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The intestinal microbiota plays important roles in animal health and growth. We investigated the efficacy and mechanisms of fecal microbiota transplantation (FMT) from adult SPF chickens against Salmonella Enteritidis (SE) infection in chicks. We transplanted 160 recipient SPF chicks (1-day-old) that were randomly divided into four groups, Ca (challenge), Cb (non-challenge), Fa (FMT and challenge) and Fb (FMT without challenge). The experiment lasted 40 days. We found that FMT reduced mortality as well as liver inflammatory lesions, promoted weight gain, improved immunity, ameliorated the digestion and absorption ability and inhibited SE colonization in the liver of challenged chicks. 16S rRNA gene high-throughput sequencing indicated that SE challenge caused a significant increase in the relative abundance of Parasutterella in the cecal microbiota of the recipient chicks (P < 0.05). FMT led to the maturation of the intestinal flora of recipients and the relative abundance of the Bacteroides, Rikenellaceae&#95; RC9&#95; gut&#95; group, Prevotellaceae&#95; UCG&#95; 001, Prevotellaceae&#95; Ga6A1&#95; group and Parabacteroides was significantly increased (P < 0.05). FMT from adult SPF chickens regulated the intestinal microbiota of chicks and increased resistance to SE infection., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Riemerella anatipestifer (RA) is the significant pathogen of septicemia and duck infectious serositis, diseases which can result in high mortality for ducklings. However, these diseases are difficult to treat because of the bacteria's broad resistance to multiple drugs. The purpose of this study was to produce a specific egg yolk immunoglobulin Y (IgY) targeted to RA, and to evaluate the protective efficacy of this IgY against RA infection. An RA-inactivated vaccine was produced via centrifugation and formalin treatment, using the most predominant serotype 2 wild-type strains in terms of worldwide prevalence. Anti-RA IgY was produced by immunizing Beijing Red No.1 hens with the inactivated vaccine. Enzyme-linked immunosorbent assays showed that the titer levels of anti-RA IgY antibodies increased significantly after exposure. Specific IgY isolated and purified from yolks effectively inhibited the growth of RA in the antibacterial activity assay, which revealed an 80 % reduction of bacteria populations. Animal experiments showed that duckling survival rates were able to reach up to 100 % after the ducklings were treated with 10 mg intramuscular injections of anti-RA IgY from 1 to 12 h after infection. However, the survival rates of ducklings treated with 30 mg of nonspecific IgY at 1 h after infection were 0%. Additionally, ducklings injected once with anti-RA IgY received complete protection in the first week, but the efficacy of this protection almost entirely disappeared after two weeks. The results suggested that specific anti-RA IgY has the potential to improve the degree of protection and responsiveness of ducklings to RA infections and provide them with passive immunity to RA. With further study, this is expected to become a new method for controlling RA infections., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Salmonella Enteritidis remains a leading cause of human foodborne disease, mostly associated with the consumption of contaminated poultry products. To more strategically implement a phage therapy scheme for S. Enteritidis control in broilers, a cocktail containing three wild-type lytic bacteriophages (LBs) previously isolated from chickens was evaluated shortly and later after a challenge. Genomic characterization, lytic spectrum and in vitro efficacy were determined for each studied LB. In independent trials, broilers challenged with S. Enteritidis on day of hatch received phage therapy from 6 to 10 days of age (early treatment), and from 31 to 35 days of age (later treatment). S. Enteritidis analyses were performed before treatment and at 1, 4, 7 and 10 days post-treatment (dpt) in both trials. Partial DNA sequence analysis of each LB revealed close similarity to the Ackermannviridae family. LBs lysed different Salmonella enterica serovars, while other tested bacteria were refractory. An in-vitro reduction of 1.49, 0.65 and 0.58 log 10 CFU/mL in S. Enteritidis number was obtained after co-incubation for 3 h with each LB. Both in vivo trials showed a significant reduction in the average number of intestinal S. Enteritidis calculated after phage therapy compared with controls. However, the highest efficiency was found in the later therapy, which resulted in a reduction of 1.08 log 10 CFU/g in the average from 4 to 10 dpt, showing potential for future use as a pre-harvest strategy to reduce the S. Enteritidis intestinal colonization in broilers on farms., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Anti-phage activity of serum is of importance in repeated phage therapy. Higher serum anti-phage activity has been associated with greater susceptibility of phages to neutralisation and phage therapy failure. In this study, in vivo and in vitro survivability and immunogenicity of four coliphages (TM1, TM2, TM3 and TM4) were investigated in naive chickens and chickens pre-immunised with phage TM1. Furthermore, two phages that displayed different survivability and immunogenicity (TM1 and TM3) were compared with respect to their efficacy in treating naive or pre-immunised (TM1) chickens suffering from colibacillosis. The efficacy of the treatments was evaluated based on body weight, relative organ weights, mortality, E. coli counts in the lungs as well as severity and frequency of internal organ lesions. At the end of the experiment, both naive and pre-immunised chickens treated with TM3 showed significantly lower mortality and higher body weights than untreated chickens and those treated with TM1. The same trend was observed in incidence and severity of organ lesions as well as relative spleen weight. However, naive chickens treated with TM1 also showed a shortened inflammation period as indicated by spleen weights. E. coli counts in the lungs of chicken treated with TM3 were lower than those of chickens treated with TM1 on days 3 and 10 post challenge. These data indicate that the outcome of phage therapy and the impact of serum anti-phage activity are highly phage-type dependent in broilers., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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 10 6 50% egg infectious dose (EID 50 )), 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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Avian pathogenic Escherichia coli (APEC) causes colibacillosis in poultry, leading to important economic losses worldwide. To cure APEC-infected chickens, a cocktail of four different APEC-specific bacteriophages (phages) was composed and tested. Specific phages were selected from a collection of phages isolated in Belgium. The selection was based on their obligate lytic infection cycle, a broad host range, low cross-resistance and low frequency of development of resistant APEC mutants. Genome analysis of the phages indicated they were close relatives of T4 and N4, considered to be safe in vivo. Chickens were intratracheally infected with APEC strain CH2 (serogroup O78), causing a mortality of about 50% during the seven days following the infection. The phage cocktail was administered 2h after the infection, via three different ways: intratracheally, intra-esophageally or via the drinking water. Treated groups did not show a significant decrease in mortality, lesion scores or weight loss compared to untreated groups, although the APEC-specific phages could be re-isolated from the lung and heart of chickens that were euthanized. Moreover, the re-isolated bacteria from infected chickens had remained sensitive to the phage cocktail. Our results indicate that the efficiency of the phage cocktail used in treating CH2-infected chickens in vivo is negligible, even though in vitro, the phages in the cocktail were able to efficiently lyse the APEC strain CH2. Our results emphasize that the 'traditional' pathway of isolation, followed by phenotypical and genotypical characterization of phages composing the cocktail, does not lead to success in phage therapy in all cases., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Infectious bursal disease virus (IBDV) causes a highly contagious and immunosuppressive disease in young chickens and results in considerable economic losses for the poultry industry. To suppress the replication of IBDV, two short hairpin RNAs (shRNAs) were designed for targeting the VP1 and VP2 genes of IBDV. Recombinant plasmids carrying each shRNA or two shRNAs were constructed based on vector pSilencer2.1-U6 in which the human U6 promoter was replaced with chicken U6 promoter. In chicken embryo fibroblasts, transfection with these shRNA plasmids 24h before infection with IBDV B87 reduced 50% tissue culture infectious doses (TCID(50)) from 10(8.75) TCID(50)/0.1 mL to 10(3.75)-10(1.0) TCID(50)/0.1 mL. In 10-day old specific pathogen-free (SPF) chicken embryos, incubation with a mixture of IBDV B87 and a shRNA plasmid via the allantoic cavity resulted in 100% mortality and high IBDV virus titer in the control group but 25-0% mortality and near normal embryo development in the specific shRNA groups; additionally, IBDV VP1 and VP2 mRNA levels were reduced by 72-95% in the shRNA groups as compared with the control groups. When challenged with a virulent strain IBDV GX8/99, 14-day-old chickens pre-treated with the single shRNA plasmids or the dual shRNA plasmid showed approximately 70% or 90% survival at 5 days post-challenge while those pre-treated with control plasmid or saline had less than 5% survival. The current study suggests that two IBDV shRNAs expressed by a plasmid under chicken U6 promoter could effectively and synergistically reduce IBDV replication in vitro and in vivo., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Infections caused by avian pathogenic Escherichia coli (APEC) cause important economic losses to poultry industry. The studies presented herein, aimed at investigating the in vivo performance of a cocktail of three phages in treating severe respiratory E. coli infections in experimentally contaminated birds and naturally infected flocks. Three lytic coliphages, phi F78E (Myoviridae), phi F258E (Siphoviridae) and phi F61E (Myoviridae) were combined in a 5.0 × 10(7)PFU/ml cocktail to be used in naturally APEC infected flocks (refractive to antibiotherapy). Experimentally infected birds were treated with phi F78E at two different titres (10(7)PFU/ml and 10(9)PFU/ml). Phage administration was performed orally and by spray, in a single application. The morbidity, mortality and pathology scores were compared with control birds not receiving phage therapy. The results revealed that the success of phage therapy in experimental rooms was dosage dependent, being 10(7)PFU/ml not enough to treat the infected chickens whereas a concentration of 10(9)PFU/ml of phi F78E allowed a decrease of 25% and 43% in chickens' mortality and morbidity, respectively. In the large scale experiments, the results obtained showed a remarkable efficacy of the low titre phage cocktail (10(7)PFU/ml) in decreasing the flocks' mortality to levels below 0.5% in no more than 3 weeks, with no recidivism. Based on the results we can conclude that phage treatment is a valuable alternative to control APEC infections in poultry., (Copyright © 2010 Elsevier B.V. All rights reserved.)

The effect of phage therapy in the control of Campylobacter jejuni colonization in young broilers, either as a preventive or a therapeutic measure, was tested. A prevention group was infected with C. jejuni at day 4 of a 10-day phage treatment. A therapeutic group was phage treated for 6 days, starting 5 days after C. jejuni colonization of the broilers had been established. Treatment was monitored by enumerating Campylobacter colony forming units (CFU) and phage plaque forming units (PFU) from caecal content. Counts were compared with control birds not receiving phage therapy. A clear 3 log decline in C. jejuni counts was initially observed in the therapeutic group, however, after 5 days bacterial counts stabilized at a level 1 log lower than that of the control group. Colonization of C. jejuni in the prevention group was delayed by the treatment and after an initial 2 log reduction, colonization stabilized within a week at levels comparable to the therapeutic group. The CFU and PFU counts displayed opposing highs and lows over time, indicative of alternating shifts in amplification of bacteria and phages. There were no adverse health effects from the phage treatment. Two different phages were combined as therapeutic treatment of Campylobacter positive chickens challenged at the age approaching broiler harvest. This again resulted in a significant decrease in Campylobacter colonization. We conclude that phage treatment is a promising alternative for reducing C. jejuni colonization in broilers.