Poster presented at the First International Congress of CiiEM - From Basic Sciences to Clinical Research. 27-28 November 2015, Egas Moniz, Caparica, Portuga

Moutinho, M. G. M.

Nascimento, T.

Neves, V.

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	  	  	  Sample	  characteriza.on:	  §  The	   individuals	   aged	   18	   to	   80	   years	   were	   grouped	   according	   to	   the	   criterion	  “diabetic	  vs	  non-­‐diabetic”,	  and	  it	  was	  found	  56,4%	  (n=66)	  were	  diabetic	  and	  43,6%	  (n=51)	  weren’t	  diabetic.	  §  The	  average	  age	   in	  the	  study	  group	  (diabetics)	  was	  64,2	  years	  and	   in	  the	  control	  group	  was	  36,1.	  §  In	  the	  study	  group:	  •  65,2%	  (n=43)	  were	  male	  and	  34,8%	  (n=23)	  were	  female.	  	  	  Presence	  of	  Candida	  sp.:	  §  The	   oral	   colonization	   by	  Candida	   sp.	  was	   detected	   in	   43,9%	   (n=29)	   in	   the	   study	  group	  and	  in	  17,6%	  (n=9)	  in	  the	  control	  group	  (figure	  2).	  §  The	  diversity	  of	  the	  isolated	  yeast	  species	  in	  the	  study	  group	  was	  as	  follows:	  33,3%	  (n=22)	   of	   C.	   albicans,	   3%	   (n=2)	   C.	   tropicalis,	   4,5%	   (n=3)	   mixed	   cultures	   of	   C.	  albicans	  and	  C.	  tropicalis	  and	  3%	  (n=2)	  of	  C.	  krusei	  (figure	  3).	  §  C.	  albicans	  was	  the	  most	  prevalent	  species.	  §  The	   chi-­‐square	   test	   showed	   significant	   difference	   between	   being	   diabetic	   and	  being	  colonized	  when	  compared	  to	  the	  control	  group	  (ρ	  =	  0,003).	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  Virulence	  factors:	  expression	  of	  proteases	  and	  phospholipases	  §  In	   the	   38	   isolates	   of	   Candida	   sp.,	   63.2%	   (n=24)	   showed	   no	   production	   of	  proteinases,	  26.3%	  (n=10)	  showed	  production	  positive	  (+)	  and	  10.5%	  (n=4)	  showed	  strong	  positive	  output	  (++)	  according	  calculating	  Pz	  (figure	  4).	  §  In	  the	  38	  isolates	  of	  Candida	  sp.,	  12	  strains	  (31.6%)	  did	  not	  express	  phospholipases	  and	   26	   (68.4%)	   produced	   these	   enzymes,	   in	   which	   10.5%	   (N	   =	   4)	   were	   weak	  positive	   (+),	  36.8%	  (N	  =	  14)	  were	  positive	   (+++)	  and	  21.1%	  (N	  =	  8)	  were	  strongly	  positive	  (++++),	  according	  to	  results	  of	  the	  calculation	  of	  the	  Pz	  (figure	  5).	  	  	  §  As	  for	  HbA1c	  ≥	  6,5%	  vs	  colonization	  by	  Candida	  sp.,	  67,9%	  had	  HbA1c	  ≥	  6,5%	  and	  were	  colonized	  and	  72,4%	  had	  HbA1c	  ≥	  6,5	  but	  were	  not	  colonized	  by	  these	  yeast	  fungi.	  Oral Candida colonization in diabetic individuals non-carriers of dental prosthesis 	  	  The	  men's	  oral	  cavity's	  flora	  is	  a	  complex	  ecosystem,	  composed	  by	  several	  microorganisms	   that	  may	   cause	   infections	   when	   the	   	   immune	   system	   is	  weakened.1,2	  The	  presence	  of	  Candida	   is	  very	  common	  and	  although	  they	  are	   commensal,	   they	   can	   become	   pathogenic.	   This	   yeast-­‐like	   fungus	   is	  responsible	   for	   the	   appearance	  of	   candidiasis	   in	  man,	  which	   arise	  due	   to	  nutrient	   imbalances,	   malignant	   diseases	   or	   metabolic	   disorders	   such	   as	  Diabetes	  Mellitus	   (DM)1,3.	   DM	   is	   a	   disease	   associated	   with	   uncontrolled	  blood	  glucose	  that	  leads	  to	  an	  increase	  of	  glucose	  in	  oral	  fluid	  and	  causes	  xerostomia,	   creating	   an	   environment	   conducive	   to	   the	   development	   of	  fungal	  infections.1,2,3	  	  	  Evaluate	   the	  prevalence	  of	  Candida	   sp.	   in	   the	  oral	   cavity	  of	  diabetics	  and	  non-­‐diabetics,	  both	  non-­‐carriers	  of	  dental	  prosthesis;	  assess	   the	  virulence	  factors	   of	  Candida	   sp.	   associated	   to	   single	   species	   and	   bearer	   group	   and	  relate	  HbA1c	  values	  with	  colonization	  by	  Candida	  sp.	  OBJECTIVES	  	  Sample:	   The	   study	   group	   included	   117	   individuals	   of	   both	   genders,	   both	  non-­‐carriers	  of	  dental	  prosthesis,	  of	  which	  66	  were	  diabetic	  (study	  group)	  and	  51	  non-­‐diabetic	  patients	  (control	  group).	  	  Collec.on	   of	   samples:	   The	   collection	   of	   samples	   was	   performed	   at	   the	  University	  Dental	  Clinic	  of	   ISCSEM	  and	  at	   the	  clinical	   analyzes	   laboratory-­‐Lumilabo.	  After	  reading	  and	  signing	  the	   informed	  consent	  by	  the	  pacients	  included	  in	  the	  study,	  a	  sample	  of	  the	  oral	  mucosa	  was	  collected	  with	  the	  aid	  of	  sterile	  swabs	  and	  included	  in	  transport	  medium.	  At	  the	  microbiology	  laboratory	   of	   ISCSEM,	   the	   samples	   were	   inoculated	   in	   specific	   mediums	  and	  identified	  according	  to	  the	  phenotypic	  tests.	  Iden.ﬁca.on	   of	   Candida	   sp.:	   The	   isolates	   were	   identified	   based	   on	  phenotypic	   methods:	   macroscopic	   and	  microscopic	   characteristics	   of	   the	  colonies	   on	   Sabouraud	   agar	   culture	   medium	   and	   chromogenic	   medium,	  and	   the	   species	   identification	   was	   subsequently	   confirmed	   by	   the	  biochemical	  profile	  (Figure	  1).	  Sta.s.cal	  analysis:	  For	  statistical	  analysis	  we	  used	  the	  SPSS	  program.	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  METHODS	  Figure	   1.	   Laboratory	   Protocol:	   (1)	   Collection	   of	   samples	  with	   sterile	   swab;	   (2)	   Inoculation	   in	  Sabouraud	   medium;	   (3)	   Inoculation	   in	   differential	   chromogenic	   medium:	   Brilliance	   Agar	  (Presumptive	   identification	   of	   Candida	   sp,	   Green	   -­‐	   C.	   albicans;	   Blue	   -­‐	   C.	   tropicalis;	   Brown	   -­‐	  Candida	   sp);	   (4)	  micromorphological	   identification	   (Blastese	   proof)	   -­‐	   (400X	   amplification);	   (5)	  biochemical	  identification	  of	  Candida	  sp.	  (API	  Candida	  -­‐	  Biomerieux®)	  RESULTS	  Figure	  2.	  Presence	  of	  Candia	  sp.	   in	   the	  diabetic	  group	  and	  control	  group.	  Figure	   3.	   Presence	   of	   different	   species	   of	  Candida	  sp.	  in	  the	  diabetic	  group.	  1.  Boparai,	  N.,	  Amasi,	  U.,	  Patil,	  R.,	  &	  Harakuni,	  S.	  (2014).	  To	  assess	  candidal	  colonization	  and	  species	  diversity	  in	  the	  oral	  cavity	  of	  diabetic	  and	  nondiabetic	  denture	  wearers	  and	  correlation	  with	  the	  presence	  of	  candida	  on	  finger	  tips	  of	  the	  patients:	  An	  in	  vivo	  study.	  Indian	  Journal	  of	  Health	  Sciences,	  7(1),	  45.	  	  2.  Kumar,	  D.	  A.,	  Muralidhar,	  S.,	  Biswas,	  K.,	  Banerjee,	  U.,	  Basir,	  S.	  F.,	  &	  Khan,	  L.	  A.	  (2014).	  Species	  Diversity	  ,	  Antifungal	  Susceptibility	  ,	  and	  Virulence	  Attributes	  of	  Candida	  Colonising	  the	  Oral	  Cavities	  of	  Adult	  Diabetic	  Patients,	  2014	  3.  Talabani,	  A.	  N.,	  Kim,	  C.	  L.,	  Ong,	  A.,	  Kiat,	  H.,	  &	  Ismail,	  A.	  R.	  (2013).	  The	  prevalence	  of	  Candida	  spp	  .	  in	  the	  saliva	  of	  controlled	  and	  uncontrolled	  diabetes	  mellitus	  type	  II	  patients.	  J	  Bagh	  College	  DenDstry,	  25(4).	  	  BIBLIOGRAPHY	  V.	  Neves	  1,	  T.	  Nascimento	  1,	  2,	  3,	  M.G.M.	  Mou.nho	  1,2	  	  1	  ISCSEM	  (University	  Campus,	  Quinta	  da	  Granja,	  2829	  -­‐	  511	  Caparica,	  Telephone:	  (+351)	  212	  946	  700,	  E-­‐mail:	  vania.sofia.neves@gmail.com;	  tmnascimento68@gmail.com;	  mgm.moutinho@gmail.com),	  	  2	  Member	  of	  CiiEM,3	  Lumilabo,	  Clinical	  Analysis	  Laboratory	  (Av.	  Columbano	  Bordalo	  Pinheiro	  71,	  1070-­‐061	  Lisbon,	  Telephone:	  21	  721	  0480)	  	  1 2  4 3 5 Incubation	  at	  35ºC	  to	  72h	  Incubation	  at	  35ºC	  72h	  Sabouraud	  Dextrose	  Agar	   Brilliance	  Agar	  Figure	   4.	   Expression	   of	   extracellular	   enzymes:	  proteases	  Figure	   5.	   Expression	   of	   extracellular	   enzymes:	  phospholipases	  	  	  ü  Being	  diabetic	  is	  an	  important	  factor	  for	  the	  colonization	  of	  Candida	  sp.	  in	  the	  oral	  cavity.	  ü  C.	  albicans	  was	  the	  most	  prevalent	  species	  and	  showed	  a	  great	  ability	  to	  express	  proteinases	  and	  phospholipases.	  	  ü  The	  uncontrolled	  HbA1c	  does	  not	  have	  a	  statistical	  significance	  for	  colonization	  by	  Candida	  sp.	  	  CONCLUSIONS	  INTRODUCTION	  •  University	  dental	  clinic	  of	  ISCSEM;	  •  Laboratory	  analyzes	  Lumilabo	  clinics;	  •  Fernanda	  Mesquita,	  Ph.D.	  and	  José	  João	  Mendes,	  Ph.D.	  of	  ISCSEM;	  •  Sandra,	  Susana	  e	  Natacha	  of	  laboratory	  microbiology	  ACKNOWLEDGEMENTS	  

Oral Candida colonization in diabetic individuals non-carriers of dental prosthesis

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Oral Candida colonization in diabetic individuals non-carriers of dental prosthesis

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