Evaluating the effect of Spirulina platensis on the immune response of broiler chickens to various vaccines and virulent Newcastle disease virus challenge.

This study investigated the efficacy of co-administration of Spirulina platensis (SP) with vaccines on the immune response to Avian influenza (AI), Infectious bronchitis (IB), and Newcastle disease (ND), along with I/M challenging by virulent ND virus (vNDV) genotype VII. 126 one-day-old broiler chicks were allocated into six groups (21 birds/group with three replicates): G1: negative control; G2: positive control; G3: vaccinated, non-SP-supplemented; G4: vaccinated, SP-supplemented (0.1%); G5: vaccinated, SP-supplemented (0.3%); and G6: vaccinated, SP-supplemented (0.5%). G2-6 were challenged with a velogenic NDV genotype VII virus. Dietary SP administration prevented the ND-induced mortality compared to G2 (52.4%) and G3 (14.3%), in addition to alleviating the clinical disease. G3-6 showed significant improvement in body weight loss% and FCR during two weeks post vNDV challenge (pc), and the overall FCR (2.64 ± 0.28, 1.56 ± 0.03, 1.60 ± 0.05, 1.53 ± 0.04, and 1.54 ± 0.03 for G2, 3, 4, 5 and 6, respectively) (P<0.05). On the challenging day, the ND-HI titer (log2) of G3 (5.44 ± 0.24) was numerically higher than G6 (4.20 ± 0.55) and lower than G4 (6.10 ± 0.34) and G5 (6.00 ± 0.28). On the 10th day pc, ND-HI titer in G4-6 was numerically lower in a dose-dependent manner than that of G3, suggesting an antiviral efficacy of SP. G4-6 had lower viral shedding titer than G2 and G3 (P<0.05). In G3-6, viral shedding was reduced by 15, 27, 24, and 33.6%, respectively. In addition, the histopathological lesions in the trachea, lung, and spleen were severe in G2, moderately reduced in G3, and more relieved in G4-6. At three weeks after vaccination, the HI antibody titer of AIH5 was significantly higher after SP administration, especially at the 0.3% level, compared to the vaccine alone (P<0.05), demonstrating an immune-stimulating effect. In conclusion, dietary administration of SP, particularly a dose of 0.3%, for vaccinated chickens against NDV exerted an antiinflammatory and antiviral effects by preventing deaths, alleviating clinical disease and weight loss, and decreasing viral shedding post heterologous NDV challenge.

Examination of the protective efficacy of two avian influenza H5 vaccines against clade 2.3.4.4b H5N8 highly pathogenic avian influenza virus in commercial broilers.

The highly pathogenic avian influenza (HPAI) H5N8 virus of clade 2.3.4.4 was detected in 2017 in Egypt, which is one of the few countries using vaccination as a control strategy in poultry farms. This study was conducted to evaluate the efficacy of the commercial recombinant turkey herpes virus-H5 (rHVT-H5) vaccine (clade 2.2), alone or in combination with commercial inactivated reverse genetically engineered H5N1 vaccine (rgH5N1) (clade 2.2), in preventing the genetically distinct HPAI H5N8 virus of clade 2.3.4.4b in commercial broiler chickens. Four experimental groups of chickens were used as follows: G1, non-vaccinated and non-challenged; G2, non-vaccinated and challenged; G3, vaccinated with rHVT-H5; and G4, prime-boost vaccinated with rHVT-H5/rgH5N1. For challenge with the Egyptian HPAI H5N8 (2.3.4.4b) virus, the groups were divided into two subgroups (A and B); chickens in subgroups A were challenged at the age of 28 days, whereas those in subgroups B were challenged at the age of 35 days. Results showed that a protective efficacy (survival rate) of 40%-50% was obtained in the vaccinated subgroups A. By delaying challenge for 1 week (subgroups B), a single rHVT-H5 vaccination provided 80% protection, whereas prime-boost vaccination induced full protection and reduced viral shedding very efficiently (1/10 birds and only detected on the 3rd day post challenge) against HPAI H5N8 virus (2.3.4.4b). Moreover, body weight loss improved from 31.39% and 43.65% in G3A and G4A, respectively, to 16.34% and 7.7% in G3B and G4B, respectively. The HI titers obtained in G3A and G4A on the challenge day (28th d) using H5N8 antigen were 3 and 3.75 log2 (p > 0.05), respectively, whereas those in G3B and G4B on the challenge day (35th d) were 6.25 and 6 log2 (p > 0.05), respectively, which increased post-challenge in all vaccinated subgroups. Therefore, the dual use of vectored rHVT-H5 and inactivated rgH5N1 vaccines in the vaccination schedule in poultry farms is the most efficient tool for preventing the disease (mortality and viral shedding) caused by the genetically distinct virus (clade 2.3.3.4b HPAI H5N8) in combination with strict biosecurity and sanitary measures.

Incidence, Pathotyping, and Antibiotic Susceptibility of Avian Pathogenic Escherichia coli among Diseased Broiler Chicks.

A total of 54 broiler flocks during the first two weeks of life was used to investigate the incidence of avian pathogenic E. coli in Egypt; 28 isolates (51.85%) were revealed by colony morphology and biochemical identification which then investigated for their serogroups and only 18/28 isolates were serotyped. The most prevalent serotypes were O115, O142, O158, O55, O125, O114, O27, O20, and O15. By application of polymerase chain reaction (PCR), 83.3% (15/18) of the serotyped isolates were confirmed to be E. coli, and 93.3% (14/15), 46.6% (7/15), and 20% (3/15) of isolates harbored the iss, iutA, and fimH genes, respectively. Virulence testing of the selected 13 APEC isolates on the specific-pathogen-free (SPF) chicks revealed them to be highly virulent (15.4%), moderately virulent (23.1%), and avirulent (61.5%); however, all isolates (100%) were extremely virulent towards SPF embryonated chicken eggs. Antibiotic resistance (100% of isolates (n = 13)) was observed for ampicillin, amoxycillin-clavulanic acid, and tetracyclines, colistin (92.31%; 12/13), doxycycline and spiramycin (84.62%; 11/13), florfenicol (69.23%; 9/13), cefotaxime (61.54%; 8/13), and ciprofloxacin (53.85%; 7/13). The highest percentage of sensitivity (53.85% of isolates; 7/13) was recorded for ofloxacin and enrofloxacin followed by gentamycin (46.15%; 6/13). The results suggest that the diagnosis of APEC with PCR is rapid and more accurate than traditional methods for E. coli identification; moreover, the presence or absence of iss, iutA, and/or fimH genes is not an indicator of in vivo pathogenicity of APEC. Thus, further studies, including a wider range of virulence genes and gene sequencing, are required. In addition, serotyping has no effect on the virulence of APEC.

Isolation, conventional and molecular characterization of Salmonella spp. from newly hatched broiler chicks.

Salmonella is an important pathogen for poultry production as well as for human due to zoonotic importance. It has more than 2600 identified serovars despite of this identification and classification of Salmonella isolates into different serovars is critical for study of incidence and surveillance. This study investigates the epidemiology and molecular characterization of Salmonella isolates in broiler chicks during 1st week of life. A total of (n = 1000) samples including liver, intestine, yolk sac, spleen and heart blood were collected from El-Gharbia, El-Behera, Kafr-Elshikh, Alexandria, Marsamatroh Provinces in Egypt and tested through bacteriological, biochemical, serological and molecular examinations. Incidence of Salmonella was demonstrated on 75 positive samples from 1000 samples and the predominance of Salmonella that isolated from internal organs of newly hatched chicks was highest from yolk sacs (10%), liver and intestines (9%) followed by the spleen (7.5%) then heart blood (2%). Serotyping of the isolated strains using slide agglutination test revealed that 24 isolates belonging to S. enteritidis (1,9,12 g.m 1,7), while, 14 isolates belonging to S. virchow (6,7 r 1,2), in addition to, 12 isolates belonging to S. typhimurium (1,4,5,12.i.1,2) and 8 isolates belonging to S. kentucky (6,8.I,z). Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR revealed that two S. enteriditis isolates were identical and one isolate differ by 40%, while two S. typhimurium isolates were identical by 80% and one isolate was similar by 20% to the other two isolates, in addition, two S. virchow isolates were identical by 80% and the two S. kentucky isolates were different.

Comparative Evaluation of HVT-IBD Vector, Immune Complex, and Live IBD Vaccines against vvIBDV in Commercial Broiler Chickens with High Maternally Derived Antibodies.

Infectious bursal disease (IBD) causes increased mortality and severe immunosuppression in commercial chickens. Currently, vaccination mainly used to control IBD. In this study, Group A (n = 30) received the HVT-IBD vector vaccine (Vaxxitek®) s/c and Group B (n = 30) received the immune complex vaccine (Bursa-Plex®) s/c at 1 day of age. Group C (n = 30) received a single dose of intermediate plus vaccine (228E) through the eye-drop route at 14 days of age. Group D (n = 30) was vaccinated twice with the intermediate vaccine (D78) at 12 and 22 days of age by eye-drop. Group E (n = 30) had the same treatment as group D along with the IBD killed vaccine (Nobilis G®) at 5 days of age. The PC (n = 20) and NC (n = 20) groups were non IBD vaccinated birds either challenged or not with vvIBDV, respectively; 20 chicks from each group were challenged with vvIBDV at 4 weeks of age. Based on clinical signs, postmortem gross lesions, histopathological changes, mortality rate, feed conversion rate, serology, bursal and spleen indices, the HVT-IBD vector vaccine administered was found to be safer and provided better protection against the vvIBDV challenge. The use of a killed IBD vaccine at an earlier age in broilers strengthened the protection induced by double doses of intermediate vaccines in broilers with high maternally derived antibodies against the vvIBDV challenge.

The Simultaneous Administration of a Probiotic or Prebiotic with Live Salmonella Vaccine Improves Growth Performance and Reduces Fecal Shedding of the Bacterium in Salmonella-Challenged Broilers.

Salmonellosis is one of the most important bacterial diseases in poultry, causing heavy economic losses, increased mortality and reduced production. The aim of this study was the comparative efficacy of a commercial probiotic and/or prebiotic with a live attenuated Salmonella Enteritidis (SE) vaccine on the protection of broiler chickens from SE challenge. The efficacy of probiotic or prebiotic products, as well as a live Salmonella Enteritidis (SE) vaccine at the 7th day of age, administered via drinking water, were evaluated for clinical protection and effects on growth performance of broiler chickens experimentally challenged with SE at the 28th day of age. The use of probiotic or prebiotic simultaneously with the live Salmonella vaccine can diminish the negative effect of live vaccine growth performance, reducing mortality rate, fecal shedding, and re-isolation of SE from liver, spleen, heart and cecum. The use of probiotic or prebiotic simultaneously with the application of the live Salmonella vaccine is a good practice to diminish the negative effect of the harmful bacteria and improve the growth performance of broilers. Thus, further studies may be carried out with layers and breeders.

In vitro inactivation of two Egyptian A/H5N1 viruses by four commercial chemical disinfectants.

The highly pathogenic H5N1 avian influenza virus (A/H5N1) devastated the poultry industry and posed a serious health threat. Cleaning and disinfection are essential parts of preventative and postoutbreak management of A/H5N1 infections in poultry. In this preliminary study, we used suspension and carrier tests to evaluate the impact of concentration, time of exposure, surface porosity, and organic matter on the ability of four commercial chemical disinfectants to inactivate two A/H5N1 viruses of clade 2.2.1 isolated in 2006 and 2010 from broiler flocks in Egypt. Viruses were incubated with 0.5%, 1%, and 2% of formalin, glutaraldehyde, TH4, and Virkon S for 15, 30, 60, and 120 min at room temperature (22 +/- 2 C). In suspension tests, in the absence of organic matter, all disinfectants, at each concentration, except Virkon S 0.5%, effectively inactivated virus suspensions after a 15-min exposure time. In the presence of organic matter, the use of low concentrations of formalin (0.5%), glutaraldehyde (0.5%), or Virkon S (0.5%) was not sufficient to inactivate the viruses after 15 min. In gauze carrier tests, only formalin at any concentration for 15 min was sufficient to inactivate the viruses, whereas different concentrations or exposure times were required for glutaraldehyde (0.5% for 60 min), TH4 (0.5% for 30 min), and Virkon S (0.5% for 60 min or 1% for 30 min). In wood carrier tests, total inactivation of the virus was obtained at concentrations of 0.5% for 30 min (formalin and TH4) or 60 min (glutaraldehyde and Virkon S). This study emphasizes the need to use high concentrations of and/or extended time of exposure to disinfectants for efficient inactivation of A/H5N1, particularly in the presence of organic matter or different surfaces, which are common in poultry operations. In addition, it seemed that the virus isolated in 2010 was more resistant to disinfectants than the isolate from 2006 when wood was used as a carrier.