Identification of cholix toxin gene in Vibrio cholerae non-O1/non-O139 isolated from diarrhea patients in Bushehr, Iran
-
Marziyeh Gholizadeh Tangestani
,
Jafar Alinezhad
,
Abdolmohammad Khajeian
,
Somayyeh Gharibi
,
Mohammad Ali Haghighi
Abstract
Background and Objectives: Cholixin (cholix toxin) is a novel exotoxin in Vibrio cholerae identified as an elongation factor II specific ADP-ribosyltransferase which inhibits protein synthesis in the eukaryotic cell. Previous researches have suggested that cholixin probably is an important virulence factor in non-O1/non-O139 V. cholerae (NAG) serotypes that could be related to extra-intestinal rather than intestinal infections. This study was aimed to investigate the frequency and genetic diversity of colixin gene (chxA) in clinical V. cholerae NAG isolates.
Materials and Methods: The presence of chxA gene in 44 clinical V. cholerae NAG isolates were screened using PCR through specific primers designed for the receptor-binding domain (RBD) of chxA gene. The five PCR products of chxA gene were sequenced.
Results: This study showed that chxA gene presented in 19 V. cholerae NAG isolates. The sequences analysis of 5 out of 19 the partial chxA genes amplicon showed that 4 of them belonged to chxA I and the other one belonged to chxA II subtypes. Two distinct clusters were revealed for these isolates by phylogenic analysis, too.
Conclusion: The chxA gene contained high frequency among V. cholerae NAG isolates in Bushehr, Iran. The polymorphism study on RBD of cholixin gene is suggested as an appropriate method for phylogenic characterization of the various chxA gene subtypes.
1. Madhusudana RB, Surendran PK. Detection of ctx gene positive non-O1/non-O139 V. cholerae in shrimp aquaculture environments. J Food Sci Technol 2013; 50: 496-504.
2. Rui H, Ritchie JM, Bronson RT, Mekalanos JJ, Zhang Y, Waldor MK. Reactogenicity of live-attenuated Vibrio cholerae vaccines is dependent on flagellins. Proc Natl Acad Sci U S A 2010; 107: 4359-4364.
3. Ceccarelli D, Chen A, Hasan NA, Rashed SM, Huq A, Colwell RR. Non-O1/non-O139 Vibrio cholerae carrying multiple virulence factors and V. cholerae O1 in the Chesapeake Bay, Maryland. Appl Environ Microbiol 2015; 81: 1909-1918.
4. Ramamurthy T, MukhopadhyayAK, Nandy RK, Balakrish Nair G (2013). Molecular Typing of Vibrio cholerae: Imprints in the Epidemiology of Cholera. In: Molecular typing in bacterial infections. Ed, De Filippis I, Mckee ML. Humana Press, 1St ed. New York, pp. 53-72.
5. Dalsgaard A, Forslund A, Hesselbjerg A, Bruun B. Clinical manifestations and characterization of extra-intestinal Vibrio cholerae non-O1, non-O139 infections in Denmark. Clin Microbiol Infect 2000; 6: 625-627.
6. Hao Y, Wang Y, Bi Z, Sun B, Jin Y, Bai Y, et al. A case of non-O1/non-O139 Vibrio cholerae septicemia and meningitis in a neonate. Int J Infect Dis 2015; 35: 117-119.
7. Shin OS, Tam VC, Suzuki M, Ritchie JM, Bronson RT, Waldor MK, et al. Type III secretion is essential for the rapidly fatal diarrheal disease caused by non-O1, non-O139 Vibrio cholerae. mBio 2011; 2(3): e00106-11.
8. Rajpara N, Vinothkumar K, Mohanty P, Singh AK, Singh R, Sinha R, et al. Synergistic effect of various virulence factors leading to high toxicity of environmental V. cholerae non-O1/ non-O139 isolates lacking ctx gene: comparative study with clinical strains. PLoS One 2013; 8(9):e76200.
9. Purdy A, Rohwer F, Edwards R, Azam F, Bartlett DH. A glimpse into the expanded genome content of Vibrio cholerae through identification of genes present in environmental strains. J Bacteriol 2005; 187: 2992-3001.
10. Awasthi SP, Asakura M, Neogi SB, Hinenoya A, Ramamurthy T, Yamasaki S. Development of a PCR-restriction fragment length polymorphism assay for detection and subtyping of cholix toxin variant genes of Vibrio cholerae. J Med Microbiol 2014; 63: 667-673.
11. O'Brien AD, Chen ME, Holmes RK, Kaper J, Levine MM. Environmental and human isolates of Vibrio cholerae and Vibrio parahaemolyticus produce a Shigella dysenteriae 1 (Shiga)-like cytotoxin. Lancet 1984; 1: 77-78.
12. Ogura K, Terasaki Y, Miyoshi-Akiyama T, Terasaki M, Moss J, Noda M, et al. Vibrio cholerae cholix toxin-induced HepG2 cell death is enhanced by tumor necrosis factor-alpha through ROS and intracellular signal-regulated kinases. Toxicol Sci 2017; 156: 455-468.
13. Lugo MR, Merrill AR. The father, son and cholix toxin: the third member of the DT group mono-ADP-Ribosyltransferase toxin family. Toxins (Basel) 2015; 7: 2757-2772.
14. Jørgensen R, Purdy AE, Fieldhouse RJ, Kimber MS, Bartlett DH, Merrill AR. Cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae. J Biol Chem 2008; 283: 10671-10678.
15. Awasthi SP, Asakura M, Chowdhury N, Neogi SB, Hinenoya A, Golbar HM, et al. Novel cholix toxin variants, ADP-ribosylating toxins in Vibrio cholerae non-O1/non-O139 strains, and their pathogenicity. Infect Immun 2013; 81: 531-541.
16. Purdy AE, Balch D, Lizarraga-Partida ML, Islam MS, Martinez-Urtaza J, Huq A, et al. Diversity and distribution of cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae. Environ Microbiol Rep 2010; 2: 198-207.
17. Chatterjee S, Ghosh K, Raychoudhuri A, Chowdhury G, Bhattacharya MK, Mukhopadhyay AK, et al. Incidence, virulence factors, and clonality among clinical strains of non-O1, non-O139 Vibrio cholerae isolates from hospitalized diarrheal patients in Kolkata, India. J Clin Microbiol 2009; 47: 1087-1095.
18. Martinez RM, Megli CJ, Taylor RK. Growth and laboratory maintenance of Vibrio cholerae. Curr Protoc Microbiol 2010; Chapter 6:Unit 6A.1.
19. Dua P, Karmakar A, Ghosh C. Virulence gene profiles, biofilm formation, and antimicrobial resistance of Vibrio cholerae non-O1/non-O139 bacteria isolated from West Bengal, India. Heliyon 2018;4(12): e01040.
20. Islam A, Labbate M, Djordjevic SP, Alam M, Darling A, Melvold J, et al. Indigenous Vibrio cholerae strains from a non-endemic region are pathogenic. Open Biol 2013; 3: 120181.
21. Octavia S, Salim A, Kurniawan J, Lam C, Leung Q, Ahsan S, et al. Population structure and evolution of non-O1/non-O139 Vibrio cholerae by multilocus sequence typing. PLoS One 2013; 8(6): e65342.
22. Al-Ouqaili MTS, Khalaf EA, Al-Kubaisy SH. Molecular detection and sequencing of SHV gene encoding for extended-spectrum β-lactamases produced by multidrug resistance some of the Gram-negative bacteria. Int J Green Pharm 2019; 12: 910.
23. Schirmeister F, Dieckmann R, Bechlars S, Bier N, Faruque SM, Strauch E. Genetic and phenotypic analysis of Vibrio cholerae non-O1, non-O139 isolated from German and Austrian patients. Eur J Clin Microbiol Infect Dis 2014; 33: 767-778.
24. Simon NC, Aktories K, Barbieri JT. Novel bacterial ADP-ribosylating toxins: structure and function. Nat Rev Microbiol 2014; 12: 599-611.
25. Bakhshi B, Barzelighi HM, Adabi M, Lari AR, Pourshafie MR. A molecular survey on virulence associated genotypes of non-O1 non-O139 Vibrio cholerae in aquatic environment of Tehran, Iran. Water Res 2009; 43: 1441-1447.
26. Siriphap A, Leekitcharoenphon P, Kaas RS, Theethakaew C, Aarestrup FM, Sutheinkul O, et al. Characterization and genetic variation of Vibrio cholerae isolated from clinical and environmental sources in Thailand. PLoS One 2017; 12(1): e0169324.
27. Saidi SM, Chowdhury N, Awasthi SP, Asakura M, Hinenoya A, Iijima Y, et al. Prevalence of Vibrio cholerae O1 El Tor variant in a cholera-endemic zone of Kenya. J Med Microbiol 2014; 63: 415-420.
28. Ogura K, Yahiro K, Tsutsuki H, Nagasawa S, Yamasaki S, Moss J, et al. Characterization of Cholix toxin-induced apoptosis in HeLa cells. J Biol Chem 2011; 286: 37207-37215.
2. Rui H, Ritchie JM, Bronson RT, Mekalanos JJ, Zhang Y, Waldor MK. Reactogenicity of live-attenuated Vibrio cholerae vaccines is dependent on flagellins. Proc Natl Acad Sci U S A 2010; 107: 4359-4364.
3. Ceccarelli D, Chen A, Hasan NA, Rashed SM, Huq A, Colwell RR. Non-O1/non-O139 Vibrio cholerae carrying multiple virulence factors and V. cholerae O1 in the Chesapeake Bay, Maryland. Appl Environ Microbiol 2015; 81: 1909-1918.
4. Ramamurthy T, MukhopadhyayAK, Nandy RK, Balakrish Nair G (2013). Molecular Typing of Vibrio cholerae: Imprints in the Epidemiology of Cholera. In: Molecular typing in bacterial infections. Ed, De Filippis I, Mckee ML. Humana Press, 1St ed. New York, pp. 53-72.
5. Dalsgaard A, Forslund A, Hesselbjerg A, Bruun B. Clinical manifestations and characterization of extra-intestinal Vibrio cholerae non-O1, non-O139 infections in Denmark. Clin Microbiol Infect 2000; 6: 625-627.
6. Hao Y, Wang Y, Bi Z, Sun B, Jin Y, Bai Y, et al. A case of non-O1/non-O139 Vibrio cholerae septicemia and meningitis in a neonate. Int J Infect Dis 2015; 35: 117-119.
7. Shin OS, Tam VC, Suzuki M, Ritchie JM, Bronson RT, Waldor MK, et al. Type III secretion is essential for the rapidly fatal diarrheal disease caused by non-O1, non-O139 Vibrio cholerae. mBio 2011; 2(3): e00106-11.
8. Rajpara N, Vinothkumar K, Mohanty P, Singh AK, Singh R, Sinha R, et al. Synergistic effect of various virulence factors leading to high toxicity of environmental V. cholerae non-O1/ non-O139 isolates lacking ctx gene: comparative study with clinical strains. PLoS One 2013; 8(9):e76200.
9. Purdy A, Rohwer F, Edwards R, Azam F, Bartlett DH. A glimpse into the expanded genome content of Vibrio cholerae through identification of genes present in environmental strains. J Bacteriol 2005; 187: 2992-3001.
10. Awasthi SP, Asakura M, Neogi SB, Hinenoya A, Ramamurthy T, Yamasaki S. Development of a PCR-restriction fragment length polymorphism assay for detection and subtyping of cholix toxin variant genes of Vibrio cholerae. J Med Microbiol 2014; 63: 667-673.
11. O'Brien AD, Chen ME, Holmes RK, Kaper J, Levine MM. Environmental and human isolates of Vibrio cholerae and Vibrio parahaemolyticus produce a Shigella dysenteriae 1 (Shiga)-like cytotoxin. Lancet 1984; 1: 77-78.
12. Ogura K, Terasaki Y, Miyoshi-Akiyama T, Terasaki M, Moss J, Noda M, et al. Vibrio cholerae cholix toxin-induced HepG2 cell death is enhanced by tumor necrosis factor-alpha through ROS and intracellular signal-regulated kinases. Toxicol Sci 2017; 156: 455-468.
13. Lugo MR, Merrill AR. The father, son and cholix toxin: the third member of the DT group mono-ADP-Ribosyltransferase toxin family. Toxins (Basel) 2015; 7: 2757-2772.
14. Jørgensen R, Purdy AE, Fieldhouse RJ, Kimber MS, Bartlett DH, Merrill AR. Cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae. J Biol Chem 2008; 283: 10671-10678.
15. Awasthi SP, Asakura M, Chowdhury N, Neogi SB, Hinenoya A, Golbar HM, et al. Novel cholix toxin variants, ADP-ribosylating toxins in Vibrio cholerae non-O1/non-O139 strains, and their pathogenicity. Infect Immun 2013; 81: 531-541.
16. Purdy AE, Balch D, Lizarraga-Partida ML, Islam MS, Martinez-Urtaza J, Huq A, et al. Diversity and distribution of cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae. Environ Microbiol Rep 2010; 2: 198-207.
17. Chatterjee S, Ghosh K, Raychoudhuri A, Chowdhury G, Bhattacharya MK, Mukhopadhyay AK, et al. Incidence, virulence factors, and clonality among clinical strains of non-O1, non-O139 Vibrio cholerae isolates from hospitalized diarrheal patients in Kolkata, India. J Clin Microbiol 2009; 47: 1087-1095.
18. Martinez RM, Megli CJ, Taylor RK. Growth and laboratory maintenance of Vibrio cholerae. Curr Protoc Microbiol 2010; Chapter 6:Unit 6A.1.
19. Dua P, Karmakar A, Ghosh C. Virulence gene profiles, biofilm formation, and antimicrobial resistance of Vibrio cholerae non-O1/non-O139 bacteria isolated from West Bengal, India. Heliyon 2018;4(12): e01040.
20. Islam A, Labbate M, Djordjevic SP, Alam M, Darling A, Melvold J, et al. Indigenous Vibrio cholerae strains from a non-endemic region are pathogenic. Open Biol 2013; 3: 120181.
21. Octavia S, Salim A, Kurniawan J, Lam C, Leung Q, Ahsan S, et al. Population structure and evolution of non-O1/non-O139 Vibrio cholerae by multilocus sequence typing. PLoS One 2013; 8(6): e65342.
22. Al-Ouqaili MTS, Khalaf EA, Al-Kubaisy SH. Molecular detection and sequencing of SHV gene encoding for extended-spectrum β-lactamases produced by multidrug resistance some of the Gram-negative bacteria. Int J Green Pharm 2019; 12: 910.
23. Schirmeister F, Dieckmann R, Bechlars S, Bier N, Faruque SM, Strauch E. Genetic and phenotypic analysis of Vibrio cholerae non-O1, non-O139 isolated from German and Austrian patients. Eur J Clin Microbiol Infect Dis 2014; 33: 767-778.
24. Simon NC, Aktories K, Barbieri JT. Novel bacterial ADP-ribosylating toxins: structure and function. Nat Rev Microbiol 2014; 12: 599-611.
25. Bakhshi B, Barzelighi HM, Adabi M, Lari AR, Pourshafie MR. A molecular survey on virulence associated genotypes of non-O1 non-O139 Vibrio cholerae in aquatic environment of Tehran, Iran. Water Res 2009; 43: 1441-1447.
26. Siriphap A, Leekitcharoenphon P, Kaas RS, Theethakaew C, Aarestrup FM, Sutheinkul O, et al. Characterization and genetic variation of Vibrio cholerae isolated from clinical and environmental sources in Thailand. PLoS One 2017; 12(1): e0169324.
27. Saidi SM, Chowdhury N, Awasthi SP, Asakura M, Hinenoya A, Iijima Y, et al. Prevalence of Vibrio cholerae O1 El Tor variant in a cholera-endemic zone of Kenya. J Med Microbiol 2014; 63: 415-420.
28. Ogura K, Yahiro K, Tsutsuki H, Nagasawa S, Yamasaki S, Moss J, et al. Characterization of Cholix toxin-induced apoptosis in HeLa cells. J Biol Chem 2011; 286: 37207-37215.
| Files | ||
| Issue | Vol 12 No 4 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v12i4.3929 | |
| Keywords | ||
| Vibrio cholerae; Cholix toxin; Polymerase chain reaction | ||
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How to Cite
1.
Gholizadeh Tangestani M, Alinezhad J, Khajeian A, Gharibi S, Haghighi MA. Identification of cholix toxin gene in Vibrio cholerae non-O1/non-O139 isolated from diarrhea patients in Bushehr, Iran. Iran J Microbiol. 2020;12(4):273-280.
