Kind: captions
Language: en
we will have a webinar but i will share
uh just the information for uh for today
so i would like uh so today is a
part of the
kurita overseas research grant so
but before the information session about
the kurita
grant would like to
have the guest lectures by
dr kitajima so
the lecture will be about one hour
and it probably a little bit more it's
probably depend on the qna
and so this uh webinar
uh then uh after the the lecture
and then q a we have the uh breakout
rooms for
participants from uh indonesia and
partition from
vietnam so bill so the information on
the
kurita overseas grant will be in the
two languages so that by indonesians by
myself
and uh professor vietnam
uh will uh give a information
on on the grant
in vietnamese so that will be a breakout
rooms
so room ones is for indonesian and then
for vietnamese
room two so that this webinar also
uh part of the also uh
that one is the project with the asia
joint research program
here asia grp with the brain
with the topic of our our research
between
itb research and also with the hokkaido
and yamanasi
and also with the vietnamese
institutions
that one regarding the visual inform
early warning system to minimize impact
of coffee 19
and also this it might be other dishes
so i would like
just uh briefly introduce
professor masaki masaaki kitajima
he is got a doctoral engineering degree
in the field of urban and practical
engineering in the university of tokyo
and also he has been in a number of the
office's
engagement either with the university of
the arizona he did the
postdoc and also in the singapore
mit alliance for research and technology
technology center in singapore
now he is now an associate professor at
hokkaido university
he wrote a lot of publications
with the edge index currently last week
was
about 33 and historical area is covered
broad aspect of the health-related water
microbiology from
an air pump to an impromptu engineering
perspective
so so that one is a two-day lecture by
the
uh dr masaki kitajima and
i think uh in the in the
uh next week at the end of next week
there will be another lecture
i think so uh now this person
yeah i don't think very well it's the
same
from the same uh i graduated from the
same university
yes uh dr rio honda from kanazawa
but they thought that the topic is still
tentative about the race
of the uh coffee uh 19
via virus in the natural water bodies
receiving measure but
that one is really like to discuss the
different definitive topic so that one
uh next year in the in thursday uh
may 6 uh in the morning
time so uh without further ado
uh i would like to welcome
uh professor doctor kitajima to to
present you lecture i think sir
dr pitcher has been assigned as a
co-host
yes already yes okay thank you
okay thank you very much for your kind
introduction uh
professor setiadi so let me start
my presentation uh sorry
okay so um i'm very
um honored to be here and
um so i'm massaging
from hokkaido university i'm a associate
professor at the department of
environmental engineering
and today i'm going to be talking about
a
wastewater based epidemiology and to
monitor kovite 19. so
i guess you all know about kobit 19
chronovirus disease 2013 right
so uh let me begin with an explanation
of kobit 19
and the causative viral agent
so the causative viral agent of kobe 19
is called novel chronovirus right
as many of you know and the
official name of this virus is severe
acute respiratory syndrome coronavirus
too
um the acronym is sas cover two so
if i mention scissors called two uh
please understand that
this is uh the name of the novel
coronavirus official name of coronavirus
so this is the name of the virus of the
uh kovit-19
and and the kovite 19 is
not the virus but the name of the
disease
caused by cells granola virus too
so cov19 means coronavirus disease 2019.
so uh for example what we detect
from wastewater is saskov2
rather than kovit 19. okay so this is
the difference between south korea
and kovite 19. and
we call the sas cavity as novel
chronovirus
currently but there was some novel
chronovirus
in the past there are two examples
the first example is sus chronovirus
this is the virus that caused severe
acute respiratory syndrome
in 2003 which caused outbreaks
in china and hong kong
and this may be called south konawa's
plan to distinguish between
chronoverse and the current novel
chronological cells polarimers too
and so that the disease caused by this
virus is sas
and the second example is mass
chronovirus so mass
stands for uh middle east respiratory
syndrome
and this uh caused outbreaks in 2013
in middle eastern countries so the
disease caused by this virus is
mars and so these
are uh corona uh noble corona
viruses and the current coronavirus is a
south korean very steep
so the uh why the reason why we call it
susquehanna virus too is that this virus
is closely related to
cells chronovirus which was uh which
emerged in 2003.
so this is a phylogenetic tree and as
you can see here
uh susquehanna rest which is current uh
normal chronoverse is closely
related to south chronovirus and most
chronovirus is here
and there are four human
common cold chronoviruses which uh
indicated with uh orange color
here and the nucleotide
identities between susquehanna verse 2
and other chronoviruses
are shown here and the south coronal
virus
ii uh is closely related to both
chronovirus
uh with 96 percent nucleated identities
pangolin chronovirus 91 percent south
chronovirus 1
original corner price with uh eighty
percent identity
with months uh only 55 percent
and common called coronavirus which are
indicated here
uh only 50 percent so this is
the phylogeny of cells chronovirus too
so please remember south chronovirus 2
is genetically closely related to
sus chronovirus which emerged in 2003.
now let me talk about waste water based
epidemiology so this is an innovative
um disease surveillance too
uh for cells granovirus or cobit 19
other infants viral infectious diseases
so as you may know um current
epidemiological approach is based on
clinical diagnosis
basically with a pcr tests
targeting individuals right so
but uh pcr test has a few
limitations or issues the first issue is
limitations of pcr
capacity due to the cost uh
like capac experimental capacity
and technician and so on the second exam
second issue is
only symptomatic infections can be
identified
by clinical diagnosis because these
tests
usually targets only symptomatic
individuals
and the third one is a stigma issue for
positive
individuals there will be some
discriminations
for positive individuals this is uh
like a major social issue for corby 19.
on the other hand wastewater-based
epidemiology can overcome
multiple issues of clinical diagnosis
because
wastewater contains viruses excreted
from asymptomatic individuals in
addition to
symptomatic individuals and wastewater
sample
contains viruses excreted
from multiple people many people
there are some comments here
so i think that we will uh we will
uh if there is any question i will i
will read it
okay okay okay okay okay okay okay okay
i will go ahead
and thank you please okay okay so
the third uh the third issue
or the stigma for positive individuals
can be overcome
by wastewater-based epidemiology because
wastewater based females
can um understand epidemiology without
personal information so and
uh our goal is to establish early
warning
and mass diagnosis mass diagnosis
infrastructure based on wastewater based
epidemiology
and this can be applicable or to
municipal wastewater
this is a wastewater treatment plant to
understand the disease prevalence in the
city
and second facility wastewater is second
example
and by applying acidity wastewater
uh by applying this technology to waste
capacity wastewater
we can understand facility level uh
disease prevalence and this can be a
very efficient bus diagnosis system for
uh
facility and then we can also apply this
technology
to international flight toilet
wastewater
to understand the infection among
immigrants
there how many people are in coming
to our country with kobe 19.
in future uh we can apply a metagenomics
approach
uh to um to identify the virus
from wastewater uh to help
uh early uh findings of um
viruses all the identification of of the
pandemic virus from wastewater samples
actually this wastewater diagnosis
is not the replacement of the ongoing
clinical diagnosis but this will provide
information to complement the
clinical diagnosis which will
enhance the pcl testing efficiency
and uh wastewater data will
complement the epi data epidemiological
data obtained from clinical diagnosis
so this is the concept of this study
and so
actually uh we are the first ones
uh who proposed wastewater based
epidemiology as
a peer of peer-reviewed
uh publication uh last year
so this uh we wrote a review paper pew
review review paper
on the south carolina verse 2 in
wastewater last year
this is the paper and we started
drafting this paper
last year march so more than one a year
ago
at that time there was no report on
detection of
this virus in wastewater and actually
this
paper was accepted uh just one year ago
on april 26th today is 27th
so more than one year ago and then
at that time uh there were only five
papers including
including preprints uh reporting the
detection of
susquehanna verse 2 or in wastewater
from all over the world so
then at that at this time we our paper
got accepted
on our paper on review paper or review
was
accepted in this review paper we
proposed the
importance and potential of wastewater
based epidemiology wbe
to better understand the disease
prevalence
for this paper we published press
release from hokkaido university
so this was what happened uh last year
and this is the contents of the paper
uh our paper is a comprehensive review
uh covering the aspects from occurrence
in wastewater
uh to risk assessment and
first we reviewed uh gastrointestinal
symptoms
of cobit 19 and shedding of this virus
in excreta
in uh in
feces and urine and we reviewed evidence
for the
presence of south korean verse 2 and
related qualifiers
in wastewater and third we reviewed
what we proposed uh that we can
understand kovit 19 epidemiology through
wastewater surveillance then we reviewed
methods
for susquehanna rise to detection in
wastewater
which is very useful or for the current
research
because we are we have to use some
method to detect this virus from
wastewater
and we also reviewed survival in india
inauguration of quantifiers and other
enveloped
surrogate viruses in water and
wastewater matrices
we also reviewed
risk analysis or risk assessment
on related viruses like respiratory
viruses in wastewater
and the occupational risks and
maybe some of you may know qmra
quantitative microbial risk assessment
for relevant viruses and we also
reviewed uh those response
of uh sas chronoversity although the lim
the data for those response of sas one
virus two is limited
and relevant uh respiratory viruses
now um let me
explain the details of the advantages of
wbe
for covid19 so the presence of
the viral rna in wastewater means many
things
first we can trace the circulation of
viruses in the community
which provides opportunities to estimate
their prevalence
genetic diversity and geographic
distribution
and this makes it possible to monitor
the epidemiology of virus infections
even if they are not
evident by clinical surveillance this
means that
uh including uh kovic 19
this kind of disease causes asymptomatic
as well
infections as well so in such
disease including kobit 19
wastewater surveillance is very
effective because we can understand the
actual prevalence of the disease
including asymptomatic
individuals infections
and this provides an unbiased method of
evaluating the spread of
infection this is especially
important for like
southeast asian countries uh like
developing countries uh because uh in
such area uh resources for clinical
diagnosis like
pcr machines or diagnosis diagnostic
systems are limited
and in some reasons reporting systems
are
available so wastewater based
surveillance is very effective
in such regions because this wastewater
surveillance method is very
resource efficient and cost effective
and we can identify variations in the
circulating strains through
phytogenetic analysis for example now um
variants are prevalent uh all over the
world right
but uh we can understand the prevalence
of the variance
through a genetic analysis of cells
called analysis
and we can compare
between regions and we can assess the
evolution of the virus genome over time
so this is kind of like
uh impressive isn't it because last year
we kind of predicted the
prevalence of variance very variants
and we propose that through genomic
analysis we can trace
the evolution and we can compare the
prevalence between regions
in this first review paper
and the wbe is also useful as an
early warning system to determine if
reintroduction of south colorado still
has occurred in a community
or conversely use this is useful as an
assessment of
whether exposures have decreased
sufficiently following
public health interventions such as
lockdown social isolation and social
distancing
and this also provides anonymous
epidemiological information on disease
prevalence in the community so
uh wbe can avoid individual ceo
stigma which often results from the
clinical diagnosis in convict 19
outbreak
so combined use with clinical testing
which is basically a pc of tests or
anticipates
could contribute to significant cost
savings
so this is the key message
from our paper now
let me talk about why um
south korea ii can be present in
wastewater
first this virus can be
shed in faces because this virus can
replicate
in our gut human gut there are two
scientific evidence of the replication
of this virus in human gods
the first exam evidence is that
the receptor of this virus is expressed
on human intestinal cells
the receptor is called angiotensin
converting enzyme to
a s2 so this is a receptor for cells
granola stew
cells and cells chronovirus original
cells pronounced
and that's what that's why this virus
can attach
to uh the cells on our html
are intestinal cells
the second evidence is that this virus
can replicate in human gut
enterocytes so this is in
vitro study so in outside our body
so the researchers uh inoculated
this virus into cell culture
of human gut endocytes
and they observe the replication of the
virus in
the vitro and they
um so
they think that this uh virus can
replicate
in vivo in our body as well
so actually this virus can be shed in
phases there are many reports on south
chronovirus two rna detection in faces
one example is here and
the viral load or viral concentration
can be
up to 10 to the eighth copies per liter
per sorry per gram feces this is quite
high concentration
and the shedding of the virus into feces
can continue for
approximately four weeks
and approximately a two-thirds of the
patients
uh shed the virus to the cells so
like 66 percent on the other hand
uh approximately only six percent of the
patients
shaped the virus in urine this means
that
uh probably the majority of the virus in
sewage or wastewater uh is originated
from
feces rather than urine okay
so and another thing is uh
i keep saying uh through wastewater
surveillance
we can understand the infection status
of uh
asymptomatic individuals as well
actually uh
susquehanna verse 2 already was detected
in theses of asymptomatic individuals
there is scientific evidence
and the ratio of a simple asymptomatic
infection
is up to 30 percent so this is not
negligible
so quite high percentage of the infected
individuals
are asymptomatic they don't show
symptoms and another concern is
whether an infectious virus can be
present
in wastewater actually
uh infectious such coronary stew was
detected in feces
so it is suggested that a fecal oral
transmission is possible
but we don't have any uh scientific
evidence on the presence
of infectious uh subscribers too
in wastewater at the moment
so this is the methodology of scovit19
a wastewater-based epidemiology
if the kovit-19 is prevalent in the city
the wastewater contains viruses
uh south korea ii and
uh we can detect sales computer in
wastewater
after we concentrate the virus in
wastewater
we use qpcr technique to detect the
virus in wastewater but
the concentration of the virus in
wastewater is usually very low
so we have to concentrate the virus so
we can detect
the virus genome by qpcr so we
can concentrate the virus using various
methods
and we extracted rna we extract rna
and then we detect and quantify the rna
of
cytoscopy so rna is the genetic material
of the cytoskeletons too
so in order to detect
virus in wastewater we have to
concentrate the virus as i mentioned
earlier
but there was no standardized protocol
for susquehanna still detection from
wastewater
so we needed to explore the methods
to detect saskatoon 2 from wastewater
efficiently
and so the reason why
we needed to explore the new method was
that
was the difference due to the difference
between
subscriber 2 and enteric viruses which
were
a major major target
for environmental biologists before
pandemic era uh covet 19 pandemic era
and specifically
enteric virus has a capsid protein this
is the
outermost layer of the virus particles
on the other hand
susquehanna virus too has lipid envelope
that's why so these two
types of viruses have different in size
hydrophobicity so as you can imagine
envelope is more hydrophobic
and stability under extreme ph
may be different as well probably the
cells cover two
is more fragile as compared to enteric
virus
like norovirus so
because of these reasons there was uh
there was difficulty
uh to apply the existing virus
concentration methods
for science cover 2. that's why we
evaluated the chronovirus recovery
efficiency from wastewater
for this experiment we inoculated urine
hepatitis virus
mhv which is not infectious to humans
but are infectious to mice
and then um yeah we
seeded this virus into wastewater
adequates
and we recovered the um
inaugurated virus using seven different
methods based on electronegative
membrane
ultrafiltration head precipitation
pollutant growth glycol presentation
and centrifugation we and that we
enumerated uh mhv
by rtqpcr and we calculated recovery
efficiency
i'm going to skip the results and from
now uh
let me show you some examples of the
actual detection results
of science cover 2 rna from wastewater
samples this is the first
study that reported the detection of
susquehanna virus
ii rna from wastewater in the world as
the peer-reviewed paper this study
was reported from australia and i also
contributed to this study
in this study we collected wastewater
samples from
three wastewater treatment plants
in brisbane australia and we
concentrated
the virus using aha electronegative
membrane
and we detected the virus using two
different qpcr assays and this is a
result
and this is kind of uh a busy
uh figure but um uh in
the summary of the result is that we
detected
uh sales cover 2 rna from wastewater
samples
during outback outbreak period in this
region
and the viral rna concentration was up
to 120 copies per liter
and we confirmed qpcr product sequence
so this was indeed the signal was
indeed originated from or derived from
south carolina and based on this
concentration
we estimated the infection prevalence
and the median
value was 0.096 percent
so this means roughly
out of 1000 habitants or population
one person got infected so one out of
1000 people got infected with this virus
this is
our calculation
and so we also reported the recovery
efficiency of coronavirus in wastewater
sorry um so this this was associated
with the
this slide for it but uh yeah we also
reported the recovery efficiency and we
published press release from our
university
in last august
we also detected this virus
rna from wastewater samples in japan as
well
so this is the paper and we i also
contributed to this study
in this study we collected wastewater
samples
uh from influence uh secondary treated
switch and river water samples uh from
wastewater treatment plant in the river
and we concentrated the virus and we
detected
suscob2 using six different assays
so here's the summarism of the results
and we detected
the virus from the wastewater samples
collected from
april 14 last year
and in this period the number of
infected people increased in this region
so this also indicates that wastewater
based epidemiology can
like grasp the prevalence of the disease
in
a certain region and this may be
applicable
to japan as well
okay and then we also detected
the virus in wastewater in north america
for the first time and this is
in this study we collected wastewater
samples from
two wastewater treatment plants in
louisiana
which is located in southern united
states
and for this result we also published
press release from our
university so this is a brief summary
of the results
looking at this figure uh we
so the fuel circle
circles indicates that detection of the
virus positive
samples from infinite ways for examples
this also indicates that a virus can be
detected
from wastewater when the number of
infected people
increases in each region in
the region actually louisiana it was one
of the most
significantly affected states in the
united states
in the first epidemic period
and this is the first detection of
south konawa still rna in wastewater in
north america
including united states so actually i
i showed you the first detection in
japan right
so this was earlier than
this report which means that our report
from japan preceded the first report
from the united states
and i was associated with both
publications
and subscribers to rna was detected in
wastewater immediately after kobe 90
outbreak in this region
and so
until now i've been talking about
application of
wastewater based epidemiology to
municipal wastewater samples
but this approach can be applicable to
aircraft and cruise ship so like
mass transportation systems um so this
wastewater surveillance can be a
potential tool
to understand import imported uh kobit
infections for when we apply
this approach to international flight
and international cruise ship
so that basically the method to detect
virus from those wastewater samples
is the same as that is used for
uh useful ways for examples but um
the main message here is that this can
be one of the tools
uh to understand how many people are in
coming
uh to a specific country uh
[Music]
with kobit-19 infections
then we also um evaluated the
persistence
of south chronologists too in wastewater
using
culture meth subculture method we
conducted
this experiment using uh infectious
sunscreen numbers too
in dsl4 facility in the united states
and we all we already we have already
published this paper
and in this paper we inaugurated
infectious granovirus 2
in tap water and wastewater and we
incubated these inoculated samples
at different temperatures and we had
two uh different titles inoculation
fighters
and we titled the
virus concentration using cell culture
and we estimated the decay rate constant
on the other hand we also quantify the
viral rna
using rdq pcr and we also
estimated the first order rna decay
constant
so these are the results and we i'm not
uh
going to have time to uh walk you
through the
details of the results but uh this
result
this paper uh research determined the
activation rates of such collaboration
in wastewater under different
temperatures
so temperature is the major determinant
of the survival
of the virus in wastewater samples and
this study revealed the persistence of
sunscreen stood in wastewater and this
is an important data for risk assessment
and we also evaluate the decay of rna
signal
in wastewater we call it rna signal
because
rn viral rna is indicative
of viral disease prevalence in
a specific region that's why we call it
rna signal and we are detecting
rna rather than infectious rna
infectious
virus for
wastewater based epidemiology studies
wbe studies
in this study we determine decay rate
constants
of sun scholars 2 and building hepatitis
fires rna in water and wastewater
and this is the results on this on
in these panels
we showed decay rates of viral rna
under different temperature for sun's
coronavirus 2
and mhv for untreated wastewater
autoclave wastewater which doesn't have
biological activity
and top water samples and the summary of
the result is here
these two figures uh summarizes the
viral rna decay constants
and in their environmental factors that
may
impact the survival or not survival but
rna decay rates
so looking at this figure so the major
determinants
of the rna decay rate
is temperature rather than the
biological activity
or a sample matrix type
tap water or wastewater temperature is
the measured
determinant that's what we obtained from
this study
so from now on uh let me talk about
our collaboration uh with shiongi
a company uh in japan shionogi is the
major pharmaceutical company in japan
and we are now actively collaborating
with this company and we are
uh conducting uh technological
development
and societal implementation of
wastewater based epidemiology
of kovit 19 in japan
and this is one of the publications that
we already
released through foreign media
please look at this site if you are
interested
and uh there are a few uh upcoming
articles from this
news media as well
and through this collaboration we
developed a highly sensitive method
for virus science code 2 or detection in
wastewater samples
let me explain some
details so in this study
we developed a new method but in
conventional methods
we used to use uh pre-treatment
so i guess many of you are environmental
or engineer or environmental scientist
and many
some of you may be doing some chemical
analysis or microbial analysis of water
quality
but in water quality analysis
we usually remove the particles or
suspended solids in wastewater
because they inhibit the analysis
so in conventional method we used
pre-treatment
that means we used to remove the solids
from wastewater before we concentrate
viruses
and we concentrate the virus from water
fraction
and we detect it with qpcr but we
realize that this method is not very
effective for
uh cells cover two although this is
effective for
other uh viruses like uh human
enteroviruses
including norovirus and other viruses
but um for or
normal coronavirus or suscover2 this
method is not
efficient that's why we developed a new
method
uh which does not include pre-treatment
so we concentrated the virus with solids
and we invented a
method to extract rna efficiently from
the solids
and we do a pre-amplification
of the genome before we
quantify the genome using qpcr so
basically we
quantify qp we quantify
the amplified dna fragments for
sensitive detection so this allows
sensitive detection
but we can still uh quantify
the number of the rna in the original
wastewater samples
and then uh we also developed a qpcr
assay for
variance identification
so we showed that we can detect
sales coverage variants from wastewater
using next generation sequencing
or energy s so this is
the map of the sunscope 2 genome
the sascom2 has length
of around 30 000 base pairs
and uh this has like many open
reading frames the important reason for
variant analysis is s or
spike region which encodes
spike protein of the viral
particle so spike protein
is associated with attachment
of the virus particle to receptor
h2 so if the amino acid
mutates uh to another amino acid
on the spike protein this may change the
phenotype of the virus or
another ones this may increase the
infectivity or
transmissibility or uh this may reduce
the vaccine efficiency
so there are uh some like important
amino acid mutations as listed here e4
454k which reduces waxing efficiency or
escape from
immunity n501 y
is a unique amino acid mutation of the
variants
which increases infectivity
and the 614g
a is associated with
increased replication efficiency and
transmissibility so this is the very
important reason that's why
we wanted to analyze this region
but the existing uh qpcr
sorry existing nested pcr methods
did not target this region so we
couldn't
analyze the genome sequence in this
region
so we developed a pcr for
variant identification so this is an
original assay
which allows identification of amino
acid mutations
among variants and this is suitable for
illumina parent sequencing
and this is the result of the
next generation sequencing library
preparation
so uh so this is a result of the
um library prep we obtained pcr product
and we quantified the pcr product and we
did library preparation
and we sequenced the genome using bisec
from illumina and we did in sql analysis
to identify the mutations and this is
the result of
bioanalyzer and we as
you can see here library preparation was
successful
for ngs analysis
and this is a result we obtain
high quality energy as reads of
approximately
this number uh like
up to closely one million reads per
sample so this is a root or deep
sequencing
and we detected uh n501y
d6144g and these are commonly observed
among variants and
so this is a result of the currently
hotel
and wastewater influence in japan
for quarantine hotel we detected
variants in
december but this variant was not
detected from
october and november which is reasonable
because uh we in japan we identify
this variant in december not
in october and november
and then we applied this technique to uh
domestic
or sorry or municipal wastewater
and this is a result and
in municipal wastewater we detected this
variant
from the samples collected in december
4th
which is kind of surprising because
japanese government first reported the
detection or identification of
this variance on december 25th
so three weeks before the first report
from the government
we detected this variant from wastewater
samples
and then the proportions of the reads
of the n501y variants
increased from december to january
so this is a significant findings
and uh yes and
501 why mutation detected in municipal
wastewater earlier than the first
confirmed case in japan
so we showed the evidence
of the application of this virus
detection in
laboratory so now the time to implement
this technology to the society and we
achieved
full automation of viral detection and
this is a press release
on the establishment of an automated
system for the analysis of suspicious
wastewater
and this system involves robotics
and high performance or high fluid
next generation sequencing system we
actually use
a lab droid maholo or which is a
versatile humanoid
robot made in japan so rbi
robotic biology institute has this robot
and this robot does everything from
virus concentration
to a pcr plate preparation
and also this robot does uh library
preparation
for next generation sequencing and we
also collaborate with ilac which has
illumina novasig
a platform which allows high throughput
sequencing analysis
for this societal implementation uh
this is uh picked up by uh or
featured by international media
innovation news network
last month and we also started
monitoring in osaka prefecture
to monitor kovite 19
through wastewater surveillance and so
this
uh monitoring activity was
for this study we started this month
and so this
is the last topic actually so
we are going to hold uh
the summer olympic games in tokyo as you
may know
right but probably the kobe 19
uh will be still there and we have to
have com
olympic games with kobe 19.
so risk and management of kobe 19 during
the
tokyo olympic games is very important
and this is
primary responsibility for relevant
japanese researchers
we think and i think i'm one of the
responsible
or researcher in japan and wasteful
testing can be used as one of the tools
to manage risks
when we apply this testing tool onto the
olympic athlete village
in tokyo to manage the risks
so this is summary of my uh
talk today so wbe
is very useful for viral disease
viral infectious diseases and wbe was
previously applied
to norovirus poliovirus and so on
basically enteric viruses
and applicability of wbe
to covet 19 was proposed by
our paper for the first time so this
uh paper was the first review paper
proposed the concept
of wbe currently uh there are a number
of
reports on the detection of south korean
virus 2
including variants from wastewater so
now the concept that we proposed
in our first paper review paper has been
proven so now we have to implement this
and i'd like to highlight that uh
the wbe can add the value of
wastewater infrastructure
specifically in the textbook i guess
many of you are environmental engineer
so if you open the textbook of
environmental engineering
it is written that the wastewater
infrastructure was constructed primarily
to treat and remove wastewater
and storm water right but
in this time the role the new role
which is monitoring of disease
prevalence
is being added to
the value or role of wastewater
infrastructure
so this is a very important time that
changes
the image and the value of the
wastewater infrastructure
now
what we need is to implement
uh this uh infrastructure or technique
uh to the society so social societal
implementation is urgently
needed with that i'd like to thank you
for your kind attention and i'll be very
happy to take any questions
okay thank you doctor so that
thank you very much so if you have
uh i think if you have any question you
you may
uh write in the chat box or
you can open you you might
and then uh raise your question directly
so we still have uh some time
so please if you have
any any question and comment or
so i think so the question from myself
so that if uh you mentioned that
the variant can be detected earlier
uh before the cases
has been uh identified in japan
so that one that one was
you realize uh after
the cases of occur right that's also in
the beginning
you already realize there's something uh
because in the review you have mentioned
that
the variant will be can be detected
but that that in the in the cases in the
real cases
when your analysis so that one is uh
you realize that it is happen what you
have predicted
yeah yeah let me clarify if i
understand your question uh uh correctly
so we actually uh detected um
variants from archive samples so
we we so start started
uh viral analysis in this year and so we
another analyze store example or frozen
samples
and we went back to the old dates all
the dates and then we
successfully identified the variance
from the
december fourth sample
which was which was preceded to the
first detection
or first report from the japanese
government on the
variance yeah so i hope this answers
your question
yes yes so this one is uh you
you you store the the sample and then
that's right
that's right we froze the sample and
we analyze the frozen samples okay
okay okay okay please if you have
uh uh any other question you may
raise directly to your kitajima
please there is one question professor
chandra
oh one question okay okay uh
uh could you help me because i have yes
because i have to read so i can read
the question maybe
please kindly share your thoughts on how
far we can develop and apply that with
my
automatic monitoring station for
um
oh yeah yeah okay so automatic
monitoring station
actually we have achieved
yes so we we are
currently uh very uh
close uh to the position where we can
develop
uh automatic monitoring station because
now
uh japanese um
labdroid uh like robot uh versatile
humanoid robot is available for
wastewater uh sample processing
and detection so well
if we can install this to
uh like many stations then uh we can
yeah you use this technique so
technically this is
feasible but the
issue is the cost actually
this is this was 1 billion
dollars from one
medium so sorry what one million dollars
per
robot so this is quite expensive
quite expensive uh but but we have
already technically we have already
established
this monitoring station so if we can
reduce the cost
probably this is feasible
yeah this probable this automatic
monitoring station
can be realized and
how far in in japan in japan
we are now we have already
such station now but this is only
available only
one station in japan so my goal
is to expand this station or make
duplicates or triplicate copies of this
station in
all over japan and all over the world
this is my goal and
another thing we we
were able to establish automatic
monitoring station
because the robot is versatile
right so this is not the robot specific
so special robot for a certain process
but this robot can do anything
that humans can do
okay so this this robot has two arms
and this robot has the freedom
of the pre-degree of seven
seven freedom of degrees so this can do
anything that we can do like pipetting
or like uh
yeah or open the tubes or open
micro tubes this robot can do anything
so that's why we we have
been successful in developing automatic
system
otherwise otherwise this may take
the robotic robotic expert says this may
take
like five years if we if
the versatile robot is not applicable or
is not available
so yeah i think how far
in in japan we have already established
and we will provide a service
of the wastewater analysis next month
actually
which is impressive isn't it using
using using the robot so this robot
can work 24 hours right
of course and 30 uh
yeah uh 365 days
all day around the year so this may
speed up
the wastewater analysis and
this will provide very high throughput
of the
wastewater analysis yeah i hope this
answers
your question okay
so don is there this
yeah yeah okay okay okay so
let me answer uh mr miss uh
gunther's question so yes
actually there's no experimental
evidence that detected
infectious cysts called two normal
quantum virus from
wastewater but there's no experimental
evidence but there is
there is epidemiological evidence which
means that
which means that in china
so this is a one example in china uh
there was an outbreak in uh apartment
complex
abutment complex right and in that
apartment complex
uh there is a failure or failure in
drainage system
wastewater drainage system okay and then
uh the wastewater spilled
from the fader and the spill out
and probably the aerosol was generated
and this the people got exposed to the
um drainage drain a wastewater
and then the people who got infected was
associated with exposure to wastewater
which means that probably wastewater
was the source of the virus transmission
from
epidemiological evidence so there is one
so this is one example but um there is
one
uh there are some uh epidemiological
evidence of
transmission of the virus through
wastewater but there is no
experimental evidence that successfully
cultured infectious sunscreen
from wastewater but so what i would like
to say is
if you get exposed to fresh wastewater
with the virus you may get infected
from the wastewater but
there is no evident risk of the
wastewater
exposure at the wastewater through
treatment plants so there is no example
of the
exposure and establishment of
infection at wastewater treatment plant
so
that means probably the risk of the
exposure to wastewater
at the wastewater treatment plant is
very low
i i think this is negligible
uh probably because like you said here
the lifetime of the virus in wastewater
is very short
so the virus doesn't survive uh in waste
for a very
very long so this was indicated by our
study so
in our in my presentation slide uh
yeah let me yeah
i i i shall let me show the slide
i i hope this answers you your question
here
so we so the latter part of your
question so how long the
virus survives in wastewater right
so in this paper you can find the
information of the survival
of the virus in wastewater
so this is the the tcid which
uh is associated with
infixes virus title results the up
upper panel so
the virus title decrease very rapidly
at room temperature and this is
the yeah
yeah this is the rna this is rna this is
the the infectious title but the
infection
tighter decreases very rapidly
so uh the virus doesn't survive in
wastewater very much
and this is temperature dependent the
higher the temperature
the higher the inactivation speed
so yeah yeah so
oh this is i guess one
example and you can find uh this paper
and this
this is downloadable so you can download
this paper pdf
from the environmental science and
technology letters website
okay yeah if you don't have a
pdf i can send you the copy of the pdf
i hope this answers your question yeah
okay uh is there any uh other question
from
the audience oh
there's a few dart tune please
um good afternoon this is professor
masat masala
thank you very much for the presentation
uh very
yeah useful information um i'm frederica
based in
rabbi with its i just have a quick
question related to the technical
uh part of the sampling of the
wastewater
so we understand that this corona virus
we
we could eventually consider this virus
as a pathogen so that means do we treat
um the people who take the wastewater
sample
um like like we normally take the
sampling like
it's just a normal protection or is
there any specific
uh consideration for that that the first
one and then the second
related to because in in your
presentation you mentioned
regarding the the needs of the
concentration
uh to be concentrated uh the sample um
is there any specific time range of the
day like the best time
that we could um take the sample from
the
for example for from the west water
treatment plant
uh for example in the morning where most
of the people use the toilet
um that the second and then
if we take the sample in the morning do
we still
need to um concentrate it at the sample
again
even though we understand that we
probably will find more swatch in the
morning
okay okay thank you for your question
very good questions so the first
question
uh so the ppe right personal protective
equipment
how we have to um
causes about the infection when we
collect wastewater samples
um basically the
normal um ppe
so is is uh enough
for wastewater sampling because uh in
municipal wastewater
uh uh subscribe to concentration is
relatively low
and uh yeah normal pp is enough but i i
would suggest
you wear gloves um goggles
and the gowns at least when you collect
samples but other than that
that's it yeah so the second
question is um
sampling right yeah the time range
regarding that was interesting yeah yeah
that's right yeah okay okay
uh so yeah yeah as you may imagine
uh the virus concentration fluctuates
over time
within a day right in the wastewater
treatment plant
if you have automated sampler you can
collect
a composite sample or you can do a
24-hour
uh continuous sampling to identify the
best time
of the day to collect uh for
efficient wastewater yeah
uh testing um
so i remember your question so whether
we can detect
virus without concentration right so in
the united states for example
uh they can detect virus um
they could actually they could detect
virus without concentration when
that means they directly extracted viral
rna
from the wastewater samples without
concentration and they could detect
the viral rna using qpcr when
the kovit 19 was very prevalent in the
united states but the number of the
cases
is is decreased rapidly decreasing in
the united states now
the deduction rate is decreasing so now
the
concentration of the virus level of
virus concentration
uh uh like uh is below
the detection threshold where they can
detect
uh virus without concentration but a
certain period of
time they could detect virus detection
so uh
depending on the prevalence you could
detect via subscribe to rna without
concentration but in many cases
including japan you will need to
concentrate the virus
uh where to detect viral rna
from wastewater and uh for the time
variation
we propose that
you collect composite sample 24-hour
composite sample
or if the automated sampler is
auto-sampler is available
but if a sampler is not available and
you need to collect graph sample
probably uh the first
uh the morning is the best time
i agree with you and actually we are
collecting grab samples as well
and we are yeah collecting around 10 a.m
considering the time that when people go
to the toilet
and residence time are in sewer and
we also monitored the virus level
uh in wastewater and uh
we observed that in the morning virus
concentration is
relatively high but in
like uh in major cities the catchment
area is very high uh very very very
wide and there are certain like several
hours residence time so
the virus concentration is kind of
normalized
over time so you can collect
in the morning but it doesn't
yeah well it may affect but it depends
on the size of the catchment
and depends on the yeah uh treatment
plant
so can we add something yeah
is it also related to the temperature
for example in japan because
i think uh if i'm not mistaken that you
need to try uh treat the sample below
for
degree celsius for the handling before
you analyze the sample or we could um
like uh what is your suggestion how we
transport the sample do we need to
read the xbox and something like that
yeah so when we transport the
uh the samples uh we transport on ice
or keep refrigerated
if the virus uh the sample arrives
within a day or two but for
longer transportation time we froze the
sample
and we freeze uh we freeze the sample
and then
transport and then defrost uh
upon arrival okay yeah thank you very
much professor message
yeah yeah no problem okay i think so
uh the last one because of the time uh
uh gunther from here this one uh one in
the chat box
is from oh
okay select on the last one from
uh okay okay lagging time of addiction
oh this is good point
also yeah a very good point okay so let
me explain
how the early detection is possible
through baseball surveillance
so when one person get infected with
virus
there is certain uh integration period
right
integration period is called on average
like two weeks you may have heard uh
within two weeks
uh there is a certain point where the
virus
shedding starts in
in feces and then uh
for after a few days to several days
the person uh start exhibiting symptoms
so by pcr tests uh
in other words a clinical diagnosis
we can identify the infection after the
person the infected individual
um exhibit symptoms right
fever or cough or and so on and and then
they'll get reported but through
wastewater surveillance we can
identify the virus through uh
from wastewater uh before the person
uh exhibit symptoms
so before they exhibit symptoms
uh they start shedding the virus like
uh several days earlier than when they
uh exhibit symptoms that's why we can
if we routinely and continuously monitor
the wastewater for
cells cover two we can uh we have the
possibility
that we can identify the virus
before the before the
increase in the reported cases
i hope this answers your question
okay thank you so much would like to end
our house our session uh we'd like to
thank to
uh dr kitajima for uh you very
uh interesting uh presentation
i think hopefully this information will
be
useful explains some of uh
the the audience here is the young
researchers
so hopefully they'll encourage the young
researcher to do the research
either on the epidemiology or it might
be in the
visual treatment wastewater environment
as uh
general so goodwill probably you
you could take a picture of all of us
before yeah yeah yeah good idea
yeah so that things are
uh in the meantime i would like to uh
give the information that for the young
researchers
so there will be two rooms uh
room one it is for the indonesian and
room two
is for uh the vietnamese so they
uh the the indonesian will goes to the
room one
and then the vietnamese to the room
two that one is for the young researcher
who interested into the
kurita grant so uh
me and will be in the room one and
uh for sure vietnam will be in the room
too
probably