Virulence Variation of Salmonella Gallinarum Isolates through SpvB by CRISPR Sequence Subtyping, 2014 to 2018

Kim, Koeun; Yoon, Sunghyun; Kim, Yeong Bin; Lee, Young Ju

doi:10.3390/ani10122346

Cited by 3 publications

(4 citation statements)

References 32 publications

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“…10). These findings are in accordance with the earlier research works reported by Awad et al, 2020 [8] , Tarabees et al, 2017 [38] , Das et al, 2012 [14] , Borges et al, 2013 [11] , Ammar et al, 2016 [43] , Zhang et al, 2018 [44] , Shanmugasamy et al, 2011 [34] , Suresh et al, 2019 [37] and Kim et al, 2020 [26] , who revealed 100% prevalence for invA gene in the Salmonella isolates recovered from poultry. Similarly, Chakroun et al, 2018 [13] and Zou et al, 2011 [42] also revealed 100% prevalence of 16S rRNA gene.…”

Section: Molecular Confirmation Of the Isolatessupporting

confidence: 91%

Prevalence of Salmonella species isolated from poultry farms and chicken meat

Gaurav,

Sharma

et al. 2024

Int. J. Adv. Biochem. Res.

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To assess the genetic diversity of strains, genotypic characterization of foodborne bacteria such as Salmonella is very significant. Salmonella is a complex bacterial species with a wide number of serotypes and strains in different host populations, including animals, birds and humans. In total, 304 samples from poultry farms (water, feed, poultry faeces and carcass) and retail chicken meat sold in market were collected randomly from Udaipur district. Isolation of Salmonella strain from the samples collected from poultry farms and chicken meat sold in retail market was done according to IS 5887 (Part 3):1999. Isolation of DNA from pure culture of presumptive Salmonella isolates was done by using the Nucleo-pore ® gDNA fungal/bacterial mini kit (Genetix) as per the instruction mentioned in the kit manual. All the Salmonella isolates were screened for the presence of genus specific genes by using the PCR. Out of the 304 samples from poultry farms and retail chicken meat including water (n= 29), feed (n= 29), poultry feces (n= 106), carcass (n= 42), and retail chicken meat (n= 98), a total of four samples were found to be positive for Salmonella spp. giving a prevalence rate of 3.44%, 0%, 1.89%, 0% and 1.02% respectively. These 4 isolates were further confirmed through PCR by targeting the genus specific primers (invA and 16S rRNA gene) giving the overall prevalence to be 1.32% (4/304).

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Section: Molecular Confirmation Of the Isolatessupporting

confidence: 91%

Prevalence of Salmonella species isolated from poultry farms and chicken meat

Gaurav,

Sharma

et al. 2024

Int. J. Adv. Biochem. Res.

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“…SpvB and C genes have been reported to cause virulence in S. typhimurium in a mouse infection model ( Roudier et al, 1992 ; Matsui et al, 2001b ). A recent study has shown that the length of the linker connecting N and C terminal domains of SpvB increases pathogenicity of S. gallinarum in chickens ( Kim et al, 2020 ). Nevertheless, there is no evidence available of SpvB’s role in causing FT in poultry.…”

Section: Resultsmentioning

confidence: 99%

“…( Skyberg et al, 2006 ). Moreover, 9R strain of S. gallinarum (live attenuated) has been used worldwide as a vaccine since 2001; however, due to the persistent evolution of the pathogen, the vaccine strain may not provide sufficient protection against S. gallinarum ( Lee et al, 2007 ; Kim et al, 2020 ). Therefore, the current study aimed to produce an avirulent strain by deleting the SpvB gene from S. gallinarum virulent plasmid through the CRISPR/Cas9 system.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken

Basit

Tahir

Haider

et al. 2022

Front. Bioeng. Biotechnol.

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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.

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“…A protospacer, which is a short external sequence at a specific location, is inserted as a spacer in the CRISPR loci of bacteria during an infection [ 18 ]. Recently, CRISPR arrays have been applied as a novel typing method for isolates because they are useful in discriminating the pathogenicity of Salmonella [ 19 ], E. coli [ 20 , 21 , 22 , 23 , 24 , 25 ], and Pseudomonas aeruginosa [ 26 ]. This study aimed to investigate the virulence potential of E. coli isolated from bulk tank milk, not from mastitis, and to analyze the pathogenic characterization of E. coli using the CRISPR typing method.…”

Section: Introductionmentioning

confidence: 99%

Distribution of CRISPR in Escherichia coli Isolated from Bulk Tank Milk and Its Potential Relationship with Virulence

Kang

Lee

2022

Animals

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Escherichia coli is one of the most common causes of mastitis on dairy farms around the world, but its clinical severity is determined by a combination of virulence factors. Recently, clustered regularly interspaced short palindromic repeat (CRISPR) arrays have been reported as a novel typing method because of their usefulness in discriminating pathogenic bacterial isolates. Therefore, this study aimed to investigate the virulence potential of E. coli isolated from bulk tank milk, not from mastitis, and to analyze its pathogenic characterization using the CRISPR typing method. In total, 164 (89.6%) out of 183 E. coli isolated from the bulk tank milk of 290 farms carried one or more of eighteen virulence genes. The most prevalent virulence gene was fimH (80.9%), followed by iss (38.3%), traT (26.8%), ompT (25.7%), afa/draBC (24.0%), and univcnf (21.9%). Moreover, the phylogenetic group with the highest prevalence was B1 (64.0%), followed by A (20.1%), D (8.5%), and C (7.3%) (p < 0.05). Among the four CRISPR loci, only two, CRISPR 1 and CRISPR 2, were found. Interestingly, the distribution of CRISPR 1 was significantly higher in groups A and B1 compared to that of CRISPR 2 (p < 0.05), but there were no significant differences in groups C and D. The prevalence of CRISPR 1 by virulence gene ranged from 91.8% to 100%, whereas that of CRISPR 2 ranged from 57.5% to 93.9%. The distribution of CRISPR 1 was significantly higher in fimH, ompT, afa/draBC, and univcnf genes than that of CRISPR 2 (p < 0.05). The most prevalent E. coli sequence types (EST) among 26 ESTs was EST 22 (45.1%), followed by EST 4 (23.2%), EST 16 (20.1%), EST 25 (19.5%), and EST 24 (18.3%). Interestingly, four genes, fimH, ompT, afa/draBC, and univcnf, had a significantly higher prevalence in both EST 4 and EST 22 (p < 0.05). Among the seven protospacers derived from CRISPR 1, protospacer 163 had the highest prevalence (20.4%), and it only existed in EST 4 and EST 22. This study suggests that the CRISPR sequence-typing approach can help to clarify and trace virulence potential, although the E. coli isolates were from normal bulk tank milk and not from mastitis.

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Virulence Variation of Salmonella Gallinarum Isolates through SpvB by CRISPR Sequence Subtyping, 2014 to 2018

Cited by 3 publications

References 32 publications

Prevalence of Salmonella species isolated from poultry farms and chicken meat

Prevalence of Salmonella species isolated from poultry farms and chicken meat

CRISPR/Cas9-Based Deletion of SpvB Gene From Salmonella gallinarum Leads to Loss of Virulence in Chicken

Distribution of CRISPR in Escherichia coli Isolated from Bulk Tank Milk and Its Potential Relationship with Virulence

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