Transcript
ddJaphfHuDk • Kurita Overseas Research Grant May Webinar
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Kind: captions Language: en you will have a presentation about 45 or 50 minutes and then after that is a q a so i will i will uh if it is ready or hospital probably the other will join uh in it's a bit okay i think that there's some feedback okay we may start uh i will introduce you uh now i will in the uh how to help this yes is there any uh there is no problem in me uh sorry the problem maybe change the microphone speaker or the speed yes right speaker oh sorry sorry sorry okay okay i think i have my problem with it okay okay i will introduce you through the i think i will share my screen first [Music] okay uh good morning uh everybody so uh today uh our uh our programs is the uh webinar uh and uh today lecture uh everybody just okay so should be uh or changing the speed uh changes besides the mute button there are the upward arrow choose the speaker towards the headphone or the microphone towards the headset microphone yes both of them already in headphone or micro or headset okay you can hear my my voice yes clearly okay it's already clear right okay sorry sorry for for a very uh just i will i would like uh to introduce uh uh for today webinar so our program is uh the big winner by dr honda sorry okay so that uh dr honda is from the karnataka uh university he is a lecturer now in the university is an associate professor and then he graduated from the university of uh tokyo so that our program for today after the webinar there will be a call for for proposal for the kurita offices grant but let's restart uh with the webinar given by the dr honda dr honda your time use please okay thank you very much for your kind introduction from the chandra so i rested my lecture can i see my slides okay so thank you very much uh for giving me this kind of good opportunity to talk about under microbial resistance so i think you some of you may already know about the antibiotic resistance so i want to talk about i have been the down the field survey in japan also some asian countries like thailand india sri lanka uh about the microbial resistance in water environment in waste border so i want to talk about my experience and including some literature review and i want to give you some perspectives so now copy 19 is the really good big issue and in the world but mr is also it's a long time long term problems in the future so this is the very famous on your reports which which is says that's the antimicro resistance becoming will predict to become the major uh cause of the deaths up to the 10 million cases and it will be more than the cancer in 2050 and this decimated deaths an eerie estimated year annual death due to anti-microbial resistance is is less than 1 million in 2013 but they feel predicted reaching 10 million in 2050 and the report estimates that the major major impacted the area is the africa and asian countries so ngo predicted to have almost 5 million tests because of the antimicrobial resistance so and one important another important issue in mr is the mri is not a problem is the one countries but also the kind of world issues so this is a case of the one superbug called ndm1 which has the resistance to the most beta-lactam antibiotics and it was this was found in swedish patients who traveled india and but after a couple of years then this nd1 was defined in the at every continent so yeah currently this uh people doesn't have the international trouble because of coffee situation but when we reopen the gates for international trouble so amr is spread all over the world because of the people trouble and also possibly by the food trade so asia and africa are now concerned the potential site of new mr emergence so once the new mri emergent emerged then it spread to the whole of the countries probably in the two or three years so concerning this situation the whole unap and the many written international organizations the proposes advocate to talk with them or issue the words and now now it is said it is important that one health approach for mr mitigation the one health is the approach to of the probability for the multiple disciplines of environment animal and human health so because this is our health approach is not only for mr but every kind of disease infectious diseases as you know copenhagen is originated from the uh bat coronavirus or the pangolin coronavirus and the infected human and spread into the human society and amr is also spreads among these disciplines domains it spreads human to human but also the animal to humor and humor human discharge mr into the environment and that's the environment destroying a mr district environment possibly transmitted to the animal or the humans again so we need the comprehensive approach yeah to control mr not only the human only in the animals not in the environment but to include all of them and we are in bioengineers so uh we can we now we work for mr issues in water environment and in water environment wastewater is a major source of ammo when we have the drugs antibiotics or the the animals the livestock animals like cows pig also chickens also heads fed with antibiotics so then the bird resistant bacteria is enriched in our body or the this livestock bodies and they are discharged in human waste like feces then as a wastewater and some may be treated but the wastewater treatment plant doesn't have the reduction lamar but not complete reduction i would talk about this later and someone there could be discharged in a war environment or in some area which doesn't have the wastewater treatment the wastewater containing antibiotic resin bacteria is discharged to a water environment directory so then when the drinking water source or the the water is used for bathing or swimming or fishing these antibiotics are possibly transmitted to humans or the animals again and possibly cause disease so when you are infected the antibiotic resistance infection to this you you cannot cure treated by the antibiotic drugs so you may suffer from the longer uh duration of disease sickness or maybe you will get more severe sickness so i want to talk about our experience of the antibiotics bacteria asia water environment so it is already uh i think almost 10 years ago maybe just six or seven years ago uh we did river catchment-based catchment scale field survey in thailand about antibiotic bacteria in river water so we target the chopper river and also uh the four major tributaries ping yum now one reverse and we took the sample maybe about 30 30 or 40 sites and we isolated the e coli from the river water and we tested the resistance to the six antibiotics [Music] of each e coli isolates from the river and what we found is the resistance bacteria is present not only the urban area but also rural area but near the urban area like in chiang mai or here's a bangkok this is japan river so there are high abundance of multiple resistance which includes the clinically important antibiotics like quinolones so mainly chopper river basin so urban drainage is its impact to increase the mr to the clinically important antibiotics but also in the rural areas the classic antibiotics like tetracycline or suffer drug is also abundant so already number is spreads all over the basin and the higher abundance near the urban area [Music] and this is a case in the sri lanka we did in the maybe two or three years ago and we took the sample from the kalani river which is the river flowing from the center of the uh ceiling island to the capital of sri lanka colombo city and this river is used for drinking water source of colombo city and we took the sample from the four sides named k1 to k4 and isolate the e cli and abundance of five antibiotics was analyzed by this diffusion methods so from k1 to k4 it's k1 the upmost upstream so for levels versions the ciprofloxacin they are they are quinolons which is clinically important antibiotics and they have have the high abundance they should have on since the upstream area because optionally i also have the the populated area uh the the along the calibre there's there are uh operator area so probably the in the upstream is too much affected by the upstream water optimal drainage and classic antibiotics it is really highly abundant in any place in the river so these reasons these drugs canamycin tetracyclines suffer drug they are already abundant is all over the river and this is the case in the india in the guwahati in the assam area it's north east of india and we took the samples from the the gandhinagar river this is a really big river b1 to b4 and also the urban drainage in guwahati gohari city doesn't have the wastewater treatment plants so wastewater is is discharged into the urban channels and we analyze the antivirus genes from these samples so b1 b2 is the upstream of the guaca city and the b3 b4 is the downstream of the quahazi city so in the downstream of the quahog city we found that asc this is the aminoglycoside uh research family glucoside and also the uh blood theme this is beta lactam and i'm sitting resistance genes and these are also found in the urban drainage so for these controversial genes the urban drainage from the guatemala city is probably the source of the sauce in the downstream river sites so antibiotics against queen confirmed drugs will often get in the urban drainage in guahana city so so there's one important questions when we study about uh antibiotic bacteria in wastewater treatment plant the wastewater plants redu treat the wastewater but also the plant keeps the sludge in the sludges the the complex uh matrix of the the microorganisms so the more question is does the wastewater in the plant works as a reservoir or barrier of arv so it is known that strategy also contains some antibiotic resistant bacteria so my question is both which works when it works is both reservoir and barrier and i want to introduce the fate of antibiotics and bacteria in wastewater treatment so let's think about the fate behavior of the resistance bacteria in the worst world treating plants but this is this is this is not about the antibiotics and bacteria but about the total bacteria population in wastewater treatment plant wastewater treatment process in nature have can reduce the population bacteria so this is data is taken from the literatures and this is the logarithmic value of the total heterotrophs and total coliform and e coli so it depends height it depends on the wastewater plant but as as average total heterotrophs is to reduce like about 1.5 log and so the conform is though about 2 lakh and the equalize is 2.203 logs so why plant can reduce the total bacteria population and in case of e coli and this is fecal indicator bacteria it reduces two log that means the equal population become one percent in the treated effluent than will influence wastewater so when we discuss about the removal performance reduction performance of arb it's the poor question is the are the air be reduced more than the total bacteria or the lesser total bacteria and the conclusion is the reduction of the antibiotic bacteria is almost equal to the total bacteria reduction so that means there's no specifically larger or smaller removal of arb compared to the other bacteria non-resistant bacteria so this this chart is created from the uh study of the novel manager and this x-axis is the population of the bacteria in the fluent and y-axis the population in the effluent and i put that so this black one is the total bacteria preparation it includes resistant bacteria and also non-resistant bacteria so this black one so if we put the block once for example in one data the influence is about six lock and this length is like about five block so when we draw the line we this the dotted line it shows the logarithmic value at one point three so total bacteria is you know the work is about 1.3 and this collat plot is the resistant bacteria blue is oxycetine reds and bacteria and orange wine the tetracycline the red one is ciprofloxacin one of the quinolones resistance bacteria so of course this the population is less than the total bacteria in most cases and they have of the fluctuated population in the fluent but reduction is almost on the same line showing there will be 1.3 so that means the total bacteria and the resin bacterias have the similar removal value so logan valve arb is equal to every total market so there's no specific removal of arb in the wastewater treatment plant and this is this is the change of abundance of resistance e coli from effluent to influential effluent at each step of the wastewater training process so this is the population of the e coli so vibration equalize uh it's about the about the five block and it doesn't change much until the aeration tank but the reduced after the secondary treatment because it's second treatment they remove the bacteria by sedimentation so you can reduce it like maybe two points five long and abundance of resistance e coli is the influence is about forty percent and if lentils are 40 percent so it was similar so this is supports the previous results the removal of the arb is doesn't change it's not different from therapy of total bacteria so that means the ratio ratio of proportion of the mr in the bacteria is is same so and but in this slash it slightly increases so something happens in the in the slides but this is the this is taken in japan based on this diffusion methods we take the sample and cultivate the e coli and isolate the e coli and put the antibiotic drug tablet on the media and [Music] check the resistance to each antibiotics but when we did it in the meta genomic survey will be quite different it should quite different results so this is the abundance of total or resistance genes is for 16s based population so this data takes but we took the samples and we exact dna and the the metagenomic sequence read the sequence of the dna and to get the number of reasonable resistance genes and we also get the uh bacteria preparation as the number of reusable 16s ribosomal energy so it's kind of the abundance gene-based abundance of antibiotic resistance so we did this survey at the five plants we took three times in summer three times in winter and we made a composite sound force then we found the interesting results and this is influence slash and the treated effluent so in influence it's total arg per 16s your population is about or 40 percent but it decreased at the slash tank and this setting is almost same so now your one plant every plant has the influence wastewater has the highest the gene-based abundance of resistance and it decreased a lot in the sauce tank and in the fluent it's a slightly increase almost same so abundance about the resource genes decreased in sludge so it's really inconsistent with the results where we got from the culture-based abundance of antibiotic resistance and so i suspect it whether it is will be true or not so but in there we we surveyed the other rituals but there it is also reported the similar results this is the metagemic and gene based abundance over adding antibiotic resistance the influence is the highest and slowest influence it has a lower abundance and this is the reactant pcl based study of the target adverse genes to the sixth senior revolutionary gene copies so influence this is a slash this is effluent but this plant has this similar order will increase but this and this plants the influent is the highest region based abundance and the decrease in sludge and the fluent this plant also so but it is likely true in the many plants that the gene based abundance about the resistance is highest in influence and decrease the sludge influence sorry so if we cultivate the e coli or the coliform and the studied abundance mr and the culture based then influence and the fluent have similar abundance but if we study the gene-based abundance email using the methodology analysis or the qpcr of the target energies this is decrease in this after slash aeration tank and the affluent so this looks really inconsistent but i think the key is the culture-based analysis target the fecal bacteria like e coli or coliform or enterococcus enterobacter but gene-based abundance target the whole bacteria committee not only the euclid not on the fecal bacteria but also the every bacteria which presents in this sludge so my hypothesis and to explain this phenomenon these results is the difference in the fecal bacterial and non-fecal bacteria in the wastewater is mostly consists of fecal bacteria and resistance maybe about 40 percent of one divergence bacteria but in slash you get this free abundance of antibiotic resistance if you can possibly increase by horizontal gene transfer or the resistance induction by the stresses like oxidative storage or whatever in the slash but let's contest a lot of non-fecal bacteria because it's arabic conditions temperatures like like 20 15 to 30 degrees celsius thicker butter grow well in the anaerobic conditions and temperature at like 35 to 40 degrees celsius so slash bacteria communities is quite different from the wastewater community so it contains many unknown fecal bacteria and my speculation is probably the norfolk but there's a small abundance resistance so when we see the total abundance resistance it looks decrease and the affluence well maybe it's speculated to have this similar ratio of the vectors discharged so they have this also has a smaller abundance of resistance and but this is uh so this is supporting data and also we have some difference from the hypothesis so this we took the five plants we compared the antibiotic gene profile of each samples and compare with the principal component analysis and this blue plot is the wastewaters of the fire plants and the orange prod is a sludge samples so wastewater sludge has a really different uh antibody resistant gene profile so this supports the the high places that's because of different micro community or different theory profile it's quite different but what is interesting is that this is effluent so wasteful in the aeration tank the slush constant lacing amongst ss is about typical ms concentrates 1500 to 2000 and the ss consensus in the wastewater after primary sedimentation it's it's about like 100 or or sometimes less so that means the slush bacteria and only one per only less than 10 percent of wastewater bacteria is flows into such bacteria so if this large bacteria is equally i mean slush bacteria and the fecal bacteria wastewater equally discharged after primary ceremonial tank maybe effluent microbial committing effluent erg profile different is quite similar with the sludge because in the sludge most of them is is not the fico bacteria but interestingly the effect has the profile like between the wastewater sludge so it's not it's not like sludge it's not a wastewater just look between them so that is another the weird and the interesting point so in the final sedimentation process this means finite function process the wastewater bacteria is has maybe remove the less than the sludge bacteria such bacteria is the it's kind of enriched bacteria community which can settle well but wastewater bacteria does not does not enriched just due to digestion from our feces so maybe some of them is is out here on the slash particle and the settle but some of them does not the merge with the sludge the flux or such bacteria community but just a float swim in the liquid phase and maybe this bucket cannot be reduced by the sediment the final ceremony so probably my hypothesis is that some of the irb in the wastewater bypass the wastewater treatments and carried over to equivalent so yeah we are trying to study about this but anyhow so this is current hypothesis so so what total total reduction of wastewater treatment has this the reduction of the bucket operation and there will be also at the erb but the sum of the rb cannot be reduced in bypass effluent so that will affect the output resistance profile in the affluent uh which is different from the wastewater difference and another important source is not the fluent but untreated wastewater especially discharged from the storm water so yeah this is the photograph of the the big flood in thailand or in 2000 or 16 sorry yeah so as you marry though the waste water is is sometimes stumped without treatment because of sugar overflow a combined shoe overflow so even when you have the waste for treating plants so in dry weather all the wastewater collecting the same to the wastewater plant but it when you have the heavy rains the in the combined shower which collects the rainwater wastewater in one pipeline the wastewater is diluted with a storm water the rainwater and if rain is really heavy so we cannot treat all of the quantity in the wastewater in the plants so when the quantity exceed the capacity of treating plants we discharged into the river water or surface water directory without treatment and in the united states it started with stole data but in 2004 the u.s ep reports that 20 percent of our near discharge of combustion this is untreated cso so in the asian context asia has a monsoon weather so in the the rainy seasons you have a lot of heavy rain so this the csa happens uh i think more frequently than then temperate with the conditions and but so what motivated this fact i invest i try to quantify the discharge of antibiotics and bacteria from combined shiva overflow in japan and in this plants all the combined shiver is go to the pumping station and pumping stations they usually pump the all the waste water to the the waste treated plants but if the shoe waste quantity exceeds the front capacity they pump up to the sum of them into the river and but this bar chart shows the amount of uh sorry it's 45 minutes i rushed to finish it so what this is data in canada canada i have the heavy snow in this winter so we have a lot of cs in the winter and mine in this plant 23 percent combined shoe which put these charges on traditionally as well and we calculate the arb discharge into the river from the 3d defluent and also under the cs4 so this plant covered publisher 30 thousands and annually it risks plant receives i'm sorry the sure combined sugar received about 18 log of the log cfu of e coli per year and really a lot of equalized discharge onto the cso so as amount of waste water is about 27 000 sorry sorry 27 million cubic meter per year and about 75 percent this is treated affluent but total amount of equalize much less but in andrea cs world distress only do one force of the annual combined sure to which but it has really a lot of equal eye continuation so amr in this review is mainly discharged from not from 3df and but also from the untreated cs4 and i want to introduce some data about the rainy season in the asian countries so this is data in the sri lanka i showed in the in the beginning of this this talk and this is the rainy season and d means a dry season so rainy season helps the higher abundance of adverse bacteria in the upstream so that means there are in the this upstream area the in rainy season is this quantity of urban drainage increased i mean it's rainwater flushes the wastewater urban drainage and pollute the water with antibiotics and bacteria and this is in case of uh and rg re detection so in the anterior resistance genes basis so as i see the e coli abundance and mr in e coli is increasing in the rainy season but the concentration of alg is decreasing radiuses so but this is because the when the rainy season is the energy is diluted so concentration becomes less but the proportion resistance has become high so that means the loading i mean the total quantity of mr discharge is increased but as as we see as a concentration decrease because of dilution by rainwater so sorry i have so now i'm doing the uh com comparing the android zone among the countries so japan now i finished the comparing japan usa and china so different country has a different era profile and but actually the this data of tropical asian country is missing so i'm gonna try to study but i think young researchers also study about the ergonomic in the tropical ageing countries to cover these facts okay so it's already times uh this is a take out home message i always support the major social model and uh stonewall has much more therapy roles and in the concepts a context of the asian countries so mr and the water made much effective monsoon it's the rain rain uh season rainy season and there are many water diluting the rainy season but they were loading from wastewater probably higher in the rainy season and the ml profile in the wastewater divide some countries they're more soviets and tropicalizing countries and necessary yeah acknowledgements thank you very much so i quickly rush i want to have questions if it's not clear for you thank you okay uh thank you honda thank you so that uh we come up with the uh q a question uh session if you have a question you may open if you are in the zoom link so you may open your mic you may directly ask the question to dr honda or if there is in the chat box it may be going to may help us that uh what will be the the question in the chat more so that's uh things up probably uh uh you are in the in the in the zoom could if you have any question you may raise your question please uh i think there are already question questions from the chat okay okay i think so in the chat box but probably uh uh dr honda you may uh turn off your share please probably uh uh we will share the questions i think so okay okay so now i see the question yeah i think so google will help right so probably you can uh you have to turn off the share first and then uh for i think it might be uh okay i'm sorry okay okay so that is for from me so that this is a or you have already mentioned about the they are represented especially in the different countries you know that most of the foreign countries they don't have the uh facilities so how serious uh from your opinion uh okay thank you very much sandra says so i think it could be i think more serious in some countries uh in the level of country also so because i think upstream as as i show in the sri lanka case sometimes upstream urban drainage distribution river water and in somewhere in the in the developed countries the rival order is you used without treatments yeah some may bathing directly in the river water or the maybe some of you may catch the fish in the river so the i think in such countries this the higher risk than the the we use the river ward after treatment in the top order so that that is really i think the exactly the one health issues because the mr in the environment directory cost suppose the risk of the humans and also when you use the feeder vivo water to the animals maybe animals can also possibly get more so that is i think really important issue in the top collision countries uh i think so that's uh is there any it might be this uh because you you also uh studying about the shark of two virus is this any relationship or what we have been working on with the shark of two viruses presence in the water environment yeah thank you very much yeah the first chandra since they just suggested me to talk about the south country in west water but uh yeah i already talked about this i choose a different topic anyhow well i think the suspect when south coast is is contained in wastewater it could be also discharged in the river water water environment so what we know is that probably probably the subscriber is not very persistent in the wastewater drone it's i think it's vulnerable than the norovirus adenovirus which is classically studied viruses but so in the treated affluent probably the susceptibility is not become the big problem mostly are reduced or the rules the infectivity but the untreated wastewater we don't know probably it's the risk is much lower than the norovirus and other viruses but it depends on the situation i think it's there are many many patients in the in the in the area and it's and then possibly has this some amount of infectious subscribe too so yeah but i guess it's it's the risk is smaller than the other pathogens wastewater content is not only the subscription but the other many viruses other pathogens so my my speculation is maybe the risk is is higher in the other viruses of pathogens in west water okay thank you i think the uh the second one this one is from tuan uh so that is about the method each straight line do you follow when you carry out this diffusion we follow the clsi methodology yeah of course css also does something revised the criteria so it's i use the most updated one at the timing of the analysis yeah okay from return is from intern so that one is the third one they also observe or is there any evidence of gene transfer or antibiotic gene uptake in turin or in the racial retreatment plan yeah this is this is a yeah it's really important questions and i do not study about that but i know some papers about the possibility probability of gene transfer in the sludge so it happens actually it happens but not very high probability but it happens when we consider about the amount of sorry i don't remember the the exact value but it happens in the sum probability so and when we consider the concentration bacteria in the sludge it must slightly increase the possibility try to increase the population of antibiotics and bacteria yes okay thank you so uh so that one is probably kundalini is another question i think probably you can have to show it yes just just come in okay okay for me to make uh everybody can uh see the creation probably guinta will help us to add the additional but uh if anybody from in the in the in the room may i would like to ask the question directly to dr honda please you open your mic yeah dr honda okay okay please yeah i thank you uh i i'm not uh microbiologist something like that but i'm uh really interested with your statement or a hypothetical statement uh that in the uh there's no not much difference between the the inlet and not light isn't it so something like that so what do you think in the uh uh the fate of that uh anti uh uh out of the the resistance micro uh microbes what is the fate of uh that kind of microbes in the anaerobic situation because i i'm not more interested in anaerobic system thank you thank you very much so uh yeah there are some studies uh about dharmic digestion it's effective in the microbiologist so basically in the anaerobic digestion it's a little bit heated so it's reduces anymore i don't know which level but they can reduce some but not for the complete removal so it possibly remains some um heating uh system when there is uh normal just normal no heating uh treatment and so on it says the anaerobic situation uh has impacted or no no effect at all to the that kind of bacteria well minor exactly temperature effects now so i think they are studying the mesophilic mesophage condition is like or so maybe you don't need a heating in the tropical weather condition and so um so if antibiotics come from the fecal bacteria they probably grow well in the anaerobic and mesophyll conditions but i didn't i didn't see the paper like that probably it's it's it's rather reduced in dynamic digestion process okay but i i don't i don't see the many papers on that maybe it's the point that we need more studies uh yeah it's a good point thank you okay still vietnam so hospital resources might contain more rp than common one common uh municipal wastewater uh okay when monitor which is not fully treated so especially in the asian countries uh did you think the street control of hospital uh by onsite racial treatment plan it's one of the make sense to control the irv yeah i think sorry yeah this is very important point so there are several points of discussion the professor vietnam's questions one is this about the kind of onsite treatment possibility treatment and that is my probably the situation is different among the countries among cities but the situation i know in bangkok is the there's a the canal network is and canal is works kind of the the open sewage channel especially when it rained so in the way it rains many entry to wastewater is just in the canal so this is probably in the in the pipelines there's many of the pollutants is it stay as a sediment when it dries and when it rains it flushes the sediments and to the canal so in that sense so it is in in monsoon area it's the rainy season it's really heavy lane [Music] flashes still the pollutants in the pipeline so to prevent that is important to have the on-site treatment good on-site treatments so yeah hospital was water could be the important source of arb so the onside treatment of hospital wastewater could be i think important uh to reduce discharge from the source and another point is that what is a good treatment technology so actually i showed the difference the biology profile of the slash affluent wastewater and we took there most of them are from the conventional active slash process but we also took the data from the membrane block process and in the membrane bug process it the its area profile is more closer to the sludge i mean the it contains the less wastewater sludge with less wastewater bacteria community so and so the memory background could be reduced more i think more reliable reduction of the the amr because waste water bacteria is somewhere that does not merge with the slush and they just float on the liquid phase and the sediment attack just go out or through the same tank and go go out in the fluent but if membrane bioreactant like every all the water should pass the membrane so that that kind of the floating bypass of the bacteria will not happen so probably it will have the better reduction of the mr okay uh things up that's one of this uh oh okay i think some gunpowder will show some more questions yeah from the parent song yeah yeah it's very technical one for me yeah this is this is important a difficult question actually yeah so it is currently is hard to uh estimate the risk i mean the quantitative estimation the risk of mr uh so it is hard to decide the risky or dangerous level so actually the its mr is the existing the natural water in the world i mean the wild animal also have the antibodies and bacteria it's natural sources so i think what we have to consider is that there are many kind of antibiotics and what and about we i think what impact most is the resistance the clinically important drugs whether important drugs is the the drugs which is used when when patients are really in a critical conditions and when we don't have the other drug options so this is definitely recent clinically important drugs should be reduced differently as much as possible or to prevent the spread uh and and to make the lifetime of the the drug uh as long as possible and actually i also tried the risk assessment anymore but it's not easy actually so when we think about the risk of the mr it's not the risk of probability of infection i mean the probability is the same but the effective mirror is though we it is harder to treat that so so probably it is better to assess the risk with using a dolly not with the chance of infection so delhi can consider the duration of the sickness so if we infected the disease with the resistance maybe we have we need a longer duration for treatment so that may be considered by risks but so could be so when we have the when we decided dangerous level we used the kind of acceptable risk risk level so we when we access the risk with delhi and when we have the access acceptable risk level as delhi before we can we can decide the sort of the standards of to reduce the amount okay i think some the next question from uh petrocity so what is the effect of the psychochemical and mineralogical characters of the disorder on the diversity of the art irb so that is a is there any effect of the psycho chemical and mineralogical characteristics yeah this is i think uh yes antibiotic resistance is sometimes related to the heavy metal resistance because it's kind of defensive mechanisms of bacteria so there's several defensive mechanisms one of them is a flux pump that is the when the garden because they pump up the bodies outside of the cell and that is sometimes common with the heavy metal resistance mechanisms so i i don't know i don't know the the specific papers on that but probably the heavy metal high heavy metal or high metal or stress in the wastewater treatment possibly enhanced the anti-resistance actuator in the sludge slash bacteria in the anterior resistance genes causing a flux pump is really abundant so could be possible and what is the best technical major bacteria it is there's no kind of best techniques i think this depends on the purposes culture depend techniques is good to know the you know the the fecal bacteria of ecobook is often often relates to the pathogens human pathogens and genomic gene based techniques can cover the old bacteria but it's hard to link the resistance to the species because when we break the cell to extra dna we lost the information link of species information and resistance information so i think it depends on the purpose so uh the last question about the your the uh that one is i think the last question have you been addressed yeah uh yeah i think so okay so that uh things are all is from the sun yeah this is i think yeah similar to the previously uh questions yeah so there are in the drinking water i think the you i know maybe doubleh or guideline has the uh proposed suggests the acceptable level of delhi so maybe it could be the one of the stun i mean the criteria we we can use but what we don't know is the how much of the horizontal transfer happens in our guts so when we intake the drinking water has the resin bacteria which is not pathogens but in our body maybe it's the resistance genes transfer to non-partisan pathogens which may pose the risk we don't know that so it's not easy yet i guess it is not we need we need a research or we need more discussion about how to assess the risk of a mr community or in other others like for the fungal or might be resistant to the antibiotic as well okay so in the culture basis that we study the main eco yeah so it's bacteria and the metagemic studied we we don't we don't separate it we just take through dna and they read all sequential genes so it includes all of them but probably most of them from originated from the bacteria fungal possibly have resistance but for its the mechanism could be different yeah so i mean that we we read a sequence of the in the wastewater and we compare it with the database research and the way the database includes many kinds of resource genes knowing the bacteria but the energy we found is probably most for the bacteria uh origin allergies okay thank you so uh thanks winter still have one more question before we are closing because there's still one okay i think there is no one more i think so one more question i think okay okay okay yes you have something for a question so this is a really important study so it is known that amr has the low quality so different cities different countries different catchment has a different erg profiles probably there's several possible the reasons of actors i guess the one is the most important is the history of antibiotic use in that area in the catchments that affect much and also i think that what i think about fact is a transportation between the different cities of countries so as i talk in the the talk my talk that's the trouble people's trouble or may carry the mr in the different countries and also food trade the livestock also the career of the mr so when we import exports like the meat or shrimp for alcohol culture or the livestock products that may cause the transformation and another thing is it's like a ship ship means uh means that like like a car big ship it's it's carried a water sea water uh when when unload the payloads they will load the ballast water to balance the ship so blast water is carried to the country's countries and discharged in the basketball district in the destination so basswood is possibly the carrier of the mr and uh especially in the ocean and yeah of course the airplane also in the long flight plane has yeah we eat a lot of foods they served a lot of fruits and ice creams uh we got the toilets uh that carries them are so yeah i think it is very important so it may be not in international travel but but inside the countries there is the import export of food race some area is very produced a lot of livestock farming and it's sold into the every places many places in the country so that may transfer the rb in the whole country so that is the important factors okay okay thank you thank you uh probably that one is the the last question but before closing will be you have you have you have mentioned about the technical mission but it might be you have something to say in a few words to the the audience for the cruising doctor honda please see if you have yeah okay so i think yeah i think that there are many questions so i think i don't have much to add about that but this is the this is a seminar without krita overseas research grant right so i think mr is the really important issue in the in the indian countries so yeah it's not in the wastewater issues but also drinking water issues and also the kind of social is like like i looks under questioned me like that the trouble and transportation warriors yeah i think yeah yeah many of many of the applicants propose about that topic to the cork yeah okay thank you so i think so uh because uh this session will be is already finished so that uh we will take the photo station first maybe you may have before okay before before after after this session we will have a short session on the uh call for professor for the young uh researcher on the water environment so that one is uh i will talk in bahasa latin will be a breakout room so that