Study of ex-ovo embryonic development of Gallus gallus domesticus / Estudio inicial del desarrollo embrionario ex-ovo de Gallus gallus domesticus

SUMMARY: The domestic chicken is a species of bird that has been extensively studied in regard to its biology and as a model organism for science. The reproduction of the species is by the laying of fertilized eggs, which in a period of 21 days will develop a chick inside. Several methods have been described to develop embryos ex-ovo, allowing the observation and manipulation of the organism. This work has the propose to standardize a method that allows the development of the embryos inside the artificial incubation system, which has a low cost and is easy to make. In this work, 100 chicken eggs were used to study the effects of humidity, mineral supplementation, and the preincubation time of the egg on the incubation ex-ovo of the embryos. Embryo development was documented through the different days. Pulverized eggshell was selected as an optimal source to provide calcium, magnesium, phosphorus, and other minerals to the developing embryo. By providing 900-1200 mg of pulverized eggshell, 40 mL of the 0.001 % solution of benzalkonium chloride, and a preincubation time of approximately 56 h, the embryos were able to develop until 19 days, and even though they did not reach hatching, the incubation conditions that allowed the survival and development of embryos until late stages were achieved. Thus, due to the conditions established for calcium, humidity and preincubation time, in the present work, the chicks reached 19 days of development.

El pollo doméstico es una especie de ave que ha sido ampliamente estudiada en cuanto a su biología y como organismo modelo para la ciencia. La reproducción de la especie es por la puesta de huevos fecundados, que en un período de 21 días desarrollarán un polluelo en su interior. Se han descrito varios métodos para desarrollar embriones ex-ovo, permitiendo la observación y manipulación del organismo. Este trabajo tuvo como objetivo estandarizar un método que permita el desarrollo de los embriones dentro del sistema de incubación artificial, el cual tiene un bajo costo y es fácil de realizar. En este trabajo se utilizaron 100 huevos de gallina para estudiar los efectos de la humedad, la suplementación mineral y el tiempo de preincubación del huevo sobre la incubación ex-ovo de los embriones. El desarrollo embrionario se documentó a través de los diferentes días. Se seleccionó la cáscara de huevo pulverizada como una fuente óptima para proporcionar calcio, magnesio, fósforo y otros minerales al embrión en desarrollo. Al suministrar 900-1200 mg de cáscara de huevo pulverizada, 40 mL de la solución de cloruro de benzalconio al 0.001 % y un tiempo de preincubación de aproximadamente 56 h, los embriones lograron desarrollarse hasta los 19 días, y aunque no llegaron a eclosionar, los embriones lograron desarrollarse hasta los 19 días. Se lograron condiciones de incubación que permitieron la supervivencia y desarrollo de los embriones hasta etapas tardías. Así, debido a las condiciones establecidas de calcio, humedad y tiempo de preincubación, en el presente trabajo los pollitos alcanzaron los 19 días de desarrollo.

Poultry egg incubation: integrating and optimizing production efficiency

Due to its central position in the production chain, in-ovo development is influenced by pre-incubation factors that affect the quality of embryonated eggs and incubation conditions themselves, and both may influence egg hatchability and chick quality, as well as bird survival, growth performance, and phenotype in the field. The evolution of the incubation process over the years is characterized by significant scientific and technological development. Presently, the main current focuses of research are the manipulation of thermal incubation conditions, eggshell temperature, and the integrated effects of factors that influence incubation. In this context, one of the questions that needs to be asked is how effective are the current physical conditions of incubation to promote greater hatchability and better quality chicks, and higher survival and better performance in the field under adverse conditions or not. What are the new and future prospects for incubation? The purpose of this paper was to review the role of the physical agents of incubation, such as temperature, relative humidity, O2 and CO2 concentration, and egg turning and position from an integrated perspective, considering egg incubation as the transitional link between egg and poultry production.(AU)

Poultry egg incubation: integrating and optimizing production efficiency

Due to its central position in the production chain, in-ovo development is influenced by pre-incubation factors that affect the quality of embryonated eggs and incubation conditions themselves, and both may influence egg hatchability and chick quality, as well as bird survival, growth performance, and phenotype in the field. The evolution of the incubation process over the years is characterized by significant scientific and technological development. Presently, the main current focuses of research are the manipulation of thermal incubation conditions, eggshell temperature, and the integrated effects of factors that influence incubation. In this context, one of the questions that needs to be asked is how effective are the current physical conditions of incubation to promote greater hatchability and better quality chicks, and higher survival and better performance in the field under adverse conditions or not. What are the new and future prospects for incubation? The purpose of this paper was to review the role of the physical agents of incubation, such as temperature, relative humidity, O2 and CO2 concentration, and egg turning and position from an integrated perspective, considering egg incubation as the transitional link between egg and poultry production.

Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis / Influência das condições de incubação nos meios de cultura para otimizar o primo isolamento de Mycobacterium bovis

The isolation of Mycobacterium bovis is critical to a surveillance system for bovine tuberculosis based on detection of lesions in abattoirs. Thus, four solid culture media and three incubation conditions were investigated to elucidate which combination overcomes the others by assessing growth, time to the first appearance of colonies and their number. Ninety-seven samples of granulomatous lesions were submitted to the decontamination procedure by 1-hexadecylpyridinium chloride at 0.75% w/v, and inoculated on two egg-based media, Stonebrink’s (ST) and Lõwenstein-Jensen’s with sodium pyruvate (LJp), and two agar-based media, tuberculosis blood agar (B83) and Middlebrook 7H11 medium (7H11). Each medium was incubated at 37°C for 90 days in three incubation conditions: in air, in air containing 10% carbon dioxide (CO2), and in air in slopes closed with burned hydrophobic cotton and subsequently plugged with a cork to create a microaerophilic atmosphere. The colonies appeared faster and in higher number when incubated in air containing 10% CO2 (p < 0.01), independent of media. B83 showed a faster growth and detected more isolates at 30 days of incubation, when compared to ST (0.0178), LJp (p < 0.0001) and 7H11 (p < 0.0001), though there was no difference between B83, ST and LJp at 60 and 90 days of incubation. 7H11 presented the lowest number of isolates (p < 0.0001) and a longer period for the appearance of the first colony (p < 0.001). According to our findings, the concomitant use of ST and B83 media incubated in air containing 10% CO2 increases the isolation of M. bovis in a shorter period of time, which improves bovine tuberculosis diagnosis.

O isolamento do Mycobacterium bovis é fundamental para um sistema de vigilância para tuberculose bovina baseado na detecção de lesões em abatedouro. Assim, quatro meios de cultura sólidos e três condições de incubação foram investigados para elucidar qual combinação supera as outras através da avaliação de crescimento, tempo para o aparecimento da primeira colônia e número de colônias. Noventa e sete amostras de lesões granulomatosas foram submetidas ao processo de descontaminação por cloreto de 1-hexadecilpiridínio a 0,75%, e inoculadas em dois meios a base de ovo, Stonebrink (ST) e Lõwenstein-Jensen com piruvato de sódio (LJp), e dois meios a base de ágar, ágar sangue tuberculose (B83) e Middlebrook 7H11 (7H11). Cada meio foi incubado a 37°C por 90 dias, em três condições de incubação: em atmosfera normal, em atmosfera com acréscimo de 10% de dióxido de carbono (CO2), e em atmosfera normal em tubos fechados com algodão hidrófobo queimado e subsequentemente fechado com rolha para criar uma atmosfera microaerófila. As colônias apareceram mais rapidamente e em maior número quando incubadas em atmosfera com 10% de CO2 (p < 0,01), independente dos meios. As micobactérias cresceram em maior abundância e mais rapidamente no meio B83 aos 30 dias de incubação, comparado a ST (0,0178), LJp (p < 0,0001) e 7H11 (p < 0,0001), apesar de não ter havido diferença entre B83, ST e LJp aos 60 e 90 dias de incubação. 7H11 exibiu o número mais baixo de isolados (p < 0,0001) e um período mais longo para o aparecimento da primeira colônia (p < 0,001). De acordo com nossos resultados, o uso concomitante dos meios ST e B83, incubados em atmosfera com acréscimo de 10% de CO2, aumenta a proporção de isolados e o número de UFC de M. bovis, além de abreviar o tempo para aparecimento das primeiras colônias, melhorando o diagnóstico direto de tuberculose.

Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis / Influência das condições de incubação nos meios de cultura para otimizar o primo isolamento de Mycobacterium bovis

The isolation of Mycobacterium bovis is critical to a surveillance system for bovine tuberculosis based on detection of lesions in abattoirs. Thus, four solid culture media and three incubation conditions were investigated to elucidate which combination overcomes the others by assessing growth, time to the first appearance of colonies and their number. Ninety-seven samples of granulomatous lesions were submitted to the decontamination procedure by 1-hexadecylpyridinium chloride at 0.75% w/v, and inoculated on two egg-based media, Stonebrinks (ST) and Lõwenstein-Jensens with sodium pyruvate (LJp), and two agar-based media, tuberculosis blood agar (B83) and Middlebrook 7H11 medium (7H11). Each medium was incubated at 37°C for 90 days in three incubation conditions: in air, in air containing 10% carbon dioxide (CO2), and in air in slopes closed with burned hydrophobic cotton and subsequently plugged with a cork to create a microaerophilic atmosphere. The colonies appeared faster and in higher number when incubated in air containing 10% CO2 (p < 0.01), independent of media. B83 showed a faster growth and detected more isolates at 30 days of incubation, when compared to ST (0.0178), LJp (p < 0.0001) and 7H11 (p < 0.0001), though there was no difference between B83, ST and LJp at 60 and 90 days of incubation. 7H11 presented the lowest number of isolates (p < 0.0001) and a longer period for the appearance of the first colony (p < 0.001). According to our findings, the concomitant use of ST and B83 media incubated in air containing 10% CO2 increases the isolation of M. bovis in a shorter period of time, which improves bovine tuberculosis diagnosis.(AU)

O isolamento do Mycobacterium bovis é fundamental para um sistema de vigilância para tuberculose bovina baseado na detecção de lesões em abatedouro. Assim, quatro meios de cultura sólidos e três condições de incubação foram investigados para elucidar qual combinação supera as outras através da avaliação de crescimento, tempo para o aparecimento da primeira colônia e número de colônias. Noventa e sete amostras de lesões granulomatosas foram submetidas ao processo de descontaminação por cloreto de 1-hexadecilpiridínio a 0,75%, e inoculadas em dois meios a base de ovo, Stonebrink (ST) e Lõwenstein-Jensen com piruvato de sódio (LJp), e dois meios a base de ágar, ágar sangue tuberculose (B83) e Middlebrook 7H11 (7H11). Cada meio foi incubado a 37°C por 90 dias, em três condições de incubação: em atmosfera normal, em atmosfera com acréscimo de 10% de dióxido de carbono (CO2), e em atmosfera normal em tubos fechados com algodão hidrófobo queimado e subsequentemente fechado com rolha para criar uma atmosfera microaerófila. As colônias apareceram mais rapidamente e em maior número quando incubadas em atmosfera com 10% de CO2 (p < 0,01), independente dos meios. As micobactérias cresceram em maior abundância e mais rapidamente no meio B83 aos 30 dias de incubação, comparado a ST (0,0178), LJp (p < 0,0001) e 7H11 (p < 0,0001), apesar de não ter havido diferença entre B83, ST e LJp aos 60 e 90 dias de incubação. 7H11 exibiu o número mais baixo de isolados (p < 0,0001) e um período mais longo para o aparecimento da primeira colônia (p < 0,001). De acordo com nossos resultados, o uso concomitante dos meios ST e B83, incubados em atmosfera com acréscimo de 10% de CO2, aumenta a proporção de isolados e o número de UFC de M. bovis, além de abreviar o tempo para aparecimento das primeiras colônias, melhorando o diagnóstico direto de tuberculose.(AU)

Estimating the impact of environmental conditions on hatching results using multivariable analysis

Hatching results are directly related to environmental and biological surroundings. This research study aimed at evaluating the influence of incubation environmental conditions on hatchability and one-day-old chickling quality of five production flocks using multivariable analysis tool. The experiment was carried out in a commercial hatchery located in the state of São Paulo, Brazil. Environmental variables such as dry bulb temperature, relative humidity, carbon dioxide concentration, and number of colony forming units of fungi were recorded inside a broiler multi-stage setter, a hatcher after eggs transference, and a chick-processing room. The homogeneity of parameter distribution among quadrants inside the setter, the hatcher, and the chick room was tested using the non-parametric test of Kruskal-Wallis, and the fit analysis was applied. The multivariate analysis was applied using the Main Component Technique in order to identify possible correlations between environmental and production parameters. Three different groups were identified: the first group is represented by temperature, which was positively correlated both with good hatchability and good chick quality; the second group indicates that poor chick quality was positively correlated with air velocity and relative humidity increase. The third group, represented by carbon dioxide concentration and fungi colonies forming units, presented strong positive association with embryo mortality increase.

Estimating the impact of environmental conditions on hatching results using multivariable analysis

Hatching results are directly related to environmental and biological surroundings. This research study aimed at evaluating the influence of incubation environmental conditions on hatchability and one-day-old chickling quality of five production flocks using multivariable analysis tool. The experiment was carried out in a commercial hatchery located in the state of São Paulo, Brazil. Environmental variables such as dry bulb temperature, relative humidity, carbon dioxide concentration, and number of colony forming units of fungi were recorded inside a broiler multi-stage setter, a hatcher after eggs transference, and a chick-processing room. The homogeneity of parameter distribution among quadrants inside the setter, the hatcher, and the chick room was tested using the non-parametric test of Kruskal-Wallis, and the fit analysis was applied. The multivariate analysis was applied using the Main Component Technique in order to identify possible correlations between environmental and production parameters. Three different groups were identified: the first group is represented by temperature, which was positively correlated both with good hatchability and good chick quality; the second group indicates that poor chick quality was positively correlated with air velocity and relative humidity increase. The third group, represented by carbon dioxide concentration and fungi colonies forming units, presented strong positive association with embryo mortality increase.