Frieseomelitta varia workers are anatomically similar. Scanning electron microscopy showed that during this developmental phase the right and left ovaries are fused and form a heart-shaped structure located above the midgut. Each ovary is connected to the genital chamber by a long and slender lateral oviduct. During pupal development, the lateral oviducts of workers from both species become extremely reduced due to a drastic process of cell death, as shown by transmission electron microscopy. During the lateral oviduct shortening, their simple columnar epithelial cells show some signs of apoptosis in addition to necrosis. Cell death was characterized by cytoplasmic vesiculation, peculiar accumulation of glycogen, and dilation of cytoplasmic organelles such as mitochondria and rough endoplasmic reticulum. The nuclei, at first irregularly contoured, became swollen, with chromatin flocculation and various areas of condensed chromatin next to the nuclear envelope. At the end of the pupal phase, deep recesses marked the nuclei. At emergence, worker and queen reproductive systems showed marked differences, although reduction in the lateral oviducts was an event occurring in both castes. However, in queens the ovarioles increased in length and the spermatheca was larger than that of workers. At the external anatomical level, the reproductive system of workers and queens could be distinguished in the white- and pink-eyed pupal phase. The metamorphic function of the death of lateral oviduct cells, with consequent oviduct shortening, is discussed in terms of the anatomical reorganization of the reproductive system and of the ventrolateral positioning of adult worker bee ovaries.

Cell death in ovarioles causes permanent sterility in Frieseomelitta varia worker bees.

Frieseomelitta varia worker bees do not lay eggs even when living in queenless colonies, a condition that favors ovary development and oviposition in the majority of highly social bees. The permanent sterility of these worker bees was initially attributed to a failure in ovary morphogenesis and differentiation. Using transmission electron microscopy we found that at the beginning of the pupal phase the ovaries of F. varia workers are formed by four ovarioles, each of them composed of 1) a terminal filament at the apex of the ovarioles, containing juxtaposed and irregularly shaped cells, 2) a germarium with clusters of cystocytes and prefollicular cells showing long cytoplasmic projections that envelop the cystocyte clusters, 3) fusiform interfollicular and basal stalk precursor cells, and 4) globular, irregularly contoured basal cells with large nuclei. However, during the pupal phase an accentuated and progressive process of cell death takes place in the ovarioles. The dying cells are characterized by large membrane bodies, electron-dense apoptotic bodies, vacuoles, vesiculation, secondary lysosomes, enlarged rough endoplasmic reticulum cisternae, swollen mitochondria, pycnotic nuclei, masses of chromatin adjacent to the convoluted nuclear envelope, and nucleoli showing signs of fragmentation. Cell death continues in ovarioles even after the emergence of the workers. Once they become nurse bees, the ovaries have become transformed into a cell mass in which structurally organized ovarioles can no longer be identified. In F. varia workers, ovariole cell death most certainly is part of the program of caste differentiation.