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Aims Characterize global genomic features of 86 genomes of Salmonella Gallinarum (SG) and Pullorum (SP), which are important pathogens causing systemic infections in poultry. Methods and results All genomes harbored efflux pump encoding gene mdsA and gold tolerance genes golS and golT. Aminoglycoside (aac(6′)-Ib, aadA5, aph(6)-Id, aph(3′')-Ib, ant(2′')-Ia), beta-lactam (bla TEM-1, bla TEM-135), efflux pump (mdsB), fosfomycin (fosA3), sulfonamide (sul1, sul2), tetracycline [ tet(A) ], trimethoprim (dfrA17), acid (asr), and disinfectant (qacEdelta1) resistance genes, gyrA, gyrB , and parC quinolone resistance point mutations, and mercury tolerance genes (mer) were found in different frequencies. Additionally, 310 virulence genes, pathogenicity islands (including SPI-1, 2, 3, 4, 5, 6, 9, 10, 12, 13, and 14), plasmids [IncFII(S), ColpVC, IncX1, IncN, IncX2, and IncC], and prophages (Fels-2, ST104, 500465-1, pro483, Gifsy-2, 103 203_sal5, Fels-1, RE-2010, vB_SenS-Ent2, and L-413C) were detected. MLST showed biovar-specific sequence types, and core genome MLST showed country-specific and global-related clusters. Conclusion SG and SP global strains carry many virulence factors and important antimicrobial resistance genes. The diverse plasmids and prophages suggest genetic variability. MLST and cgMLST differentiated biovars and showed profiles occurring locally or worldwide. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Gallinarum (S. gallinarum) is an important host-specific pathogen that causes fowl typhoid, a severe systemic, septicemic, and fatal infection, in chickens. S. gallinarum causes high morbidity and mortality in chickens and poses a significant burden and economic losses to the poultry industry in many developing countries. However, the virulence factors and mechanisms of S. gallinarum-induced systemic infection in chickens remain poorly understood. In this study, we constructed a Salmonella pathogenicity island-14 (SPI-14) mutant strain (mSPI-14) of S. gallinarum and evaluated the pathogenicity of mSPI-14 in the chicken systemic infection model. The mSPI-14 exhibited the same level of bacterial growth and morphological characteristics but significantly reduced resistance to bile acids compared with the wild-type (WT) strain in vitro. The virulence of mSPI-14 was significantly attenuated in the chicken oral infection model in vivo. Chickens infected with WT showed typical clinical symptoms of fowl typhoid, with all birds succumbing to the infection within 6 to 9 days post-inoculation, and substantial increases in bacterial counts and significant pathological changes in the liver and spleen were observed. In contrast, all mSPI-14-infected chickens survived, the bacterial counts in the organs were significantly lower, and no significant pathological changes were observed in the liver and spleen. The expression of interleukin (IL)-1β, IL-12, CXCLi1, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in the liver of mSPI-14-infected chickens were significantly lower than those in the WT-infected chickens. These results indicate that SPI-14 is a crucial virulence factor in systemic infection of chickens, and avirulent mSPI-14 could be used to develop a new attenuated live vaccine to prevent S. gallinarum infection in chickens. [ABSTRACT FROM AUTHOR]

Fowl typhoid has revolved into a pathogenic hazard to the poultry industry and extensive use of antibiotic is leading to the induction of antimicrobial resistance in microorganism. Main aim of this study was to find out substitute of antibiotic against Fowl Typhoid in broiler in terms of immunological, serum biochemistry and lipid profile parameters. Broiler chicks (Age = 1 d and n = 90) were kept under uniform management conditions. Chicks were divided randomly into six groups on day ten of their age with 15 replicates in each; control negative (CN), control positive (CP) and four treatments (T1, T2, T3 and T4). Wheras challenge infection of Salmonella gallinarum was given on nineteen days of age to all experimental birds except of those in CN group. On 22nd day, infected birds in T1 were given Florfenicol antibiotic. Whereas infected birds in groups; T2, T3 and T4 were given concentration of nanoparticles zinc oxide and copper oxide at different rate; 25+10, 37.5+15 and 50+20 mg/kg/d, respectively. Collected data were analyzed using complete randomized design through ANOVA technique. Mortality percent was found minimum 13.3% in birds under T3 group. Effect of all nanoparticle levels was not different (P>0.05) to that of antibiotic in terms of total serum protein and lipid profile; total glycerides, very low-density lipids, high density lipids and total cholesterol. It was concluded on the basis of findings of current study that nanoparticles zinc oxide and copper oxide mixture (37.5 + 15 mg/Kg/d) was found optimum alternate to Florfenicol antibiotic against Salmonella gallinarum infection in broiler birds. Hence, Zinc oxide and copper oxide nanoparticles could be an adequate alternative treatment replacing antibiotics against Fowl Typhoid in broilers. [ABSTRACT FROM AUTHOR]

One of the leading diseases affecting laying poultry in Argentina is fowl typhoid, caused by the pathogenSalmonella entericaserovar Gallinarum (S.Gallinarum). Although this disease was eradicated fromindustrial hatcheries in several developed countries, it is still present in commercial farms in LatinAmerica, causing substantial economic losses due to high mortality rates, limited production of qualityeggs, high veterinary costs, and the need to renovate infected facilities. To develop a newS.Gallinarumbiocontrol method, in the present study, we characterized a bacteriophage in terms of host specificity,growth kinetics, and stability under different environmental conditions. The phage could infect anample range of bacterial hosts and had no lytic activity against the vaccine strain 9R. Moreover, thephage was stable at pH 3, chloroform presence did not significantly reduce its titer, and its activity wasnot affected by antibiotics or various chemical agents often used in aviculture. Application of a phagesuspension onS.Gallinarum-contaminated surfaces resulted in a significant reduction of bacterial load(p < 0.05). These results show that lytic bacteriophages such as the one studied here are innovativealternatives with potential applications to controlS.Gallinarum in laying poultry farms.

The effects of Salmonella gallinarum infection on the haematology and oxidative stress (OS) in pullets were evaluated in this study. Fifty 7-week-old pullets were randomly assigned to two groups of 25 pullets per group. The infected pullets were inoculated orally with S. gallinarum (109 S. gallinarum colony forming units/mL), while uninfected pullets were the controls. Haematological and OS parameters were determined following standard protocols. There was a significant loss (p<0.05) of body weight. The packed cell volume (PCV), haemoglobin (Hb) concentration, red blood cell (RBC) count and mean corpuscular haemoglobin concentration (MCHC) were significantly lower (p<0.05). In contrast, the mean corpuscular volume (MCV) was significantly higher (p<0.05) in the infected pullets when compared to the controls. The total white blood cell (TWBC), absolute heterophil, lymphocyte, eosinophil and monocyte counts, of the infected pullets, were significantly higher than that of the controls. Serum malondialdehyde (MDA) level, glutathione peroxidase (GPx) and catalase (CAT) activities of the infected pullets were significantly higher (p<0.05) when compared to the uninfected controls. Absolute heterophil count correlated strongly, positively and significantly with MDA. An association was established between significantly elevated GPx and CAT, which have antioxidant properties, and survival/health improvement indices such as improved weight gain and self-recovery. It was concluded that S. gallinarum infection of pullets caused significant alterations in the haematology, induced OS, and stimulated the body's antioxidant defence mechanism to elaborate GPx and CAT. This may suggest the use of antioxidants in the treatment of fowl typhoid. [ABSTRACT FROM AUTHOR]

ABSTRACT: An effective vaccine strategy is indispensable against infectious bronchitis virus (IBV) and fowl typhoid (FT), both of which threaten the poultry industry. This study demonstrates a vector system, pJHL270, designed to express antigens in prokaryotic and eukaryotic cells. The vector system stimulates immune responses via synchronized antigen presentation to MHC class-I and -II molecules to produce balanced Th1/Th2 responses. The vaccine antigens were crafted by selecting the consensus sequence of the N-terminal domain of the spike protein (S1-NTD) and a conserved immunogenic region of the nucleocapsid protein (N321-406 aa) from IBV strains circulating in South Korea. The vaccine antigen was cloned and transformed into a live-attenuated Salmonella Gallinarum (SG) strain, JOL2854 (∆lon, ∆cpxR, ∆rfaL, ∆pagL, ∆asd). Western blot analysis confirmed concurrent antigen expression in Salmonella and eukaryotic cells. Oral immunization with the SG-based IBV vaccine construct JOL2918 induced IBV antigen and Salmonella-specific humoral and cell-mediated immune responses in chickens. PBMCs collected from immunized chickens revealed that MHC class-I and -II expression had increased 3.3-fold and 2.5-fold, respectively, confirming MHC activation via bilateral antigen expression and presentation. Immunization induced neutralizing antibodies (NAbs) and reduced the viral load by 2-fold and 2.5-fold in the trachea and lungs, respectively. The immunized chickens exhibited multifaceted humoral, mucosal, and cell-mediated responses via parallel MHC class-I and -II activation as proof of a balanced Th1/Th2 immune response. The level of NAbs, viral load, and gross and histological analyses provide clear evidence that the construct provides protection against IBV and FT.

Fowl typhoid is a septicemic disease caused by Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum. It is a host-specific disease primarily affecting chickens and turkeys, although it has been reported in various animal species and sporadically in humans. Here, we present a case of a fowl typhoid outbreak on a turkey poult farm where the source of infection was the hatchery. The birds started showing symptoms of growth retardation at 21 days of age, after which the mortality rates gradually started to increase. Post mortem examination revealed that the main lesions were granulomatous proliferations in the small intestines. The results of the histopathological examination indicate that the severity of the infection was alleviated by the application of phytogenic mixtures and probiotics as a supportive treatment, even though the affected flock was eventually culled at 60 days of age. The farmer was advised to apply more strict biosecurity measures to prevent the spread of the disease on the farm and try to eradicate the pathogen from the barn. Since the outbreak, there have been no recurrent infections. [ABSTRACT FROM AUTHOR]

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Simple Summary: Fowl typhoid is an infectious bacterial disease of poultry that causes significant economic losses in the poultry sector. Antimicrobial therapy, biosecurity practices and vaccination programs are used to prevent and treat fowl typhoid. Misuse of antibiotics generally results in emergence of multidrug resistance in Salmonella Gallinarum. Although vaccination reduces flock losses, disease is still prevailing in many developing countries including Pakistan. To develop an alternative to these approaches, the present study was designed to explore Lactobacillus spp. of poultry origin which can inhibit Salmonella Gallinarum in vitro. Twenty one lactobacilli isolated from poultry have antibacterial activity against Salmonella Gallinarum. These isolates were further selected for the characterization of in vitro probiotic properties. Out of 21, two Limosilactobacillus fermentum strains isolated in this study have the ability to survive physicochemical barriers of the gut such as tolerance to low pH, bile salts, auto-aggregation and co-aggregation activity with Salmonella Gallinarum, and caused significant reduction in growth of Salmonella Gallinarum. These isolates may be used for the development of anti-Salmonella Gallinarum probiotics after in vivo study in broilers. Fowl typhoid, a septicaemic disease of poultry, is caused by Salmonella Gallinarum and leads to severe economic losses. The aim of the present study was to isolate, select and characterize indigenous probiotic lactobacilli with anti-Salmonella Gallinarum activity. A total 55 lactobacilli were isolated from the caeca and ileum parts of healthy chickens and identified to species level by 16S rDNA sequencing. All the isolates were initially screened for antimicrobial activity and selected isolates were further subjected to in vitro evaluation of probiotic properties. Lactobacilli isolates (n = 21) showed varying levels of activity (08–18 mm) against Salmonella Gallinarum. These selected isolates also showed tolerance to acidic conditions (pH 3 and 4). Out of these 21 isolates, 13 showed growth (>0.5 OD at 600 nm) 0.3% bile salts. Moreover, these isolates also had the ability of auto-aggregation (20.05 ± 0.62%–50.70 ± 1.40%), and co-aggregation with Salmonella Gallinarum (5.22 ± 0.21%–42.07 ± 0.70%). Results revealed that lactobacilli had a higher level of resistance to vancomycin (100%), streptomycin (100%), ciprofloxacin (95%), gentamicin (90%), doxycycline (90%), oxytetracycline (85%), and bacitracin (80%), and a lower level of resistance to penicillin (33%), erythromycin (28%), chloramphenicol (23%), fusidic acid (23%) and amoxicillin (4%). The Limosilactobacillus fermentum PC-10 and PC-76 were sensitive to most of the antibiotics. The overall results revealed that two Limosilactobacillus fermentum strains (PC-10 and PC-76) fulfill the in vitro selection criteria of probiotics, i.e, tolerance to low pH, resistance to bile salts, auto-aggregation, co-aggregation with Salmonella Gallinarum, and absence of acquired antibiotic resistance. The Limosilactobacillus fermentum PC-10 and PC-76 also inhibited the (>5 log10) growth of Salmonella Gallinarum in co-culture assay. It is concluded that Limosilactobacillus fermentum PC-10 and PC-76 may be further investigated and developed as anti-Salmonella Gallinarum probiotics for poultry. [ABSTRACT FROM AUTHOR]

Fowl typhoid (FT) is an economically significant bacterial disease of layers leading to a drastic drop in egg production. Due to increased public health concerns about antibiotics in poultry feed, a search for new safe antimicrobials for treating fowl typhoid is crucial. The antimicrobial effect of cinnamaldehyde essential oil (CnEO) against fowl typhoid in layers was investigated in this experiment. The 60-week-old BV300-layer birds (n = 100) were divided into five groups: the non-challenged control group A, only cinnamaldehyde-treated group B (CnEO @ 1:8000 dilutions through drinking water for 60 days), the challenged group C, challenged plus cinnamaldehyde therapy group D (CnEO @ 1:8000 dilutions through drinking water from 16 to 30 dpi), and challenged plus antibiotic therapy group E (chloramphenicol @ 1 gm/5lit through drinking water from 16 to 30 dpi). Hens from all challenged groups were challenged with Salmonella Gallinarum (VTCCBAA588) @ 1 × 108 CFU/ml orally. Various parameters such as clinical signs, mortality, egg production and egg weight, colony-forming unit (CFU) count of cecal content, eggshell surface, and egg yolk were evaluated all through 60 days of an experimental trial. Results indicated that, in the case of the cinnamaldehyde therapeutic group, there was a significant improvement in egg production, mild clinical signs, lower feed conversion ratio (FCR), and a significantly lower bacterial count in ceca and on the eggshell surface compared to the control challenge group. Thus, CnEO @ 1:8000 dilutions through drinking water can be a potential antimicrobial for controlling fowl typhoid. [ABSTRACT FROM AUTHOR]

IntroductionSalmonella enterica subspecies enterica serovar Gallinarum biovar Gallinarum (SG) is associated with fowl typhoid fever, and the attenuated rough strain SG9R is widely used as a vaccine in many regions. Reversion to virulence of vaccine strains was suspected as the cause during recent fowl typhoid fever outbreaks in poultry in South Africa and Eswatini.MethodsTo compare nine field isolates with global wild-type SG9 strains and the two commercial SG9R vaccines in use, Nobilis® SG9R and Cevac®-SG, we used whole-genome comparison with single-nucleotide polymorphism (SNP) detection.ResultsSNP phylogenic analysis showed that all the southern African field isolates were more closely related to the vaccine strains than wild-type SG9 strains. Furthermore, SNPs in the pyruvate dehydrogenase (aceE) and/or lipopolysaccharide 1,2-glucosyltransferase (rfaJ) genes, which are known markers of attenuation, were found in four of the field isolates along with intact spv, SPI-1, and SPI-2 gene clusters, providing conclusive evidence that these four isolates were originally vaccine strains that reverted to virulence. Five other field isolates lacked the SG9R attenuation markers, but variant analysis identified an SNP in the yihX gene, insertions in the ybjX and hydH genes, and deletions in the ftsK and sadA genes that were shared between the field isolates and vaccine strains but absent in wild-type SG9, indicating that these field isolates were also likely revertant vaccines.DiscussionOverall, this study highlights different mechanisms of reversion of two commercial vaccines, where virulence caused by field isolates closely related to the Nobilis® SG9R vaccine was associated with the restoration of intact virulence gene clusters, and those derived from the Cevac®-SG vaccine were characterized by point mutations resulting in restored aceE and rfaJ genes. A possible new marker of attenuation was identified as a point mutation in the yihX gene, as well as four new candidate genes that could potentially be used to distinguish current vaccine strains from wild-type strains using PCR assays.

Fowl typhoid is a septicemic disease caused by Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum. It is a host-specific disease primarily affecting chickens and turkeys, although it has been reported in various animal species and sporadically in humans. Here, we present a case of a fowl typhoid outbreak on a turkey poult farm where the source of infection was the hatchery. The birds started showing symptoms of growth retardation at 21 days of age, after which the mortality rates gradually started to increase. Post mortem examination revealed that the main lesions were granulomatous proliferations in the small intestines. The results of the histopathological examination indicate that the severity of the infection was alleviated by the application of phytogenic mixtures and probiotics as a supportive treatment, even though the affected flock was eventually culled at 60 days of age. The farmer was advised to apply more strict biosecurity measures to prevent the spread of the disease on the farm and try to eradicate the pathogen from the barn. Since the outbreak, there have been no recurrent infections.

Salmonella Gallinarum (SG) is a host-restricted enterobacteria and the causative agent of fowl typhoid in poultry. Here, we report the complete genomes of two strains belonging to this serotype. SA68 is a field strain isolated from the livers of dead hen carcasses of a commercial layer farm presenting high mortality located in São Paulo city, Brazil, in 1990. Strain 9R corresponds to a live attenuated SG commercial vaccine. DNA was extracted from pure cultures and subjected to whole genome sequencing (WGS) using the Ion Torrent PGM System. The assemblies reached lengths of 4,657,435 (SA68) and 4,657,471 (9R) base pairs. Complete genomes were deposited in GenBank under the accession numbers CP110192 (SA68) and CP110508 (9R). Both genomes were analyzed and compared in terms of molecular typing, antibiotic resistance genes, virulence genes, Salmonella pathogenic islands (SPIs), insertion sequences and prophages. The data obtained show many similarities in the genetic content, with the exception of the SPI-12 and CS54 pathogenic islands, which are exclusive to the field strain. The information generated will help to understand the virulence differences of field and vaccinal SG strains and can be used to perform evolutionary and epidemiologic studies.

ABSTRACT: Fowl typhoid is a severe disease caused by Salmonella Gallinarum with considerable mortality and morbidity in laying hen farms. The current study has focused on controlling the infection in laying hens using anti-Salmonella spp. bacteriophage. The treatments included, PC, without challenge; NC, S. Gallinarum challenged (SGC); B5, 5 mg bacteriophage/kg + SGC; B10, 10 mg bacteriophage/kg + SGC. The Salmonella shedding, inflammatory responses, and gene expression of pro-inflammatory cytokines, toll-like receptor (TLR), and heat shock protein (HSP) in the jejunum, liver, and thigh muscle were tested in laying hens. Supplementation of bacteriophage reduced the abundance of S. Gallinarum in the excreta at d 3, 7, and 14. The abundance of S. Gallinarum was lower in the B10 than the B5 at d 7. Supplementation of bacteriophage decreased the abundance of S. Gallinarum in the oviduct, spleen, and cecum at d 14. The laying hens in the NC group showed an increased relative spleen weight compared with the PC and B10 treatments. Among the SGC treatments, the NC treatment showed higher gene expressions of IL-4 compared with the B5, higher gene expressions of interferon (IFNγ), TLR4, and tumor necrosis factor-α (TNF-α) compared with the B5 and B10, and higher gene expressions of HSP27 compared with the B10 in the jejunum. Dietary supplementation of B10 decreased the mRNA expressions of TLR4 and TNF-α compared with the B5 treatment in the jejunum. The NC treatment showed the highest gene expressions of HSP27, TLR4, and TNF-α in the liver. Dietary supplementation of B10 showed lower mRNA expressions of HSP27 compared with the B5 treatment in the liver. Moreover, the IFNγ and HSP27 were upregulated in the NC treatment compared with the B5 and B10 in the muscle. In conclusion, it can be suggested that bacteriophage is an effective supplement to control S. Gallinarum infection in laying hens and possibly lower horizontal contaminations in laying hen flocks.

Salmonellosis is a bacterial zoonosis causing an array of health conditions. Non-typhoidal salmonellosis (NTS) has a discrete adaptation to certain animals; in poultry, pullorum and fowl typhoid are its primary disease manifestations. The diseases are prevalent in Nigerian poultry and have been well-studied in Nigeria, but less so in North Central Nigeria (NCN). Using field sampling, laboratory methods and a semi-structured questionnaire for 1000 poultry farms in NCN, we explored the incidence and risk factors for the persistence of NTS infection in poultry. Approximately 41.6% of the farms had experienced NTS over the last 18 months. Farm experience of NTS moderately predicted awareness of salmonellosis. Increasing stock in smallholder farms, self-mixing of concentrate on the farm, usage of stream water, pen odour, non-adherence and partial adherence of farms to recommended poultry vaccination against pullorum and fowl typhoid and lack of and non-adherence to biosecurity were identified risk factors that increased the odds of NTS infection in poultry. Antibiotic use practice may have reduced the isolation rate of NTS, yet NTS continues to challenge poultry farms in Nigeria. Identified risk practices must be mitigated intentionally and biosecurity and hygiene must be improved to reduce the burden of NTS. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Gallinarum (S. Gallinarum) is a host-specific pathogen causing fowl typhoid, a severe systemic infection in poultry, which leads to substantial economic losses due to high morbidity and mortality in many developing countries. However, less is known about the pathogenic characteristics and mechanism of S. Gallinarum-induced systemic infection in chickens. In this study, we deleted the S. Gallinarum UDP- N -acetylglucosamine-1-phosphate transferase gene, which contributes to the biosynthesis of enterobacterial common antigen (ECA), and studied the pathogenicity of this wecB ::Cm strain in a chicken model of systemic infection. The wecB ::Cm mutant strain showed comparable growth but lower resistance to bile acid and nalidixic acid than the wild-type strain in vitro. In the oral infection model of chickens, the virulence of the wecB ::Cm strain was significantly attenuated in vivo. Chickens infected with wild-type strain showed typical clinical signs and pathological changes of fowl typhoid and died between 6 and 9 days post-infection, and the bacteria rapidly disseminated to systemic organs and increased in the livers and spleens. In contrast, the wecB ::Cm mutant strain did not cause chicken death, there were no significant clinical changes, and the bacterial numbers in the liver and spleen of the chickens were significantly lower than those of the chickens infected with the wild-type strain. In addition, the expression of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and CXCLi1 in the livers of wecB ::Cm-infected chickens was significantly lower than that of the chickens infected with the wild-type strain. Furthermore, the attenuated wecB ::Cm strain could persistently colonize the liver and spleen at low levels for up to 25 days post-infection and could induce a protective immune response in the chickens. These results indicate that the wecB gene is an important virulence factor of S. Gallinarum in the chicken model of systemic infection, and the avirulent wecB ::Cm mutant could possibly be used as a live-attenuated vaccine strain for controlling fowl typhoid. [ABSTRACT FROM AUTHOR]

Salmonella Gallinarum causes fowl typhoid in poultry leading to a huge economic loss to the poultry industry. The large virulence plasmid of S. gallinarum has been associated with various systemic infections in poultry. A five-gene spanning region (spvRABCD) of 7.8 kb on the large plasmid mainly confers virulence to the bacteria. However, the exact role of these genes in virulence has not been elucidated yet. SpvB exhibits delayed cell death by preventing actin polymerization followed by apoptosis during intracellular infection. The specific role of SpvB in causing the disease is not known yet. In the current study, the SpvB gene was deleted through CRISPR/Cas9 method from a large virulent plasmid of locally isolated S. gallinarum strain (SG18). The homology-directed repair method was used for complete deletion of SpvB gene using the modified pCas9 plasmid. The SpvB-deleted S. gallinarum strain (ΔSpvB_SG18), when tested for its virulence in broiler chicken showed no diseases signs and mortality. In addition, the avirulent strain does not affect the bird’s weight and was rapidly cleared from the liver after infection. However, it cleared from the intestine only after 4–5 days, which suggests that the ΔSpvB_SG18 strain is unable to invade from the intestine to the liver. This is the first study to report a complete gene deletion from the S. gallinarum virulent plasmid and its effect. This method will be useful for the deletion of virulent genes from S. gallinarum, to study their role in pathogenesis, and to prepare an effective vaccine strain for controlling fowl typhoid in poultry.

Salmonella enterica serovar Gallinarum (S. Gallinarum) is a host-specific pathogen causing fowl typhoid, a severe systemic infection in poultry, which leads to substantial economic losses due to high morbidity and mortality in many developing countries. However, less is known about the pathogenic characteristics and mechanism of S. Gallinarum-induced systemic infection in chickens. In this study, we deleted the S. Gallinarum UDP-N-acetylglucosamine-1-phosphate transferase gene, which contributes to the biosynthesis of enterobacterial common antigen (ECA), and studied the pathogenicity of this wecB::Cm strain in a chicken model of systemic infection. The wecB::Cm mutant strain showed comparable growth but lower resistance to bile acid and nalidixic acid than the wild-type strain in vitro. In the oral infection model of chickens, the virulence of the wecB::Cm strain was significantly attenuated in vivo. Chickens infected with wild-type strain showed typical clinical signs and pathological changes of fowl typhoid and died between 6 and 9 days post-infection, and the bacteria rapidly disseminated to systemic organs and increased in the livers and spleens. In contrast, the wecB::Cm mutant strain did not cause chicken death, there were no significant clinical changes, and the bacterial numbers in the liver and spleen of the chickens were significantly lower than those of the chickens infected with the wild-type strain. In addition, the expression of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and CXCLi1 in the livers of wecB::Cm-infected chickens was significantly lower than that of the chickens infected with the wild-type strain. Furthermore, the attenuated wecB::Cm strain could persistently colonize the liver and spleen at low levels for up to 25 days post-infection and could induce a protective immune response in the chickens. These results indicate that the wecB gene is an important virulence factor of S. Gallinarum in the chicken model of systemic infection, and the avirulent wecB::Cm mutant could possibly be used as a live-attenuated vaccine strain for controlling fowl typhoid.

An effective vaccine strategy is indispensable against infectious bronchitis virus (IBV) and fowl typhoid (FT), both of which threaten the poultry industry. This study demonstrates a vector system, pJHL270, designed to express antigens in prokaryotic and eukaryotic cells. The vector system stimulates immune responses via synchronized antigen presentation to MHC class-I and -II molecules to produce balanced Th1/Th2 responses. The vaccine antigens were crafted by selecting the consensus sequence of the N-terminal domain of the spike protein (S1-NTD) and a conserved immunogenic region of the nucleocapsid protein (N 321 - 406 aa) from IBV strains circulating in South Korea. The vaccine antigen was cloned and transformed into a live-attenuated Salmonella Gallinarum (SG) strain, JOL2854 (∆ lon , ∆ cpxR , ∆r faL , ∆ pagL , ∆ asd). Western blot analysis confirmed concurrent antigen expression in Salmonella and eukaryotic cells. Oral immunization with the SG-based IBV vaccine construct JOL2918 induced IBV antigen and Salmonella -specific humoral and cell-mediated immune responses in chickens. PBMCs collected from immunized chickens revealed that MHC class-I and -II expression had increased 3.3-fold and 2.5-fold, respectively, confirming MHC activation via bilateral antigen expression and presentation. Immunization induced neutralizing antibodies (NAbs) and reduced the viral load by 2-fold and 2.5-fold in the trachea and lungs, respectively. The immunized chickens exhibited multifaceted humoral, mucosal, and cell-mediated responses via parallel MHC class-I and -II activation as proof of a balanced Th1/Th2 immune response. The level of NAbs, viral load, and gross and histological analyses provide clear evidence that the construct provides protection against IBV and FT. [ABSTRACT FROM AUTHOR]

Fowl typhoid, a septicaemic disease of poultry, is caused by Salmonella Gallinarum and leads to severe economic losses. The aim of the present study was to isolate, select and characterize indigenous probiotic lactobacilli with anti-Salmonella Gallinarum activity. A total 55 lactobacilli were isolated from the caeca and ileum parts of healthy chickens and identified to species level by 16S rDNA sequencing. All the isolates were initially screened for antimicrobial activity and selected isolates were further subjected to in vitro evaluation of probiotic properties. Lactobacilli isolates (n = 21) showed varying levels of activity (08–18 mm) against Salmonella Gallinarum. These selected isolates also showed tolerance to acidic conditions (pH 3 and 4). Out of these 21 isolates, 13 showed growth (>0.5 OD at 600 nm) 0.3% bile salts. Moreover, these isolates also had the ability of auto-aggregation (20.05 ± 0.62%–50.70 ± 1.40%), and co-aggregation with Salmonella Gallinarum (5.22 ± 0.21%–42.07 ± 0.70%). Results revealed that lactobacilli had a higher level of resistance to vancomycin (100%), streptomycin (100%), ciprofloxacin (95%), gentamicin (90%), doxycycline (90%), oxytetracycline (85%), and bacitracin (80%), and a lower level of resistance to penicillin (33%), erythromycin (28%), chloramphenicol (23%), fusidic acid (23%) and amoxicillin (4%). The Limosilactobacillus fermentum PC-10 and PC-76 were sensitive to most of the antibiotics. The overall results revealed that two Limosilactobacillus fermentum strains (PC-10 and PC-76) fulfill the in vitro selection criteria of probiotics, i.e, tolerance to low pH, resistance to bile salts, auto-aggregation, co-aggregation with Salmonella Gallinarum, and absence of acquired antibiotic resistance. The Limosilactobacillus fermentum PC-10 and PC-76 also inhibited the (>5 log10) growth of Salmonella Gallinarum in co-culture assay. It is concluded that Limosilactobacillus fermentum PC-10 and PC-76 may be further investigated and developed as anti-Salmonella Gallinarum probiotics for poultry.

Although it has rapidly decreased since the early 2000s, fowl typhoid still occurs in commercial layer chickens, causing a significant economic loss in Korea. There is growing concern about the emergence of new pathogenic strains of the causative agent, Salmonella Gallinarum, which is able to overcome vaccine immunity. It has also been suspected that the poultry red mite, Dermanyssus gallinae, which is commonly found in layer chicken farms, may be an important cause of the recurrence of fowl typhoid in the farms. This study was conducted to examine changes in the virulence of recent isolates of S. Gallinarum obtained from layer farms and estimate the potential of the disease transmission of D. gallinae in the farms. Clinical and environmental samples and mites collected from layer farms affected by fowl typhoid between 2013 and 2018 were tested for S. Gallinarum. The isolates were characterized by genotypic analyses and in vitro virulence assays with chicken-derived cell lines. Vaccine protection against recent isolates was examined in the chickens. A total of 45 isolates of S. Gallinarum were collected and there was no evidence of changes in their virulence. It has also been demonstrated that the S. Gallinarum 9R vaccine strain widely used in Korea is still effective in controlling fowl typhoid if the susceptibility of birds to the disease is not increased by stress. Salmonella Gallinarum isolated from the outer and inner parts of D. gallinae, environmental dust, and dead birds of the same farm showed the same or closely related genotypes. Consequently, the present study indicated that the horizontal transmission and environmental persistence of S. Gallinarum and the increased disease susceptibility of chickens in layer farms could be mediated by D. gallinae, causing persistent outbreaks of fowl typhoid.

Abstract Background The poultry red mite Dermanyssus gallinae (De Geer, 1778) is a major ectoparasite of poultry. Infestations are found in most laying hen farms in Europe, and breeder flocks have also been reported to be affected. Mite infestation has detrimental effects on animal welfare, it causes significant economic losses, and, additionally, D. gallinae is often considered as a vector for pathogens. Despite suspicion of a close relationship between the poultry red mite and Salmonella enterica enterica serovar Gallinarum biovar Gallinarum (serovar Gallinarum), the causative agent of fowl typhoid disease (FT), there has been no definitive proof of mite-mediated transmission. Therefore, an investigation was conducted to determine if D. gallinae-mediated transmission of serovar Gallinarum could be demonstrated among four different hen groups. Methods Two groups of 8 hens (A and B) were experimentally infected with serovar Gallinarum in two isolators. After 7 days, when birds showed signs of FT, about 25,000 mites were introduced. After 3 days, mites were harvested and used to infest two other hen groups of 8 (C and D), in two separate isolators. The health status of hens was constantly monitored; detection and quantification of serovar Gallinarum were performed by PCR and qPCR from mites and organs of dead hens. The maximum likelihood estimation of the infection rate and mite vectorial capacity were calculated. Results Clinical disease was observed in groups infected with serovar Gallinarum (A and B) and in hens of groups C and D infested with mites harvested from the isolators containing groups A and B. In all four groups, serovar Gallinarum was detected from liver, spleen, ovary, and cecum of hens, thus confirming the diagnosis of FT. Mite analysis demonstrated the presence of the pathogen, with an estimated infection rate ranging between 13.72 and 55.21 infected per thousand mites. Vectorial capacity was estimated to be 73.79. Conclusions Mites harvested from birds infected with serovar Gallinarum were shown to carry the mite, and then to transfer serovar Gallinarum to isolated groups of pathogen-free birds that subsequently showed signs of FT. Mite vectorial capacity was high, demonstrating that D. gallinae should be considered an effective vector of FT.

SUMMARY: Salmonella is a bacterium usually associated with mild gastroenteritis in chickens, but strains of the serotype Gallinarum can result in 2 severe systemic chicken diseases: fowl typhoid (by biovar Gallinarum) and pullorum disease (by biovar Pullorum). These diseases can be controlled by immunization with a specific vaccine (SG 9R). The aim of this study was to characterize recent Salmonella Gallinarum isolates by molecular analyses, as well as detect field and vaccine strains in clinical and environmental samples from poultry flocks. Field isolates of S. Gallinarum (n = 18) and other serotypes (n = 117) were obtained from Brazilian poultry farms between 2012 and 2015. In addition, clinical and environmental samples of poultry flocks (vaccinated or not) with fowl typhoid were obtained in a 3-year period (2013 to 2015). The results demonstrated that specific PCR targeted on speC and glgC genes and PCR-restriction fragment length polymorphisms based on fliC and sefA genes detected all the 18 isolates, 10 identified as field isolates of biovar Gallinarum and 8 as biovar Pullorum. All isolates of other serotypes presented negative results. In addition, PCR tests were performed directly from the pre-enrichment broth of the bacteriological procedures. Vaccine SG 9R and field S. Gallinarum strains were detected in samples with (n = 19) and without (n = 59) vaccination. These results demonstrate the use of PCR tests for detection of S. Gallinarum in clinical and environmental Brazilian poultry samples.

Salmonella enterica serovar Gallinarum ( S. gallinarum ) is an important host-specific pathogen that causes fowl typhoid, a severe systemic, septicemic, and fatal infection, in chickens. S. gallinarum causes high morbidity and mortality in chickens and poses a significant burden and economic losses to the poultry industry in many developing countries. However, the virulence factors and mechanisms of S. gallinarum -induced systemic infection in chickens remain poorly understood. In this study, we constructed a Salmonella pathogenicity island-14 (SPI-14) mutant strain (mSPI-14) of S. gallinarum and evaluated the pathogenicity of mSPI-14 in the chicken systemic infection model. The mSPI-14 exhibited the same level of bacterial growth and morphological characteristics but significantly reduced resistance to bile acids compared with the wild-type (WT) strain in vitro . The virulence of mSPI-14 was significantly attenuated in the chicken oral infection model in vivo . Chickens infected with WT showed typical clinical symptoms of fowl typhoid, with all birds succumbing to the infection within 6 to 9 days post-inoculation, and substantial increases in bacterial counts and significant pathological changes in the liver and spleen were observed. In contrast, all mSPI-14-infected chickens survived, the bacterial counts in the organs were significantly lower, and no significant pathological changes were observed in the liver and spleen. The expression of interleukin (IL)-1β, IL-12, CXCLi1, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in the liver of mSPI-14-infected chickens were significantly lower than those in the WT-infected chickens. These results indicate that SPI-14 is a crucial virulence factor in systemic infection of chickens, and avirulent mSPI-14 could be used to develop a new attenuated live vaccine to prevent S. gallinarum infection in chickens., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Hu, Ojima, Zhu, Yu, Sugiyama, Haneda, Okamura, Ono and Hu.)

Salmonella enterica subsp. enterica serovar Gallinarum (SG) is a common pathogen in chickens, and causes an acute systemic disease that leads to high mortality. The live attenuated vaccine 9R is able to successfully protect chickens older than six weeks by activating a robust cell-mediated immune response, but its safety and efficacy in young chickens remains controversial. An inactivated SG vaccine is being used as an alternative, but because of its low cellular immune response, it cannot be used as a replacement for live attenuated 9R vaccine. In this study, we employed gamma irradiation instead of formalin as an inactivation method to increase the efficacy of the inactivated SG vaccine. Humoral, cellular, and protective immune responses were compared in both mouse and chicken models. The radiation-inactivated SG vaccine (r-SG) induced production of significantly higher levels of IgG2b and IgG3 antibodies than the formalin-inactivated vaccine (f-SG), and provided a homogeneous functional antibody response against group D, but not group B Salmonella. Moreover, we found that r-SG vaccination could provide a higher protective immune response than f-SG by inducing higher Th17 activation. These results indicate that r-SG can provide a protective immune response similar to the live attenuated 9R vaccine by activating a higher humoral immunity and a lower, but still protective, cellular immune response. Therefore, we expect that the radiation inactivation method might substitute for the 9R vaccine with little or no side effects in chickens younger than six weeks. [ABSTRACT FROM AUTHOR]

Salmonellosis is a bacterial zoonosis causing an array of health conditions. Non-typhoidal salmonellosis (NTS) has a discrete adaptation to certain animals; in poultry, pullorum and fowl typhoid are its primary disease manifestations. The diseases are prevalent in Nigerian poultry and have been well-studied in Nigeria, but less so in North Central Nigeria (NCN). Using field sampling, laboratory methods and a semi-structured questionnaire for 1000 poultry farms in NCN, we explored the incidence and risk factors for the persistence of NTS infection in poultry. Approximately 41.6% of the farms had experienced NTS over the last 18 months. Farm experience of NTS moderately predicted awareness of salmonellosis. Increasing stock in smallholder farms, self-mixing of concentrate on the farm, usage of stream water, pen odour, non-adherence and partial adherence of farms to recommended poultry vaccination against pullorum and fowl typhoid and lack of and non-adherence to biosecurity were identified risk factors that increased the odds of NTS infection in poultry. Antibiotic use practice may have reduced the isolation rate of NTS, yet NTS continues to challenge poultry farms in Nigeria. Identified risk practices must be mitigated intentionally and biosecurity and hygiene must be improved to reduce the burden of NTS.

Salmonella enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum) is a hostspecific pathogen causing systemic infection in poultry, which leads to significant economic losses due to high mortality. However, little is known about the dynamic process of systemic infection and pathogenic characteristics of S. Gallinarum in chickens. In the present study, we developed an oral infection model that reproduces the pathology of S. Gallinarum and clarified the host immune response of the infected chickens. Chickens at 20 days of age orally inoculated at a dose of 108 colony forming unit (CFU) showed typical clinical signs of fowl typhoid and died between 6 and 10 days post infection. The inoculated S. Gallinarum rapidly disseminated to multple organs and the bacterial counts increased in the liver and spleen at 3 days post infection. Pathological changes associated wirh inflammation in the liver and spleen became apparent at 4 days post infection, and increased expression of interferon (IFN)-γ and interleuikin (IL)-12 in the liver and spleen did not observed until 3 days post infection. These results indicate that S. Gallinarum rapidly spread to entire body through intestine, and the low-level of inflammatory responses in the liver during the early stage of infection may contribute to rapid, systemic dissemination of the bacteria. Our infection model and findings will contribute to the better understanding of the pathogenic mechanism of S. Gallinarum, and provide new insights into the prevention and control of fowl typhoid. [ABSTRACT FROM AUTHOR]

Salmonella Gallinarum (SG) is an avian-restricted pathogen that causes fowl typhoid in poultry. Although it has been reported frequently over many decades in poultry flocks worldwide, the microorganism is more commonly associated with poultry in developing countries, particularly those with high ambient temperatures, where the acute form of the disease results in considerable economic losses. A more detailed investigation of environmental factors that affect the course of disease may assist in identifying effective prevention and control measures. Heat stress is known to impair the immunological response to a variety of pathogens and clearly may be an important contributory factor in the prevalence of disease in countries with warm or hot climates. Thus, the objective of the present study was to evaluate the effects of heat stress on chickens infected with SG. For this, light and semi-heavy commercial laying hens were distributed randomly within four groups as follows: infected and non-infected groups in rooms held at ambient temperature, and infected and non-infected groups under heat stress. Clinical signs, egg production, and mortality were recorded daily. Bacteriological counts in liver and spleen samples were estimated at 2, 5, 7, and 14 days post-infection. The results showed that both SG infection and heat stress had similar effects on egg production and a synergistic effect of the two stressors was observed. The data show an interaction between disease and heat stress which could point towards environmental and biosecurity approaches to resolving the possible 30% fall in production observed in such countries. [ABSTRACT FROM AUTHOR]

1. Salmonella Gallinarum (SG) infections cause fowl typhoid, which leads to important economic losses. Multidrug resistance (MDR) and the capacity for bacteria to form biofilms could play an important role in the persistence of SG in poultry flocks resulting in intermittent disease outbreaks. The aim of the following study was to assess the lytic activity of two new bacteriophages (Salmonella phages UPF_BP1 and UPF_BP2) against MDR and biofilm-forming SG. 2. Forty-six strains of SG, isolated in 2015, were characterised by antimicrobial resistance, biofilm formation profiles and susceptibility to two new bacteriophages. 3. Of these strains, 24% were multidrug resistant and more than 80% formed biofilm, with no statistical difference between incubation temperatures (42°C or 22°C). With regard to the lytic activity of the phages, 85% of strains were susceptible to at least one phage. Of these, 74% were lysed by both phages, including MDR and biofilm producing strains. 4. The use of salmonella phages UPF_BP1 and UPF_BP2 were shown to be promising alternatives for the biological control of fowl typhoid. [ABSTRACT FROM AUTHOR]

Background: The poultry red mite Dermanyssus gallinae (De Geer, 1778) is a major ectoparasite of poultry. Infestations are found in most laying hen farms in Europe, and breeder flocks have also been reported to be affected. Mite infestation has detrimental effects on animal welfare, it causes significant economic losses, and, additionally, D. gallinae is often considered as a vector for pathogens. Despite suspicion of a close relationship between the poultry red mite and Salmonella enterica enterica serovar Gallinarum biovar Gallinarum (serovar Gallinarum), the causative agent of fowl typhoid disease (FT), there has been no definitive proof of mite-mediated transmission. Therefore, an investigation was conducted to determine if D. gallinae-mediated transmission of serovar Gallinarum could be demonstrated among four different hen groups. Methods: Two groups of 8 hens (A and B) were experimentally infected with serovar Gallinarum in two isolators. After 7 days, when birds showed signs of FT, about 25,000 mites were introduced. After 3 days, mites were harvested and used to infest two other hen groups of 8 (C and D), in two separate isolators. The health status of hens was constantly monitored; detection and quantification of serovar Gallinarum were performed by PCR and qPCR from mites and organs of dead hens. The maximum likelihood estimation of the infection rate and mite vectorial capacity were calculated. Results: Clinical disease was observed in groups infected with serovar Gallinarum (A and B) and in hens of groups C and D infested with mites harvested from the isolators containing groups A and B. In all four groups, serovar Gallinarum was detected from liver, spleen, ovary, and cecum of hens, thus confirming the diagnosis of FT. Mite analysis demonstrated the presence of the pathogen, with an estimated infection rate ranging between 13.72 and 55.21 infected per thousand mites. Vectorial capacity was estimated to be 73.79. Conclusions: Mites harvested from birds infected with serovar Gallinarum were shown to carry the mite, and then to transfer serovar Gallinarum to isolated groups of pathogen-free birds that subsequently showed signs of FT. Mite vectorial capacity was high, demonstrating that D. gallinae should be considered an effective vector of FT. [ABSTRACT FROM AUTHOR]

Salmonella is a bacterium usually associated with mild gastroenteritis in chickens, but strains of the serotype Gallinarum can result in 2 severe systemic chicken diseases: fowl typhoid (by biovar Gallinarum) and pullorum disease (by biovar Pullorum). These diseases can be controlled by immunization with a specific vaccine (SG 9R). The aim of this study was to characterize recent Salmonella Gallinarum isolates by molecular analyses, as well as detect field and vaccine strains in clinical and environmental samples from poultry flocks. Field isolates of S. Gallinarum (n = 18) and other serotypes (n = 117) were obtained from Brazilian poultry farms between 2012 and 2015. In addition, clinical and environmental samples of poultry flocks (vaccinated or not) with fowl typhoid were obtained in a 3-year period (2013 to 2015). The results demonstrated that specific PCR targeted on speC and glgC genes and PCR-restriction fragment length polymorphisms based on fliC and sefA genes detected all the 18 isolates, 10 identified as field isolates of biovar Gallinarum and 8 as biovar Pullorum. All isolates of other serotypes presented negative results. In addition, PCR tests were performed directly from the pre-enrichment broth of the bacteriological procedures. Vaccine SG 9R and field S. Gallinarum strains were detected in samples with (n = 19) and without (n = 59) vaccination. These results demonstrate the use of PCR tests for detection of S. Gallinarum in clinical and environmental Brazilian poultry samples. [ABSTRACT FROM AUTHOR]

Fowl typhoid (FT), which is caused by Salmonella enterica serovar Gallinarum (S. Gallinarum), leads to high morbidity and acute or subacute mortality in chickens of all ages. Although a live S. Gallinarum 9R vaccine was introduced in 2001 for commercial layer chickens in Korea, until recently, a variety of antimicrobials were widely used to prevent or treat FT. In this study, we investigated antimicrobial resistance in S. Gallinarum strains isolated from 2014 to 2018 and characterized the multidrug-resistant (MDR) strains to better understand the resistance trends in recent isolates. A total of 130 S. Gallinarum isolates were collected from chickens with FT, and the isolates showed highest rates of resistance to nalidixic acid (78.5%), followed by gentamicin (52.3%), ciprofloxacin (26.9%), and ampicillin (14.6%). Particularly, significant increases (P < 0.05) in the frequencies of resistance to the following antimicrobials were observed: ampicillin (from 7.7 to 28.6%), amoxicillin-clavulanate (from 0.0 to 10.7%), nalidixic acid (from 69.2 to 100.0%), ciprofloxacin (from 15.4 to 50.0%), chloramphenicol (from 0.0 to 17.9%), and colistin (from 0.0 to 14.3%). The prevalence of MDR isolates also rapidly increased from 23.1% in the 2014 to 60.7% in the 2018 (P < 0.05). The distribution of antimicrobial resistance genes in the 39 MDR S. Gallinarum isolates was as follows: ant (2") -I gene (22 isolates), bla TEM-1 gene (13 isolates), sul1 (9 isolates), sul2 (3 isolates), cmlA (3 isolates), and qnrB (3 isolates). Of 39, 25 (64.1%) MDR S. Gallinarum isolates also carried class 1 integrons, and these showed 5 types of resistance gene cassettes: dfr A12+ aad A2 (36.0%), aad A2 (36.0%), aad A1- aad A2 (20.0%), dfr A12+ cat B3+ aad A2 (4.0%), and dfr A12 (4.0%). Among the plasmid replicons, B/O (33.3%) was more prevalent than the other replicon types, followed by Frep (25.0%), FIIA (19.4%), FIB (13.9%), and I1 (8.3%). Antimicrobial resistance may become a serious problem because many drugs are likely ineffective for the treatment of FT. Therefore, these data support the critical need for comprehensive surveillance of antimicrobial resistance in poultry. [ABSTRACT FROM AUTHOR]

Salmonella Gallinarum (SG) provokes fowl typhoid, an infectious disease of acute clinical course that affects gallinaceous of any age and leads to high mortality rates. During the typhoid-like systemic infection of S. Typhimurium (STM) in mice, the bacterium expresses the mgtC gene, which is encoded in the Salmonella Pathogenecity Island – 3 (SPI-3). In this serovar, the function is linked to bacterial replication within macrophages, and its absence attenuates the pathogen. We hypothesized that deleting mgtC from SG genome would alter the microorganism pathogenicity in susceptible commercial poultry in a similar manner. Thus, the present study sought to elucidate the importance of mgtC on SG pathogenicity. For this, a mgtC -mutant lacking S. Gallinarum mutant was constructed (SG Δ mgtC). Its ability to replicate in medium that mimicries the mgtC -related intracellular environment of macrophages as well as in primary macrophages from chicken was evaluated. Moreover, the infection of susceptible chickens was performed to elucidate its pathogenicity and the elicited immune responses by measuring key interleukins by qRT-PCR and the population of macrophages and lymphocytes T CD4+ and CD8+ by means of immunohistochemistry. It was observed that mgtC was required for S. Gallinarum replication in acidified low-Mg2+ media and survival within macrophages. However, unlike its requirement for initial phase of STM infection in mice, lower bacterial counts were only observed at the late stage of macrophage infection without affecting the citotoxicity. Experiments showed that knocking-out the mgtC gene neither altered bacterial uptake by macrophages nor affects bacterial counts in liver and spleen and total chicken mortality. However, plotting a survival curve and analyzing the clinical-pathologic conditions, it was observed a slower progression of the disease in chickens infected by SG Δ mgtC compared to those challenged by the wild-type strain. Furthermore, the mRNA expression of IFN-γ and LITAF were similar between the infected chickens, but higher than in the uninfected group. The same was observed in macrophages and lymphocytes T CD4+ populations. On the other hand, the presence of lymphocytes T CD8+ was increased in the initial phase of the disease provoked by the wild-type strain over the mutant strain. We concluded that the role of mgtC in Fowl Typhoid in susceptible chickens differs from the role in typhoid-like infections in mammals. Thus, the deletion of mgtC gene from S. Gallinarum genome does not affect the overall pathogenicity, but slightly alters the pathogenesis. [ABSTRACT FROM AUTHOR]

The H9N2 avian influenza virus significantly affects the health of poultry and humans. We identified a prokaryotic and eukaryotic dual-expression vector system, pJHL270, that can provide simultaneous MHC class I and II stimulation of the host immune system, and we designed vaccine antigens by selecting the consensus HA1 sequence and M2e antigens from H9N2 virus circulating in South Korea from 2000 to 2021. The genes were cloned into the pJHL270 vector, and the cloned plasmid was delivered by a live-attenuated Salmonella Gallinarum (SG) strain. The immunity and protective efficacy of the SG-based H9N2 vaccine construct, JOL2922, against avian influenza and fowl typhoid (FT) were evaluated. The Ptrc and CMV promoters conferred antigen expression in prokaryotic and eukaryotic cells to induce balanced Th-1/Th-2 immunity. Chickens immunized with JOL2922 yielded high antigen-specific humoral and mucosal immune responses. qRT-PCR revealed that the strain generated polyfunctional IFN-γ and IL-4 secretion in immunized chickens. Furthermore, a FACS analysis showed increased CD3CD4+ and CD3CD8+ T-cell subpopulations following immunization. Peripheral Blood Mononuclear Cells (PBMCs) harvested from the immunized chickens significantly increased MHC class I and II expression, 3.5-fold and 2.5-fold increases, respectively. Serum collected from the immunized groups had an evident hemagglutinin inhibition titer of ≥6 log 2. Immunization reduced the lung viral titer by 3.8-fold within 5 days post-infection. The strain also generated SG-specific humoral and cellular immune responses. The immunized birds all survived a virulent SG wild-type challenge. In addition, the bacterial burden was reduced by 2.7-fold and 2.1-fold in spleen and liver tissue, respectively, collected from immunized chickens. Our data indicate that an attenuated SG strain successfully delivered the dual-expression vector system and co-stimulated MHC class I and II antigen presentation pathways via exogenous and endogenous antigen presentation, thereby triggering a balanced Th-1/Th-2-based immune response and conferring effective protection against avian influenza and FT. • The pJHL270 vector system could express recombinant H9N2 antigens in both prokaryotic and eukaryotic systems. • Exogenous and endogenous antigens stimulated the host MHC class I and II. • Vaccine construct, JOL2922 induced protective humoral and cellular immune responses against avian influenza and fowl typhoid. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Gallinarum causes devastating outbreaks of fowl typhoid across the globe, especially in developing countries. With the use of antimicrobial agents being reduced due to legislation and the absence of licensed vaccines in some parts of the world, an attractive complementary control strategy is to breed chickens for increased resistance to Salmonella. The potential for genetic control of salmonellosis has been demonstrated by experimental challenge of inbred populations. Quantitative trait loci (QTL) associated with resistance have been identified in many genomic regions. A major QTL associated with systemic salmonellosis has been identified in a region termed SAL1. In the present study, two outbreaks of fowl typhoid in 2007 and 2012 in the United Kingdom were used to investigate the genetic architecture of Salmonella resistance in commercial laying hens. In the first outbreak 100 resistant and 150 susceptible layers were genotyped using 11 single nucleotide polymorphism (SNP) and 3 microsatellite markers located in the previously identified SAL1 region on chromosome 5. From the second outbreak 100 resistant and 200 susceptible layers, belonging to a different line, were genotyped with a high-density (600 K) genome-wide SNP array. Substantial heritability estimates were obtained in both populations (h2 = 0.22 and 0.26, for the layers in the first and second outbreak, respectively). Significant associations with three markers on chromosome 5 located close to AKT1 and SIVA1 genes, coding for RAC-alpha serine/threonine protein kinase, and the CD27-binding protein SIVA1, respectively, were identified in the first outbreak. From analysis of the second outbreak, eight genome-wide significant associations with Salmonella resistance were identified on chromosomes 1, 6, 7, 11, 23, 24, 26, 28 and several others with suggestive genome-wide significance were found. Pathway and network analysis revealed the presence of many innate immune pathways related to Salmonella resistance. Although, significant associations with SNPs located in the SAL1 locus were not identified by the genome-wide scan for layers from the second outbreak, pathway analysis revealed P13K/AKT signaling as the most significant pathway. In summary, resistance to fowl typhoid is a heritable polygenic trait that could possibly be enhanced through selective breeding.

Dermanyssus gallinae (Mesostigmata: Dermanyssidae, De Geer, 1778) is an ectoparasite of poultry, suspected to play a role as a vector of Salmonella enterica subsp. enterica ser. Gallinarum. Despite an association between them being reported, the actual dynamics in field remain unclear. Therefore, the present study aimed to confirm the interactions among mites, pathogen and chickens. The study was carried out in an industrial poultry farm infested by D. gallinae, during an outbreak of fowl typhoid. The presence of S. Gallinarum in mites was assessed and quantified by a semi‐nested polymerase chain reaction (PCR) and real‐time PCR, respectively, in mites collected during two subsequent productive cycles and the sanitary break. The anti‐group D Salmonella antibodies were quantified by an enzyme‐linked immunosorbent assay. During the outbreak and the sanitary break, S. Gallinarum was constantly present in mites. In the second cycle, scattered positivity was observed, although hens did not exhibit signs of fowl typhoid, as a result of the vaccination with BIO‐VAC SGP695 (Fatro, Ozzano Emilia Bo, Italy). The data strongly suggest that D. gallinae acts as reservoir of S. Gallinarum, thus allowing the pathogen to persist in farms. Furthermore, the present study has highlighted the interactions among D. gallinae, S. Gallinarum and hens with respect to enhancing the mite‐mediated circulation of S. Gallinarum in an infested poultry farm. The association between Dermanyssus gallinae and Samonella enterica subsp. enterica ser. Gallinarum has been confirmed in the field during a fowl typhoid outbreak in an infested industrial poultry farm.The infection rate of Dermanyssus gallinae and the microbial load in mites were numerically related to the circulation of Salmonella Gallinarum in the flock.Preventive measures, such as a proper vaccination programme, may prevent the infection and the common reinfection of the new flocks after the sanitary break. [ABSTRACT FROM AUTHOR]

Poultry farming in the rural areas of Nigeria plays a major role in the sustenance of the dwellers and contributes to the total national income of the country. Protecting the birds from fowl typhoid is of dire importance. Vials of a marketed fowl typhoid vaccine were subjected to variable storage conditions namely;:daily freeze-thaw cycles, room temperature, 40 °C, 85% relative humidity (RH) and refrigeration (with fluctuating power) for three weeks. Biochemical tests were carried out on the vaccine samples before storage to confirm the presence of live 9R viable Salmonella before commencement of the stability studies. The stability of the vaccines was assessed by performing these studies: percent (%) viability, pH, FTIR spectroscopy and ninhydrin test. For the % viability, the daily freeze-thaw cycle samples had the highest number of viable cells, while the refrigerated samples had the least, after three weeks of storage. The pH showed a slight increases for all the extreme storage conditions. The FTIR spectroscopy showed slight shifts in the peaks of all the spectra after three weeks indicating imminent loss of viability. The results show a gradual loss of viability of the vaccines kept under unfavorable conditions. Stringent measures need to be taken to ensure adequate storage of the vaccines to prevent loss of viability that may lead to infection with fowl typhoid. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Gallinarum causes devastating outbreaks of fowl typhoid across the globe, especially in developing countries. With the use of antimicrobial agents being reduced due to legislation and the absence of licensed vaccines in some parts of the world, an attractive complementary control strategy is to breed chickens for increased resistance to Salmonella. The potential for genetic control of salmonellosis has been demonstrated by experimental challenge of inbred populations. Quantitative trait loci (QTL) associated with resistance have been identified in many genomic regions. A major QTL associated with systemic salmonellosis has been identified in a region termed SAL1. In the present study, two outbreaks of fowl typhoid in 2007 and 2012 in the United Kingdom were used to investigate the genetic architecture of Salmonella resistance in commercial laying hens. In the first outbreak 100 resistant and 150 susceptible layers were genotyped using 11 single nucleotide polymorphism (SNP) and 3 microsatellite markers located in the previously identified SAL1 region on chromosome 5. From the second outbreak 100 resistant and 200 susceptible layers, belonging to a different line, were genotyped with a high-density (600 K) genome-wide SNP array. Substantial heritability estimates were obtained in both populations (h 2 = 0.22 and 0.26, for the layers in the first and second outbreak, respectively). Significant associations with three markers on chromosome 5 located close to AKT1 and SIVA1 genes, coding for RAC-alpha serine/threonine protein kinase, and the CD27-binding protein SIVA1 , respectively, were identified in the first outbreak. From analysis of the second outbreak, eight genome-wide significant associations with Salmonella resistance were identified on chromosomes 1, 6, 7, 11, 23, 24, 26, 28 and several others with suggestive genome-wide significance were found. Pathway and network analysis revealed the presence of many innate immune pathways related to Salmonella resistance. Although, significant associations with SNPs located in the SAL1 locus were not identified by the genome-wide scan for layers from the second outbreak, pathway analysis revealed P13K/AKT signaling as the most significant pathway. In summary, resistance to fowl typhoid is a heritable polygenic trait that could possibly be enhanced through selective breeding. [ABSTRACT FROM AUTHOR]

Salmonella Gallinarum is a Gram-negative bacteria that causes fowl typhoid, a septicemic disease with high morbidity and mortality that affects all ages of chickens. Although vaccines and antimicrobials have been used nationwide to eradicate the disease, the malady is still prevalent in Korea. In this study, we investigated the virulence and genetic variation of 116 S. Gallinarum isolates from laying hens between 2014 and 2018. A total of 116 isolates were divided into five Gallinarum Sequence Types (GST) through clustered regularly interspaced short palindromic repeats (CRISPR) subtyping method. The GSTs displayed changes over time. The 116 isolates showed no difference in virulence gene distribution, but the polyproline linker (PPL) length of the SpvB, one of the virulence factors of Salmonella spp., served as an indicator of S. Gallinarum pathogenicity. The most prevalent PPL length was 22 prolines (37.9%). The shortest PPL length (19 prolines) was found only in isolates from 2014 and 2015. However, the longest PPL length of 24 prolines appeared in 2018. This study indicates that PPLs of S. Gallinarum in Korea tend to lengthen over time, so the pathogenic potency of the bacteria is increasing. Moreover, the transition of GST was associated with PPL length extension over time. These results indicate that surveillance of changing GST and PPL length are necessary in the monitoring of S. Gallinarum isolates.

Our study was undertaken to determine the best samples and selective-differential plating media to be used for Salmonella spp. isolation. We also compared hematological and serum biochemical values, Salmonella biovar Gallinarum (SG) detection (isolation and serological test), and inflammatory intestinal response (fecal leukocyte) in laying hens with naturally occurring fowl typhoid outbreaks. Furthermore, we looked for a biomarker of SG infection. Spleen, liver, ovarian follicle content, and bone marrow were found to be the best samples for SG isolation and the agreement between MacConkey- Salmonella Shigella agar was slight to excellent. The laying hens with SG isolation and rapid serum plate agglutination positive results showed a higher percentage of heterophils, heterophil/lymphocyte ratio and total white blood cells, and a lower percentage of lymphocytes than those with negative results. Furthermore, the positive fecal leukocyte samples had a higher percentage of heterophils, gamma-glutamyl transferase, total protein and globulin values than negative samples. Five biomarkers' cut-offs are proposed to distinguish between laying hens positive and negative to SG isolation. [Display omitted] • Organs and bone marrow were the best samples for S. ser. Gallinarum isolation. • Diverse blood parameters were increased in positive S. ser. Gallinarum laying hens. • Agreement between two agar media in organs and bone marrow samples was between slight and excellent. • Cut-offs of 5 biomarkers of S. ser. Gallinarum infection in laying hens were proposed. [ABSTRACT FROM AUTHOR]

From a disease outbreak among Vanaraja fowl, an indigenous Indian Breed reared by backyard system in Jhargram, West Bengal, Salmonella Gallinarum was isolated and characterised. The outbreak occurred among 6-8 day old chicks. A total of 150 birds died in a span of 5 days. Salmonella Gallinarum were identified and confirmed by standard bacteriological methods and presence of invasion ( invA) gene was detected by PCR. The isolates were susceptible to 15 common antimicrobials in vitro. Although chemotherapy may be effective, outbreaks of fowl typhoid in backyard poultry warranted precise control policy

Salmonella enterica serovar Gallinarum causes Fowl Typhoid in poultry, and it is host specific to avian species. The reasons why S. Gallinarum is restricted to avians, and at the same time predominately cause systemic infections in these hosts, are unknown. In the current study, we developed a surgical approach to study gene expression inside the peritoneal cavity of hens to shed light on this. Strains of the host specific S. Gallinarum, the cattle-adapted S. Dublin and the broad host range serovar, S. Enteritidis, were enclosed in semi-permeable tubes and surgically placed for 4 h in the peritoneal cavity of hens and for control in a minimal medium at 41.2 °C. Global gene-expression under these conditions was compared between serovars using tiled-micro arrays with probes representing the genome of S. Typhimurium, S. Dublin and S. Gallinarum. Among other genes, genes of SPI-13, SPI-14 and the macrophage survival gene mig-14 were specifically up-regulated in the host specific serovar, S. Gallinarum, and further studies into the role of these genes in host specific infection are highly indicated. Analysis of pathways and GO-terms, which were enriched in the host specific S. Gallinarum without being enriched in the two other serovars indicated that host specificity was characterized by a metabolic fine-tuning as well as unique expression of virulence associated pathways. The cattle adapted serovar S. Dublin differed from the two other serovars by a lack of up-regulation of genes encoded in the virulence associated pathogenicity island 2, and this may explain the inability of this serovar to cause disease in poultry., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest to declare., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In poultry, host-specific Salmonella infections cause fowl typhoid and pullorum disease, which are severe systemic diseases of chickens that result in severe problems and high mortality and produce economic losses in developing countries. Fowl typhoid and pullorum disease have reemerged in recent years in developing countries, and birds of any age can be infected. Developing countries have established sanitary measures and official programs to prevent and control both diseases; however, there are cyclic or seasonal outbreaks related to disease management. An overview of the current situation and future perspectives regarding live attenuated vaccines against fowl typhoid and pullorum disease are discussed. [ABSTRACT FROM AUTHOR]

Oral fowl typhoid (FT) vaccine is necessary for improved flock vaccinations and economic growth. This study was undertaken to evaluate the immune responses of birds given oral fowl typhoid vaccine coated with chitosan/alginate microparticles and comparing it with the conventional subcutaneous route of administration. Preliminary studies were done to evaluate the particle size, encapsulation efficiency and agglutination. Sixty day-old chicks were divided into three groups of twenty birds each. This comprised a negative control group NEG 451 (non-vaccinated and non-challenged used as control for cytokine quantification), SC 634 (live 9R vaccine by the injection route) and OCV 567 (live 9R vaccine coated with chitosan/alginate microparticles). Vaccination was done at 10 weeks and 14 weeks of age followed by challenge at 16 weeks of age. IgG was measured using ELISA. mRNA fold expression of IFN-γ in spleen was calculated using qRT-PCR. Particle sizes ranged between 0.55 µm and 10 µm. Encapsulation efficiency was above 60%. ELISA showed E-values of 0.10 ± 0.14, 0.07 ± 0.01 and 0.02 ± 0.01 for OCV 567, SC 634 and NEG 451 respectively after primary vaccination. Also E-values were 0.25 ± 0.16, 0.19 ± 0.04 and 0.0008 ± 0.005 for SC 634, OCV 567 and NEG451 respectively after boost vaccination. The expression of IFN- γ in spleen using 2 −ΔΔ CT calculation was upregulated with values of 1.97 and 0.75 for OCV 567 and SC 634 respectively. After challenge with the 85-kb virulence plasmid SG9, there was 100% protection of the birds in both OCV 567 and SC 634 groups with no mortality. In conclusion, there was no significant difference at p < 0.05 of the means ± SD in immune responses between the oral fowl typhoid vaccine coated with chitosan/alginate microparticles and the subcutaneous route of administration. However, it is noteworthy to mention that the protective efficacy of the oral route is due to the chitosan/alginate biopolymers which coated the vaccine preventing destruction in the gastrointestinal tract. [ABSTRACT FROM AUTHOR]