Zoonotic Diseases

Human Animals

Environmental

Figure 1. Flow chart of the pathway from introduction of a zoonotic pathogen to public health impact, represented by 7 criteria

(C1–C7) from which the risk to public health of emerging zoonoses was derived.

Havelaar AH, van Rosse F, Bucura C, Toetenel MA, Haagsma JA, et al. (2010) Prioritizing Emerging Zoonoses in The Netherlands. PLoS ONE 5(11): e13965..

Brucella spp* Brucellosis

Francisella tularensis*

Tularemia

Leptospira Leptospirosis

Borrelia spp Lyme disease

B.pseudomallei*

melioidosis

Coxiella burnetii*

Q fever

Y.pestis Plague etc.

*Biotorrism

• Brucella is a fastidious,

aerobic, small, gram-

negative coccobacillus.

• Biosafety Level 2/3 agent

(once Brucella is suspected,

work should only be done

in a certified Class II

Biosafety Cabinet).

• Biotorrism :

Wimol Petkanchanapong, Thai NIH

Brucella spp*/Brucellosis โรคบรูเซลโลซิส /โรคแท้งติดต่อ

B.suis B.melitensis

B.abortus

B.canis

Symptoms and signs %

Fever 93

Chills 82

Sweats 87

Aches 91

Lack of energy 95

Joint and back pain 86

Arthritis 40

Spinal tenderness 48

Headache 81

Loss of appetite 78

Weight loss 65

Constipation 47

Abdominal pain 45

Diarrhoea 7

Cough 24

Testicular pain/epididymo-orchitis 21a

Rash 14

Sleep disturbance 37

Ill appearance 25

Pallor 22

Lymphadenopathy 32

Splenomegaly 25

Hepatomegaly 19

Jaundice 1

Central nervous system abnormalities 4

Cardiac murmur 3

Pneumonia 1 Wimol Petkanchanapong, Thai NIH

Culture :Specimens

•Blood/ Castaneda’s media

•Urine, sputum, breast milk

•Lymph node biopsy

•Bone marrow aspirate

• EDTA blood • Tissue/BM/spleen/liver

NA detection /PCR

Specimens : clotted blood/

serum •single serum > 1 ml • paired serum >14 day

Antibody detection : IC/LA/RB/ELISA/IFA-IgM-IgG

Brucellosis is a commonly acquired laboratory infection; all work on suspect Brucella spp. cultures should be performed at a minimum under BSL2 conditions with BSL3 practices.

Lab testing

Wimol Petkanchanapong, Thai NIH

สถาบันวิจยัวทิยาศาสตร์สาธารณสขุ •เพาะเชือ้ จากสิ่งสง่ตรวจและตรวจยืนยนั • ตรวจสารพันธกุรรม ด้วยวธิ ีPCR • ตรวจแอนตบิอดดีว้ยวธิ ีRose Bengal Test และ IgG/ IgM-ELISA ให้บริการทุกวนั

Persons at Risk Exposure Activities

PEP Recommendations

Follow up/Monitoring

Person performing

activity and any

person within a 5 ft.

radius

•Work with a

Brucella isolate

•Sniffed or opened

culture plate

•Mouth pipetted

specimen material

•Worked in Class

II biosafety

cabinet or on open

bench without

using BSL-3

precautions

•Doxycycline 100mg

twice daily and

rifampin 600mg once

daily for 3 weeks

•TMP-SMZ should be

considered for patients

with contraindications

to doxycycline

•Persons with

contraindications to

rifampin should consult

with their HCP

•Pregnant women

should consult with

obstetrician

•Sequential

serologic testing at

0, 6, 12, 18 and 24

weeks post exposure

•Symptom watch

(e.g. weekly) and

daily self fever check

for 24 weeks

•No serological

monitoring available

for RB51 and B.

canis exposures

All persons present in laboratory room

Occurrence of widespread aerosol generating procedures*

Brucellosis @CDC.org

Risk Level = Low Persons at Risk

Exposure Activities PEP

Recommendations

Follow

up/Monitoring

All persons present

in laboratory room

at distance greater

than 5 ft. from

activity

Present in the lab at

the time of

manipulation of

Brucella isolate on

an open bench, but

who do not have

high risk exposures

as defined above

• Discuss with HCP

•.May consider if

immunocompromis

ed or pregnant

• Sequential

serologic testing at

0, 6, 12, 18 and 24

weeks post exposure

• Symptom watch

(e.g. weekly) and

daily self fever

check for 24 weeks

• No serological

monitoring available

for RB51 and B.

canis exposures

Brucellosis @CDC.org

Risk Level = None

Persons at Risk Exposure Activities

PEP Recommendations

Follow up/Monitoring

None Handling and testing of Brucella isolate in a Class II biosafety cabinet using BSL-3 precautions

None N/A

Brucellosis @CDC.org

Brucella as a Biological Weapon Aerosolized B. melitensis

City of 100,000 people

Inhale 1,000 cells (2% decay per min)

Case-fatality rate of 0.5%

50% hospitalized for 7 days Outpatients required 14 visits

5% relapsed

Results 82,500 cases requiring extended therapy

413 deaths

$477.7 million economic impact

Center for Food Security and Public Health, Iowa

State University, 2012

Q fever/ Coxiella burnetii

• A small obligate intracellular

gram-negative bacterium that is

distributed globally.

• Primary reservoirs; cattle,

sheep, and goats

• Acute and chronic illness in

humans with case fatality rate

is 25-60%. • group B bioterrorism.

Wimol Petkanchanapong, Thai NIH

Signs and symptoms

• Chronic patients may have initially had an asymptomatic and

present months or years after bacterial infection.

• About 80% cases present as blood culture negative

endocarditis.

• Other chronic of infection;

• Vascular infections

• Infections after pregnancy

• Hepatitis

• Osteomyelitis

Wimol Petkanchanapong, Thai NIH

Q fever/ Coxiella burnetii

Tularemia/ F. tularensis

Wimol Petkanchanapong, Thai NIH

Tick –borne disease

Tularemia : -Ulceroglandular/Glandular/Oculoglandular/Oropharyngeal/pneumonic/typhoid form.

Fever/chill/

Risk factor: working closely with animals/ living in forested areas/gardening/

Sources: Rabbit and deer ticks/deerflies/rodents/cats

Route: skin exposure/inalation/eye /ingestion/

Tularemia/ F. tularensis

Wimol Petkanchanapong, Thai NIH

Lab testing

•Antibody detection :

•Skin biopsy/LN biopsy /BM

• PCR •Culture :

เลปโตสไปโรสิส Hardjo or Pomona

เชื้อไปสะสม ที่ไต และออก มากับปัสสาวะ

เชื้อเขา้สูก่ระแสโลหติ และไปอวัยวะอื่นๆ

การติดตอ่ เชื้อเขา้สูร่า่งกายคนโดย การไชเขา้ทางผวิหนงั

ที่มีบาดแผล หรือไชผา่น เยื่อเมอืก เยื่อบตุา

เยือ่บทุางเดนิอาหาร และทางเดนิหายใจ

การก่อโรค ติดเชือ้ในกระแสโลหติ เกิดการอกัเสบทีต่บั ไต สมองและอวยัวะอื่น ท าลายหลอดเลอืด และเนือ้เยือ่

Wimol Petkanchanapong, National Institute of Health, Thailand

ไข้ 90-100%

ปวดเมือ่ยกล้ามเนื้อ 40-93.8%

ปวดศรษีะ 55-74.6%

Conjunctival suffusion 25-60%

Jauandice 16-92.7%

Figure 1 Global annual incidence of human leptospirosis. Colors reflect

incidence, in declining order: red, pink, green, yellow. Gold reflects areas with probable, but not estimated, high incidence. White reflects absence of data.

Wimol Petkanchanapong, Thai NIH

18/07/2013 W. Petkanchanapong, Leptospirosis Laboratory, National Institute of Health. 16

Laboratory investigations / PUO

Different approaches of laboratory diagnosis

Culture*

Isolation

Microscopy

•Dark Field Microscopy

•Immunofluorescence

Microscopy

•Silver Impregnation techniques

Immunology

Latex*, ICT*

ELISA, IHA, IFA*

Microscopic Agglutination Test (MAT)*

Molecular

•Polymerase

Chain Reaction

(PCR)*

• LAMP**

•MALDI TOP

mass

spectrophotometr

y

•Next generation sequencing

Emerging Zoonosis Research Project

Determine the incidence of

leptospirosis, brucellosis,melioidosis,

murine and scrub typhus, parasites in

animals with close contact to human

(dogs, cats, rats)

Development of new techniques for lab

identification of zoonoses using

multiplex approach; multiplex real-time

PCR, multi-dot IFA

Development incidence of bartonellosis,

Q fever and tularemia in wildlife and risk groups

Sample collection : 1. Identify rodents and ectoparasites 2. Specimen collection: clotted

blood, EDTA blood, blood smear

3. kidney/spleen/liver: culture,

PCR for ---------------

ตารางที ่1 ผลการตรวจหาการติดเชื้อโรคในกลุม่ Zoonoses *

ชนิดสัตว์

จ านวน

สัตว์ (ตัว)

Leptospirosis/ MAT

+/ToTal (%)

Brucellosis/ Agglutination

+/ToTal (%)

Melioidosis/IHA

+/ToTal (%)

Q fever/ IFA /ELISA

+/ToTal (%)

Bartonellosis /IFA

+/ToTal (%)

Scubtyphus/ IFA+/ToTal

(%)

Murine typhus/

IFA+ /ToTal (%)

Trypanosome * +/Total (%)

หนู 238 2/112 (1.8) 0/28 (0)

12/125 (9..6)

1/85 ( 1.2 )

1/85 (1.2)

0/44 (0)

0/44 (11.8)

10/85 (11.8)

สุนัข 255 20/255 (7.8)

31/167 (18.6)

15/160 (9.4)

0/17 (0)

0/167 50/82 (60.9)

3/82 (3.7)

8/167 (4.8)

แมว 93 1/70 (1.4)

2/33 (6.1)

2/57 (3.5) 1/33 (3.0 )

0/33 (0)

8/11 (72.7)

0/11 (0)

0/35 (0)

รวม 586 23/420 (6.1)

Bandicota

indica R. norvegicus Rattus rattus Rattus exulans

Suncus

murinus Total

ภูเกต็ - 5 23 11 39

+ (%) 5 (12.8) A 5 (12.8) ระนอง - 56 21 2 35 114 + (%) 17 (14.9) B 8 (7.0) C 25 (21.9)

นครพนม -- 3 7 14 44 + (%)

ตาก 4 1 1 25 10 41 + (%)

รวม 4 65 52 52 45 238 (+ %) ของชนดิหนู 0 17 (26.2) 13 (25.5) 0 (0) 0 (0)

(+%) ของทัง้หมด 0 (7.1) (5.5) 0 0 30 (12.6)

serogroup ท่ีพบ A B C %

Bataviae 1 12 5 18 (7.6) Canicola 2 2 1 5(2.1)

Celledoni 2 3 2 7(2.9)

ตารางท่ี 2 ผลการตรวจหาการติดเช้ือโรคในกลุ่ม Zoonoses

Test kit

Multiplex real-time PCR for detection of 9

vector-borne diseases and zoonoses;

1. leptospirosis, brucellosis, melioidosis, and tularemia

2. bartoneloses, scrub typhus, murine typhus, and rickettsial

spotted fever

3. Q fever, leptospirosis, scrub typhus, and brucellosis

Thanks for your attention

Plague/ Yersinia pestis Very severe disease in humans, with 30-60% case-fatality

ratio

Develop ‘flu- like” symptoms after an incubation period

of 3-7 days.

3 forms of plague infection:

Bubonic/septicemic/pneumonic

783 cases and 126 deaths reported worldwide in 2013

Most endemic countries : Madagascar, Congo, Peru

Wimol Petkanchanapong, Thai NIH

ตารางที ่2 ประเมินความถกูต้องของวิธ ีMultidot IFA เปรียบเทยีบกบัวิธอี้างองิ Brucellosis

(%) Leptospirosis

(%) Melioidosis

(%) Murine typhus

(%) Scrub typhus

(%)

Sensitivity 93.1 (27/29)

89.6 (26/29)

100 (17/17)

92 (23/25)

100 (25/25)

Specificity 82.5 (137/166)

92.2 (153/166)

88.2 (157/178)

94.7 (161/170)

95.3 (162/170)

Blood donors

97.1 (68/70)

100 (70/70)

95.7 (67/70)

98.6 (69/70)

98.6 (69/70)

Other infection

71.9 (69/96)

86.4 (83/96)

83 (90/108)

92.0 (92/100)

93.0 (93/100)