Aim: This study aimed to investigate the prevalence of different bacterial species affecting ducks as well as demonstrating the antimicrobial susceptibility and molecular typing of the isolated strains. Materials and Methods: A total of 500 samples were randomly collected from different duck farms at Ismailia Governorate, Egypt. The collected samples were subjected to the bacteriological examination. Polymerase chain reaction (PCR) was applied for amplification of Kmt1 gene of Pasteurella multocida and X region of protein-A (spA) gene of the isolated Staphylococcus aureus strains to ensure their virulence. The antibiotic sensitivity test was carried out. Results: The most common pathogens isolated from apparently healthy and diseased ducks were P. multocida (10.4% and 25.2%), Escherichia coli (3.6% and 22.8%), Staphylococcus epidermidis (10% and 8.8%), Pseudomonas aeruginosa (2% and 10%), and Proteus vulgaris (0.8% and 10%), respectively. In addition, S. aureus and Salmonella spp. were isolated only from the diseased ducks with prevalence (12.2%) and (2.8%), respectively. Serotyping of the isolated E. coli strains revealed that 25 E. coli strains were belonged to five different serovars O1, O18, O111, O78, and O26, whereas three strains were untypable. Salmonella serotyping showed that all the isolated strains were Salmonella Typhimurium. PCR revealed that four tested P. multocida strains were positive for Kmt1 gene with specific amplicon size 460 bp, while three strains were negative. In addition, all the tested S. aureus strains were positive for spA gene with specific amplicon size 226 bp. The antibiotic sensitivity test revealed that most of the isolated strains were sensitive to enrofloxacin, norfloxacin, and ciprofloxacin. Conclusion: P. multocida is the most predominant microorganism isolated from apparently healthy and diseased ducks followed by E. coli and Staphylococci. The combination of both phenotypic and genotypic characterization is more reliable an epidemiological tool for identification of bacterial pathogens affecting ducks.
Newcastle disease virus ( NDV ) is a major threat to the poultry industry worldwide, with a diversity of genotypes associated with severe economic losses in all poultry sectors. Class II genotype VII NDV are predominant in the Middle East and Asia, despite intensive vaccination programs using conventional live and inactivated NDV vaccines. In Egypt, the disease is continuously spreading, causing severe economical losses in the poultry industry. In this study; the protective efficacy of a commercial, inactivated recombinant genotype VII NDV–matched vaccine (KBNP-C4152R2L strain) against challenge with the velogenic NDV strain (Chicken/USC/Egypt/2015) was evaluated in commercial layers. Two vaccination regimes were used; live NDV genotype II (LaSota) vaccine on days 10, 18, and 120, with either the inactivated NDV genotype II regime or inactivated NDV genotype VII–matched vaccine regime on days 14, 42, and 120. The 2 regimes were challenged at the peak of egg production on week 26. Protection by the 2 regimes was evaluated after experimental infection, based on mortality rate, clinical signs, gross lesions, virus shedding, seroconversion, and egg production schedule. The results show that these 2 vaccination regimes protected commercial layer chickens against mortality, but some birds showed mild clinical signs and reduced egg production temporarily. However, the combination of live NDV genotype II and recombinant inactivated genotype VII vaccines provided better protection against virus shedding (20% and 0% vs. 60% and 40%) as assessed in tracheal swabs and (20% and 0% vs. 20% and 20%) in cloacal swabs collected at 3 and 5 D post challenge ( dpc ), respectively. In addition, egg production levels in birds receiving the inactivated NDV genotype VII–matched vaccine regime and in those given inactivated genotype II vaccines were 76.6, 79, 82, and 87.4% and 77.7, 72.5, 69, and 82.5% at 7, 14, 21, and 28 dpc, respectively. The results of this study indicate that recombinant genotype-matched inactivated vaccine along with a live attenuated vaccine can reduce virus shedding and improve egg production in commercial layers challenged with a velogenic genotype VII virus under field conditions. This regime may ensure a proper control strategy in layers.
The current study was carried out to evaluate efficiency of different vaccination programs in protecting broiler chicken against Virulent Viscerotropic Strain of Newcastle disease virus. Different live and inactivated NDV vaccines were used throughout the experiment including; ND-HB1,Elite, LaSota, Inactivated GII and Inactivated GVII. Broiler chicks were divided into 8 groups; 6 groups undergo different vaccination programs against NDV while two groups were kept without vaccination to be the control groups. Challenge was done at day 30 th of age via intranasal administration of NDV velogenic GVII (NDV/CK/Egypt/567F/2012). Abs titers were determined at days 1, 7, 14, 21, 28 and 35 days of the experiment. The obtained results of shedding titers of NDV clarified that the lowest shedding titer was recorded in group vaccinated by HB + double shots of Inactivated GVII + Elite + LaSota and HB + double shots of Inactivated GII + Elite + LaSota, then group vaccinated by HB + one shot of Inactivated GVII + Elite + LaSota and HB + one shot of Inactivated GII + Elite + LaSota at days 3, 5, 7 and 9 post challenge day at 30 th , respectively compared to those vaccinated by live vaccines HB + LaSota only and HB + Elite + LaSota only. Also, no mortalities (100% protection rate) were recorded in groups vaccinated with both live and inactivated NDV vaccines compared to low mortality rates recorded in groups vaccinated with live vaccines only. Based on the recorded results, it was concluded that application of ND vaccination programs containing both live and double inactivated vaccines (either GII or GVII) was found to be more effective than those depending on one shot of inactivated vaccine (either GII or GVII) plus live vaccines and more effective than program including live vaccines only.
Toll-like receptors (TLRs) are the best understood of the innate immune receptors that detect infections in vertebrates. However, the fish TLRs also exhibit very distinct features and a large diversity, which is likely derived from their diverse evolutionary history and the distinct environments that they occupy. Little is known about the fish immune system structure. Our work was aimed to identify and clone the Nile tilapia TLR-3 as a model of fresh water fish species; we cloned the full-length cDNA sequence of Nile tilapia (Oreochromis niloticus) TLR-3 and according to our knowledge, it is the first report illustrating tilapia TLR-3. The complete cDNA sequence of Nile tilapia TLR-3 was 2736 pair base and it encodes a polypeptide of 912 amino acids. Analysis of the deduced amino acid sequence indicated that Nile tilapia TLR-3 has typical structural features and main component of proteins belonging to the TLR family. Our results illustrate a complete and functional Nile tilapia TLR-3 and it is considered an ortholog of the other vertebrate's receptor.
Egyptian poultry suffer from frequent respiratory disease outbreaks associated with Infectious Bronchitis Virus (IBV) variant 2 strains (Egy/VarII). Different vaccination programs using imported vaccines have failed to protect the flocks from field challenge. Recent studies confirmed a successful protection using homologous strains as live attenuated vaccines. In this study, a newly developed live attenuated IB-VAR2 vaccine representing the GI-23 Middle East IBV lineage was evaluated in day-old commercial broilers in an IBV-endemic area. A commercial broiler flock was vaccinated with the IB-VAR2 vaccine at day-old age followed by IB-H120 at day 16. The vaccinated flock was monitored on a weekly basis till the slaughter age. The health status and growth performance were monitored, and selected viral pathogen real-time RT-PCR (rRT-PCR) detection was conducted on a weekly basis. Finally, the flock was compared to a nearby farm with only the classical IB-H120 vaccination program. Results showed that the IB-VAR2 vaccine was tolerable in day-old broiler chicks. The IBV virus rRT-PCR detection was limited to the trachea as compared to its nephropathogenic parent virus. Respiratory disease problems and high mortalities were reported in the IB-H120-only vaccinated flock. An exposure to a wild-type Egy/VarII strain was confirmed in both flocks as indicated by partial IBV S1 gene sequence. Even though the IB-VAR2-vaccinated flock performance was better than the flock that received only IB-H120, the IBV ELISA (enzyme-linked immunosorbent assay) and log2 Haemagglutination inhibition (HI) antibody mean titers remained high (3128 ± 2713 and ≥9 log2, respectively) until the 28th day of age. The current study demonstrates the safety and effectiveness of IB-VAR2 as a live attenuated vaccine in day-old commercial broilers. Also, the combination of IB-VAR2 and classical IBV vaccines confers a broader protective immune response against IBV in endemic areas.
Low pathogenic avian influenza virus is one of the major threats that has been affecting the poultry industry in the Middle East region for decades. Attempts to eradicate this disease have failed. Currently, there are commercial vaccines that are either imported or produced locally from recently circulating isolates of H9N2 in Egypt and Middle Eastern countries. This present work focused on comparing the effectiveness of two vaccines belonging to these categories in Egypt. Two commercial broiler flocks (Cobb-500 Broiler) with maternally derived immunity (MDA) against H9N2 virus were employed and placed under normal commercial field conditions or laboratory conditions. Immunity was evaluated on the basis of detectable humoral antibodies against influenza H9N2 virus, and challenge was conducted at 28 days of life using a recent wild H9N2 virus. The results showed that vaccination on the 7th day of life provided significantly higher immune response in both vaccine types, with significantly lower virus shedding compared to vaccination at day 1 of life, regardless of field or laboratory conditions. In addition, the vaccine produced from a recent local H9N2 isolate (MEFLUVAC-H9-16) provided a significantly higher humoral immune response under both field and laboratory conditions, as measured by serology and virus shedding (number of shedders and amount of shedding virus), being significantly lower following challenge on the 28th day of life, contrary to the imported H9 vaccine. In conclusion, use of H9N2 vaccine at 7 days of life provided a significantly higher protection than vaccination at day 1 of life in birds with MDA, suggesting vaccination regimes between 5–8-days of life for broiler chicks with MDA. Moreover, use of a vaccine prepared from a recently circulating H9N2 virus showed significantly higher protection and was more suitable for birds in the Middle East.
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