Silica nanoparticles alleviate the immunosuppression, oxidative stress, biochemical, behavioral, and histopathological alterations induced by Aeromonas veronii infection in African catfish (Clarias gariepinus).

In the aquaculture industry, silica nanoparticles (SiNPs) have great significance, mainly for confronting diseases. Therefore, the present study aims to assess the antibacterial efficiency of SiNPs as a versatile trial against Aeromonas veronii infection in African catfish (Clarias gariepinus). Further, we investigated the influence of SiNPs in palliating the immune-antioxidant stress biochemical, ethological, and histopathological alterations induced by A. veronii. The experiment was conducted for 10 days, and about 120 fish were distributed into four groups at random, with 30 fish each. The first group is a control that was neither exposed to infection nor SiNPs. The second group (SiNPs) was vulnerable to SiNPs at a concentration of 20 mg/L in water. The third group was experimentally infected with A. veronii at a concentration of 1.5 × 107 CFU/mL. The fourth group (A. veronii + SiNPs) was exposed to SiNPs and infected with A. veronii. Results outlined that A. veronii infection induced behavioral alterations and suppression of immune-antioxidant responses that appeared as a clear decline in protein profile indices, complement 3, lysozyme activity, glutathione peroxidase, and total antioxidant capacity. The kidney and liver function biomarkers (creatinine, urea, alkaline phosphatase, and alanine aminotransferase) and lipid peroxide (malondialdehyde) were substantially increased in the A. veronii group, with marked histopathological changes and immunohistochemical alterations in these tissues. Interestingly, the exposure to SiNPs resulted in a clear improvement in all measured biomarkers and a noticeable regeneration of the histopathological changes. Overall, it will establish that SiNPs are a new, successful tool for opposing immunological, antioxidant, physiological, and histopathological alterations induced by A. veronii infection.

Robust dense-field based copy-move forgery localization using generic radial harmonic Fourier moment invariants.

As the importance of reliable multimedia content increases in today's society, image forensics is a growing field of research. The act of copying and pasting specific parts of an image, known as copy-move forgery, may be utilized for illegal or unethical purposes. Just as with other vision-related technologies, the accuracy of forensic analysis depends on having an appropriate image representation. Most existing feature extraction techniques do not accurately reflect the underlying image content leading to reduced performance. In this article, to detect the copy-move forgery attack, the Generic Radial Harmonic Fourier Moment (GRHFM) is proposed for reliable and distinctive image representation. The algorithm has the ability to effectively manipulate the distribution of zeros to emphasize certain image regions. Additionally, the relationships between complex exponentials and trigonometric functions are exploited to efficiently compute and easily implement the transform kernels. The efficacy of the algorithm is illustrated through experiments on dense-domain-based matching patterns. Experimental results on five benchmarking databases prove the effectiveness of the proposed approach compared with the state-of-the-art methods. According to the average scores, the proposed method demonstrates superior accuracy in overall localization performance. The F1 score, precision, and recall percentage values obtained are 92.5, 95.44, and 91.96, respectively. Robustness experiments on more challenging attacks are also conducted on FAU dataset. Results show that the proposed framework satisfies invariance to the various image variations, and thus an enhanced robustness compared to the previous methods. Moreover, the advantage of reasonable computational cost implies its potential use in real-world forensic applications.

Effects of dietary nano-selenium supplementation on Riemerella anatipestifer vaccinated and challenged Pekin ducklings (Anas platyrhynchos).

Riemerella anatipestifer (RA) is a common disease causing economic losses to duck farms worldwide. Novel supplements are crucially needed to control this bacterium, enhance poultry performance, and produce synergistic effects with vaccines in stimulating the immune system. This study investigated the effect of nano-selenium (Nano-Se) on the vaccinated (VAC) and challenged (Ch) Pekin ducklings (Anas platyrhynchos) with RA. Five experimental groups (G1-G5) were included in this study: G1 was the control group, G2 was the RA-challenged group, G3 was the Nano-Se+Ch group, G4 was the VAC+Ch group, and G5 was the Nano-Se+VAC+Ch group. The Nano-Se (0.3 mg/kg diet) was supplemented for 5 weeks post-vaccination (PV). The ducklings were vaccinated subcutaneously with the RA vaccine at 7 days of age and challenged with RA at the 3rd week PV. Blood, pharyngeal swabs and tissue samples were collected at the 3rd week PV and at different times post-challenge (PC). The growth performance (weight gain and feed conversion ratio), clinical signs, gross lesions, mortality, bacterial shedding, haematological, immunological, and biochemical parameters, cytokines production, and histopathological lesion scores showed significant differences (P < 0.05) between the challenged (G2) group and the supplemented (G3 & G5) groups. G5 showed the highest (P < 0.05) growth performance, phagocytic activity, IgM and IgG, splenic interleukin-2 (IL-2), IL-10, and interferon-gamma (IFN-γ) gene expressions, and the lowest mortality, bacterial shedding, hepatic and renal damage, heterophil/lymphocyte ratio and lesion scores compared to the other groups. In conclusion, the supplementation of nano-selenium for five weeks in the diet can improve the growth performance, immune status, and cytokines production in ducklings vaccinated and challenged with RA.

Antimicrobial, immunological and biochemical effects of florfenicol and garlic (Allium sativum) on rabbits infected with Escherichia coli serotype O55: H7.

Florfenicol (FFC) is a synthetic broad-spectrum antibiotic and garlic has a bactericidal action against coliforms. This study was carried out to compare the antimicrobial, immunological and biochemical effects of florfenicol and garlic, for their ability to treat enteropathogenic Escherichia coli serotype O55: H7 infection in rabbits. Four groups (G1-G4) were included. G1 group was the negative control; G2 group was the infected with a field-isolated strain of E. coli and untreated; G3 group was the infected+treated with FFC for 5 days; and G4 group was the infected+treated with garlic tablets for 14 days. The rabbits were observed for clinical signs, growth performance and mortality rates. Garlic-infused disks had a larger clear zone of inhibition than other antibiotic disks. Garlic treatment improved growth performance, biochemical parameters, and immunological response and reduced the fecal shedding and histopathological lesions in E. coli O55: H7 infected rabbits compared to the other groups. Colonization of E. coli more rapidly declined in G3 & G4 than in G2. Hepatic and intestinal gene expressions; tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were significantly elevated in G2 compared to the other groups, and their levels were elevated more in G3 than in G4. Serum interferon-gamma (IFN-γ) and phagocytic activity were significantly elevated in G4 compared to G3. G3 revealed macrocytic hypochromic anaemia that was confirmed histopathologically by moderate haematopoiesis of the bone marrow. In conclusion, garlic powder can reduce rabbit colibacillosis, like FFC, and can enhance the immune status of rabbits.

Molecular characterization and pathogenicity of very virulent infectious bursal disease virus isolated from naturally infected turkey poults in Egypt.

This study was conducted to investigate the molecular characterization and pathogenicity of very virulent infectious bursal disease virus (vvIBDV) isolated from naturally infected turkey poults and possible spread to chickens. Thirty samples were collected from turkey poults in the vicinity or in the same backyards with chickens suspected to be infected with IBDV and from live bird markets from different localities in Dakahlia governorate, Egypt. There were no obvious clinical signs in tested turkey poults except dehydration and whitish diarrhoea in some birds with no mortality, and post-mortem lesions were observed in few birds as atrophied bursae, nephritis and petechial haemorrhages on thigh muscles. Reverse transcription polymerase chain reaction (RT-PCR), histopathological examination and immunohistochemistry were used for identification of the IBDV. Out of 30 tested samples, 17 samples (56.7%) were positive by RT-PCR. Phylogenetic analysis of VP2 gene of two selected IBDV strains (turkey 1 and turkey 2) showed a close genetic relationship to vvIBDV strains (serotype 1) isolated from chickens in Egypt and other countries with 93.1 to 95.99% identity for turkey 1 strain and 95.54 to 98.51% for turkey 2 strain. Both turkey 1 and turkey 2 strains were closely related to the Nigerian vvIBDV strain isolated from turkeys with 95.78% and 96.37% identity, respectively. Sequence analysis of both strains demonstrated that they have conserved amino acid residues of vvIBDV (I242, I294 and S299) and Y220F amino acid substitution which is very common in Egyptian vvIBDV chicken strains, while Turkey 1 strain has amino acid substitutions at A222P and I256V. Histopathological examination showed marked depletion of bursal lymphoid tissue. In conclusion, for the first time in Egypt, the molecular characterization and pathogenicity confirmed the presence of natural infection of turkey poults with vvIBDV (serotype 1) with possible spread to chickens causing severe economic losses.

Immunostimulant effect of a mixed herbal extract on infectious bursal disease virus (IBDV) vaccinated chickens in the context of a co-infection model of avian influenza virus H9N2 and IBDV.

This study was conducted to assess the comparative effects of a mixed herbal extract (MHE) containing Ocimum sanctum, Withania somnifera, Emblica officinalis, Tinospora cordifolia, Mangifera indica, and Asphaltum (shilajit) on infectious bursal disease virus (IBDV)-vaccinated (VAC) chickens infected with IBDV and avian influenza virus (AIV) H9N2. The experiment included three groups (G1-G3): G1, the negative control group; G2, the VAC + challenged (Ch) group; and G3, the VAC + Ch + MHE group. MHE was orally administered continuously for 5 weeks post-vaccination (PV) with IBDV at 12 days of age, and the chicks were simultaneously challenged with virulent IBDV (intraocularly) and AIV H9N2 (intranasally) at 21 days PV. Blood and tissue samples as well as tracheal and cloacal swabs were gathered at different times PV and post-challenge. Immunological and haematological parameters, histopathological lesions, relative organ weights and final live weights revealed significant differences (P ≤ 0.05) between G2 and G3 groups. Furthermore, in the G3 group, the protection rates, ELISA and HI titers and CD4+/CD8+ ratio were significantly increased, whereas viral shedding titers and the heterophil/lymphocyte ratio were decreased. In conclusion, the oral administration of the mixed herbal extract for 5 weeks can stimulate the immune response to IBDV vaccination and relieves the pathogenicity of an AIV H9N2 and IBDV co-infection in chickens.

Effect of omega-3 rich diet on the response of Japanese quails (Coturnix coturnix japonica) infected with Newcastle disease virus or avian influenza virus H9N2.

This study was performed to evaluate the effects of omega-3 supplementation on growth performance, clinical signs, post-mortem lesions, haemagglutination inhibition (HI) antibody titres, gene expression and histopathology in quails (Coturnix coturnix japonica) infected with Newcastle disease virus (NDV) and avian influenza virus (AIV) H9N2. One hundred, 40-day-old male quails were divided into 5 groups: G1, fed a control basal diet; G2A, infected with NDV; G2B, infected with H9N2; G3A, infected with NDV and given omega-3, and G3B, infected with H9N2 and given omega-3. The dietary omega-3 supplementation was continued for 4 weeks: two weeks before infection and two weeks after intranasal infection with virulent NDV and AIV H9N2. Our results revealed significant differences (P < 0.05) in growth performance, HI antibody titres, clinical signs, post-mortem lesions, mortality, viral shedding rates, immunological parameters, and histopathological lesions between the treated (G3A and G3B) and untreated (G2A and G2B) groups. In conclusion, dietary omega-3 supplementation for 4 weeks can improve growth performance and alleviate the deleterious immunological and pathological effects of NDV and AIV H9N2 infection in quails.

Effect of colibacillosis on the immune response to a rabbit viral haemorrhagic disease vaccine.

Viral haemorrhagic disease (VHD) and colibacillosis are common diseases in rabbits that cause economic losses worldwide. The effect of colibacillosis on the immune response of vaccinated rabbits against rabbit haemorrhagic disease virus (RHDV) was studied. Four groups (G1-G4) were included. G1 was the negative control group; G2 was the RHDV vaccine group; G3 was the E. coli-infected group; and G4 was the E. coli-infected + RHDV vaccine group. The E. coli infection and RHDV vaccination were simultaneously performed, with another previous infection, 3 days before vaccination. At 28 days post-vaccination (PV), the rabbits (G2-G4) were challenged intramuscularly with 0.5 ml of RHDV at a dose of 103 50% median lethal dose (LD50)/rabbit. The rabbits were observed for clinical signs, body weight gain and mortality rates. Tissue, blood, serum, and faecal samples and rectal swabs were collected at 3, 5, 7, 14, 21 and 28 days PV. Significant clinical signs and mortality and a decrease in BW were observed in the infected + RHDV vaccine group. On the 3rd day post-infection (PI), compared with all the other groups, the vaccinated group (G2) had significantly upregulated hepatic tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels; however, the infected + RHDV vaccine group had significantly higher intestinal levels of TNF-α and IL-6 than the other groups. Furthermore, E. coli infection in vaccinated rabbits led to immunosuppression, as shown by significant decreases (P < 0.05) in heterophil phagocytic activity, the CD4+/CD8+ ratio, and HI antibody responses to RHDV and a significant increase in the heterophil to lymphocyte (H/L) ratio. In conclusion, colibacillosis leads to immunosuppression involving a shift in the equilibrium of cytokines and reduced weight gain and mortality in vaccinated rabbits and could be a contributing factor in RHDV vaccination failure in rabbit farming.

Comparative molecular characterization, pathogenicity and seroprevalence of avian influenza virus H9N2 in commercial and backyard poultry flocks.

This study was conducted to perform the comparative molecular characterization of avian influenza virus (AIV) H9N2, pathogenicity and seroprevalence in commercial and backyard poultry flocks. Fifty commercial poultry flocks were investigated between 2012 and 2015. Eighteen flocks (36%) out of 50 were positive HA. Seven (38.9%) out of 18 were positive by chromatographic strip test for AI common antigen. By Real-time RT-PCR, only two flocks were positive H9. The molecular characterization of two different AI-H9N2 viruses, one isolated from a broiler flock (A/chicken/Egypt/Mansoura-18/2013) and the other from a layer flock (A/chicken/Egypt/Mansoura-36/2015) was conducted on HA gene. Moreover, a higher seroprevalence, using the broiler strain as a known antigen, was shown in backyard chicken flocks 15/26 (57.7%) than duck flocks 9/74 (12.2%). Interestingly, the pathogenicity index (PI) of the H9N2 broiler strain in inoculated experimental chickens ranged from 1.2 (oculonasal route) to 1.9 (Intravenous route). The PI indicated a highly pathogenic effect, with high mortality (up to 100%) in the inoculated chickens correlated with the high mortality (80%) in the flock where the virus was isolated. The firstly recorded clinical signs, including cyanosis in the combs and wattles and subcutaneous haemorrhages in the leg shanks and lesions, as well as histopathology and immunohistochemistry, revealed a systemic infection of the high pathogenicity with the H9N2 virus. Conversely, the H9N2 layer strain showed a low pathogenicity. In conclusion, as a first report, the molecular analysis and pathogenicity of the tested strains confirmed the presence of a high pathogenicity AIV-H9N2 with systemic infections.

Immunological and pathological effects of vitamin E with Fetomune Plus® on chickens experimentally infected with avian influenza virus H9N2.

Avian influenza virus (AIV) H9N2 infection causes economic losses on poultry farms, and immunostimulants are essential for improving chicken immunity. This study evaluated the immunological and pathological effects of vitamin E with Fetomune Plus® (a commercial product based on a yeast extract and vitamins) on chickens experimentally infected with AIV H9N2. Three groups of white Hy-Line chicks were included. The G1 group was kept as an uninfected untreated control, the G2 group was intranasally infected with the AIV H9N2 strain (0.5 ml of 106 50% egg infectious dose (EID50)), and the G3 group was infected and treated with vitamin E (200 mg/kg of diet) and Fetomune Plus® (1 ml/liter of drinking water) for four weeks. The gene expression of interferon-gamma (IFN-γ), interleukin (IL)-6, and IL-2 was determined at 3, 5 and 7 days post-infection (PI). Virus shedding titers and rates and haemagglutination inhibition (HI) antibody titers were detected. Clinical signs, mortalities and post-mortem lesions were recorded. The birds were weighed, and relative organ weights were calculated. Tissue specimens were taken for histopathological examination and immunohistochemistry (IHC). The expression of IFN-γ in the duodenum revealed a significant increase in G2 compared to G3 at 3 days PI, while the duodenal and splenic expression of IL-6 was significantly increased in G2 compared to G3 at 5 days PI. IL-2 was overexpressed in the duodenum in G3 compared to G2 at 3 and 5 days PI. A significant decrease (P ≤ 0.05) in the virus shedding titer and an increase in the HI titers were detected in G3 compared to G2. The clinical signs and the mortality rate were clearly appeared in G2 than in G3. By IHC, lower H9N2 staining intensity was observed in the examined organs from G3 than in those from G2. In conclusion, as a first report, vitamin E with Fetomune Plus® supplementation for four weeks could improve the immunological and pathological effects of H9N2 infection on chickens.