Ariya Chindamporn, Ph.D. Dept. of Microbiology, Fac. of Medicine, Chulalongkorn University, Bangkok, Thailand. Taipei, Taiwan Nov. 16, 2018; 11:45 am.
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1 Mimic fungal infections in Asia Ariya Chindamporn, Ph.D. Dept. of Microbiology, Fac. of Medicine, Chulalongkorn University, Bangkok, Thailand Outline: Introduction Taipei, Taiwan Nov. 16, 2018; 11:45 am. Clinical manifestation human & animals Diagnosis Treatment Epidemiology Message 1
2 3 Rare Fungal infections in Asia Yeast o Fereydounia khargensis o Pichia anomala o Kodamaea ohmeri o Trichosporon inkin o T. mucoides o Rhodotorula mucilaginosa o Saccharomyces cerevisiae o Blastoschizomyces capitatus Septate mould o Alternaria spp. o A. alternate o A. malorum o Chaetomium globosum o Exserohilum spp. o Paecilomyces formosus o Pyrenochaeta romeroi o Scedosporium apiospermum o S. prolificans Cause of emergence Non-septate mould o Conidiobolus coronatus o Cunninghamella b ertholletiae o Rhizomucor spp. o Saksenaea erythrospora Dimorphic fungi o Emergomyces Fungus-like microbes o o o o Lagenidium albertoi Prototheca wickerhamii Rhinosporidium seeberi Pythium insidiosum Fungi adapting higher temperature and acquire virulence factors Advancement of medical devices and management Broad-spectrum and steroid use International travel and natural disasters Challenges Epidemiology not well understood with regard to environmental reservoirs, modes of transmission, & ways to detect them Their relative rarity, laboratory diagnosis of these potential pathogens is challenging.. Specific identification requires expertise Antifungal susceptibility testing challenging because reliable methodology & antifungal breakpoints not available. Quality-assured diagnosis requires reference laboratories. Reference laboratory facilities are not available in majority of Asian countries. 4 2
3 Rare yeast infections in Asian countries with misdiagnosis issues Organisms Specimens (n) Disease spectrum (n) Underlying Misdiagnosed Microbiological Laboratory Diagnosis conditions/history (n) by common identification methods Fereydounia khargensis 5 Organisms Blood (1), Bloodstream pleural fluid (1) infection (1), respiratory track conditions, DM and (1) Low CD4 count (1), complicated medical hepatitis B (1) Macro: dry, slightly wrinkled and fringed F. khargensis can be misdiagnosed margins colony at 48h on SDA then turn by API 20C (Cr. neoformans) and darker after h Micro: vegetative cells W or W/O blastospores produced by polar budding on short stalks. Pseudohyphae is occasionally observed. The MICs v alues (μg/ml)* VITEK 2 (Cr. laurentii) F. khargensiscan be misdiagnosed by API 20C Cr. neoformans VITEK 2 Cr. laurentii Azoles Polyenes Echinocandins FLC VOR POS ITR AMB CAS MIC ANI Fereydounia khargensis ND >32 4->32 ND >32 Fungus-like infections in Asian countries with less effective of antifungal treatment 6 3
4 Vet Micro EID Vol. 11, No. 3, 2005 Fig. 1. GOAT: Extensive and ulcerative cutaneous lesion in the metatarsal-phalangeal region of the left hind limb Equine, Bovine, Canine, Beef Calve, Tiger, Sheep, Goat, Bird The First Human Case J Comp Pathol 2015 r;152 : Courtesy from Dr. Leonel Mendoza, MSU Prof. M. Thianprasit, 1985 Thai male Painful subcutaneous granulomatous infiltrative lumps with ulcer Treated SSKI Courtesy from Sathapatayavongs B., Mahidol U. 4
5 Type of Infection in Animal & Human (Sub) cutaneous Type Animals/Human Note Equine (most) Dog, Cat, Cattle, Sheep, Bird, Bear, Camel / 5% Tissue containing eosinophil and broad non-setate hyphae 10 Vascular type 59% Abnormality of RBC Ocular type 33% No underlying disease Disseminated pythiosis, GI Equine (rarely for GI) Dog (most), Bear, Bengal tiger (GI), Jaguar tiger (Pulmonary) / 3% Vomiting, w eight loss, diarrhea, some mass in abdomen Modified from Krajaejun T et al,. CID 2006;43:569-76, Wim Gaastra et al., 2010 Localized Type: Cutaneous / Subcutaneous Pythiosis 1 st human subcutaneous case in Thailand (Thianprasit,1985) 1 st human subcu. in Brazil (Marques, SA. et al. 2006) Onset 1-3 months An Bras Dermatol. 2006;81(5):483-5 Present as lump/ulcer H&E: rare septate hyphae Might be misdiagnosed as zygomycotic infection Suppurative granulomatous inflammation 5
6 Cutaneous pythiosis in a goat. do Carmo PM 1 et al. J Comp Pathol 2015: Subcu. Pythiosis vs Zygomycosis Splendore-Hoeppli phenomenon (H&E) Human Pythiosis, Brazil Fig. 2. Section of the lesion showing a central area with neutrophils, eosinophils and hyphae (arrows), surrounded by a granulomatous exudate with some multinucleate giant cells (arrowheads). HE. Bar, 50 mm. J Comp Pathol Feb-Apr;152(2-3): doi: /j.jcpa Epub 2014 Dec 31 Pathological finding: EID. V.11 (5), 715-8, 2005 Cutaneous/ Subcutaneous infection Granuloma with palisade-like Arrangement (H&E 200x) Wide, distorted, rare septate hyphae Non-septated hyaline filaments, in PDA agar (cotton blue 40x) Marques S el al, An Bras Dermatol 2006;81 6
7 Clinical manifestation in Human Localized type: Cornea - Immunocompetent - Corneal ulcer - Hypopyon - Endomophthalmitis - Corneal perforation Clinical manifestation in Human J Infect Dis 1989; 159:274 Vascular type Angiogram Pupaibun P. et al., 20 7
8 - 27 yr Thai male, w orker in Forestry department - beta-thalassemia haemoglobin E - 3 mo. before: toothache left upper molars, nasal congestion w ith w hitish discharge from left nostril - 3 w k prior adm, high grade fever - 1 w k later, occipital headache & funny feelings, focal seizure lasting for a min. & 2 nd generalized tonic-conic seisure for a min. Figure 1. Showing MRI, left carotid angiography and Pus Wright stain. 1A: Axial FLAIR MRI brain demonstrates - Abt treatment, referred to another university hospital hypersignal intensity lesion over the left frontoparietal region (arrow) and extensively involved corpus collosum (arrow C fever, drowsiness, significant left carotid bruit, head). 1B: Saggital T1W MRI brain with gadolinium rt. facial palsy & spastic rt. Hemiparesis enhancement reveals a large irregularly rim enhancing lesion (arrow) over the left cerebral hemisphere with satellite - Initial diag: maxillary sinus --MRI surrounding small lesions (arrow head). 1C: Left carotid - Pythium antibody, culture isolation and PCR : angiography shows multiple aneurysms (arrow) and a dissection (arrow head) over the left common and proximal Pythium insidiosum internal carotid arteries. 1D: Pus Wright s stain highlights multiple thin-walled hyphae with infrequently septate and branching (arrow). 15 Trakulhun T. et al., J Infectious Diseases and Antimicrobial Agents. V28:2, May-Aug2011 Serological approach: Antibody detection Im m unodiffusion Im m unoperoxidase Lateral flow Hemaglutination test FA ELISA W estern Blot Laboratory Diagnosis Conventional approach Direct examination KOH-preparation GMS stain H&E, PAS Calcoflour Cultivation Temperature Moisture Media : any Identification: Zoospore production Molec. identification Molecular approach PCR (Diagnosis & Identification) 8
9 How to due with suspected Vascular type?? Host : - symptom limb: gangrene - failure from antifungal treatment - location: tropical / subtropical - history & occupation : swampy / agricultural area - abnormal red blood cells: thalassemia, PNH Physician : Angiogram, CTA - aneurysm Serology : antibody Specimens & Diag. : serum : antibody testing Skin, artery, thrombus : muscle KOH, staining, culture KOH-preparation: rare septate hyphae KOH-Calcofluor preparation Histopathology: GMS Conventional approach 9
10 Treatment NO standard treatment Amphotericin B, Itraconazole & terbinafine SSKI Surgery Immunotherapy with P. insidiosum antigen (PIA) % successful rate in human (Wanachiwanawin W. et al., 2004, Permpalung N et. al., 2015) 10
11 Treatment outcome of surgery, antifungal therapy and immunotherapy in ocular & vascular human pythiosis: a retrospective study of 18 patients JAC 2015, Jan 27. Environmental isolated P. insidiosum: Northern part,thailand samples - 92 sites - Positive in 59 samples 11
![Mycol Res, Kammarnjessadakul et al. Med Mycol. 2010,49:289-295 ] - Jan.](/docs-images/96/127056769/images/12-3.jpg "1985-June 2003; 102 cases/18 yr - 9 tertiary care hospitals - 1-13 case (Mean")
12 Phylogenetic distribution of P.insidiosum isolated from different geographic regions based on ITS Ref: S N C Clade A TH E NE Clade C TH CladeB TH [Schurko et al Mycol Res, Kammarnjessadakul et al. Med Mycol. 2010,49: ] - Jan.1985-June 2003; 102 cases/18 yr - 9 tertiary care hospitals case (Mean 5.5 case/yr.) - Age year - Male : Female 2.4:1 - Agricultural occupation 75% - Type of infection: Localized, Systemic 12
13 Yr : KCMH (N =123) - Vas. (75); Ocular(48) By gender Vascular Ocular By Year 25 Female 44%,21) Male (56%,27) By month Male (77 %,58) By Hometown 26 13
14 Factors relates to survival rate Criteria of 1 year survival Survival group (45) Deceased group (5) P- value* Patient related parameters Age (years) 33.6 ± ± Male gender 25 (56%) 4 (80%) 0.29 Occupation Agriculture related 39 (86.7%) 5 (100%) - Non-agriculture related 6 (13.3%) - History of water exposure within 3 months Exposed water 41 (91.1%) 5 (100%) - Non exposed water 4 (8.9%) - Underlying disease α-thalassemia 3 (6.7%) - - β-thalassemia 3 (6.7%) - - β-thalassemia Hemoglobin E disease 32 (71.1%) 4 (80%) - H-constant spring 2 (4.4%) - - Hemoglobin H disease 5 (11.1%) 1 (20%) Serum ferritin (ng/ml) Duration from disease onset to 1 st medical attention (mo.) Duration from diagnosis to first definitive surgery (mo.) Factors relates to survival rate Criteria of 1 year survived Survival group (45) 1,388.40± 653 1,676 ± Duration time (disease 1.9 ± ± 0.7 < onset to surgery) 0.6 ± ± 1.2 < Deceased group (5) P-value Disease and treatment related parameters Lesions Brachial artery 1 (2.2%) - - Radial artery 11 (24.4%) - - Ulnar artery 1 (2.2%) - - Femoral artery 17 (37.8%) 4 (80%) - Anterior tibial artery 9 (20%) - - Posterior tibial artery 3 (6.7%) - - Iliac artery 3 (6.7%) - - External carotid artery - 1 (20%) Surgery < Amputation 45 (100%) 2 (40%) - Debridement - 3 (60%) Antif ungal agent Itraconazole alone 5 (11.1%) 2 (40%) - Itraconazole + terbinaf ine 34 (75.6%) 3 (60%) - SSKI + terbinaf ine 6 (13.3%) - Duration of antif ungal treatment (months) 5.9 ± ± Iron chelation drug 45 (100%) 5 (100%) - * By chi-square or Fisher s exact test Age Type of Surgery 14
15 Factors relates to survival rate Survival Criteria of 1 year survived group (45) Clinical sign/symptoms post treatment initiation Deceased group (5) P-value - Fever > 38.2 o C - 3 (60%) Arterial insufficiency syndrome (claudication, paresthesia, gangrenous ulceration) - 2 (40%) Mass at surgical sites (arterial aneurysm) - 1 (20%) New skin lesions Inflammation/infection at surgical sites - 3 (60%) Biomarker (1,3)-β-D-glucan profile; serum (pg/ml) The presence of Pi-Ab Clinical symptoms post treatment initiation 30 15
16 31 Temperature of Southeast Asia in August September, 2015 Ref: Climate prediction center, NOAA 16
17 - King Chulalongkorn Memorial Hospital (Thai Redcross Hospital) - Faculty of Medicine, Chulalongkorn University Our team + 12 Hospitals enroll 34 17
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