2. Antimicrobial Resistance. 1. AMR: the global issues for health 2. AMR in E. coli of the endangered bird in Okinawa Island, Japan

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1 2. Antimicrobial Resistance 1. AMR: the global issues for health 2. AMR in E. coli of the endangered bird in Okinawa Island, Japan

2 AMR: the global issues for health Pathway of antibiotics for human and veterinary use in the environment Berkner et al. (2014) Pathogen flow among wild and domestic host-pathogen systems: spill-over & spill-back (Thompson et al., 2009)

3 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

4 AMR in E. coli of the endangered bird in Okinawa Island, Japan Okinawa rail (Gallirallus okinawae)

5 The evergreen laurel forest zone called Yanbaru located in northern part of Okinawa Island where many endangered endemic species inhabit

6 Development threats to natural environments

7 Some endangered species use stock farms as a new habitat

8 Surveillance of wildlife in a stock farm 60m Camera trapping :The installation site of cameras and their direction 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

9 Surveillance of AMR in wildlife Areas of surveillance Kunigami village, Okinawa Main Island, located at N and 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

10 Sites of collecting fecal samples FA : Forest area : Human habitation : Stock farm : Pref. road 70 : Pref. road 58 : Sampling site HH Google maps of sampling points at the northern part of Okinawa Island Feces of the Okinawa rail on the Pref. road

11 Detection of AMR E. coli from feces Forest area 20.0% 3 SPL 42.3% 11 SPL 57.7% 15 SPL Prevalence of E. coli 80.0% 12 SPL Prevalence of AMR E. coli Human habit. 27.3% 6 SPL 32.1% 72.7% 16 SPL Prevalence of E. coli 5 SPL 68.8% 11 SPL Prevalence of AMR E. coli : Forest area : Human habitation : Stock farm : Pref. road 70 : Pref. road 58 : E. coli : AMR E. coli

12 Sites of collecting environmental samples HH : 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 HH

13 Detection of AMR bacteria from environmental samples Forest area 16.6% 25.5% 2 SPL 12 SPL 74.5% 66.7% Prevalence of E. coli Prevalence of AMR E. coli 35 SPL 10 SPL Human habit. 36.4% 63.4% 12 SPL 21 SPL 8.3% 1 SPL 91.7% 11 SPL Prevalence of E. coli Prevalence of AMR E. coli : : : : : : Human habitation Stock farm Pref. road 70 Pref. road 58 E. coli AMR E. coli

14 Genotype of E. coli from samples : Fecal sample collected at the forest area : Fecal sample collected at the stock farm : Environmental sample collected at the forest area Same genotype btw two sites 5km apart

15 Resistance to antimicrobial agents Number of AMR E. coli detected from FA and HH Abbreviations of antimicrobial agents ABPC PIPC KM OTC NA OFLX CP ST CET FA HH Minimum inhibitory concentration (MIC) score MIC (µg/ml) Max. Min. OTC NA OFLX CP ST CET >256 - >32 - >256 - >32 - Red: synthetic antibacterial agents 32 - MIC (µg/ml) Max. Min. ABPC PIPC KM OTC OFLX CP NA ST CET >256 - > >256 - > >32 - >256 - >256 - >32 - >256 8

16 3. Avian Influenza 1. Avian Influenza in Asian countries 2. Avian Influenza in wild birds of Japan 3. Molecular surveillance of avian influenza virus, using waterfowl fecal samples

17 High & Low Pathogenic Avian Influenza outbreak The status of the outbreaks in poultries of HPAI & LPAI incidents of the world As of 22 May 2017 HPAI LPAI

18 High & Low Pathogenic AI in Asian Countries The status of occurrence of HPAI/LPAI in Asian regions from October 2016 to May 2017 In Indonesia 18, avian influenza outbreak has continuously occurred. As of 17 May 2017 chickens wild birds Red: HPAI Blue: LPAI

19 Avian Influenza in wild birds of Japan Background 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 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 and No HPAI in wild birds H5N8 outbreaks 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 cases of wild birds in 22 prefectural and city governments - the highest number in 9 years 12 of cases of poultry farms in 9 prefectural and city governments : wild birds : poultry farm

20 Molecular surveillance of avian influenza virus using waterfowl fecal samples by courtesy of Dr. Onuma of National Institute for Environmental Studies

21 Material and Methods Fecal sample collection in 53 sites covering all over Japan ( ) Total: 17,540 samples AIV gene detection Species identification 221 samples RT-LAMP method (EIKEN) AIV gene Positive 335 samples (1.9%) DNA barcoding and information of direct observation

22 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

23 Species identification using fecal samples

24 Risk analysis for avian influenza in wild birds Independent 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 (Moriguchi et. al., 2013) Risk Index Not analyzed

25 Risk species Mallard plays important roll to introduce AIV into Japan and maintain AIV in Siberia Northern Pintail may bring North American lineage of AIV Migration route of the mallard duck Migration route of the northern pintail Biodiversity Center, Web GIS Atlas of Birds (Bird Banding Survey, Data of recovery records

26 Fecal sample is very valuable to get a lot of useful information of wildlife infectious diseases

27 Conclusion Regarding the surveillance for wildlife diseases, it is needed; 1. To record and report for sharing information of wildlife diseases internationally 2. To understand interface and interaction among live stocks, wildlife and human 3. To understand Ecosystem and Ecology, and also keep the image of One Health in mind

28 Thank you for your attention! The Sumatran tiger /Harimau (Panthera tigris sumatrae)

29 Regional Workshop: Training of OIE National Focal Points for Wildlife (4th Cycle) Bali, Indonesia, 4-6 July 2017

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