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Home / Equine Research Bank / J Clin Microbiol / Regulated expression of the beta2-toxin gene (cpb2) in Clost...
Journal of clinical microbiology2005; 43(8); 4002-4009; doi: 10.1128/JCM.43.8.4002-4009.2005

Regulated expression of the beta2-toxin gene (cpb2) in Clostridium perfringens type a isolates from horses with gastrointestinal diseases.

Abstract: Recent epidemiological studies suggested that cpb2-positive Clostridium perfringens isolates are associated with gastrointestinal (GI) diseases in horses. These putative relationships, indicated by PCR genotyping, were tested in the present study by further genotyping and phenotyping of 23 cpb2-positive C. perfringens isolates from horses with GI disease (referred to hereafter as horse GI disease isolates). Our beta2-toxin (CPB2) Western blot analyses demonstrated that all of the tested isolates were unable to produce detectable levels of CPB2. However, Southern blot and nucleotide sequencing analyses identified intact cpb2 open reading frames in all of our surveyed horse GI disease isolates. Furthermore, reverse transcriptase PCR and Northern blot analyses showed that cpb2 genes in all of our surveyed horse GI disease isolates were transcriptionally active, i.e., an approximately 1.2-kb cpb2-specific mRNA was identified in total RNA from our surveyed isolates. The levels of cpb2 mRNA in CWC245 (a high-CPB2-producing pig strain) and our surveyed horse GI disease isolates differed to such an extent (35-fold) that this difference could be considered as a major cause of the difference in levels of CPB2 production by CWC245 and horse GI disease isolates. This finding received further support from our observation that the complementing strain 106902(pMRS140), which produced significantly higher levels of mRNA than strain 106902, produced high levels of CPB2. Collectively, our results indicated that there is a positive correlation between cpb2 transcription levels and the amount of CPB2 produced by a C. perfringens cell and that decreased transcription and/or message instability may be involved, at least in part, in the low CPB2 production noted for horse GI disease isolates in comparison to that noted for pig GI disease isolate CWC245.
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Publication Date: 2005-08-06 PubMed ID: 16081942PubMed Central: PMC1233996DOI: 10.1128/JCM.43.8.4002-4009.2005Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study explores the link between Clostridium perfringens, a bacterium, and gastrointestinal diseases in horses. It finds that while the bacteria are present in the horses and have the potential to produce a toxin that could cause the disease, they are not producing it at detectable levels, though they are still active on a genetic level.

Objective of the Research

  • This research was conducted to test the previously suggested association between cpb2-positive Clostridium perfringens isolates and gastrointestinal diseases in horses. Various genotyping and phenotyping techniques were used to analyze 23 such isolates from horses with GI disease.

Testing Method and Analyses

  • The researchers used Western blot analyses to determine if the tested bacterial isolates can produce a specific toxin (CPB2). However, no detectable levels of this toxin were found. An absence of this toxin suggests that these bacteria may not be directly causing the observed GI diseases in horses.
  • Despite the inability of the isolates to produce the toxin, the researchers found intact cpb2 genes in all of the tested samples via Southern blot and nucleotide sequencing, showing that the potential is there.
  • Through the use of reverse transcriptase PCR and Northern blot analyses, the researchers found that the cpb2 genes were transcriptionally active, meaning they were still producing a reticular type of RNA.

Comparison of cpb2 transcription levels

  • The levels of the cpb2 gene varied between the CWC245 (a strain from pigs known to produce high levels of CPB2 toxin) and the horse GI disease isolates. This suggests that the difference in levels of CPB2 production might come from how active the cpb2 gene is in these bacteria.
  • This possibility is further supported by a strain that produced higher levels of mRNA (a type of RNA) and a high level of the CPB2 toxin.

Conclusion

  • From this study, a positive correlation between cpb2 transcription levels and the amount of CPB2 produced became evident. Moreover, decreased transcription and/or message instability might be involved, at least in part, in the low CPB2 production observed for horse GI disease isolates as compared to the pig GI disease isolate CWC245.

Cite This Article

APA
Waters M, Raju D, Garmory HS, Popoff MR, Sarker MR. (2005). Regulated expression of the beta2-toxin gene (cpb2) in Clostridium perfringens type a isolates from horses with gastrointestinal diseases. J Clin Microbiol, 43(8), 4002-4009. https://doi.org/10.1128/JCM.43.8.4002-4009.2005

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 43
Issue: 8
Pages: 4002-4009

Researcher Affiliations

Waters, Michael
  • Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA.
Raju, Deepa
    Garmory, Helen S
      Popoff, Michel R
        Sarker, Mahfuzur R

          MeSH Terms

          • Animals
          • Bacterial Toxins / biosynthesis
          • Bacterial Toxins / chemistry
          • Bacterial Toxins / genetics
          • Clostridium perfringens / genetics
          • Electrophoresis, Gel, Pulsed-Field
          • Gastrointestinal Diseases / microbiology
          • Gastrointestinal Diseases / veterinary
          • Gene Expression Regulation, Bacterial
          • Genotype
          • Horse Diseases / microbiology
          • Horses
          • Open Reading Frames
          • Polymorphism, Restriction Fragment Length
          • Reverse Transcriptase Polymerase Chain Reaction

          References

          This article includes 29 references
          1. Bacciarini LN, Pagan O, Frey J, Gröne A. Clostridium perfringens beta2-toxin in an African elephant (Loxodonta africana) with ulcerative enteritis.. Vet Rec 2001 Nov 17;149(20):618-20.
            pubmed: 11761293doi: 10.1136/vr.149.20.618google scholar: lookup
          2. Bacciarini LN, Boerlin P, Straub R, Frey J, Gröne A. Immunohistochemical localization of Clostridium perfringens beta2-toxin in the gastrointestinal tract of horses.. Vet Pathol 2003 Jul;40(4):376-81.
            pubmed: 12824509doi: 10.1354/vp.40-4-376google scholar: lookup
          3. Bannam TL, Rood JI. Clostridium perfringens-Escherichia coli shuttle vectors that carry single antibiotic resistance determinants.. Plasmid 1993 May;29(3):233-5.
            pubmed: 8356117doi: 10.1006/plas.1993.1025google scholar: lookup
          4. Brynestad S, Sarker MR, McClane BA, Granum PE, Rood JI. Enterotoxin plasmid from Clostridium perfringens is conjugative.. Infect Immun 2001 May;69(5):3483-7.
            pmc: PMC98316pubmed: 11292780doi: 10.1128/iai.69.5.3483-3487.2001google scholar: lookup
          5. Bueschel DM, Jost BH, Billington SJ, Trinh HT, Songer JG. Prevalence of cpb2, encoding beta2 toxin, in Clostridium perfringens field isolates: correlation of genotype with phenotype.. Vet Microbiol 2003 Jul 1;94(2):121-9.
            pubmed: 12781480doi: 10.1016/s0378-1135(03)00081-6google scholar: lookup
          6. Collie RE, McClane BA. Evidence that the enterotoxin gene can be episomal in Clostridium perfringens isolates associated with non-food-borne human gastrointestinal diseases.. J Clin Microbiol 1998 Jan;36(1):30-6.
            pmc: PMC124802pubmed: 9431915doi: 10.1128/jcm.36.1.30-36.1998google scholar: lookup
          7. Cornillot E, Saint-Joanis B, Daube G, Katayama S, Granum PE, Canard B, Cole ST. The enterotoxin gene (cpe) of Clostridium perfringens can be chromosomal or plasmid-borne.. Mol Microbiol 1995 Feb;15(4):639-47.
            pubmed: 7783636doi: 10.1111/j.1365-2958.1995.tb02373.xgoogle scholar: lookup
          8. Czeczulin JR, Collie RE, McClane BA. Regulated expression of Clostridium perfringens enterotoxin in naturally cpe-negative type A, B, and C isolates of C. perfringens.. Infect Immun 1996 Aug;64(8):3301-9.
            pmc: PMC174222pubmed: 8757868doi: 10.1128/iai.64.8.3301-3309.1996google scholar: lookup
          9. Fisher DJ, Miyamoto K, Harrison B, Akimoto S, Sarker MR, McClane BA. Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene.. Mol Microbiol 2005 May;56(3):747-62.
            pubmed: 15819629doi: 10.1111/j.1365-2958.2005.04573.xgoogle scholar: lookup
          10. Garmory HS, Chanter N, French NP, Bueschel D, Songer JG, Titball RW. Occurrence of Clostridium perfringens beta2-toxin amongst animals, determined using genotyping and subtyping PCR assays.. Epidemiol Infect 2000 Feb;124(1):61-7.
            pmc: PMC2810884pubmed: 10722131doi: 10.1017/s0950268899003295google scholar: lookup
          11. Gibert M, Jolivet-Reynaud C, Popoff MR. Beta2 toxin, a novel toxin produced by Clostridium perfringens.. Gene 1997 Dec 5;203(1):65-73.
            pubmed: 9426008doi: 10.1016/s0378-1119(97)00493-9google scholar: lookup
          12. Hansen MV, Elliott LP. New presumptive identification test for Clostridium perfringens: reverse CAMP test.. J Clin Microbiol 1980 Oct;12(4):617-9.
            pmc: PMC273650pubmed: 6252247doi: 10.1128/jcm.12.4.617-619.1980google scholar: lookup
          13. Herholz C, Miserez R, Nicolet J, Frey J, Popoff M, Gibert M, Gerber H, Straub R. Prevalence of beta2-toxigenic Clostridium perfringens in horses with intestinal disorders.. J Clin Microbiol 1999 Feb;37(2):358-61.
            pmc: PMC84307pubmed: 9889218doi: 10.1128/jcm.37.2.358-361.1999google scholar: lookup
          14. Huang IH, Waters M, Grau RR, Sarker MR. Disruption of the gene (spo0A) encoding sporulation transcription factor blocks endospore formation and enterotoxin production in enterotoxigenic Clostridium perfringens type A.. FEMS Microbiol Lett 2004 Apr 15;233(2):233-40.
            pubmed: 15063491doi: 10.1016/j.femsle.2004.02.014google scholar: lookup
          15. Jolivet-Reynaud C, Popoff MR, Vinit MA, Ravisse P, Moreau H, Alouf JE. Enteropathogenicity of Clostridium perfringens beta toxin and other clostridial toxins. Zentbl. Bakteriol. S15:145-151.
          16. Jost BH, Billington SJ, Trinh HT, Bueschel DM, Songer JG. Atypical cpb2 genes, encoding beta2-toxin in Clostridium perfringens isolates of nonporcine origin.. Infect Immun 2005 Jan;73(1):652-6.
            pmc: PMC538998pubmed: 15618211doi: 10.1128/iai.73.1.652-656.2005google scholar: lookup
          17. Katayama S, Matsushita O, Minami J, Mizobuchi S, Okabe A. Comparison of the alpha-toxin genes of Clostridium perfringens type A and C strains: evidence for extragenic regulation of transcription.. Infect Immun 1993 Feb;61(2):457-63.
            pmc: PMC302750pubmed: 8423073doi: 10.1128/iai.61.2.457-463.1993google scholar: lookup
          18. Klaasen HL, Molkenboer MJ, Bakker J, Miserez R, Häni H, Frey J, Popoff MR, van den Bosch JF. Detection of the beta2 toxin gene of Clostridium perfringens in diarrhoeic piglets in The Netherlands and Switzerland.. FEMS Immunol Med Microbiol 1999 Jul;24(3):325-32.
            pubmed: 10397318doi: 10.1111/j.1574-695x.1999.tb01301.xgoogle scholar: lookup
          19. Manteca C, Daube G, Jauniaux T, Linden A, Pirson V, Detilleux J, Ginter A, Coppe P, Kaeckenbeeck A, Mainil JG. A role for the Clostridium perfringens beta2 toxin in bovine enterotoxaemia?. Vet Microbiol 2002 May 1;86(3):191-202.
            pmc: PMC7117356pubmed: 11900954doi: 10.1016/s0378-1135(02)00008-1google scholar: lookup
          20. McClane, B. A. 2001. Clostridium perfringens, p. 351-372. In M. P. Doyle, L. R. Beuchat, and T. J. Montville (ed.), Food microbiology: fundamentals and frontiers, 2nd ed. ASM Press, Washington, D.C.
          21. Mcdonel, J. L. 1986. Toxins of Clostridium perfringens type A, B, C, D, and E, p. 477-517. In F. Dorner and H. Drews (ed.), Pharmacology of bacterial toxins. Pergamon Press, Oxford, United Kingdom.
          22. Ohtani K, Kawsar HI, Okumura K, Hayashi H, Shimizu T. The VirR/VirS regulatory cascade affects transcription of plasmid-encoded putative virulence genes in Clostridium perfringens strain 13.. FEMS Microbiol Lett 2003 May 16;222(1):137-41.
            pubmed: 12757957doi: 10.1016/s0378-1097(03)00255-6google scholar: lookup
          23. Sarker MR, Carman RJ, McClane BA. Inactivation of the gene (cpe) encoding Clostridium perfringens enterotoxin eliminates the ability of two cpe-positive C. perfringens type A human gastrointestinal disease isolates to affect rabbit ileal loops.. Mol Microbiol 1999 Sep;33(5):946-58.
            pubmed: 10476029doi: 10.1046/j.1365-2958.1999.01534.xgoogle scholar: lookup
          24. Sarker MR, Shivers RP, Sparks SG, Juneja VK, McClane BA. Comparative experiments to examine the effects of heating on vegetative cells and spores of Clostridium perfringens isolates carrying plasmid genes versus chromosomal enterotoxin genes.. Appl Environ Microbiol 2000 Aug;66(8):3234-40.
            pmc: PMC92139pubmed: 10919775doi: 10.1128/aem.66.8.3234-3240.2000google scholar: lookup
          25. Sparks SG, Carman RJ, Sarker MR, McClane BA. Genotyping of enterotoxigenic Clostridium perfringens fecal isolates associated with antibiotic-associated diarrhea and food poisoning in North America.. J Clin Microbiol 2001 Mar;39(3):883-8.
            pmc: PMC87845pubmed: 11230399doi: 10.1128/jcm.39.3.883-888.2001google scholar: lookup
          26. Songer JG. Clostridial enteric diseases of domestic animals.. Clin Microbiol Rev 1996 Apr;9(2):216-34.
            pmc: PMC172891pubmed: 8964036doi: 10.1128/cmr.9.2.216google scholar: lookup
          27. Thiede S, Goethe R, Amtsberg G. Prevalence of beta2 toxin gene of Clostridium perfringens type A from diarrhoeic dogs.. Vet Rec 2001 Sep 1;149(9):273-4.
            pubmed: 11558663doi: 10.1136/vr.149.9.273google scholar: lookup
          28. Tsutsui K, Minami J, Matsushita O, Katayama S, Taniguchi Y, Nakamura S, Nishioka M, Okabe A. Phylogenetic analysis of phospholipase C genes from Clostridium perfringens types A to E and Clostridium novyi.. J Bacteriol 1995 Dec;177(24):7164-70.
            pmc: PMC177596pubmed: 8522524doi: 10.1128/jb.177.24.7164-7170.1995google scholar: lookup
          29. Waters M, Savoie A, Garmory HS, Bueschel D, Popoff MR, Songer JG, Titball RW, McClane BA, Sarker MR. Genotyping and phenotyping of beta2-toxigenic Clostridium perfringens fecal isolates associated with gastrointestinal diseases in piglets.. J Clin Microbiol 2003 Aug;41(8):3584-91.
            pmc: PMC179868pubmed: 12904359doi: 10.1128/jcm.41.8.3584-3591.2003google scholar: lookup

          Citations

          This article has been cited 12 times.
          1. Khan MZ, Li F, Huang X, Nouman M, Bibi R, Fan X, Zhou H, Shan Z, Wang L, Jiang Y, Cui W, Qiao X, Li Y, Wang X, Tang L. Oral Immunization of Chickens with Probiotic Lactobacillus crispatus Constitutively Expressing the α-β2-ε-β1 Toxoids to Induce Protective Immunity. Vaccines (Basel) 2022 Apr 29;10(5).
            doi: 10.3390/vaccines10050698pubmed: 35632454google scholar: lookup
          2. Kao WM, Chang CR, Chang TJ, Li SY, Chen WJ, Chau CF. Inclusion of Fructooligosaccharide and Resistant Maltodextrin in High Fat Diets Promotes Simultaneous Improvements on Body Fat Reduction and Fecal Parameters. Molecules 2018 Aug 28;23(9).
            doi: 10.3390/molecules23092169pubmed: 30154352google scholar: lookup
          3. Shaw SD, Stämpfli H. Diagnosis and Treatment of Undifferentiated and Infectious Acute Diarrhea in the Adult Horse. Vet Clin North Am Equine Pract 2018 Apr;34(1):39-53.
            doi: 10.1016/j.cveq.2017.11.002pubmed: 29426709google scholar: lookup
          4. Serroni A, Magistrali CF, Pezzotti G, Bano L, Pellegrini M, Severi G, Di Pancrazio C, Luciani M, Tittarelli M, Tofani S, De Giuseppe A. Expression of deleted, atoxic atypical recombinant beta2 toxin in a baculovirus system and production of polyclonal and monoclonal antibodies. Microb Cell Fact 2017 May 25;16(1):94.
            doi: 10.1186/s12934-017-0707-8pubmed: 28545467google scholar: lookup
          5. Mehdizadeh Gohari I, Parreira VR, Nowell VJ, Nicholson VM, Oliphant K, Prescott JF. A novel pore-forming toxin in type A Clostridium perfringens is associated with both fatal canine hemorrhagic gastroenteritis and fatal foal necrotizing enterocolitis. PLoS One 2015;10(4):e0122684.
            doi: 10.1371/journal.pone.0122684pubmed: 25853427google scholar: lookup
          6. Uzal FA, Vidal JE, McClane BA, Gurjar AA. Clostridium Perfringens Toxins Involved in Mammalian Veterinary Diseases. Open Toxinology J 2010;2:24-42.
            pubmed: 24511335
          7. Mendez M, Huang IH, Ohtani K, Grau R, Shimizu T, Sarker MR. Carbon catabolite repression of type IV pilus-dependent gliding motility in the anaerobic pathogen Clostridium perfringens. J Bacteriol 2008 Jan;190(1):48-60.
            doi: 10.1128/JB.01407-07pubmed: 17981974google scholar: lookup
          8. Gurjar AA, Yennawar NH, Yennawar HP, Rajashankar KR, Hegde NV, Jayarao BM. Expression, crystallization and preliminary X-ray diffraction studies of recombinant Clostridium perfringens beta 2-toxin. Acta Crystallogr Sect F Struct Biol Cryst Commun 2007 Jun 1;63(Pt 6):484-7.
            doi: 10.1107/S1744309107020313pubmed: 17554168google scholar: lookup
          9. Fernandez-Miyakawa ME, Fisher DJ, Poon R, Sayeed S, Adams V, Rood JI, McClane BA, Uzal FA. Both epsilon-toxin and beta-toxin are important for the lethal properties of Clostridium perfringens type B isolates in the mouse intravenous injection model. Infect Immun 2007 Mar;75(3):1443-52.
            doi: 10.1128/IAI.01672-06pubmed: 17210666google scholar: lookup
          10. Johansson A, Aspan A, Bagge E, Båverud V, Engström BE, Johansson KE. Genetic diversity of Clostridium perfringens type A isolates from animals, food poisoning outbreaks and sludge. BMC Microbiol 2006 May 31;6:47.
            doi: 10.1186/1471-2180-6-47pubmed: 16737528google scholar: lookup
          11. Harrison B, Raju D, Garmory HS, Brett MM, Titball RW, Sarker MR. Molecular characterization of Clostridium perfringens isolates from humans with sporadic diarrhea: evidence for transcriptional regulation of the beta2-toxin-encoding gene. Appl Environ Microbiol 2005 Dec;71(12):8362-70.
            doi: 10.1128/AEM.71.12.8362-8370.2005pubmed: 16332823google scholar: lookup
          12. Talukdar PK, Alnoman M, Sarker MR. Identification of Germinants and Expression of Germination Genes in Clostridium perfringens Strains Isolated from Diarrheic Animals. Pathogens 2024 Feb 22;13(3).
            doi: 10.3390/pathogens13030194pubmed: 38535537google scholar: lookup
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