No abstract

Background The applications of Cu and CuNPs based on the earth-abundant and inexpensive Cu metal have generated a great deal of interest in recent years, including medical applications. A novel, specific, precise, accurate and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method with UV detection has been developed and validated to quantify copper (Cu) and copper nanoparticles (CuNPs) in different biological matrices and pharmaceutical products. Methods The developed method has been validated for linearity, precision, sensitivity, specificity and accuracy. Cu concentration was detected in pharmaceutical products without an extraction process. Moreover, liver, serum and muscle tissues were used as biological matrices. High Cu recovery in biological samples was afforded by using citric acid as a green chelating agent, exact extraction time and pH adjustment. Cu pharmaceutical and biological samples were eluted by acetonitrile: ammonium acetate (50 mM) with 0.5 mg/ml EDTA (30:70 v:v) as an isocratic mobile phase. EDTA reacted with Cu ions forming a Cu-EDTA coloured complex, separated through the C18 column and detected by UV at 310 nm. Results The developed method was specific with a short retention time of 4.95 min. It achieved high recovery from 100.3% to 109.9% in pharmaceutical samples and 96.8–105.7% in biological samples. The precision RSD percentage was less than two. The method was sensitive by achieving low detection limits (DL) and quantification limits (QL). Conclusion The validated method was efficient and economical for detecting Cu and CuNPs by readily available chemicals as EDTA and Citric acid with C18 column, which present the best results on RP-HPLC.

Background Newcastle disease virus (NDV) is a severe disease that affects domestic and wild birds. Controlled antibiotics derived from probiotics have been examined as prospective solutions for preserving seroconversion in NDV-vaccinated fowl. In this study, the secondary metabolite “telomycin” was extracted from Streptomyces coeruleorubidus (S. coeruleorubidus) isolated from Egypt's cultivated soil. The structure of telomycin was determined by the elucidation of spectroscopic analysis, including nuclear magnetic resonance (NMR) and mass spectrometry (MS) spectra, and comparison with the literature. The antiviral activity of the secondary metabolite was tested by checking its effect on NDV hemagglutination activity (HA). Moreover, HA of NDV was tested after inoculation of NDV (control) and a combination of telomycin and NDV in 10- days- specific pathogen-free embryonated chicken eggs (SPF-ECE) daily candling. Histopathological examination was performed for chorioallantoic membranes and liver of SPF-ECE. Results S. coeruleorubidus secondary metabolite “telomycin” showed complete hemagglutination inhibition (HI) activity of NDV strain (MN635617) with log106 infectivity titers (EID50/mL). The HA of NDV strain was 8 log2 and 9 log2 with 0.5% and 0.75% of chicken RBCs, respectively. Preserved structures of chorioallantoic-membranes (CAM) with dilated capillary networks were observed in the treated group inoculated with telomycin and NDV. Histological changes in SPF-ECE liver were examined after inoculation in ova to further characterize the telomycin effect. Telomycin and NDV mixture inoculated group showed preserved cytoarchitecture of hepatocytes with the presence of perivascular foci of lymphocytes. The group that was inoculated with telomycin alone showed normal histology of hepatic acini, central veins, and portal triads. Conclusion S. coeruleorubidus telomycin is a promising bioactive agent that might be a biological weapon against a deadly chicken NDV that costs farmers a lot of money.

Chicken infectious anemia is one of the most economic immunosuppressive problems facing commercial poultry sector worldwide. Present study demonstrated the viral load in body organs and viral specific antibody titres of the SPF chicks experimentally infected with chicken anemia virus (CAV) strain CAV/Kal.2 and its contact group in vivo. Also, followed the emergence of lysozyme activity and nitric oxide (NO) levels, pro-inflammatory {IL-1β, IL-6 and CXCLi2 (IL-8 like chemokine)}, type I IFN (IFN-α and IFN-β) and IFN-γ cytokines. The obtained data illustrated that CAV-specific antibody development started after 7 days post infection (dpi) and reached its maximum level at 21 dpi in infected group and contact group. The CAV genome was detected in tissues of the chicks of both infected and contact groups at 7 dpi and continued for 21 dpi with 7 days intervals between sampling. Lysozyme activity and NO levels were greatly impaired in CAV infected chicks. The relative mRNA expression levels of most examined cytokines in the infected group were increased on 14 dpi compared to 7 and 21 dpi. On the other hand, CXCLi2 was generally not altered by CAV infection. The contact group showed undetermined changes in all examined cytokines (IL-1β, IL-6, CXCLi2, IFN-α, IFN-β and IFN-γ). The obtained data revealed cytokine imbalances after infection with CAV as a result of hindrance of transcription of the most of the examined cytokines. As the immunosuppressive viruses of chickens may confuse with transcription for several cytokines (IL-1β, IL-6, IFN-α, IFN-β and IFN-γ), so we suggest using this confliction in order to evaluate the immune status of chickens.

Objective: Rabbit viral hemorrhagic disease (VHD) is a transmittable and lethal viral illness of rabbits. In this study, genetic identification and genetic analysis of the rabbit hemorrhagic disease virus (RHDV) was made in three governorates in Egypt from 2014 to 2019. Materials and Methods: Livers from 18 freshly dead rabbits, which was guessed to be VHD epidemics in Egypt (Giza, Menofia, and Fayoum governorates) from 2014 to 2019, were examined for RHDV. The examination was based on the hemagglutination assay (HA) test against different mammalian (human O-type and sheep) and avian (chicken and pigeon) erythrocytes, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequencing of the segment of VP60. Results: 33% of the examined samples’ virus titers were 5 log 2 to 8 log 2 hemagglutination of human O-type erythrocytes when compared to 28%, 11%, and 28% of sheep, chicken, and pigeon erythrocytes, respectively. Four RHDV isolates out of eight RT-PCR positives were sequenced and phylogenetically analyzed. Sequenced isolates were designed and submitted to GenBank with accession numbers MN904506, MN904507, MN904508, and MN904509. These four RHDV isolates were related to classic G3 (GI.1d/RHDV). Twelve amino acid differences were detected between the vaccine strain sequence (Giza-2006) and RHDV isolates. Amino acid differences at 416, 423, and 476 positions seem interesting as they changed polarity that could change the protein structure and affect host interaction. Conclusions: There is antigenic variation between circulating RHVD strains and the vaccinal strain. This may be the leading cause of vaccination failure and may increase the need to check out the vaccination program against RHVD.

Background The Newcastle Disease Virus (NDV) is present throughout the world, and outbreaks in Egypt caused serious economic losses in the poultry industry. Actinobacteria are a phylum of bacteria known for their potential in producing structurally diversified natural products which promising natural compounds used to combat viruses are presented and evaluated. Streptomyces Misakimycin isolated from Egyptian soil, and evaluated for their efficacy in controlling NDV. On the basis of the biochemical characteristics S. Misakimycin metabolites, identified by infrared (IR) and nuclear magnetic resonance (NMR) spectroscopy. Results In this investigation, NDV was found to have been isolated in February from a chicken farm in the Dakahlia Governorate of Egypt by the Animal Health Research Institute in Dokki, Giza. Diethylpthalate (DEP), a secondary metabolite of S. misakimycin, completely inhibited the hemagglutination (HI) activity of the NDV strain (MN635617) at log107 infectivity titers (EID50/mL). With 0.5 percent and 0.75 percent of chicken RBCs, the HA of the NDV strain was 2 log2 and 5 log2, respectively. In the treated group that received DEP and NDV inoculations, chorioallantoic-membranes (CAM) structures were preserved along with dilated capillary networks. Histological changes in SPF-ECE liver were examined after inoculation in ova to further characterize the DEP effect. Diethylpthalate and NDV mixture inoculated group showed preserved cytoarchitecture of hepatocytes with the presence of perivascular foci of lymphocytes. The group that was inoculated with telomycin alone showed normal histology of hepatic acini, central veins, and portal triads. Conclusion A potentially bioactive substance called diethylpthalate has the potential to be a biological weapon against a fatal chicken NDV that quickly increase cost losses for farmers.

Aflatoxin B1 (AFB1) is a metabolic product of the Aspergillus spp. of molds, which grow on several feedstuffs stored in hot moist conditions. It is one of the immunosuppressive agents that might influence the pathogenesis of avian influenza virus (AIV) subtype H9N2 in broilers, which can exacerbate the disease outcomes. The immunological, biochemical and pathological adverse health effects of an interaction between low levels of dietary aflatoxins (AFs) and H9N2 infection in broiler chickens were investigated. One hundred and eighty of unvaccinated 1-day-old COBB chicks were, therefore, raised for 35 days in the following treatment groups: control, AFs, AFs+H9N2, and H9N2. AFs in the basal diet was added at 200 ppb starting from the first day of age, while H9N2 virus was intra-nasally installed at a dose of 100 μl of 10 6 EID 50 /bird of allantois fluid at 23rd day. Humoral and cell-mediated immune responses were evaluated. Evidence of H9N2-AIV viral shedding was also detected. It has been observed that concurrent exposure of AFs and H9N2 virus negatively affected chicken performance traits i.e. lowered feed intake and body weights with exaggerated respiratory and digestive disturbances, and 20% mortality rate. Ten days' post H9N2 infection, significant (p≤ 0.05) increment in serum transaminases (AST and ALT) and falling in cell-mediated immunity i.e. total leukocyte count, lymphocyte transformation activity and macrophage phagocytic activity were detected. Additionally, AFs+H9N2 significantly (p≤ 0.05) lowered H9N2-HI titers (5.5 Log2) than H9N2 alone (6.3 Log2). Pathologically, aflatoxicated chickens showed hydropic degeneration, hepatocytic vacuolation and necrosis of liver tissues with nephrosis and urates deposition in ureters, as well as bursal and thymic lesions, which were potent in H9N2-inoculated chickens. AFs exposure increased the incidence and titer of H9N2 viral shedding. It could be concluded that dietary contamination with AFs even at very low levels has explanatory effect in H9N2-inoculated broilers, and vice versa. (0) no lesions; (1) mild lesions; (2) moderate lesions; (3) severe lesions and (4) very severe lesions http://epublishing.ekt.gr | e-Publisher: EKT | Downloaded at 08/07/2020 07:18:50 |

No abstract