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ABSTRACT: Selenium is a heat-stress-reducing substance that improves heat resistance and is being studied for its effective application in the broiler industry. However, research on feed additives is labor-intensive and time-consuming because of the need for feeding experiments. We aimed to compare the effects of selenium under heat stress in vitro and in ovo, specifically examining the gene expression of heat shock proteins (HSP) and inflammatory markers. Two groups were included in the in-vitro study: in-vitro control (TC; selenium 0 μg/ml) and in-vitro selenium (TS; selenium 5 μg/ml). The satellite cells were cultured at 42°C for 48 h after selenium treatment. The in-ovo study comprised 4 groups: in-ovo control and in-ovo selenium 1-3 (OC, OS1, OS2, and OS3; selenium 2.5, 5, and 10 μg/egg, respectively). Selenium was injected on the 18th day after hatching, and heat treatment was performed at 32–34°C from the 14th to the 21st day after hatching, and the leg muscles of the chicks were collected on the 21st day. The gene expression of heat shock proteins (HSP), caspase3, nuclear factor kappa light-chain enhancer of activated B cells (NF-kB), and IL-8 was analyzed in in-vitro and in-ovo experiments, respectively. In-vitro results showed significant increases in HSP90, HSP60, and HSP40 in TS compared to TC, with a significant decrease in HSP70. In the in-ovo study, HSP70, caspase3, NF-kB and IL-8 were significantly increased in OS1. HSP90, HSP60, HSP40, HSP27 and NF-kB were significantly decreased in in-ovo OS2 compared to in-vitro TS, implying a trend in ratio compared to control. Selenium appeared to enhance heat resistance in-vitro and in-ovo by modulating HSPs and inflammation. However, differences in mRNA expression were observed depending on the concentration of selenium. These findings suggest that selenium modulates heat resistance through different mechanisms in-vitro and in-ovo, likely due to the complexity of whole-organism interactions in-ovo compared to the single-cell-type environment in-vitro. Therefore, to directly apply in-vitro results to in-ovo, a concentration comparison study for each additive is necessary.

Boron is reported to cause fertility disorders in birds. We investigated the potential adverse effects of boric acid administered during embryogenesis on testicular development and spermatogenesis after hatching and puberty. The fertilized eggs were divided into four groups. Three groups were injected with different concentrations of boric acid (1000, 1500, or 2000 ppm) dissolved in 9% NaCl into the egg yolk, and the control group received only a 9% NaCl injection. The evaluations were conducted at two time points: post-hatching day 1 and 26 weeks of age. Testes of oneday-old- and 26-week-old birds were subjected to morphological and stereological evaluations to assess the potential effects of boric acid on testicular development. Effects of boric acid on spermatogenesis and semen quality parameters were also investigated in 26-week-old birds. Finally, plasma FSH, LH, and testosterone levels were analyzed in all birds. No statistically significant differences were noted between one-day-old chicks and 26-week-old birds regarding stereological parameters. Likewise, no significant differences were detected among groups concerning Sertoli, germ, and Leydig cells. Semen quality parameters did not reveal statistically significant differences, and similarly, plasma LH and testosterone levels did not statistically differ between the study and the control groups. Comb height and plasma FSH levels were increased in experimental groups, compared to the control in 26-week-old birds. A single administration of different boric acid doses did not impact the testicular morphological features after hatching and puberty. In-ovo boric acid exposure quantitatively altered the numbers of the Sertoli, germ, and Leydig cells, respectively, but revealed no adverse effect on spermatogenesis. This study might contribute to further research targeting boric acid's multifaceted effects on the dynamics of spermatogenesis. [ABSTRACT FROM AUTHOR]

ABSTRACT: The aim of the study was to determine the effect of in ovo administration of zinc glycine chelate (Zn-Gly), and a multistrain probiotic on the hatchability and selected parameters of the cellular and humoral immune response of chickens. The study was conducted on 1,400 fertilized eggs from commercial broiler breeders (Ross x Ross 708). Material for the study consisted of peripheral blood and spleens of chicks taken 12 h and 7 d after hatching. The results showed that both combined and single in ovo administration of the multistrain probiotic and zinc glycine chelate significantly reduced hatchability of chicks. The flow cytometry study showed that the highest percentage of CD4+ T cells, CD4+CD25+, and high expression of KUL01 in the serum were obtained in the group supplemented with probiotic and Zn-Gly both 12 h and 7 d after hatching. In birds supplemented with probiotic and zinc chelate, a high percentage of TCRγδ+ cells was found in serum and spleen 12 h after hatching and in serum after 7 d. The percentage of Bu-1A+ lymphocytes in serum and spleen 12 h and 7 d after hatching was the highest in the group supplemented with probiotic and Zn-Gly. The highest expression of CD79A was observed in the group supplemented only with zinc chelate. There were no significant differences in the percentage of CD4+ cells in the spleens of birds in the groups receiving the multistrain probiotic at 12 h after hatching, and after 7 d, the percentage of CD4+ T cells was lower in the experimental groups than in the control group. The percentage of CD8+ cells in the serum of birds after hatching was lower in the group supplemented with multistrain probiotic and Zn-Gly than in the control group, but reached the highest value on d 7 after hatching. The obtained results confirm the strong effect of the combined administration of a multistrain probiotic and Zn-Gly chelate on lymphocyte proliferation and stimulation of cellular immune mechanisms in birds.

This is a preliminary study evaluating the effect of different fractions of Concord grapes (Vitis labrusca L.) on the brush border membrane (BBM) morphology, duodenal gene expression, and specific gut bacterial populations. For this study, we utilized a unique intraamniotic approach, wherein, the test substances are administered into the amnion of the Gallus gallus egg (on day 17). The embryo orally consumes the amniotic fluid along with the injected test substance before the hatch. We randomly divided ~50 fertilized eggs into 5 groups including 6% grape (juice, puree, and pomace) along with controls (no injection and diluent—H2O). The grape juice was prepared by crushing the grapes; the grape residues were used as pomace. The grape puree included the grape skin, endocarp, mesocarp, and juice but not the seeds. On day 21, the hatch day, the blood, pectoral muscle, liver, duodenum, and large intestine were harvested. Our results showed no significant differences in blood glucose, pectoral glycogen level, or body weight. However, significant (p < 0.05) differences in duodenal and liver gene expression were observed between the treatment groups. The grape puree treatment resulted in higher Clostridium numbers and lower Bifidobacterium numbers when compared to all other groups. In summary, the dietary consumption of grape polyphenols has the potential to beneficially modulate aspects of intestinal health provided their concentration is limited. [ABSTRACT FROM AUTHOR]

[Display omitted] • A handheld NIR was evaluated to predict fertility and sex in-ovo. • The NIR method was useful to distinguish between fertile and infertile egg samples. • The classification results are not sufficient for industrial in ovo sexing of chicken eggs. The objective of this study was to evaluate the ability of a handheld near-infrared device (900–1600 nm) to predict fertility and sex (male and female) traits in-ovo. The NIR reflectance spectra of the egg samples were collected on days 0, 7, 14 and 18 of incubation and the data was analysed using principal component analysis (PCA), linear discriminant analysis (LDA) and support vector machines classification (SVM). The overall classification rates for the prediction of fertile and infertile egg samples ranged from 73 % to 84 % and between 93 % to 95 % using LDA and SVM classification, respectively. The highest classification rate was obtained on day 7 of incubation. The classification between male and female embryos achieved lower classification rates, between 62 % and 68 % using LDA and SVM classification, respectively. Although the classification rates for in-ovo sexing obtained in this study are higher than those obtained by chance (50 %), the classification results are currently not sufficient for industrial in-ovo sexing of chicken eggs. These results demonstrated that short wavelengths in the NIR range may be useful to distinguish between fertile and infertile egg samples at days 7 and 14 during incubation. [ABSTRACT FROM AUTHOR]

Diatoms are single cell eukaryotic microalgae; their surface possesses a porous nanostructured silica cell wall or frustule. Diatomaceous earth (DE) or diatomite is a natural siliceous sediment of diatoms. Since silica has been proved to have adjuvant capabilities, we propose that diatoms and DE may provide an inexpensive and abundant source of adjuvant readily available to use in livestock vaccines.In a first experiment, the safety of diatoms used as an adjuvant for in-ovo vaccination was investigated. In a second experiment, we assessed the humoral immune response after one in-ovo vaccination with inactivated Newcastle Disease Virus (NDV) and DE as adjuvant followed by 2 subcutaneous boosters on d 21 and 29 of age. In both experiments, results were compared to Freund's incomplete adjuvant and aluminum hydroxide.No detrimental effects on hatchability and chick quality were detected after in-ovo inoculation of diatoms and DE in experiments 1 and 2 respectively. In experiment 2 no humoral responses were detected after the in-ovo vaccination until 29 d of age. Seven d after the second subcutaneous booster an antibody response against NDV was detected in chickens that had received vaccines adjuvanted with Freund's incomplete adjuvant, aluminum hydroxide, and DE. These responses became significantly higher 10 d after the second booster. Finally, 15 d after the second booster, the humoral responses induced by the vaccine with Freund's incomplete adjuvant were statistically higher, followed by comparable responses induced by vaccines containing DE or aluminum hydroxide that were significantly higher than DE+PBS, PBS+INDV and PBS alone. From an applied perspective, we can propose that DE can serve as a potential adjuvant for vaccines against poultry diseases.

This study was designed to compare immunopathological effects of in ovo vaccination with post-hatch vaccination against IBD in White Leghorn chicks. A total of 189 embryonated eggs were divided into six groups. At day 18 of incubation, groups A–C were administered in ovo with 228E, Winterfield 2512:10/3 and 2512/90:10/2.7, respectively, group D (post-hatch vaccination) and group E as shamed control (for quality evaluation of in ovo vaccination technique), and group F as control. The results showed that antibody titers against IBD detected by ELISA on days 2, 17, and 28 were significantly higher in all in ovo groups as compared to control groups E and F. On day 17, all vaccinated groups (in ovo and post-hatch vaccinated) showed no significant differences in antibody titers among themselves; however, at day 28, only the post-hatch group showed significantly higher antibody titers followed by in ovo vaccinated groups. The cell-mediated immunity determined by PHA-P assay was significantly higher in all vaccinated groups than the non-vaccinated groups. No clinical signs of IBD infection were observed in any of the vaccinated groups. There was only increase in bursa size of groups vaccinated with intermediate plus strains (groups A, C, and D) at day 28. The histopathology showed that all the treatment groups had mild lesions induced by IBD virus in bursa. This study concluded that in ovo vaccination with live IBD vaccines provides protective immunity to the chickens even in the presence of IBD-specific MDA; therefore, the onset of immunity was much earlier than the post-hatch vaccination and in ovo groups also maintained protective immunity against IBD for longer time.

Copyright of Indonesian Journal of Animal & Veterinary Sciences / Jurnal Ilmu Ternak dan Veteriner is the property of Indonesian Center for Animal Science Research & Development and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Selenium is a heat-stress-reducing substance that improves heat resistance and is being studied for its effective application in the broiler industry. However, research on feed additives is labor-intensive and time-consuming because of the need for feeding experiments. We aimed to compare the effects of selenium under heat stress in vitro and in ovo, specifically examining the gene expression of heat shock proteins (HSP) and inflammatory markers. Two groups were included in the in-vitro study: in-vitro control (TC; selenium 0 μg/ml) and in-vitro selenium (TS; selenium 5 μg/ml). The satellite cells were cultured at 42°C for 48 h after selenium treatment. The in-ovo study comprised 4 groups: in-ovo control and in-ovo selenium 1-3 (OC, OS1, OS2, and OS3; selenium 2.5, 5, and 10 μg/egg, respectively). Selenium was injected on the 18th day after hatching, and heat treatment was performed at 32-34°C from the 14th to the 21st day after hatching, and the leg muscles of the chicks were collected on the 21st day. The gene expression of heat shock proteins (HSP), caspase3, nuclear factor kappa light-chain enhancer of activated B cells (NF-kB), and IL-8 was analyzed in in-vitro and in-ovo experiments, respectively. In-vitro results showed significant increases in HSP90, HSP60, and HSP40 in TS compared to TC, with a significant decrease in HSP70. In the in-ovo study, HSP70, caspase3, NF-kB and IL-8 were significantly increased in OS1. HSP90, HSP60, HSP40, HSP27 and NF-kB were significantly decreased in in-ovo OS2 compared to in-vitro TS, implying a trend in ratio compared to control. Selenium appeared to enhance heat resistance in-vitro and in-ovo by modulating HSPs and inflammation. However, differences in mRNA expression were observed depending on the concentration of selenium. These findings suggest that selenium modulates heat resistance through different mechanisms in-vitro and in-ovo, likely due to the complexity of whole-organism interactions in-ovo compared to the single-cell-type environment in-vitro. Therefore, to directly apply in-vitro results to in-ovo, a concentration comparison study for each additive is necessary., Competing Interests: DISCLOSURES The authors affirm that there are no conflicts of interest to declare., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The aim of the study was to determine the effect of in ovo administration of zinc glycine chelate (Zn-Gly), and a multistrain probiotic on the hatchability and selected parameters of the cellular and humoral immune response of chickens. The study was conducted on 1,400 fertilized eggs from commercial broiler breeders (Ross x Ross 708). Material for the study consisted of peripheral blood and spleens of chicks taken 12 h and 7 d after hatching. The results showed that both combined and single in ovo administration of the multistrain probiotic and zinc glycine chelate significantly reduced hatchability of chicks. The flow cytometry study showed that the highest percentage of CD4+ T cells, CD4+CD25+, and high expression of KUL01 in the serum were obtained in the group supplemented with probiotic and Zn-Gly both 12 h and 7 d after hatching. In birds supplemented with probiotic and zinc chelate, a high percentage of TCRγδ+ cells was found in serum and spleen 12 h after hatching and in serum after 7 d. The percentage of Bu-1A+ lymphocytes in serum and spleen 12 h and 7 d after hatching was the highest in the group supplemented with probiotic and Zn-Gly. The highest expression of CD79A was observed in the group supplemented only with zinc chelate. There were no significant differences in the percentage of CD4+ cells in the spleens of birds in the groups receiving the multistrain probiotic at 12 h after hatching, and after 7 d, the percentage of CD4+ T cells was lower in the experimental groups than in the control group. The percentage of CD8+ cells in the serum of birds after hatching was lower in the group supplemented with multistrain probiotic and Zn-Gly than in the control group, but reached the highest value on d 7 after hatching. The obtained results confirm the strong effect of the combined administration of a multistrain probiotic and Zn-Gly chelate on lymphocyte proliferation and stimulation of cellular immune mechanisms in birds. [ABSTRACT FROM AUTHOR]

Black seed (Nigella sativa) is an important medicinal herbal plant used to treat different ailments. In the present experiment antiviral effects of black seed was evaluated. Ethanolic extact of black seed was usd in the embryonated eggs inoculated with Newcastle Disease Virus (NDV). For this purpose Sixty (n=60) embryonated chicken eggs (14th day), were randomly divided into 5 groups (G 1-5) with 12 eggs in each group. The treatment groups were G1 (control negative) and G2 (control positive) groups were inoculated with NDV @ 0.2ml/egg of (10-10.2 EID50), G3 (NDV+ Nigella sativa, 0.2mg/0.2ml), G4 (Nigella sativa only, 0.2mg/0.2 ml) and G5 (NDV+ Ribavirin 20µg/ml). Egg candling, histopathology and gross pathology were performed after 24, 48, 72 and 96 hours post inoculation. Different survival rates were observed in different groups such as, G1 and G4 (100%), G2 (33%), G3 (75%) and G5 (83%), respectively exhibiting significant difference (P<0.05) between groups. The gross study of G2 showed degeneration of thigh muscle and severe haemorrhages, while in G3 and G5 the lesions were not intense and showing viral inhibition effects of herbal extract and Ribavirin. Degeneration in hepatocytes, mild necrosis, lymphocytosis and increased number of macrophages were also evident in G3 and G5. There were dominant population of plasma cells, lymphocytes with mild degeneration and lymphoblasts. In G2 degeneration of bursal follicles, while in G3 and G5 aggregation of lymphocytes and lymphoblasts, developed bursal follicles with clear cortex and medulla was noticed. The gross examination and histopathology of the embryonated eggs revealed that Nigella sativa extract have strong immunotherapeutic effect against NDV infcetion. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effect of in-ovo injection of Lactobacilla Rhamnosus (0, 16x10³, 32x10³ and 48x10³ cfu) as probiotic source on hatching parameters and growth performance of quails. In this, a total of 400 quail hatching eggs, were randomly distributed among 4 experimental groups. Each experimental group contained 4 replicates of 25 eggs each. In-ovo injections of Lactobacilla Rhamnosus into quail eggs did not statistically affect incubation characteristics and some tissue weights. A high amount of Lactobacilla Rhamnosus injection into the breeder quail eggs negatively affected live weight and live weight gain. As a result, it can be said that low or medium levels Lactobacilla Rhamnosus injection into quail eggs have no effect on incubation and growth parameters of quails. [ABSTRACT FROM AUTHOR]

Copyright of Iraqi Journal of Veterinary Medicine is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Sulforaphane (SFN) is a bioactive component found in broccoli that has neuroprotective and anti-cancer effects. SFN has been encapsulated in various drug carriers to improve its stability and effectiveness. Specifically, this study presents an SFN delivery system based on human Amniotic Fluid-EVs (hAFEVs) for the treatment of neurological disorders. The binding affinity between hAFEVs (<200 nm) and SFN was investigated by spectroscopic analysis, revealing a temperature-dependent binding mechanism. The binding constants (k sv , k q , and k b) were also assessed, indicating a static quenching mechanism. Additionally, the docking studies of SFN and CD9, CD81, HSP70, and TSG101 were performed using AutoDock 4.2.6 software's Lamarckian genetics algorithm to investigate the type of interaction and their effects on the targetability of hAFEVs. The obtained results showed that SFN-loaded hAFEVs had a favorable effect on increasing the survival and proliferation of human PC12 cells (137%). The results of gene expression studies showed that this release system can upregulate the expression of Nrf2 and decrease the expression of IL-6, which shows the support of nerve cells against oxidative stress. In the In-Ovo study, treatment with SFN-hAFEVs resulted in improved grey and white matter cohesion and the improved orientation of bipolar neurons. The results indicate a significant increase in synaptophysin expression and the number of doublecortin-positive cells improved neuronal development and synapse formation. The findings of this study suggest that SFN-hAFEVs have great potential as a treatment for neurological disorders. [Display omitted] [ABSTRACT FROM AUTHOR]

This study was designed to compare immunopathological effects of in ovo vaccination with post-hatch vaccination against IBD in White Leghorn chicks. A total of 189 embryonated eggs were divided into six groups. At day 18 of incubation, groups A-C were administered in ovo with 228E, Winterfield 2512:10/3 and 2512/90:10/2.7, respectively, group D (post-hatch vaccination) and group E as shamed control (for quality evaluation of in ovo vaccination technique), and group F as control. The results showed that antibody titers against IBD detected by ELISA on days 2, 17, and 28 were significantly higher in all in ovo groups as compared to control groups E and F. On day 17, all vaccinated groups ( in ovo and post-hatch vaccinated) showed no significant differences in antibody titers among themselves; however, at day 28, only the post-hatch group showed significantly higher antibody titers followed by in ovo vaccinated groups. The cell-mediated immunity determined by PHA-P assay was significantly higher in all vaccinated groups than the non-vaccinated groups. No clinical signs of IBD infection were observed in any of the vaccinated groups. There was only increase in bursa size of groups vaccinated with intermediate plus strains (groups A, C, and D) at day 28. The histopathology showed that all the treatment groups had mild lesions induced by IBD virus in bursa. This study concluded that in ovo vaccination with live IBD vaccines provides protective immunity to the chickens even in the presence of IBD-specific MDA; therefore, the onset of immunity was much earlier than the post-hatch vaccination and in ovo groups also maintained protective immunity against IBD for longer time., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zaheer, Chen, Khan, Elokil, Saleemi, Zaheer and Khan.)

Nutrient administration in-ovo could be considered as an alternative method to improve hatchability and duckling weights followed by better economic performance. On the 12th day of incubation, fertile duck eggs (n= 500) were distributed into 5 groups: uninjected control; 0.1 ml corn oil; 0.1 ml corn oil plus 10 mg vitamin E; 0.1 ml saline; and 0.1 ml saline plus 3 mg ascorbic acid. In-ovo injection of vitamin E but not ascorbic acid resulted in higher hatchability percentage compared to the uninjected control; however this was not statistically confirmed. In-ovo injection of either vitamin E or ascorbic acid resulted in significantly (P < 0.05) higher body weights at hatch, final body weights, and feed intakes than the uninjected control group. However, both male and female ducklings hatched from eggs injected with either vitamin E or ascorbic acid had better feed conversion during the starting period only. The carcass characteristics and relative lymphoid organ weights did not differ (P < 0.05) between the in ovo injected groups and the uninjected controls. In-ovo injection of either vitamin E or ascorbic acid resulted in significant increase (P < 0.05) in the geometric mean of the 1st antibodies titers of males and the 1st and 2nd antibodies titers of females compared to the uninjected control. It could be concluded that in-ovo injection of vitamin E and ascorbic acid improve the embryonic and post-hatch growth of Muscovy ducks. [ABSTRACT FROM AUTHOR]

Summary: The protection of poultry from H5N1 highly pathogenic avian influenza A (HPAI) and Newcastle disease virus (NDV) can be achieved through vaccination, as part of a broader disease control strategy. We have previously generated a recombinant influenza virus expressing, (i) an H5 hemagglutinin protein, modified by the removal of the polybasic cleavage peptide and (ii) the ectodomain of the NDV hemagglutinin–neuraminidase (HN) protein in the place of the ectodomain of influenza neuraminidase (Park MS, et al. Proc Natl Acad Sci USA 2006;103(21):8203–8). Here we show this virus is attenuated in primary normal human bronchial epithelial (NHBE) cell culture, and demonstrate protection of C57BL/6 mice from lethal challenge with an H5 HA-containing influenza virus through immunisation with the recombinant virus. In addition, in-ovo vaccination of 18-day-old embryonated chicken eggs provided 90% and 80% protection against highly stringent lethal challenge by NDV and H5N1 virus, respectively. We propose that this virus has potential as a safe in-ovo live, attenuated, bivalent avian influenza and Newcastle disease virus vaccine. [Copyright &y& Elsevier]