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Vibrio vulnificus Klinik İzolatlarının Virülens Genlerine Dayalı Olarak Selektif Zenginleştirme Ortamının Geliştirilmesi

Yıl 2019, Cilt: 8 Sayı: 2, 173 - 179, 25.12.2019
https://doi.org/10.31196/huvfd.667690

Öz

Bu çalışmanın amacı, farklı zenginleştirme besiyerlerinin V. vulnificus'un klinik ve çevresel suşlarının seçimini etkileyip etkilemediğini bulmaktı. Bu nedenle, geleneksel zenginleştirme ve koruyucu besiyerlerinin selektif özellikleri, V. vulnificus’ un klinik ve çevresel suşları için test edildi. Laboratuarda, aynı tuz konsantrasyonuna sahip Alkali Peptonlu Su (APW) ve Beyin Kalp İnfüzyon Broth (BHIB) eşit konsantrasyonlarda klinik ve çevresel suşlar ile ekimi yapıldı ve 35 °C' de 20 saat boyunca inkübe edildi. Kantitatif Polimeraz Zincir Reaksiyon (PZR) sayım sonuçları; BHIB’ de geliştirilen suşların %85’ ini klinik suşlar olduğunu göstermiştir. Bu oran vcgC geninin vvhA genine yüzdelik hesaplamalarıyla belirlenmiştir. APW' de klinik suşların (47%) çevresel suşlara (53%) oranı neredeyse eşitti. BHIB, APW ve Luria Bertani (LB) broth ile zenginleştirilen deniz suyu numunelerinin büyük kısmında, klinik suşlarının toplam suşlara oranı BHIB’de yüksek çıkarken, APW ve LB’de çıkmamıştır. Ancak, deniz suyu örneklerinde klinik veya çevresel suşların birbirine oranında mevsimsel farklılıklar zenginleşme sürecini etkilemiştir. Çalışmamız BHIB10'un virülans faktörlerini çalışmak için avantajlı olabileceğini ve APW10'un V. vulnificus'ta izolasyon ve MPN sayımı için uygun olabileceğini göstermektedir. Tuzluluk oranı %10 olan BHIB besiyerinin klinik suşlar için selektif özelliği, demirin insanlarda oluşturduğu infeksiyonlarda önemli bir rol oynadığını gösterir.

Kaynakça

  • Abbott SL, Janda M, Farmer JJ, 2011: Vibrio and related organisms. In ''Manual of Clinical Microbiology'', Ed; Versalaovic J, Carroll KC, Jorgensen JH, Funke G, Landry ML, Warnock DW Inc., 10th ed. Washington, D.C. ASM Press. pp. 666-76.
  • Alice AF, Naka H, Crosa JH, 2008: Global gene expression as a function of the iron status of the bacterial cell: Influence of differentially expressed genes in the virulence of the human pathogen Vibrio vulnificus. Infect Immun, 76, 4019-37.
  • Cam S, 2016a: Environmental Influences on Virulence Factors in Vibrio Vulnificus. PhD thesis, Texas A&M University, USA, pp. 67-99
  • Campbell MS, Wright AC, 2003: Real-time PCR analysis of Vibrio vulnificus from oysters. Appl Environ Microbiol, 69, 7137-44.
  • Chase E, Harwood VJ, 2011: Comparison of the effects of environmental parameters on growth rates of Vibrio vulnificus biotypes I, II, and III by culture and quantitative PCR analysis. Appl Environ Microbiol, 77, 4200-7.
  • Çam S, Brinkmeyer R, Schwarz JR, 2019: Quantitative PCR enumeration of vcgC and 16S rRNA type A and B genes as virulence indicators for environmental and clinical strains of Vibrio vulnificus in Galveston Bay oysters. Can J Microbiol, 65, 613-621.
  • DePaola A, Capers GM, Alexander D, 1994: Densities of Vibrio vulnificus in the intestines of fish from the U.S. Gulf Coast. Appl Environ Microbiol, 60, 984-8.
  • Elliot EL, Kaysner CA, Jackson HJ, Tamplin ML, 1995: V. cholerae, V. vulnificus, and other Vibrio spp. FDA Bacteriological Analytical Manual. Arlington, USA.
  • Gholami P, Lew SQ, Klontz KC, 1998: Raw shellfish consumption among renal disease patients. A risk factor for severe Vibrio vulnificus infection. Am J Prev Med, 15, 243-5.
  • Gordon KV, Vickery MC, DePaola A, Staley C, Harwood VJ, 2008: Real-time PCR assays for quantification and differentiation of Vibrio vulnificus strains in oysters and water. Appl Environ Microbiol, 74, 1704-9.
  • Gulig PA, Bourdage KL, Starks AM, 2005: Molecular pathogenesis of Vibrio vulnificus. J Microbiol, 43, 118-31.
  • Han F, Pu S, Hou A, Ge B, 2009: Characterization of clinical and environmental types of Vibrio vulnificus isolates from Louisiana oysters. Foodborne Pathog Dis, 6, 1251-8.
  • Han F, Wang F, Ge B, 2011: Detecting potentially virulent Vibrio vulnificus strains in raw oysters by quantitative loop-mediated isothermal amplification. Appl Environ Microbiol, 77, 2589-95.
  • Hlady WG, 1997: Vibrio infections associated with raw oyster consumption in Florida, 1981–1994. J Food Prot, 60, 353-7.
  • Huebers HA, Eng MJ, Josephson BM, Ekpoom N, Rettmer RL, Labbe RF, Pootrakul P, Finch CA, 1987: Plasma iron and transferrin iron-binding capacity evaluated by colorimetric and immunoprecipitation methods. Clin Chem, 33, 273-7.
  • Høi L, Dalsgaard I, Dalsgaard A, 1998: Improved isolation of Vibrio vulnificus from seawater and sediment with cellobiose-colistin agar. Appl Environ Microbiol, 64, 1721-4.
  • Jackson JK, Murphree RL, Tamplin ML, 1997: Evidence that mortality from Vibrio vulnificus infection results from single strains among heterogeneous populations in shellfish. J Clin Microbiol, 35, 2098-101.
  • Jones MK, Oliver JD, 2009: Vibrio vulnificus: Disease and pathogenesis. Infect Immun, 77, 1723-33.
  • Kaysner CA, DePaola A, 2004: Vibrio. In ''Bacteriological Analytical Manual'', Ed; Hammack T, Davidson MW, Feng P, Gharst G, Ge B, Jinneman K, Regan PM, Kase J, Orlandi P, Burkhardt W, U.S. Food and Drug Administration, Washington, D.C..
  • Kim CM, Chung YY, Shin SH, 2009: Iron differentially regulates gene expression and extracellular secretion of Vibrio vulnificus cytolysin-hemolysin. J Infect Dis, 200, 582-9.
  • Kobayashi T, Enomoto S, Sakazaki R, Kuwahara S, 1963: A new selective isolation medium for Vibrio group on a modified Nakanishi's medium (TCBS agar medium). Jpn J Bacteriol, 18, 387-92.
  • Lin M, Schwarz JR, 2003: Seasonal shifts in population structure of Vibrio vulnificus in an estuarine environment as revealed by partial 16S ribosomal DNA sequencing. FEMS Microbiol Ecol, 45, 23-7.
  • Masad G, Oliver JD, 1987: New selective and differential medium for Vibrio cholerae and Vibrio vulnificus. Appl Environ Microbiol, 53, 2262-4.
  • Nwafia WC, Aneke JO, Okonji CU, 2006: Serum iron and total iron binding capacity levels among the ABO blood groups in Enugu, South Eastern Nigeria. Niger J Physiol Sci, 21, 9-14.
  • Oliver JD, Warner RA, Cleland DR, 1983: Distribution of Vibrio vulnificus and other lactose-fermenting Vibrios in the marine environment. Appl Environ Microbiol, 45, 985-98.
  • Panicker G, Myers ML, Bej AK, 2004: Rapid detection of Vibrio vulnificus in shellfish and Gulf of Mexico water by real-time PCR. Appl Environ Microbiol, 70, 498-507.
  • Potasman I, Paz A, Odeh M, 2002: Infectious outbreaks associated with bivalve shellfish consumption: A worldwide perspective. Clin Infect Dis, 35, 921-8.
  • Randa MA, Polz MF, Lim E, 2004: Effects of temperature and salinity on Vibrio vulnificus population dynamics as assessed by quantitative PCR. Appl Environ Microbiol, 70, 5469-76.
  • Rosche TM, Smith DJ, Parker EE, Oliver JD, 2005: A rapid and simple PCR analysis indicates there are two subgroups of Vibrio vulnificus which correlate with clinical or environmental isolation. Microbiol Immunol, 49, 381-9.
  • Sanjuán E, Fouz B, Oliver JD, Amaro C, 2009: Evaluation of genotypic and phenotypic methods to distinguish clinical from environmental Vibrio vulnificus Strains. Appl Environ Microbiol, 75, 1604-13.
  • Starks AM, Schoeb TR, Tamplin ML, Parveen S, Doyle TJ, Bomeisl PE, Escudero GM, Gulig PA, 2000: Pathogenesis of infection by clinical and environmental strains of Vibrio vulnificus in iron-dextran-treated mice. Infect Immun, 68, 5785-93.
  • Strom MS, Paranjpye RN, 2000: Epidemiology and pathogenesis of Vibrio vulnificus. Microb Infect, 2, 177-88.
  • Tamplin ML, Martin AL, Ruple AD, Cook DW, Kaspar CW, 1991: Enzyme immunoassay for identification of Vibrio vulnificus in seawater, sediment, and oysters. Appl Environ Microbiol, 57, 1235-40.
  • Tison DL, Kelly MT, 1984: Factors affecting hemolysin production by Vibrio vulnificus. Curr Microbiol, 10, 181-4.
  • Warner E, Oliver JD, 2007: Refined medium for direct isolation of Vibrio vulnificus from oyster tissue and seawater. Appl Environ Microbiol, 73, 3098-100.
  • Warner JM, Oliver JD, 1999: Randomly amplified polymorphic DNA analysis of clinical and environmental isolates of Vibrio vulnificus and other Vibrio species. Appl Environ Microbiol, 65, 1141-4.
  • Wright AC, Hill RT, Johnson JA, Roghman M-C, Colwell RR, Morris JG, 1996: Distribution of Vibrio vulnificus in the Chesapeake Bay. Appl Environ Microbiol, 62, 717-24.

Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes

Yıl 2019, Cilt: 8 Sayı: 2, 173 - 179, 25.12.2019
https://doi.org/10.31196/huvfd.667690

Öz

The objective of this study was to find out if different enrichment media would influence the selection of clinical and environmental strains of Vibrio vulnificus. Therefore, traditional enrichment and maintenance medium for V. vulnificus were tested for selective preference for clinical or environmental strains. In the laboratory, Alkaline Peptone Water (APW) and Brain Heart Infusion Broth (BHIB), with adjusted salinity to that of APW, were inoculated with equal concentrations of clinical and environmental strains and incubated for 20 hr at 35 °C. BHIB selected for clinical strains (85%) as indicated by percentage of vcgC to vvhA genes, enumerated with quantitative PCR. In APW, the ratio of clinical (47%) to environmental strains (53%) was roughly equal. Enrichments of seawater with BHIB, APW, and Luria Bertani broth (LB) resulted in higher percentage of clinical strains in BHIB but not in APW or LB, in most samples. However seasonal differences in predominance of clinical or environmental strains in the seawater samples influenced the enrichment process. Our study suggests that BHIB10 might be advantageous for studying virulence factors and APW10 could be suitable for isolation and MPN enumeration in V. vulnificus. The selective preference of the BHIB10 medium for clinical strains indicates that iron plays an important role in the infection of the human host.

Kaynakça

  • Abbott SL, Janda M, Farmer JJ, 2011: Vibrio and related organisms. In ''Manual of Clinical Microbiology'', Ed; Versalaovic J, Carroll KC, Jorgensen JH, Funke G, Landry ML, Warnock DW Inc., 10th ed. Washington, D.C. ASM Press. pp. 666-76.
  • Alice AF, Naka H, Crosa JH, 2008: Global gene expression as a function of the iron status of the bacterial cell: Influence of differentially expressed genes in the virulence of the human pathogen Vibrio vulnificus. Infect Immun, 76, 4019-37.
  • Cam S, 2016a: Environmental Influences on Virulence Factors in Vibrio Vulnificus. PhD thesis, Texas A&M University, USA, pp. 67-99
  • Campbell MS, Wright AC, 2003: Real-time PCR analysis of Vibrio vulnificus from oysters. Appl Environ Microbiol, 69, 7137-44.
  • Chase E, Harwood VJ, 2011: Comparison of the effects of environmental parameters on growth rates of Vibrio vulnificus biotypes I, II, and III by culture and quantitative PCR analysis. Appl Environ Microbiol, 77, 4200-7.
  • Çam S, Brinkmeyer R, Schwarz JR, 2019: Quantitative PCR enumeration of vcgC and 16S rRNA type A and B genes as virulence indicators for environmental and clinical strains of Vibrio vulnificus in Galveston Bay oysters. Can J Microbiol, 65, 613-621.
  • DePaola A, Capers GM, Alexander D, 1994: Densities of Vibrio vulnificus in the intestines of fish from the U.S. Gulf Coast. Appl Environ Microbiol, 60, 984-8.
  • Elliot EL, Kaysner CA, Jackson HJ, Tamplin ML, 1995: V. cholerae, V. vulnificus, and other Vibrio spp. FDA Bacteriological Analytical Manual. Arlington, USA.
  • Gholami P, Lew SQ, Klontz KC, 1998: Raw shellfish consumption among renal disease patients. A risk factor for severe Vibrio vulnificus infection. Am J Prev Med, 15, 243-5.
  • Gordon KV, Vickery MC, DePaola A, Staley C, Harwood VJ, 2008: Real-time PCR assays for quantification and differentiation of Vibrio vulnificus strains in oysters and water. Appl Environ Microbiol, 74, 1704-9.
  • Gulig PA, Bourdage KL, Starks AM, 2005: Molecular pathogenesis of Vibrio vulnificus. J Microbiol, 43, 118-31.
  • Han F, Pu S, Hou A, Ge B, 2009: Characterization of clinical and environmental types of Vibrio vulnificus isolates from Louisiana oysters. Foodborne Pathog Dis, 6, 1251-8.
  • Han F, Wang F, Ge B, 2011: Detecting potentially virulent Vibrio vulnificus strains in raw oysters by quantitative loop-mediated isothermal amplification. Appl Environ Microbiol, 77, 2589-95.
  • Hlady WG, 1997: Vibrio infections associated with raw oyster consumption in Florida, 1981–1994. J Food Prot, 60, 353-7.
  • Huebers HA, Eng MJ, Josephson BM, Ekpoom N, Rettmer RL, Labbe RF, Pootrakul P, Finch CA, 1987: Plasma iron and transferrin iron-binding capacity evaluated by colorimetric and immunoprecipitation methods. Clin Chem, 33, 273-7.
  • Høi L, Dalsgaard I, Dalsgaard A, 1998: Improved isolation of Vibrio vulnificus from seawater and sediment with cellobiose-colistin agar. Appl Environ Microbiol, 64, 1721-4.
  • Jackson JK, Murphree RL, Tamplin ML, 1997: Evidence that mortality from Vibrio vulnificus infection results from single strains among heterogeneous populations in shellfish. J Clin Microbiol, 35, 2098-101.
  • Jones MK, Oliver JD, 2009: Vibrio vulnificus: Disease and pathogenesis. Infect Immun, 77, 1723-33.
  • Kaysner CA, DePaola A, 2004: Vibrio. In ''Bacteriological Analytical Manual'', Ed; Hammack T, Davidson MW, Feng P, Gharst G, Ge B, Jinneman K, Regan PM, Kase J, Orlandi P, Burkhardt W, U.S. Food and Drug Administration, Washington, D.C..
  • Kim CM, Chung YY, Shin SH, 2009: Iron differentially regulates gene expression and extracellular secretion of Vibrio vulnificus cytolysin-hemolysin. J Infect Dis, 200, 582-9.
  • Kobayashi T, Enomoto S, Sakazaki R, Kuwahara S, 1963: A new selective isolation medium for Vibrio group on a modified Nakanishi's medium (TCBS agar medium). Jpn J Bacteriol, 18, 387-92.
  • Lin M, Schwarz JR, 2003: Seasonal shifts in population structure of Vibrio vulnificus in an estuarine environment as revealed by partial 16S ribosomal DNA sequencing. FEMS Microbiol Ecol, 45, 23-7.
  • Masad G, Oliver JD, 1987: New selective and differential medium for Vibrio cholerae and Vibrio vulnificus. Appl Environ Microbiol, 53, 2262-4.
  • Nwafia WC, Aneke JO, Okonji CU, 2006: Serum iron and total iron binding capacity levels among the ABO blood groups in Enugu, South Eastern Nigeria. Niger J Physiol Sci, 21, 9-14.
  • Oliver JD, Warner RA, Cleland DR, 1983: Distribution of Vibrio vulnificus and other lactose-fermenting Vibrios in the marine environment. Appl Environ Microbiol, 45, 985-98.
  • Panicker G, Myers ML, Bej AK, 2004: Rapid detection of Vibrio vulnificus in shellfish and Gulf of Mexico water by real-time PCR. Appl Environ Microbiol, 70, 498-507.
  • Potasman I, Paz A, Odeh M, 2002: Infectious outbreaks associated with bivalve shellfish consumption: A worldwide perspective. Clin Infect Dis, 35, 921-8.
  • Randa MA, Polz MF, Lim E, 2004: Effects of temperature and salinity on Vibrio vulnificus population dynamics as assessed by quantitative PCR. Appl Environ Microbiol, 70, 5469-76.
  • Rosche TM, Smith DJ, Parker EE, Oliver JD, 2005: A rapid and simple PCR analysis indicates there are two subgroups of Vibrio vulnificus which correlate with clinical or environmental isolation. Microbiol Immunol, 49, 381-9.
  • Sanjuán E, Fouz B, Oliver JD, Amaro C, 2009: Evaluation of genotypic and phenotypic methods to distinguish clinical from environmental Vibrio vulnificus Strains. Appl Environ Microbiol, 75, 1604-13.
  • Starks AM, Schoeb TR, Tamplin ML, Parveen S, Doyle TJ, Bomeisl PE, Escudero GM, Gulig PA, 2000: Pathogenesis of infection by clinical and environmental strains of Vibrio vulnificus in iron-dextran-treated mice. Infect Immun, 68, 5785-93.
  • Strom MS, Paranjpye RN, 2000: Epidemiology and pathogenesis of Vibrio vulnificus. Microb Infect, 2, 177-88.
  • Tamplin ML, Martin AL, Ruple AD, Cook DW, Kaspar CW, 1991: Enzyme immunoassay for identification of Vibrio vulnificus in seawater, sediment, and oysters. Appl Environ Microbiol, 57, 1235-40.
  • Tison DL, Kelly MT, 1984: Factors affecting hemolysin production by Vibrio vulnificus. Curr Microbiol, 10, 181-4.
  • Warner E, Oliver JD, 2007: Refined medium for direct isolation of Vibrio vulnificus from oyster tissue and seawater. Appl Environ Microbiol, 73, 3098-100.
  • Warner JM, Oliver JD, 1999: Randomly amplified polymorphic DNA analysis of clinical and environmental isolates of Vibrio vulnificus and other Vibrio species. Appl Environ Microbiol, 65, 1141-4.
  • Wright AC, Hill RT, Johnson JA, Roghman M-C, Colwell RR, Morris JG, 1996: Distribution of Vibrio vulnificus in the Chesapeake Bay. Appl Environ Microbiol, 62, 717-24.
Toplam 37 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Veteriner Cerrahi
Bölüm Araştıma
Yazarlar

Sedat Çam 0000-0001-9030-6713

Robin Brınkmeyer Bu kişi benim 0000-0001-5058-8103

Yayımlanma Tarihi 25 Aralık 2019
Gönderilme Tarihi 11 Haziran 2019
Kabul Tarihi 4 Aralık 2019
Yayımlandığı Sayı Yıl 2019 Cilt: 8 Sayı: 2

Kaynak Göster

APA Çam, S., & Brınkmeyer, R. (2019). Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes. Harran Üniversitesi Veteriner Fakültesi Dergisi, 8(2), 173-179. https://doi.org/10.31196/huvfd.667690
AMA Çam S, Brınkmeyer R. Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes. Harran Univ Vet Fak Derg. Aralık 2019;8(2):173-179. doi:10.31196/huvfd.667690
Chicago Çam, Sedat, ve Robin Brınkmeyer. “Development of Selective Enrichment Medium for Clinical Isolates of Vibrio Vulnificus Based Upon Virulence Correlating Genes”. Harran Üniversitesi Veteriner Fakültesi Dergisi 8, sy. 2 (Aralık 2019): 173-79. https://doi.org/10.31196/huvfd.667690.
EndNote Çam S, Brınkmeyer R (01 Aralık 2019) Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes. Harran Üniversitesi Veteriner Fakültesi Dergisi 8 2 173–179.
IEEE S. Çam ve R. Brınkmeyer, “Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes”, Harran Univ Vet Fak Derg, c. 8, sy. 2, ss. 173–179, 2019, doi: 10.31196/huvfd.667690.
ISNAD Çam, Sedat - Brınkmeyer, Robin. “Development of Selective Enrichment Medium for Clinical Isolates of Vibrio Vulnificus Based Upon Virulence Correlating Genes”. Harran Üniversitesi Veteriner Fakültesi Dergisi 8/2 (Aralık 2019), 173-179. https://doi.org/10.31196/huvfd.667690.
JAMA Çam S, Brınkmeyer R. Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes. Harran Univ Vet Fak Derg. 2019;8:173–179.
MLA Çam, Sedat ve Robin Brınkmeyer. “Development of Selective Enrichment Medium for Clinical Isolates of Vibrio Vulnificus Based Upon Virulence Correlating Genes”. Harran Üniversitesi Veteriner Fakültesi Dergisi, c. 8, sy. 2, 2019, ss. 173-9, doi:10.31196/huvfd.667690.
Vancouver Çam S, Brınkmeyer R. Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes. Harran Univ Vet Fak Derg. 2019;8(2):173-9.