Control of Escherichia coli in Poultry Using the In Ovo Injection Technique.

Pathogens, such as Escherichia coli (E. coli), have been identified as significant causes of poultry mortality. Poultry can serve as potential sources of E. coli transmission, even when asymptomatic, posing a substantial threat to food safety and human health. The in ovo administration of antimicrobials is crucial for preventing and/or effectively combating acute and chronic infections caused by poultry pathogens. To achieve this goal, it is critical that antimicrobials are properly injected into embryonic fluids, such as the amnion, to reach target tissues and trigger robust antimicrobial responses. Several protocols based on antimicrobials were evaluated to meet these requirements. This review analyzed the impacts of antimicrobial substances injected in ovo on the control of E. coli in poultry. The reduction in infection rates, resulting from the implementation of in ovo antimicrobials, combined with efforts aimed at hygienic-sanitary action plans in poultry sheds, reinforces confidence that E. coli can be contained before causing large scale damage. For example, antimicrobial peptides and probiotics have shown potential to provide protection to poultry against infections caused by E. coli. Issues related to the toxicity and bacterial resistance of many synthetic chemical compounds represent challenges that need to be overcome before the commercial application of in ovo injection protocols focused on microbiological control.

Effect of in ovo injection of DL-methionine on hatchability, embryo mortality, hatching weight, blood biochemical parameters and gastrointestinal tract development of breeder chicks.

The current investigation was conducted to test the potential effects of in ovo feeding of DL-methionine (MET) on hatchability, embryonic mortality, hatching weight, blood biochemical parameters and development of heart and gastrointestinal (GIT) of breeder chick embryos. 224 Rhode Island Red fertile eggs were randomly distributed into seven experimental treatments: untreated egg (control), buffered saline (0.5% NaCl), and five solutions containing increased levels of MET (0.5, 1.0, 1.5, 2.0 and 2.5%) + 0.5% NaCl, being separated into four groups/replicates (each one with 8 eggs), totaling 32 eggs/treatment. All embryos submitted to in ovo injection with MET presented a decrease in the hatchability results and an increase in the results of intermediary embryonic mortality. Chicks hatched from eggs injected with until to 1.0% MET were heavier and presented better development of the heart and GIT, especially important organs and regions for digestion and nutrient absorption. Conclusively, the in ovo feeding using MET showed positive impacts on hatching weight and GIT development of breeder chicks. However, caused negative impacts on hatchability when used at high levels.

In ovo feeding of creatine monohydrate increases performances of hatching and development in breeder chicks.

The current investigation was conducted to test the potential effects of in ovo feeding of creatine monohydrate (CMH) on hatchability, embryonic mortality, hatching weight, and development of heart and gastrointestinal tract (main organs and regions) of breeder chick embryos. Rhode Island Red fertile eggs were randomly distributed into seven experimental treatments: untreated egg (control), a sterile buffered solution (0.50% NaCl), and five solutions containing increased levels of CMH (0.50, 1.00, 1.50, and 2.00%) + 0.50% NaCl, being separated in four groups/replicates (three with 15 eggs and one with 16 eggs), totaling 61 eggs/treatment and a total of 427 fertile eggs used. All-in ovo injected groups with CMH decreased the hatchability and increased the intermediary embryonic mortality. At hatching, all-in ovo injected groups with CMH also increased the hatching weight and stimulated the development of the heart and the total length of the gastrointestinal tract, especially important organs for digestion of nutrients (yolk sac, pro-ventricle and gizzard) and regions for nutrient absorption (jejunum + ileum and colon + rectum). Conclusively, the in ovo feeding using CMH showed positive impacts on hatching weight and the development of gastrointestinal tract of chicks. However, caused negative impacts on hatchability.

Effects of in ovo injection of nano-selenium and nano-zinc oxide and high eggshell temperature during late incubation on antioxidant activity, thyroid and glucocorticoid hormones and some blood metabolites in broiler hatchlings

This experiment was conducted to evaluate the effects of in ovo injection of nano-selenium (Nano-Se) and nano-zinc oxide (Nano-ZnO) and high eggshell temperature (EST) during late incubation on blood parameters of broiler hatchlings. A total of 750 fertile eggs, were weighed and randomly distributed among 5 treatment groups on each of 5 replicate tray levels. The injection was performed on 17 d of incubation. Treatments included of: 1) Eggs not injected and incubated at normal EST (control); 2) Eggs not injected and incubated at high EST; 3) Eggs injected NaCl solution and incubated at high EST (sham); 4) Eggs injected NaCl solution containing 40 µg Nano-Se and incubated at high EST; 5) Eggs injected NaCl solution containing 500 µg Nano-ZnO and incubated at high EST. EST of 37.8ºC (normal) or 38.9ºC (high) was applied from d 19 to 21 of incubation. In ovo injection of Nano-Se and Nano-ZnO significantly increased activity of GSH-Px and SOD and total protein, but decreased the levels of corticosterone, cortisol, T4 and T3 at high EST. Injection of Nano-Se and Nano-ZnO had a significant role in alleviating the negative effects of high temperature incubation and heat stress by increased antioxidant activity and reduced oxidative stress.

Effect of in-ovo feeding of iron nanoparticles and methionine hydroxy analogue on broilers chickens small intestinal characteristics

The experiment was conducted with 644 Ross fertilized egg by 7 treatments 4 replicates and 23 eggs in each. Seven treatments included two control with and without injection, iron sulfate, iron sulfate nanoparticles, Alimet, Alimet + iron sulfate, Alimet + iron sulfate nanoparticles. After hatching 2 mg iron nanoparticles were applied as new treatment. The highest increased in the intestinal relative weight (p < 0.05) was observed by iron+Alimet in late feeding at day old of age. Also similar trend was found in cecum and duodenum length by iron control 2 and late feeding (18 hours’ after hatching). The highest cecum length was found among all treatments by in ovo injection of iron nanoparticles in early feeding at 21 days of age (p < 0.05). Significantly increased the duodenum length was found by iron sulfate in early feeding at 42 days of age (p < 0.05). In ovo injection of Alimet in late feeding was resulted in decrease jejunum crypt depth at 21 days of age (p < 0.05). The results of this study have shown that the highest jejunum villi width and surface area were recorded in dietary iron sulfate nanoparticles in late feeding at 21 and 42 days of age (p < 0.05).

Effects of in ovo injection of nano-selenium and nano-zinc oxide and high eggshell temperature during late incubation on antioxidant activity, thyroid and glucocorticoid hormones and some blood metabolites in broiler hatchlings

This experiment was conducted to evaluate the effects of in ovo injection of nano-selenium (Nano-Se) and nano-zinc oxide (Nano-ZnO) and high eggshell temperature (EST) during late incubation on blood parameters of broiler hatchlings. A total of 750 fertile eggs, were weighed and randomly distributed among 5 treatment groups on each of 5 replicate tray levels. The injection was performed on 17 d of incubation. Treatments included of: 1) Eggs not injected and incubated at normal EST (control); 2) Eggs not injected and incubated at high EST; 3) Eggs injected NaCl solution and incubated at high EST (sham); 4) Eggs injected NaCl solution containing 40 µg Nano-Se and incubated at high EST; 5) Eggs injected NaCl solution containing 500 µg Nano-ZnO and incubated at high EST. EST of 37.8ºC (normal) or 38.9ºC (high) was applied from d 19 to 21 of incubation. In ovo injection of Nano-Se and Nano-ZnO significantly increased activity of GSH-Px and SOD and total protein, but decreased the levels of corticosterone, cortisol, T4 and T3 at high EST. Injection of Nano-Se and Nano-ZnO had a significant role in alleviating the negative effects of high temperature incubation and heat stress by increased antioxidant activity and reduced oxidative stress.(AU)

Effect of in-ovo feeding of iron nanoparticles and methionine hydroxy analogue on broilers chickens small intestinal characteristics

The experiment was conducted with 644 Ross fertilized egg by 7 treatments 4 replicates and 23 eggs in each. Seven treatments included two control with and without injection, iron sulfate, iron sulfate nanoparticles, Alimet, Alimet + iron sulfate, Alimet + iron sulfate nanoparticles. After hatching 2 mg iron nanoparticles were applied as new treatment. The highest increased in the intestinal relative weight (p < 0.05) was observed by iron+Alimet in late feeding at day old of age. Also similar trend was found in cecum and duodenum length by iron control 2 and late feeding (18 hours after hatching). The highest cecum length was found among all treatments by in ovo injection of iron nanoparticles in early feeding at 21 days of age (p < 0.05). Significantly increased the duodenum length was found by iron sulfate in early feeding at 42 days of age (p < 0.05). In ovo injection of Alimet in late feeding was resulted in decrease jejunum crypt depth at 21 days of age (p < 0.05). The results of this study have shown that the highest jejunum villi width and surface area were recorded in dietary iron sulfate nanoparticles in late feeding at 21 and 42 days of age (p < 0.05).(AU)

Effects of In-Ovo Rutin Injection to Fertile Japanese Quail (Coturnix Coturnix Japonica) Egg on Hatchability, Embryonic Death, Hatchling Weight, and Hatchling Liver Oxidative and Nitrosative Stress

This study was conducted with the aim of investigating the effects of the antioxidant rutin injected in fertilized quail eggs on incubation parameters and some hatchling liver biochemical parameters. The study was carried out with 6 groups including a control group and 5 different doses of rutin, and it involved 720 fresh Japanese quail (Coturnix coturnix japonica) eggs. It was observed that rutin dose did not affect the early embryo mortality, whereas intermediate and late embryo mortality rates were higher in all groups given rutin in comparison to the control group. The mean hatchability of fertile eggs and total eggs for the control, 0.25 mg, 0.50 mg, 0.75 mg, 1 mg and 1.5 mg groups were calculated as 82.06, 82.23, 64.43, 68.84, 44.08, 22.95 % and 48.10, 55.49, 34.33, 33.00, 18.03, 8.45% respectively. Compared with the control group, hatchling mortality rate was higher only in the 0.25 rutin group, and lower in all other groups receiving rutin in-ovo. The highest hatchling weight was found in the 0.25 mg rutin group, and hatchling weight decreased as rutin dose increased. Consequently, considering the mortality rates, hatchling weights, and liver antioxidant/oxidant capacities of the hatchlings, it is believed that the in-ovo injection of 0.25 mg rutin may be useful for Japanese quail production.

Effects of In-Ovo Rutin Injection to Fertile Japanese Quail (Coturnix Coturnix Japonica) Egg on Hatchability, Embryonic Death, Hatchling Weight, and Hatchling Liver Oxidative and Nitrosative Stress

This study was conducted with the aim of investigating the effects of the antioxidant rutin injected in fertilized quail eggs on incubation parameters and some hatchling liver biochemical parameters. The study was carried out with 6 groups including a control group and 5 different doses of rutin, and it involved 720 fresh Japanese quail (Coturnix coturnix japonica) eggs. It was observed that rutin dose did not affect the early embryo mortality, whereas intermediate and late embryo mortality rates were higher in all groups given rutin in comparison to the control group. The mean hatchability of fertile eggs and total eggs for the control, 0.25 mg, 0.50 mg, 0.75 mg, 1 mg and 1.5 mg groups were calculated as 82.06, 82.23, 64.43, 68.84, 44.08, 22.95 % and 48.10, 55.49, 34.33, 33.00, 18.03, 8.45% respectively. Compared with the control group, hatchling mortality rate was higher only in the 0.25 rutin group, and lower in all other groups receiving rutin in-ovo. The highest hatchling weight was found in the 0.25 mg rutin group, and hatchling weight decreased as rutin dose increased. Consequently, considering the mortality rates, hatchling weights, and liver antioxidant/oxidant capacities of the hatchlings, it is believed that the in-ovo injection of 0.25 mg rutin may be useful for Japanese quail production.(AU)

The effects of in-ovo injection of glucose on hatchability, hatching weight and subsequent performance of newly-hatched chicks

This experiment was conducted to determine the effects of an in-ovo injection of glucose on hatchability, weight and subsequent performances of newly-hatched chickens. The 720 fertile eggs used in this experiment were obtained from a Cobb-500 broiler breeder flock of 28 weeks of age. Treatments were the following: 1) control group (withou- injection), 2) group injected with 0.5 ml deionized water (sham group), 3) group injected with 0.5 ml glucose 15% in deionized water, 4) group injected with 0.5 ml glucose 20% in deionized water and 5) group injected with 0.5 ml glucose 25% in deionized water. There werh four replicates per treatment and 36 eggs per replicate. The experimental design was a completely randomized design. Upon hatch, hatchability and weight of newly-hatched chicks were measured, and 20 chicks per replicate were transferred to an experimental house and reared for 42 days. Live weight and feed intakeswere measured for each experimental unit at 21 and 42 days of age and then weight gai, and feed conversion ratio were calculated. The results of the study indicate that group that received glucose at 15%, 20% and 25% had significantly higher hatching weight as compared with the control and the sham groups,tbut,eggs injected in ovo had significantly lower hatchability than the controls (p 0.01). Also, there were no significant treatment effects on feed intake between 0 and 21 days post-hatch. Chicks that received in-ovo injection of glucose haddbetter feed intake, weight gain and feed conversion ratio compared with chicks hatched from the control and the sham groups. The obtained Data suggest that in-ovo injection may increase weight and improve the performancs of newly-hatched chickens.

The effects of in-ovo injection of glucose on hatchability, hatching weight and subsequent performance of newly-hatched chicks

This experiment was conducted to determine the effects of an in-ovo injection of glucose on hatchability, weight and subsequent performances of newly-hatched chickens. The 720 fertile eggs used in this experiment were obtained from a Cobb-500 broiler breeder flock of 28 weeks of age. Treatments were the following: 1) control group (withou- injection), 2) group injected with 0.5 ml deionized water (sham group), 3) group injected with 0.5 ml glucose 15% in deionized water, 4) group injected with 0.5 ml glucose 20% in deionized water and 5) group injected with 0.5 ml glucose 25% in deionized water. There werh four replicates per treatment and 36 eggs per replicate. The experimental design was a completely randomized design. Upon hatch, hatchability and weight of newly-hatched chicks were measured, and 20 chicks per replicate were transferred to an experimental house and reared for 42 days. Live weight and feed intakeswere measured for each experimental unit at 21 and 42 days of age and then weight gai, and feed conversion ratio were calculated. The results of the study indicate that group that received glucose at 15%, 20% and 25% had significantly higher hatching weight as compared with the control and the sham groups,tbut,eggs injected in ovo had significantly lower hatchability than the controls (p < 0.01). Also, there were no significant treatment effects on feed intake between 0 and 21 days post-hatch. Chicks that received in-ovo injection of glucose haddbetter feed intake, weight gain and feed conversion ratio compared with chicks hatched from the control and the sham groups. The obtained Data suggest that in-ovo injection may increase weight and improve the performancs of newly-hatched chickens.(AU)

EFFECT OF IN OVO VACCINATION PROCEDURES ON JAPANESE QUAIL EMBRYOS (Coturnix japonica) AND INCUBATION PERFORMANCE / EFEITO DOS PROCEDIMENTOS DE VACINAÇÃO IN OVO SOBRE EMBRIÕES DE CODORNA JAPONESA (Coturnix japonica) E DESEMPENHO DA INCUBAÇÃO

This work aimed to evaluate the effects of in ovo vaccination procedures on incubation of Japanese quail eggs. The experiment was carried out in a (4 x 3) factorial design with 12 experimental treatments (4 injection days x 3 injection protocols). The injections were tested on four incubation days: at 0, 5, 10 or 15. On each injection day, the eggs were submitted to one out of three distinct injection procedures: saline injection and Newcastle disease (ND) vaccine plus saline or industrial diluent. The eggs were incubated at 37.5º C and 60% RH. All eggs and hatched quails were weighed. Unhatched eggs were opened to classify embryo mortality. Hatched quails were raised to obtain blood to evaluate antibody responseagainst Newcastle disease virus (NDV). The injection process itself (saline) was not harmful at 10 and 15 days of incubation for Japanese quail eggs; however, in ovo vaccination with live ND vaccine (HB1 strain) is not recommended to fertile quail eggs at any incubation periods due to high levels of embryo mortality and poor post-hatch antibody titers.KEYWORDS: eggs; in ovo injection; Japanese quail; Newcastle disease; vaccination.

Esta pesquisa avaliou os efeitos dos procedimentos de vacinação in ovo na incubação artificial de codornas japonesas. Foi realizado um delineamento fatorial 4 x 3 com 12 tratamentos (4 dias de injeção x 3 protocolos de injeção). As injeções foram realizadas nos dias 0, 5, 10 ou15 de incubação. Em cada um desses dias, os ovos foram injetados com 3 protocolos diferentes: injeção de soro fisiológico e injeção da vacina vírus vivo da doença de Newcastle (DN) com soro fisiológico ou diluenteindustrial. Os ovos foram incubados a 37,5C e 60% UR. Todos os ovos e codornas nascidas foram pesados. Os ovos não eclodidos foram submetidos ao embriodiagnóstico. As codornas nascidas foram criadaspara coleta de sangue e avaliação de títulos de anticorpos contra o vírus da DN. Os resultados demonstraram que a inoculação em si (soro) não foi prejudicial para os ovos com 10 e 15 dias de incubação. Verificou-se que a vacinação in ovo com o vírus vivo da DN (cepa HB1) não é recomendada para ovos férteis de codorna em nenhuma idade do embrião, devido aos elevados índices de mortalidade embrionária e pouca resposta de anticorpos após o nascimento.PALAVRAS-CHAVE: codorna japonesa; doença de Newcastle; injeção in ovo; ovos; vacinação.

EFFECT OF IN OVO VACCINATION PROCEDURES ON JAPANESE QUAIL EMBRYOS (Coturnix japonica) AND INCUBATION PERFORMANCE / EFEITO DOS PROCEDIMENTOS DE VACINAÇÃO IN OVO SOBRE EMBRIÕES DE CODORNA JAPONESA (Coturnix japonica) E DESEMPENHO DA INCUBAÇÃO

This work aimed to evaluate the effects of in ovo vaccination procedures on incubation of Japanese quail eggs. The experiment was carried out in a (4 x 3) factorial design with 12 experimental treatments (4 injection days x 3 injection protocols). The injections were tested on four incubation days: at 0, 5, 10 or 15. On each injection day, the eggs were submitted to one out of three distinct injection procedures: saline injection and Newcastle disease (ND) vaccine plus saline or industrial diluent. The eggs were incubated at 37.5º C and 60% RH. All eggs and hatched quails were weighed. Unhatched eggs were opened to classify embryo mortality. Hatched quails were raised to obtain blood to evaluate antibody responseagainst Newcastle disease virus (NDV). The injection process itself (saline) was not harmful at 10 and 15 days of incubation for Japanese quail eggs; however, in ovo vaccination with live ND vaccine (HB1 strain) is not recommended to fertile quail eggs at any incubation periods due to high levels of embryo mortality and poor post-hatch antibody titers.KEYWORDS: eggs; in ovo injection; Japanese quail; Newcastle disease; vaccination.

Esta pesquisa avaliou os efeitos dos procedimentos de vacinação in ovo na incubação artificial de codornas japonesas. Foi realizado um delineamento fatorial 4 x 3 com 12 tratamentos (4 dias de injeção x 3 protocolos de injeção). As injeções foram realizadas nos dias 0, 5, 10 ou15 de incubação. Em cada um desses dias, os ovos foram injetados com 3 protocolos diferentes: injeção de soro fisiológico e injeção da vacina vírus vivo da doença de Newcastle (DN) com soro fisiológico ou diluenteindustrial. Os ovos foram incubados a 37,5C e 60% UR. Todos os ovos e codornas nascidas foram pesados. Os ovos não eclodidos foram submetidos ao embriodiagnóstico. As codornas nascidas foram criadaspara coleta de sangue e avaliação de títulos de anticorpos contra o vírus da DN. Os resultados demonstraram que a inoculação em si (soro) não foi prejudicial para os ovos com 10 e 15 dias de incubação. Verificou-se que a vacinação in ovo com o vírus vivo da DN (cepa HB1) não é recomendada para ovos férteis de codorna em nenhuma idade do embrião, devido aos elevados índices de mortalidade embrionária e pouca resposta de anticorpos após o nascimento.PALAVRAS-CHAVE: codorna japonesa; doença de Newcastle; injeção in ovo; ovos; vacinação.