69 research outputs found
Experimental infection of chickens by a flagellated motile strain of Salmonella enterica serovar Gallinarum biovar Gallinarum
Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (SG) causes fowl typhoid (FT), a septicaemic disease which can result in high mortality in poultry flocks. The absence of flagella in SG is thought to favour systemic invasion, since bacterial recognition via Toll-like receptor (TLR)-5 does not take place during the early stages of FT. In the present study, chicks susceptible to FT were inoculated with a wild type SG (SG) or its flagellated motile derivative (SG Fla+). In experiment 1, mortality and clinical signs were assessed, whereas in experiment 2, gross pathology, histopathology, systemic invasion and immune responses were evaluated. SG Fla+ infection resulted in later development of clinical signs, lower mortality, lower bacterial numbers in the liver and spleen, and less severe pathological changes compared to SG. The CD8+ T lymphocyte population was higher in the livers of chicks infected with SG at 4 days post-inoculation (dpi). Chicks infected with SG had increased expression of interleukin (IL)-6 mRNA in the caecal tonsil at 1 dpi and increased expression of IL-18 mRNA in the spleen at 4 dpi. In contrast, the CD4+ T lymphocyte population was higher at 6 dpi in the livers of birds infected with SG Fla+. Therefore, flagella appeared to modulate the chicken immune response towards a CD4+ T profile, resulting in more efficient bacterial clearance from systemic sites and milder infection
The influence of the mode of administration in the dissemination of three coliphages in chickens
Escherichia coli can cause severe respiratory and systemic infections in chickens, and it is often associated with significant economic losses in the poultry industry. Bacteriophages (phages) have been shown to be potential alternatives to the antibiotics in the treatment of bacterial infections. To accomplish that, phage particles must be able to reach and remain active in the infected organs. The present work aims at evaluating the effect of the route of administration and the dosage in the dissemination of 3 coliphages in the chicken’s organs. In vivo trials were conducted by infecting chickens orally, spray, and i.m. with 106, 107, and 108 plaque-forming units/mL suspensions of 3 lytic phages: phi F78E (Myoviridae), phi F258E (Siphoviridae), and phi F61E (Myoviridae). Birds were killed 3, 10, and 24 h after challenge and the phage titer was measured in lungs and air sacs membranes, liver, duodenum, and spleen. When administered by spray, the 3 phages reached the respiratory tract within 3 h. Oral administration also allowed all phages to be recovered in lungs, but only phi F78E was recovered from the duodenum, the liver, and the spleen. These differences can be explained by the possible replication of phi F78E in commensal E. coli strains present in the chicken gut, thus leading to a higher concentration of this phage in the intestines that resulted in systemic circulation of phage with consequent phage in organs. When phages were administered i.m., they were found in all of the collected organs. Despite this better response, i.m. administration is a nonpracticable way of protecting a large number of birds in a poultry unit. In general, the results suggest that oral administration and spray allowed phages to reach and to remain active in the respiratory tract and can, therefore, be considered promising administration routes to treat respiratory E. coli infections in the poultry industry.Fundação para a Ciência e a Tecnologia (FCT) - SFRH/BDE/15508/200
Safety and efficacy of a salmonella gallinarum ΔcobSΔcbiA strain with potential to prevent chicken infections by salmonella gallinarum and salmonella enteritidis
Recovery of Salmonella Gallinarum in the Organs of Experimentally-Inoculated Japanese Quails (Coturnix coturnix)
Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine
Orally administered phages to control zoonotic pathogens face important challenges, mainly related to the hostile conditions found in the gastrointestinal tract (GIT). These include temperature, salinity and primarily pH, which is exceptionally low in certain compartments. Phage survival under these conditions can be jeopardized and undermine treatment. Strategies like encapsulation have been attempted with relative success, but are typically complex and require several optimization steps. Here we report a simple and efficient alternative, consisting in the genetic engineering of phages to display lipids on their surfaces. Escherichia coli phage T7 was used as a model and the E. coli PhoE signal peptide was genetically fused to its major capsid protein (10A), enabling phospholipid attachment to the phage capsid. The presence of phospholipids on the mutant phages was confirmed by High Performance Thin Layer Chromatography, Dynamic Light Scattering and phospholipase assays. The stability of phages was analysed in simulated GIT conditions, demonstrating improved stability of the mutant phages with survival rates 102107 pfu.mL1 higher than wild-type phages. Our work demonstrates that phage engineering can be a good strategy to improve phage tolerance to GIT conditions, having promising application for oral administration in veterinary medicine.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and under the scope of the Project PTDC/BBB-BSS/6471/2014 (POCI-01-0145-FEDER-016678). Franklin L. Nobrega and Ana Rita Costa acknowledge FCT for grants SFRH/BD/86462/2012 and SFRH/BPD/94648/2013, respectively. Melvin F. Siliakus acknowledges funding from the Biobased Ecologically Balanced Sustainable Industrial Chemistry (BE-BASIC) foundation. Electron microscopy work was performed at the Wageningen Electron Microscopy Centre (WEMC) of Wageningen University
Atividade de desinfetantes frente a sorotipos de Salmonella isolados de granjas avÃcolas
Orally Administered P22 Phage Tailspike Protein Reduces Salmonella Colonization in Chickens: Prospects of a Novel Therapy against Bacterial Infections
One of the major causes of morbidity and mortality in man and economically important animals is bacterial infections of the gastrointestinal (GI) tract. The emergence of difficult-to-treat infections, primarily caused by antibiotic resistant bacteria, demands for alternatives to antibiotic therapy. Currently, one of the emerging therapeutic alternatives is the use of lytic bacteriophages. In an effort to exploit the target specificity and therapeutic potential of bacteriophages, we examined the utility of bacteriophage tailspike proteins (Tsps). Among the best-characterized Tsps is that from the Podoviridae P22 bacteriophage, which recognizes the lipopolysaccharides of Salmonella enterica serovar Typhimurium. In this study, we utilized a truncated, functionally equivalent version of the P22 tailspike protein, P22sTsp, as a prototype to demonstrate the therapeutic potential of Tsps in the GI tract of chickens. Bacterial agglutination assays showed that P22sTsp was capable of agglutinating S. Typhimurium at levels similar to antibodies and incubating the Tsp with chicken GI fluids showed no proteolytic activity against the Tsp. Testing P22sTsp against the three major GI proteases showed that P22sTsp was resistant to trypsin and partially to chymotrypsin, but sensitive to pepsin. However, in formulated form for oral administration, P22sTsp was resistant to all three proteases. When administered orally to chickens, P22sTsp significantly reduced Salmonella colonization in the gut and its further penetration into internal organs. In in vitro assays, P22sTsp effectively retarded Salmonella motility, a factor implicated in bacterial colonization and invasion, suggesting that the in vivo decolonization ability of P22sTsp may, at least in part, be due to its ability to interfere with motility… Our findings show promise in terms of opening novel Tsp-based oral therapeutic approaches against bacterial infections in production animals and potentially in humans
Antimicrobial susceptibility of Salmonella Gallinarum and Salmonella Pullorum isolated from ill poultry in Brazil
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