2. Antimicrobial Resistance1. AMR: the global issues for health2. AMR in E. coli of the endangered bird in
Okinawa Island, Japan
AMR: the global issues for health
Berkner et al. (2014)
Pathogen flow among wild and domestic host-pathogen systems: spill-over & spill-back (Thompson et al., 2009)http://www.who.int/foodsafety/areas_work/antimicrobial-resistance/tripartite/en/
Pathway of antibiotics for human and veterinary use in the environment
E. coli as an indicator of AMR Why Escherichia coli?
• Wide range of distribution: human, livestock, wildlife and environment
• Easy to detect comparing with the other bacteria• Many back ground medical data and information
http://www.ecl-lab.com/en/ecoli/index.asp
The evergreen laurel forest zone called “Yanbaru” located in northern part of Okinawa
Island where many endangered endemic species inhabit
:The installation site of cameras and their direction
Camera trapping Trail camera (Ltl Acorn LTL 5310a) Relative Abundance Index (RAI)
=(effective shooting numbers ÷ (number of cameras × operating days) × 30
Installation condition 1-2m off the ground Angle of depression: 0-30°
60m
Surveillance of wildlife in a stock farm
Surveillance of AMR in wildlife Areas of surveillance
• Kunigami village, Okinawa Main Island, located at 26°45’ N and 128°17’ E
• Forest areas (FA) and Human habitation (HH) Samples
• Fecal samples from the Okinawa rail: 48 samples• Environmental samples (soil and water) from
the forest area of “Yanbaru”: 80 samples
: Forest area (FA): Human habitation (HH): Stock farm: Pref. road 70: Pref. road 58
Sites of collecting fecal samples: Forest area: Human habitation: Stock farm: Pref. road 70: Pref. road 58: Sampling site
FA
HH
Google maps of sampling points at the northern part of Okinawa Island Feces of the Okinawa rail on the Pref. road
Detection of AMR E. coli from feces
Human habit.
: Forest area: Human habitation: Stock farm: Pref. road 70: Pref. road 58: E. coli: AMR E. coli
57.7%15 SPL
42.3%11 SPL
72.7%16 SPL
27.3%6 SPL
Prevalence of E. coli
Prevalence of E. coli
20.0%3 SPL
80.0%12 SPL
32.1%5 SPL 68.8%
11 SPL
Prevalence of AMR E. coli
Forest area
Prevalence of AMR E. coli
: HH: Stock farm: Pref. road 70: Pref. road 58: Soil sampling site: Water sampling site
Site Soil(No of SPL)
Water(No. of SPL)
Total(No. of SPL)
FA 33 14 47
HH 21 13 34
HH
Sites of collecting environmental samples
Human habit.
Forest area : Human habitation: Stock farm: Pref. road 70: Pref. road 58: E. coli: AMR E. coli
16.6%2 SPL
66.7%10 SPL
8.3%1 SPL
91.7%11 SPL
25.5%12 SPL
74.5%35 SPL
36.4%12 SPL63.4%
21 SPL
Detection of AMR bacteria from environmental samples
Prevalence of E. coli
Prevalence of E. coli
Prevalence of AMR E. coli
Prevalence of AMR E. coli
: Fecal sample collected at the forest area
: Environmental sample collected at the forest area
Genotype of E. coli from samples
: Fecal sample collected at the stock farmSame genotype btw two sites 5km apart
MIC(µg/ml) OTC NA OFLX CP ST CET
Max.Min.
1281
>256-
>32-
>256-
>32-
32-
Abbreviations of antimicrobial agentsABPC PIPC KM OTC NA OFLX CP ST CET
FA 0 0 0 3 1 1 2 1 1HH 4 3 1 9 2 2 2 4 5
MIC(µg/ml) ABPC PIPC KM OTC OFLX CP NA ST CET
Max.Min.
>256-
>25632
>256-
>256128
>32-
>256-
>256-
>32-
>2568
Number of AMR E. coli detected from FA and HH
Minimum inhibitory concentration (MIC) score
Red: synthetic antibacterial agents
Resistance to antimicrobial agents
3. Avian Influenza1. Avian Influenza in Asian countries2. Avian Influenza in wild birds of Japan3. Molecular surveillance of avian influenza virus,
using waterfowl fecal samples
High & Low Pathogenic Avian Influenza outbreak The status of the outbreaks in poultries of HPAI & LPAI incidents of the world
HPAILPAI
As of 22 May 2017
High & Low Pathogenic AI in Asian Countries The status of occurrence of HPAI/LPAI in Asian regions from October 2016 to May 2017
As of 17 May 2017●chickens ▲wild birdsRed: HPAI Blue: LPAI
In Indonesia ⑱, avian influenza outbreak has continuously occurred.
Avian Influenza in wild birds of JapanBackground of coping with HPAI
surveillance in wild birds in Japan• 2004 infected crows were confirmed around
the chicken farms which had HPAI outbreaks (secondary infection) the first time in 79 years
• 2007 One Mountain hawk-eagle• 2008 Whooper swans• 2008 Manual of technique and coping with
HPAI in wild birds for local governmental agencies was published by Ministry of Environment→ Start nation-wide surveillance
• 2010-2011 Outbreaks in wild birds (15 species, 60 birds), domestic birds, and some endangered species as Hooded cranes and White-naped cranes in Izumi City, Kagoshima Prefecture
• 2011-2012 and 2012-2013 No HPAI in wild birds
• 2014-2015 H5N8 outbreaks • 2016-2017 H5N6 detected from 218 samples
consist of wild birds (22 species), 82 zoo birds (5 species), 5 fecal samples from wild duck and 3 water samples of roosting sites
HPAI (H5N6) outbreak in wild birds and poultry farm in Japan from 2016 to 2017• 218 cases of wild birds in 22 prefectural and
city governments - the highest number in 9years
• 12 of cases of poultry farms in 9 prefectural and city governments
: wild birds: poultry farm
Molecular surveillance of avian influenza virus using waterfowl fecal samples
by courtesy of Dr. Onuma of National Institute for Environmental Studies
AIV gene Positive335 samples (1.9%)
RT-LAMP method (EIKEN)
AIV gene detectionSpecies identification
221 samples
DNA barcoding and information of direct observation
Fecal sample collection in 53 sites covering all over Japan (2008-2015)
Total: 17,540 samples
Material and Methods
Temporal changes of RT-LAMP positive rate Seasonality of HPAI virus positive in wild birds from 2008 to 2015
• High positive rate in October to December• Positive rate declined in April to May
Risk Index0.9‐1.00.8‐0.90.7‐0.80.6‐0.70.5‐0.60.4‐0.50.3‐0.40.2‐0.30.1‐0.20.0‐0.1Not analyzed
(Moriguchi et. al., 2013)
Risk analysis for avian influenza in wild birdsIndependent variables Contribution
Dabbling duck population 49.4 ± 5.5
Urban area 12.5 ± 3.5
Altitude 8.2 ± 2.3
Diving duck population 3.5 ± 3.5
Lake area 2.2 ± 1.5
Farmland area 0.9 ± 1.2
Poultry density 0.5 ± 0.5
Mean precipitation in winter 0.3 ± 1.0
Min temperature in winter 0.1 ± 0.1
Distance to lakes 0.0 ± 0.0
Sum of spatial filters 22.5 ± 5.6
Biodiversity Center, Web‐GIS Atlas of Birds (Bird Banding Survey, Data of recovery records)http://www.biodic.go.jp/birdRinging_en/index.html
Mallard plays important roll to introduce AIV into Japan and maintain AIV in Siberia
Northern Pintail may bring North American lineage of AIV
Risk species
Migration route of the mallard duck Migration route of the northern pintail
ConclusionRegarding the surveillance for wildlife diseases,
it is needed;1. To record and report for sharing information of
wildlife diseases internationally2. To understand interface and interaction among live
stocks, wildlife and human3. To understand Ecosystem and Ecology, and also
keep the image of “One Health” in mind
http://iiad.tamu.edu/about-iiad/about/one-health/