Chan, AWK

Chan, TM

Chu, FCS

Jin, LJ

Leung, WK

Tsang, PCS

Yau, JYY

Journal of Dental Research

HKU Scholars Hub

Title Predominant cultivable subgingival flora of renal transplantrecipientsAuthor(s) Leung, WK; Jin, LJ; Yau, JYY; Chan, AWK; Tsang, PCS; Chu,FCS; Chan, TMCitationThe 79th General Session and Exhibition of the InternationalAssociation for Dental Research, Chiba, Japan, 27-30 June 2001.In Journal of Dental Research, 2001, v. 80 Sp Iss, abstract no.1921Issued Date 2001URL http://hdl.handle.net/10722/53229Rights Creative Commons: Attribution 3.0 Hong Kong License1921 Predominant Cultivable Subgingival Flora of Renal Transplant Recipients.W.K. LEUNG*, L.J. JIN, J.Y.Y. YAU, A.W.K. CHAN, P.C.S. TSANG, F.C.S. CHU & T.M. CHAN1Faculties of Dentistry and 1Medicine, The University of Hong Kong, Hong Kong SAR, China.Fig 2.  Quantity (colony-forming units/paper point) and relative mean proportion of predominant cultivable bacterial types from subgingivalplaque samples of renal transplant recipients.  No growth refers to bacterial isolates that failed to survive after primary culture. Panel A:quantity of gram-positive species and no growth. Panel B: quantity of gram-negative species. Panel C: proportion (% of total) gram-positive species and no growth. Panel D: proportion (% of total) gram-negative species. Note the larger quantity of colony-forming unitsfrom Shallow Pocket and Deep Pocket samples.  Substantial amount of gram-negative rods were lost upon subculturing in Deep Pocketgroup.  A multiple comparison (Fisher’s PLSD) was performed on different bacterial types expressed as proportion of the total bacteriarecovered. Significant different (P < 0.05, Fisher’s PLSD) of data values were observed among several bacterial types (I to X) between theclinical groups: I) total gram-positive species: Healthy vs Shallow pocket and Deep Pocket groups, Deep Pocket vs Pseudo-Pocket groups;II) total obligatory anaerobic gram-positive species: Healthy vs Shallow Pocket groups; III) total facultative anaerobic gram-positivespecies: Deep Pocket vs Healthy and Pseudo-Pocket groups; IV) total gram-positive cocci: Shallow Pocket vs Healthy, Shallow Pocket vsPseudo-Pocket groups, V) facultative anaerobic gram-positive cocci: Healthy vs Shallow Pocket, Healthy vs Deep Pocket groups; VI)obligatory anaerobic gram-positive cocci: Shallow Pocket vs Deep Pocket groups;  VII) facultative anaerobic gram-positive rods: ShallowPocket vs Deep Pocket groups; VIII) obligatory anaerobic gram-positive filaments: Healthy vs Deep Pocket, Healthy vs Pseudo-Pocketgroups; IX) total gram-negative species: Deep Pocket vs Healthy and Pseudo-Pocket groups; X) total gram-negative rods: Healthy vs DeepPocket groups.REFERENCESCarpenter CB, Lazarus JM. Dialysis and transplantation in the treatment of renal failure. In: Fauci AS, Braunwald E, Isselbacher KJ et al. ed,Harrison’s principles of internal medicine, 14th ed, New York: McGraw-Hill Co, 1998:1520.Chan TM.  Chin Med J 1997: 110:431.Chu FCS, Tsang PCS, Chan AWK et al. Ann Roy Australas Coll Dent Surg 2000: 15: 286.King GN, Fullinfaw R, Higgins TJ, et al.  J Clin Periodotol 1993: 20:286.Remuzzi G, Bertani T.  New Engl J Med 1998: 339:1448.Sketris I, Yatscoff R, Keown P et al. Clin Biochem 1995: 28:195.Spratt H, Boomer S, Irwin C et al.  Oral Dis 1999: 5:27.Tang S, Lui SL, Li FK et al.  Int J Artif Organs 1999: 22:131.Thomason JM, Seymour RS, Ellis JS et al.   J Clin Periodontol 1996: 23:628.RESULTSTransplant typeSex Sample Sitesa Living Immunosuppressant usedWith gingival Age Post-transplant NotGroup Male Female Incisors/ Premolars Molars Overgrowth Probing Depth (mm) (year) Duration  (year) Cadaveric genetic Genetic Cyclosporin Tacrolimus NilCanines (Prevalence, %) (mean ± SD) (mean ± SD) (mean ± SD) related related AHealthy 2 2 4 0 0 0 2.2-3.2 30-41 0.5-6 3 1 0 3 1 0(2.5±0.5) (35.4±4.6) (2.7±2.4)Shallow 7 7 5 2 7 29 3.6-5.4 31-55 0.5-12 10 1 3 12 1 1Pocket (4.3±0.8) (41.0±6.1) (5.0±3.8)Deep 9 4 4 0 9 23b 5.8-10.0 40-67 1-13 11 0 2 12 1 0Pocket (7.1±1.4) (49.9±9.2)c (5.0±3.6)Pseudo- 4 3 2 2 3 100 5.8-7.2  21-68 1-7 5 1 1 6 1 0Pocket (6.2±0.5) (38.3±14.7) (3.3±2.5)Total 22 16 15 4 19 42d 2.2–10.0 21-68 0.5-13 29 3 6 33 4 1(5.5 ± 1.9) (42.2±11.3) (4.4±3.4)a One sample site per subject.b Out of 46% of the subgroup subjects showed gingival overgrowth clinically.c Significant different (P < 0.05, Fisher’s PLSD) of data value between: Deep Pocket vs Healthy, Deep Pocket vs Shallow Pocket and Deep Pocket vs Pseudo-Pocket groups.d Out of 50% of all subjects showed gingival overgrowth clinically.Table 1.  Demographic data, post-transplant duration and types of immunosuppressant used.INTRODUCTIONProgression of renal disease from inflammation, fibrosis and atrophy into end-stage renal failure is a multi-factorial process.  Alteration in intra-renal haemodynamics and hence increase in proteinuria andsubsequent release of vasoactive and inflammatory substances would lead to worsening of the kidney condition(Remuzzi & Bertani 1998).  Renal transplantation offers the best opportunity for rehabilitation of individualssuffering from end-stage renal failure (Carpenter & Lazarus & 1998).  Renal replacement therapy for patientswith end-stage renal failure was introduced in the Hong Kong public hospital system some 24 years ago (Chan1997).  Immunosuppression therapy by various combinations of: glucocorticoids, azathioprine, cyclosporin,tacrolimus, monoclonal antibody OKT3, or antilymphocyte globulin were often used to reduce graft rejectionin imperfectly matched donor-receptors cases.  It was shown that immunosuppression agents such as cyclosporinA could improve survival rate of  transplant recipients (Sketris et al. 1995).  A recent study reported the long-term 10-year allograft survival rate of a cohort of renal recipients in Hong Kong was 53% (Tang et al. 1999).This study implied that with continuously improving medical care and survival of this patient group, increasedoral health care demand will be a natural consequence.The present report is part of a project which focused on the oral health status and the corresponding treatmentneeds of post-operative, stable renal transplant recipients.  The prevalence rate of gingival overgrowth in thelocal sample was at a high level (53%) (Chu et al. 2000) compatible to reports from other parts of the world(King et al. 1993, Spratt et al. 1999, Thomason et al. 1996).  Including pseudo-pockets, up to 80% of the localcohort had probing depth  3.5 mm (Chu et al. 2000).  This study investigated the impact that renal transplanttherapy had on periodontal status and the subgingival microflora.MATERIALS AND METHODSSubjects:- 38  renal transplant  recipients  (16  females,  age  21 – 68  years;   6  months  post-transplant)  fromNephrology Clinic, Department of Medicine, HKU were recruited for the study.Clinical/Laboratory investigations:- Clinical examination by Florida Probe® and paralleling periapical radiography- Subgingival plaque samples (one/subject)- Gram-stain smear- Anaerobic culture on enriched Columbia blood agar- Selective culture on MacConkey and Sabouraud’s dextrose agars- Identification and quantification using:RapID ANAII, API 20 Strep, API Staph, API 20E, and API 20C AUX kitsTable 2.  Differential cell counts from Gram-stained smearsa.Healthy Shallow Pocket Deep Pocket Pseudo-PocketGram-positive microorganismsbcoccic 1.5 – 50.5 0 - 27.5 0 - 4.5 0 – 35.0(40.0, 33.0) (0, 5.4) (0.5, 0.7) (0, 5.3)rods 0 – 10.5 0 – 24.0 0 – 2.5 0 – 6.5(4.0, 4.6) (0, 3.5) (0, 0.4) (0, 0.9)Totalc 1.5 – 5.7 0 – 51.5 0 – 5.0 0 – 41.5(46.0, 37.6) (0, 8.9) (0.5, 1.2) (0, 6.2)Gram-negative microorganismscocci 0 – 11.5 0 – 10.0 0.5 – 17.5 0 – 10.5(8.0, 6.9) (2.3, 3.6) (2.5, 6.0) (2.5, 3.6)rods 25 – 86.5  26.0 – 73.5 22.0 – 88.5 47.5 – 75.0(43.3, 49.5) (56.3, 56.5) (68.0, 60.8) (70.5 – 62.6)fusiforms 0 – 7.0 0 – 5.0 0 – 2.5 0 – 3.5(1.5, 1.5) (1.3, 1.4) (1.5, 1.3) (1.0, 1.3)curved rodsd 0 – 1.5 0 – 6.0 0.5 – 4.5 0.5 – 4.5(1.0, 0.9) (3.0, 2.9) (1.5, 1.8) (2.5, 2.4)filaments 0 – 0.5 0 – 26.5 0 – 2.5 0 – 5.0(0, 0.1) (0.5, 2.4) (0.5, 0.7) (1.5, 1.6)spirochetese 0 – 5.5 0 – 66.5 0.5 – 63.0 0 – 44.0(1.3, 2.0) (26.3, 24.1) (24.0, 28.2) (16.0, 22.1)Totalc 43.0 – 98.5 48.5 – 100.0 95.0 – 100.0 58.5 – 100.0(54.0, 62.4) (99.5, 90.9) (99.5, 98.8) (100.0, 93.8)Fungifyeast form 0 0 – 2.0 0 0(0, 0) (0, 0.1) (0, 0) (0, 0)a Data shown are percentage range, median and mean (in parenthesis).b No gram-positive filaments observable.c Significant different (P < 0.05, Fisher’s PLSD) of data value between: Healthy vs Shallow Pocket, Healthy vs Deep Pocket   and Healthy vs Pseudo-Pocket groups.d Significant different (P < 0.05, Fisher’s PLSD) of data value between: Shallow Pocket vs Healthy, and Shallow Pocket vs   Deep Pocket groups.e Significant different (P < 0.05, Fisher’s PLSD) of data value between: Healthy vs Shallow Pocket, and Healthy vs Deep   Pocket groups.f No mycelial form observable.Table 3. Prevalence of microbes isolated and the corresponding median and mean percentage isolation fromsubgingival plaque samples of renal transplant recipientsa.Healthy Shallow Pocket Deep Pocket Pseudo-PocketGram-positiveFacultative anaerobic cocciGemella haemolysans 25 (0, 1.3)b 7.1 (0, 0.5) 7.7 (0, 2.6) 14.3 (0, 1.9)Gemella morbillorum 25 (0, 6.1) 50.0 (0.5, 6.3) 30.8 (0, 2.6) 28.6 (0, 1.6)Leuconostoc spp.c 0 (0, 0) 28.6 (0,4.3) 0 (0, 0) 0 (0, 0)Staphylococcus epidermidis 25 (0, 9.2) 0 (0, 0) 7.7 (0, 0.1) 0 (0, 0)Staphylococcus spp.d,e 25 (0, 7.6) 0 (0, 0) 0 (0, 0) 0 (0, 0)Streptococcus acidominimus 0 (0, 0) 21.4 (0, 0.7) 7.7 (0, 1.4) 0 (0, 0)Streptococcus constellatusf,g 25 (0, 2.6) 7.1 (0, 0.2) 0 (0, 0) 42.9 (0, 7.2)Streptococcus intermedius 25 (0, 9.5) 0 (0, 0) 7.7 (0, 1.3) 0 (0, 0)Streptococcus mitis biovar 1 50 (0.9, 3.8) 7.1 (0, 0.7) 30.8 (0, 2.0) 14.3 (0, 0.9)Streptococcus oralis 25 (0, 1.3) 7.1 (0, 0.3) 0 (0, 0) 0 (0, 0)Streptococcus pneumoniae 0 (0, 0) 0 (0, 0) 15.4 (0, 4.3) 0 (0, 0)Streptococcus sanguis 25 (0, 2.6) 7.1 (0, 0.1) 7.7 (0, 0.4) 14.3 (0, 3.3)Streptococcus salivarius salivariusf 0 (0, 0) 0 (0, 0) 15.4 (0, 1.5) 42.9 (0, 5.7)Anaerobic cocciPeptostreptococcus anaerobius 0 (0, 0) 0 (0,0) 15.4 (0, 3.2) 14.3 (0, 3.1)Peptostreptococcus micros 50 (4.1, 7.5) 7.1 (0, 0.8) 46.2 (0, 9.6) 14.3 (0, 0.5)Peptostreptococcus prevotiic 0 (0, 0) 7.1 (0, 0.2) 15.4 (0, 1.0) 42.9 (0, 3.3)Facultative anaerobic rodsActinomyces georgiae/gerencseriae 0 (0, 0) 28.6 (0, 3.2) 15.4 (0, 1.9) 0 (0, 0)Actinomyces naeslundii 0 (0, 0) 21.4 (0, 2.2) 0 (0, 0) 28.6 (0, 4.7)Actinomyces viscosus 25 (0, 3.1) 0 (0, 0) 7.7 (0, 0.1) 0 (0, 0)Arachnia propionica 25 (0, 0.9) 35.7 (0, 5.9) 15.4 (0, 2.4) 14.3 (0, 1.5)Lactobacillus acidophilus 0 (0, 0) 7.1 (0, 3.9) 15.4 (0, 1.0) 14.3 (0, 0.7)Lactobacillus jenseniic 0 (0, 0) 14.3 (0, 2.2) 23.1 (0, 1.3) 14.3 (0, 7.0)Rothia dentocariosaf 0 (0, 0) 0 (0, 0) 0 (0, 0) 28.6 (0, 5.9) Anaerobic rodsActinomyces israelii 0 (0, 0) 21.4 (0, 4.3) 15.4 (0, 3.0) 0 (0, 0)Mobiluncus spp.c,d,e 25 (0, 25.0) 0 (0, 0) 0 (0, 0) 0 (0, 0)Gram-negativeFacultative anaerobic rodsKingella kingae 0 (0, 0) 0 (0, 0) 15.4 (0, 1.1) 0 (0, 0)Pasteurella pneumotropica/haemolyticad,e 25 (0, 1.0) 0 (0, 0) 0 (0, 0) 0 (0, 0)Anaerobic rodsCampylobacter gracilis 25 (0, 1.4) 28.6 (0, 3.2) 38.5 (0, 7.4) 28.6 (0, 9.3)Campylobacter rectus 0 (0, 0) 28.6 (0, 6.1) 15.4 (0, 1.1) 0 (0, 0)Prevotella buccae 0 (0, 0) 7.1 (0, 1.0) 15.4 (0, 1.8) 0 (0, 0)Prevotella corporis 0 (0, 0) 42.9 (0, 2.3) 23.1 (0, 0.6) 14.3 (0, 5.3)Prevotella intermedia 25 (0, 1.5) 14.3 (0, 0.8) 23.1 (0, 1.4) 0 (0, 0)Prevotella loescheii 0 (0, 0) 0 (0, 0) 15.4 (0, 3.3) 0 (0, 0)Prevotella oralis 0 (0, 0) 0 (0, 0) 15.4 (0, 1.8) 14.3 (0, 1.0)Facultative fusiformshCapnocytophaga gingivalis 25 (0, 0.5) 21.4 (0, 2.5) 23.1 (0, 5.4) 14.3 (0, 2.0)Capnocytophaga ochracea 0 (0, 0) 35.7 (0, 3.8) 7.7 (0, 0.8) 0 (0, 0)Capnocytophaga sputigenad,e 25 (0, 3.5) 0 (0, 0) 0 (0, 0) 0 (0, 0)Capnocytophaga spp. 0 (0, 0) 21.4 (0, 2.8) 0 (0, 0) 0 (0, 0)Anaerobic fusiformshFusobacterium nucleatumd,e 25.0 (0, 1.8) 0 (0, 0) 0 (0, 0) 0 (0, 0)Non-orali 25.0 (0, 25.0) 57.1 (5.8, 11.7) 61.5 (3.5, 9.9) 42.9 (0, 10.3)Lost/unidentified spp. 75.0 (7.7, 9.9) 85.7 (29.2, 25.5) 69.2 (4.5, 25.7) 57.1 (7.1, 22.4)a Only species with frequency of isolation > 15% in any one group are included.b Data shown are percentage prevalence; median and mean percentage proportion (in parenthesis).c Microbes that are not normally considered as member of the oral or oropharyngeal flora.d Significantly higher prevalence of isolation in Healthy vs other groups (P < 0.05, 2 test).e Significant difference quantity of bacterial species (% proportion) between: Healthy vs Shallow Pocket; Healthy vs Deep Pocket groups (P < 0.05,Fisher’s PLSD).f Significant higher prevalence of isolation in Pseudo-Pocket vs other groups (P < 0.05, 2 test).g Significant difference quantity of bacterial species (% proportion) between Pseudo-Pocket vs Deep Pocket groups (P < 0.05, Fisher’s PLSD).h Significant lower prevalence of fusiform species isolated in Pseudo-Pocket vs other groups (P < 0.05, 2 test).i Data including species with frequency of isolation  15%.051015202530Colony-forming units (x 1e6)1 11AnaerobicFacultative anaerobicLostAnaerobicFacultative anaerobicLostAnaerobicFacultative anaerobicCocci (Column 1)Rods (Column 2)No growth (Column 4)Healthy ShallowPocketDeepPocketPseudo-Pocket(A) Gram-positiveFilaments (Column 3)01020304050Percentage1 11AnaerobicFacultative anaerobicLostAnaerobicFacultative anaerobicLostAnaerobicFacultative anaerobicCocci (Column 1)Rods (Column 2)Filaments (Column 3)No growth (Column 4)Healthy ShallowPocketDeepPocketPseudo-Pocket(C) Gram-positiveCONCLUSIONS1. Subgingival microflora of renal transplant recipients with inflammatory periodontal conditions are mainlycomprised of gram-negative rods and spirochetes;2. Non-oral microbes were highly prevalent in the subgingival plaque of the renal transplant recipients;3. A substantial amount of the renal transplant subgingival flora are not recoverable by anaerobic culture(spirochetes and various lost species);4. Based on the above, we postulate that the subgingival biofilm of renal transplant recipients would be anunique microbial entity regardless the various periodontal conditions.Fig. 1.  Photomicrograph of Gram-stained smearsspecimen prepared from a sample from the DeepPocket group.  Spirochetes of various sizes werepresent in abundance.  Other morphotypes like gram-negative rods and curved rods were easily observable.Bar = 25μm.01020304050Percentage1 11LostAnaerobicFacultativeLostAnaerobicFacultative anaerobicLostAnaerobicFacultative anaerobicCocci (Column 1)Rods/filaments (Column 2)Fusiforms (Column 3)Healthy ShallowPocketDeepPocketPseudo-Pocket(D) Gram-negative051015202530Colony-forming units (x 1e6)1 11LostAnaerobicFacultativeLostAnaerobicFacultative anaerobicLostAnaerobicFacultative anaerobicCocci (Column 1)Rods/filaments (Column 2)Fusiforms (Column 3)(B) Gram-negativeHealthy ShallowPocketDeepPocketPseudo-Pocket

Predominant cultivable subgingival flora of renal transplant recipients

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Predominant cultivable subgingival flora of renal transplant recipients

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