text:
Accessory Fuse Block and Cable Install in a 4WD – How to add 12v accessories to your vehicle
so if you want to install a secondary
fuse block this is what you'll basically
need I've got some six b&s cable I've
chosen 6b&s for a specific reason
which I'll talk about later and most
importantly I also have a circuit
breaker which is put between the battery
and this cable so in the event of
electrical fault somewhere along this
cable length this shuts off and stops a
fire after that I've got a STfuse
block holder then of course various
connectors grommets and shrink wrap and
then the lighter duty cable for running
to my accessories such as this 12 volt
socket so let's get into the
installation for your fuse block i'd
recommend something like this st fuse
block holder this one comes with the
negative bus which i'd recommend as it
allows you to trace all your earths back
to the one single source which is your
12-volt battery so your main cable lead
will come here connect to this the positive
connects to this it reduces any chance
of reliability issues being introduced
from poor earths if you try earthing to
the chassis in various points something
like a defender for instance with
corrosion in different areas dissimilar
metals cause aluminium corrosion old
rust dirt in between panels it reduces
the effectiveness of the earthing system
but by running all the earth back to one
place then you reduce or minimize
completely that reliability issue from
poor Earth's plus all the fuses are in
one place and can be labeled and easily
found these are made in six and twelve
block fuse holders so this time I
opted just for a six fuse holder as I
don't need much more but should ever
want to upgrade I can pull this out swap
it over to the 12 fuse variety and I'm
good to go again
I found a spot in the back end in my
rear storage box where I'm going to
install the fuse block it's a fairly
protected spot so I know it's safe and
it's easy to get to if I have a blown
another thing to consider for your
wiring to ensure reliability is to make
sure you don't have any places where
you've drilled a hole and run a wire
through that is bare steel it's using
this scrap steel as an example if a wire
went through a hole it would sit on the
bare steel and over millions of
corrugations it would sit there rubbing
and rubbing and rubbing until eventually
the insulation rubs through and it
earths to your bodywork which could
cause a fire or at least many blown
fuses so to stop this from happening you
should always use rubber grommets these
basically run the cable through it so
you're protecting any holes in your
firewall or any bodywork panels you've
ran wires through and this now protects
the wire so it's bouncing around and
protected by insulation another
possibility is to use split tubing if
you're running a wire on the outside of
the vehicle this is a simple way of
running it into the split tubing through
in the inside
as you see there it's nicely protected
so that could also be used if running
through a wall a hole in the in the
bodywork just again making sure you have
a reliable electrical system which isn't
gotta start a fire when you're in the
middle of nowhere. The following section
is a general guide for entertainment
purposes only. Consult an auto
electrician for your particular needs.
So let's look now at the two cables I've
chosen this is six b&s cable it has a
thirteen point five millimeter square
cross-section of copper and has a maximum
carrying capacity of eighty two point
four amps at thirty degrees Celsius this
one is six millimeter auto cable and
it's 11 b&s approximately it is a
four point five millimeter square
cross-section of copper and has a
maximum of thirty eight amps so to work
out which cable is good for what we need
to take into consideration the
voltage drop so there are three ways of
calculating voltage drop for your system
there are formulas you can use which I
have two here to show you as an
example there are tables online plenty
available online Just do a quick search
for 12 volt voltage drop calculator
or table and there are apps available as
well such as on your smartphone or as
free calculators online so there's
plenty of ways to do it if you don't
want to work out the maths itself let's
look here at a couple of these formulas
which you can work out so voltage drop
equals amps by the total length of a
positive and minus so if it takes three
meters to run to the battery to the fuse
block then it's three meters of
positive and three meters of negative
totaling six meters You times that by the
resistance the ohms per meter which you
can get from the manufacturers website
so in this case six b&s I made a
theoretical 50 amp carrying capacity
just in case I want to add a anderson
plug in the back end in the future and
run maybe a small inverter for 240
electricity or if I buy a secondary
fridge which might take another six amps
or so to run or any future additions I
want to make sure that I never have to
upgrade this again so 50 amps by 6
meters at the two lengths times the
resistance equals point four two volt
voltage drop which works out three point
five percent loss so three point five
percent is between three and four
percent which is about the maximum you
want in the 12 volt system so 6 b&s
for my particular cable run here of 3
meters from the battery to the fuse
block is excellent this will be plenty
for all future additions in reality I'm
only drawing twelve point five maximum
now with all my current accessories so
assuming I do get a second fridge or a
little camp oven or something crazy like
that then we'll say 20 amps of a maximum
current draw by the six meters by the
resistance equals 0.16 volt or 1.4
percent loss so this is more than
capable of running a highly efficient
electrical system to my
fuse block without any real power loss
let's look at the six millimeter auto
eleven b&s. this is running from the fuse
block out to the accessories the most
I'd ever draw through a 12 volt
cigarette lighter adapter it'd be about
10 amps a laptop is currently the
greatest draw I have which is at a 8
amps so assuming a 10 amps for a bit of
leeway 10 amps by the 8 meters by
resistance equals 0.33 volts or 2.7
percent loss so again this is below the
three percent recommended maximum so
this is also quite sufficient of
carrying 10 amps the 4 meters with less
than 3% voltage drop which makes
quite an effective reliable system
another calculation you can get if you
do not have the ohms per meter or ohms
per kilometer manufacturer's numbers is
to use this formula down here voltage
drop equals a total length of the
positive and minus times the amps you
want the cable to carry times point zero
one seven which is a standard rating for
resistance of copper divide that by the
copper cross-section in millimeters
squared which you get up here such as
six millimeter Auto is four point five
millimeter square you can get that from
tables online if you don't know the
exact number that works out down here
to 0.30 volt voltage drop which works out
very similar to the 0.33 volts of 2.7%
again using either formula I'm well
within a safe range for voltage drop for
this particular sized cable for the
length that I wanted to carry and the
current it must transport so hopefully
that's a bit of a rundown of why I've
chosen these particular cables another
thing to note too is I also chose to get
fully sheathed dual core positive and
negative in a single wire that
just adds an extra layer of protection
on the outside to protect the wire from
chafing I also chose marine tinned
copper cable which is a
higher spec for marine environments good
for when it's getting near water such as
a Defender going through water crossings
as this cable will be running on the
outside of the vehicle down below along
the bottom chassis rail so it's just one
extra level of protection to ensure a
long-term reliable system not necessary
but I was happy to spend the extra few
dollars to get tinned copper wire I
found a spot I'm going to run my cables
unfortunately there's a lot of wiring
there already so rather than trying to
force the cable through the grommets
already in place I'll drill a new hole
put a new grommet in and that way it
keeps everything separate and there's
less chance of wires chafing and rubbing
together or contacting bare steel and
rubbing through the insulation
and there's two of my finished cables
one end has Spade terminals for the
12-volt sockets and the other end ring
terminals for the fuse block
everything's crimped heat-shrinked so
there's very little chance of any
moisture getting into the cable and
causing corrosion down the track this
cable should give me many years of
reliable service in the outback when
running cables on the outside of the
vehicle I like to add split tubing just
as an extra layer of protection although
this cable does come in its own
protective sheath it's just one extra
layer which protects against heat for
example if it comes close to the exhaust
or from any other stones or chips or
potentially even rats or mice out in the
bush climbing in and having a nibble
which I've had happen before all the
connections were completed with the
battery and the fuse block wired up I
added a cover over the socket
connections for protection on the
passenger side I have a 12 volt socket
and a dual USB charger and the driver
side to 12 volt sockets the fuse box
also powers all the lights in the
backend I also did something crazy and
bought myself a 12 volt oven.
Did you enjoy this video? Then please
click subscribe and share it on Facebook
and Twitter. If you'd like to help
support me create new videos then please
consider becoming a Patreon. Click on the
Patreon button on the screen now. Thanks


text:
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subset: commoncrawl


text:
In 2016, the share of innovative companies in Estonia increased to 47.7 per cent from only 26.5 in 2014, while in Portugal the share increased to 67 per cent from 54, according to the latest data from Eurostat's Community Innovation Survey.
Strong increases were also reported for Finland, Croatia and Lithuania. The share of innovative companies in Finland grew from 55 to 65 per cent, helping the country, along with Portugal, to move into EU's top five member states with the largest share of innovative companies.
However, the number of innovative companies decreased in eight member states, while the EU average only increased slightly. According to the survey, 51 per cent of enterprises with more than 10 employees reported innovation activities between 2014 and 2016, while only 49 per cent did so between 2012 and 2014.
Between 2014 and 2016, the highest proportions of enterprises with innovation activity were recorded in Belgium (68 per cent), Portugal (67 per cent), Finland (65 per cent), Luxembourg (64 per cent) and Germany (64 per cent). In the same period, the share was below 30 per cent in Romania (10 per cent), Poland (22 per cent), Bulgaria (27 per cent) and Hungary (29 per cent).
The Community Innovation Survey is tracking the number of enterprises that have either introduced an innovation or have any kind of innovation activity, including enterprises with abandoned and suspended innovation activities.


text:
Get a Professional Report of Your Home Value!
It is a sellers market! It has never been a better time to sell your home! To request a Professional Report to Find Out How Much Your Home is Worth, please start filling up the form below. We provide you with a free extensive professional valuation of your home's current value.
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If you have any questions please call Oscar to (626) 340 2573. (Hablamos Espan


text:
A guide to the perfect meeting room
Meetings play an important role in business. It's here where the big decisions are made, numbers are crunched and performance scrutinised. To make your life easier, we tell you what it takes to create the perfect meeting room.
Choose your furniture wisely
A meeting room needs to impress clients and promote a relaxed atmosphere that is separate from the rest of your office.
You must fill the space efficiently. The furniture you choose should ideally be moveable, so you have the option of changing the layout whether you're hosting a meeting for two people or 20 people.
Tables with wheels are a good idea. That way you can easily position them so employees can see, hear and interact with each other more effectively. Furthermore, tables and chairs should be easy to stack away.
You also need to think about comfort when it comes to seats. After all, you could be in a meeting for hours on end.
Think carefully about light and lighting
Employees tend to work better in a room lit by natural light rather than a room illuminated by artificial light, so all blinds should be up during meetings to let as much natural light into the room as possible.
If you haven't got big windows, or any windows whatsoever, you need to be clever with your lighting. Avoid using direct lighting and typical overhead solutions, which can reflect off computer screens.
Instead, opt for more subtle and unobtrusive lighting. Indirect lighting that is angled so it fills the room is less disrupting and helps create a relaxed atmosphere. Just remember to position it away from the gaze of participants.
Get your hands on the latest technology
Having the latest technology can make a huge difference to the productivity and eventual success of a meeting.
Digital whiteboards, for instance, provide you with an effective method to engage employees or clients, while tablets remove the need for paperwork, which can clutter the room and slow down proceedings.
If you're working on a limited budget, and therefore cannot afford such luxuries, just make sure your meeting room has a good spread of power outlets and a decent Wi-Fi connection. Both features are essential.
Pay attention to décor
Going back to aesthetics, the simple process of decorating the room has the power to make or break a meeting.
Believe it or not, the colour of your walls can actually affect things like productivity. Employees are inspired and motivated by what they see around them, while clients form impressions based on what they see.
Light shades of yellow and blue are considered to be stimulating and creativity-inducing colours, pink evokes a sense of tranquillity and green is seen as smart and traditional, so mix up your walls rather than staying neutral.
Go green
Introducing plants is a way of enhancing meeting rooms with minimal effort. They soften right angles, add a splash of colour and even oxygenate the room, making them an extremely worthwhile investment.
Non-flowering plants work best, while it's better to introduce those that require little or no maintenance.
Their presence is enough to lift the spirits of employees and clients, so stick a tall fern in the corner of the room or put a potted plant on a few of the tables.
Posted by Julie Tucker


text:
Our team was engaged by the family run development group to rebrand them and create a brand platform for the properties, capital division and foundation.
#101 2020 Abbotsford Way V2S 6X8,
Abbotsford, British Columbia, Canada
Tel 1.888.427.9997
Tel 604.852.6976
Monday 8:30a.m.–5:30p.m.
Tuesday 8:30a.m.–5:30p.m.
Wednesday 8:30a.m.–5:30p.m.
Thursday 8:30a.m.–5:30p.m.
Friday 8:30a.m.–5:30p.m.
Saturday Closed
Sunday Closed
With a head-office in Abbotsford, Rally Creative also has locations in Kamloops and Victoria, serving some of British Columbia's fastest growing communities.


text:
Bran Book 2006
Paperback, 140 Pages
Price: $41.96
Prints in 3-5 business days
A Bran is a template. A canvas if you like. Think back to when you were a kid - remember he-man figures? Each one essentially the same piece of plastic with a different paint job? Bran is a bit like that. www.bran-man.com gives designers and illustrators, from first year students through to industry professionals, the opportunity to put their stamp on a simple object - this book is the result of that. 75 of the best Bran-man submissions since the website launched in the summer of 2005 are collected here, along with a few words from their creators.
Product Details
- 9781411673199
- Copyright
- Standard Copyright License
- Publisher
- Lulu.com
- Published
- January 24, 2006
- Language
- English
- Pages
- 140
- Binding
- Perfect-bound Paperback
- Interior Ink
- Full color
- Weight
- 0.84 lbs.
- Dimensions (inches)
- 9 wide x 7 tall
- Product ID
- 270436
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There are multiple challenges preventing integration of sexual and reproductive health (SRH) information into the curriculum of medical and nursing education. To achieve the end goal of having an inter-professional workforce capable of taking care of patients' sexual health in a variety of settings, integrating this information into the education of doctors and nurses is imperative.
In an effort to explore the issues that impact SRH education for both medical and nursing education, as well as IPE between these two fields as it relates to SRH curriculum, the American Sexual Health Association, Reproductive Health in Nursing, the Association of Reproductive Health Professionals, and Nursing Students for Sexual and Reproductive Health are bringing together representatives from nursing and medicine to participate in a Summit on Medical and Nursing School Education in Sexual Health.
The goals of the summit include identifying barriers to inclusion of SRH content in curriculum, and creating actionable solutions that will address these barriers. A report of the findings and recommendations will be drafted for publication following the summit.
Where: Hyatt Regency Atlanta, 265 Peachtree St NE, Atlanta, GA 30303. Tel: +1 404-577-1234
When: Tuesday, October 17, 8am – 5pm
We will be providing breakfast, lunch, and two snacks on the day of the meeting.
Hotel accommodations: The Forum on Family Planning has graciously allowed us to use their block room rate for this one-day summit. Hotel arrangements can be made at the Hyatt Regency through this booking link: https://www.regonline.com/builder/site/custom.aspx?EventID=1484387&_i=h1GBfV%2f4fNc%3d
- Check-in time: 3:00 pm
- Check-out time: 12 noon
The hotel has a 24-hour fitness center, a FedEx Business Center, and a seasonal outdoor pool.
Getting there: The Hyatt Regency is just ten miles away from the Hartsfield-Jackson International Airport and located near many public transportation options. You can take the MARTA rapid transit from either the GOLD line or the RED line to the PEACHTREE CENTER STATION and walk 5 minutes to the Hyatt Regency (~1 hour).
For a free Uber ride (first-time users only) sign up using code samanthaa770
Parking: Valet parking at the hotel is $35 for 6 hrs-24hrs. There is no self-service parking available on the property. Guests may elect to self-park at off-property locations, such as 31 Baker St. (31 Baker Street NE). This covered garage is one block from the hotel and is available at a discounted rate for Hyatt guests. Charges are $20 for 24 hours with no in/out privileges, valid Friday – Sunday.
Things to do: The Hyatt Regency is conveniently located in downtown Atlanta, close to the Georgia Aquarium, the Center for Civil and Human Rights, and the World of Coca-Cola. Check out the hotel activities page for a comprehensive list of attractions and things to do while visiting Atlanta.
Getting around: The Hyatt Regency is directly connected to the MARTA transit system. Currently, the one-way fare for MARTA costs $2.50. Here are the buses and stations that stop at the Hyatt Regency:
- Bus 10A, 10B, 100, 101, 102, 103, 413, 419, 426, 430, 432, 440, 441, 442, 453, 463, 476, 480, 490 at Baker St. & Peachtree St. Bus Station
- 16 Bus at Baker St. & Peachtree Center Ave Bus Station
- Peachtree Center Transit Station at Peachtree St. & Forsyth St.
Summit Organizer contact info:
Sarah Macrorie – email@example.com cell: 541-359-6551
Samantha Andrews – email@example.com cell: 505-401-6097


text:
The spring 2020 graduation ceremony will be held Sunday, May 3 at 11 a.m. at the Donald L. Tucker Civic Center. U.S. Representative Kathy Castor ('91) will be the commencement speaker. Immediately following the ceremony, there will be a reception at the College of Law.
To watch the 2019 graduation ceremony live on our website, click here. To watch on YouTube, click here.
For more information about graduation, please contact the Office of Student Advancement at 850-644-7338 or firstname.lastname@example.com.
View additional photos from the event on our Facebook page.


text:
No. 2D12–6460.
Thomas Erskine Ice and Amanda L. Lundergan of Ice Appellate, Royal Palm Beach, for Appellants. Nancy M. Wallace and Michael J. Larson of Akerman Senterfitt, Tallahassee, and William P. Heller of Akerman Senterfitt, Fort Lauderdale, for Appellee.
Thomas Erskine Ice and Amanda L. Lundergan of Ice Appellate, Royal Palm Beach, for Appellants.
Nancy M. Wallace and Michael J. Larson of Akerman Senterfitt, Tallahassee, and William P. Heller of Akerman Senterfitt, Fort Lauderdale, for Appellee.
SLEET, Judge.
Daniel and Aimee Wolkoff appeal the trial court's final judgment of foreclosure in favor of American Home Mortgage Servicing, Inc. (AHMSI). The Wolkoffs contend that there was insufficient evidence to support the amount of indebtedness owed to AHMSI. We agree and reverse.
On May 16, 2007, the Wolkoffs executed a promissory note and mortgage for $580,300 with American Brokers Conduit. The original note and mortgage were transferred to AHMSI via blank endorsement and assignment shortly thereafter. When the Wolkoffs defaulted on the September 2007 payment, AHMSI issued a default notice to them on October 2, 2007. The Wolkoffs failed to cure the default, and AHMSI filed for foreclosure in January 2008.
At the bench trial in November 2012, AHMSI offered the testimony of Jay Vent, a default case resolution representative for AHMSI, who authenticated the promissory note, mortgage, and payment history. However, when Vent testified about the amount of debt owed by the Wolkoffs he merely confirmed that the totals given to him on a proposed final judgment "seemed accurate"; he never openly recited the total amount of indebtedness, nor did counsel for AHMSI ask him to. AHMSI did not submit the proposed final judgment or any business records relevant to the Wolkoffs' mortgage payment history as evidence. At trial, the Wolkoffs, who are both experienced in real estate, admitted to executing the note and testified that "money was owed to somebody." The trial court ultimately entered a final judgment of foreclosure in the amount of $960,660.96, which included costs and an award of attorney's fees.
It is axiomatic that the party seeking foreclosure must present sufficient evidence to prove the amount owed on the note. Typically a foreclosure plaintiff proves the amount of indebtedness through the testimony of a competent witness who can authenticate the mortgagee's business records and confirm that they accurately reflect the amount owed on the mortgage. Thereafter, the business records are admitted into evidence. Here, AHMSI attempted to prove the amount of indebtedness by presenting Vent with a proposed final judgment, presumably prepared by AHMSI's counsel. A document that was identified but never admitted into evidence as an exhibit is not competent evidence to support a judgment. Correa v. U.S. Bank Nat'l Ass'n, 118 So.3d 952, 955 (Fla. 2d DCA 2013).
To the extent that AHMSI argues that the Wolkoffs failed to preserve a challenge to the use of the proposed final judgment through contemporaneous objection, its argument is without merit. Florida Rule of Civil Procedure 1.530(e) provides:
When an action has been tried by the court without a jury, the sufficiency of the evidence to support the judgment may be raised on appeal whether or not the party raising the question has made any objection thereto in the trial court or made a motion for rehearing, for new trial, or to alter or amend the judgment.
The Wolkoffs were not required to make a contemporaneous objection to the sufficiency of the evidence in order to preserve the issue for appeal. See Correa, 118 So.3d at 954.
During discovery, the Wolkoffs attempted to obtain copies of all business records that AHMSI intended to introduce into evidence at trial. Despite these efforts and over four years of litigation, AHMSI never provided the records that established the total amount of indebtedness. The mortgage payment history records introduced through Vent were incomplete, out of date, and failed to reflect the current debt owed on the mortgage. These records do not support the dollar amounts contained in the final judgment. There was no testimony or evidence to support the award of interest, taxes, property inspections, property evaluations, or attorney's fees. Accordingly, we must reverse the final judgment of foreclosure.
At oral argument, AHMSI argued for the first time that under Sas v. Federal National Mortgage Ass'n, 112 So.3d 778 (Fla. 2d DCA 2013), the proper remedy is for this court to reverse and remand for further proceedings so that AHMSI may properly establish the amounts allegedly due and owing under the Wolkoffs' mortgage. Although this argument may not be properly before the court, see Powell v. State, 120 So.3d 577, 593 (Fla. 1st DCA 2013) (noting that an appellate court should consider arguments raised for the first time at oral argument only in "[r]are or unusual instances"), Sas is distinguishable from the present case. In Sas, the bank submitted evidence of the amount of indebtedness through witness testimony without producing the business records on which the amount was based. 112 So.3d at 779. The trial court overruled Sas's objection to the testimony as hearsay, and Sas sought review of that ruling in this court. Id. We held that the trial court abused its discretion when it allowed the loan servicer's witness to recite over objection the amount of indebtedness as reflected in the mortgagee's business records without first admitting those records.Id.
Similarly, in Kelsey v. SunTrust Mortgage, Inc., 131 So.3d 825 (Fla. 3d DCA 2014), the Third District reviewed the trial court's evidentiary ruling. Kelsey objected to admission of the bank's records because the records custodian was unqualified and incompetent to testify. The trial court overruled the objection, and the bank submitted the business records that established the amount due on the mortgage. On appeal, the Third District held that it was error for the trial court to admit the documents over objection without proper authentication and reversed and remanded for a rehearing. Id. at 826.
Unlike the lenders in both Sas and Kelsey, AHMSI failed to submit into evidence either the amount of indebtedness or the business records on which the amount was based. Although it appears that the trial court may have relied on the proposed final judgment to enter its order, the proposed final judgment was not entered into evidence, is not a part of the record on appeal, and is not likely an admissible business record of AHMSI. The defendants in the Sas and Kelsey cases objected to the banks' evidence on hearsay grounds; however AHMSI presented no evidence detailing any amounts due and owing, and under rule 1.530(e) the Wolkoffs did not need to object to challenge the sufficiency of the evidence. In Sas and Kelsey, the focus was the admissibility of the evidence of indebtedness and the equities were balanced in favor of remand. Because we are not reviewing an evidentiary ruling, our standard of review is different than the standard applied in Sas and Kelsey. We review the trial court's findings for competent, substantial evidence supporting the verdict. Crawford Residences, LLC v. Banco Popular N. Am., 88 So.3d 1017, 1019 (Fla. 2d DCA 2012).
This court's Correa opinion is instructive. 118 So.3d 952. In Correa, the bank failed to present evidence of the terms of a lost note under section 673.3091(2), Florida Statutes (2007), or its right to enforce the lost note under section 673.3091(1)(b). Although the bank presented the testimony of a records custodian who identified a copy of the lost note, the bank did not submit the document into evidence and the custodian did not testify as to its terms. Correa, 118 So.3d at 955. Accordingly, this court reversed and remanded for the trial court to enter an involuntary dismissal of the complaint. Id. at 957. Similarly, AHMSI's failure to prove a prima facie case warrants dismissal. See Fla. R. Civ. P. 1.420(b) ; Allard v. Al–Nayem Int'l, Inc., 59 So.3d 198, 201 (Fla. 2d DCA 2011). Because there is no evidence to support the amounts contained in the final judgment, we are constrained to reverse.
"[A]ppellate courts do not generally provide parties with an opportunity to retry their case upon a failure of proof." Correa, 118 So.3d at 956 (quoting Morton's of Chi., Inc. v. Lira, 48 So.3d 76, 80 (Fla. 1st DCA 2010) ). When a party seeking monetary damages fails to establish an evidentiary basis for the damages ultimately awarded at trial, reversal for entry of an order of dismissal is warranted. Morton's, 48 So.3d at 80. AHMSI had over four years to produce business records that could prove the amount of the Wolkoffs' indebtedness, and it had the opportunity at trial to prove its case. Instead, at trial, it relied on an unauthenticated document prepared in anticipation of litigation. AHMSI failed to satisfy its burden to prove the amount of debt owed. Therefore, dismissal is appropriate. See Allard, 59 So.3d at 201.
Accordingly, we see no reason to afford AHMSI a second opportunity to prove its case. We reverse and remand with directions for the trial court to enter an order of involuntary dismissal.
DAVIS, C.J., and SILBERMAN, J., Concur.


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Nursing Home Negligence
There are several types of injuries common in nursing home abuse. Some examples include bed sores, dehydration, malnutrition, uro-sepsis, septic conditions, administration of the wrong medicine, incorrect administration of the correct medication, wrong dosage, physical abuse, failure to properly notify the doctor of a problem which could be addressed before injuring the patient, failure to notify the family of a problem being experienced by the family member, neglect of a patient, and failure to follow a care plan for a patient…ReadMore
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N-Power Final Selection Stage – What Applicants Must Do After Physical Verification
N-Power Final Selection Stage – What Applicants Must Do After Physical Verification
N-Power Final Selection Stage
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Abstract Text:
Join us as we examine how USGBC's Living Standard campaign is highlighting the power of storytelling for a more sustainable future. Creating that future requires connecting the dots between data and people—particularly in the real world and outside our environmentalist and sustainability bubbles. Most people are aware of the stakes, but say they feel distant and existential, rather than tangible and actionable. But what happens when we change the dialogue around data? What happens when we bring quantitative data to life by ensuring the stories behind the numbers are the heart of the green building narrative? In the face of unprecedented climate-related risks, we will explore how we can become more effective communicators and community partners, how we can find and share the meaning behind our measurements, and how we can leverage the shift in narrative to create both a better quality of life and higher living standard for all.
This session will be moderated by Mahesh Ramanujam, USGBC president and CEO and features a diverse group of sustainability advocates who will share their sustainability stories and discuss ways they see Living Standard take shape in their communities. We will also cover how to use various resources to inspire others to take action and expand the conversation.
Learning Objectives:
Understand the value in storytelling to inspire action for a more sustainable future
Be able to encourage others to share their sustainability stories while sharing your own
Be able to use findings from the Standard Issue reports and the Living Standard Action Toolkit to expand the conversation around Living Standard themes
Become a more effective communicator in encouraging others to act through a data-driven approach
Learning Level:
- Basic
- GBCI Credit Hours:
- AIA Credit Hours:
Education Tracks: USGBC: People|Projects|Performance
Event Type: Conference Session > Session
Mahesh Ramanujam
President and CEO at U.S. Green Building Council; Green Business Certification Inc.
Living sustainably has always been a core belief and practice of Mahesh Ramanujam. Growing up in India where much of the population around him faced economic and environmental hardships, Mahesh learned from a young age to make do and be grateful for what …
David Bluestone
Co-Founder and Partner at ClearPath Strategies
David Bluestone is a Founder and Partner of ClearPath Strategies. He has over a decade of experience providing strategic consulting and public opinion research to politicians, businesses, and organizations. His work spans five continents and over two doz …
Rami Vagal
Senior Manager of Sustainability at Mohawk Group
Rami Vagal, senior manager, sustainability at Mohawk, is a multi-disciplinary professional with a strong background in sustainability, architecture and construction. At Mohawk, Rami leads strategic sustainability initiatives that create positive impacts o …
Diana Yazzie Devine
President/CEO at Native American Connections
Phoenix, Arizona, United States
Diana Yazzie Devine, CEO of Native American Connections (NAC) since 1979 – over 40 years. NAC a Native American operated nonprofit providing health services, affordable housing, and community economic development. With Native Americans as a priority pop …
Mike Carey
Sustainability Coordinator at Orange Coast College
Costa Mesa, California, United States
Mike Carey - LEED AP - TRUE Zero Waste Advisor Sustainability Coordinator, Orange Coast College Mike Carey began his career at Orange Coast College in 1985 as an intern with the campus Recycling Program and eventually transitioned to the campus Susta …
Lisa Borders
CEO at LMB Group, LLC
Lisa Borders has three decades of leadership across the private, public and non-profit sectors. She is the CEO of LMB Group, LLC, a management consultancy firm, and recently chaired the Borders Commission, an independent and strategic task force charter …


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'Taichung Our City' gives viewers a bird's eye view of its scenic spots and recent improvements
Taiwan News
Date: 2019/09/10
By: George Liao, Taiwan News, Staff Writer
According to the introduction, Taichung City Government worked with the late director Chi Po-lin (齊柏林), who directed the award-winning documentary, "Beyond Beauty – Taiwan From Above," to produce the video, "Taichung – The Heart of Taiwan," in 2014. The video is full of beautiful Taichung scenery.
Four years later, the city government has once again cooperated with the aerial photography and filming company Above Taiwan Cinema for the production of "Taichung Our City." This gives viewers a bird's eye view of recent developments in the city, according to the introduction.
The video features Gaomei Wetland, rivers, and valleys; night views of the downtown area; and recent developments in the city such as the Taichung Metro Green Line and National Taichung Theater. [FULL STORY]


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We had two of our recipes featured in the first ever 'PlantBased: the food revolution' Magazine this month, so if you missed out, we thought we would share one of them with you. It's too good not to try!
TURMERIC PANCAKES (makes approx. 10 pancakes)
220g (1 cup) spelt flour
2 tbls coconut sugar
Pinch of Himalayan pink salt
1 tsp baking powder
250ml (1 cup) Sweet Revolution Turmeric Latte, cooled
1 tsp vanilla extract
2 tblsp coconut oil, for cooking
1. Combine flour, salt, sugar and baking powder in a bowl
2. Add latte mix and vanilla and whisk until well combined
3. Heat frying pan with a small amount of the coconut oil
4. Add 2-4 tbls of mixture per pancake to the pan and cook on each
side for a couple of minutes, adding a little more oil for each pancake
5. Serve with fresh berries, maple syrup and coconut cream


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Pre-K & Kindergarten Teacher
- Early Childhood
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- Bachelor
- 3 years of teaching experience
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Teaching Credential/License
- Prior teaching experience at the lower Elementary School level
- Furnished accommodation: allocation based upon family size and availability, AUS provides utilities and transportation to and from work
- Travel allowance: Annual home leave (summer break) cash allowance (equivalent of return economy class airfare to point-of-origin for employee and eligible dependents residing in Kuwait)
The Early Years Classroom Teacher embraces the ideals of holistic, whole child development and views the early years as a crucial time for establishing strong foundations for life. The classroom teacher is responsible for all aspects of teaching and learning in a multi-age, early years setting for students ages 4 to 6 years. The classroom teacher strives to nurture each child's development from a strengths-based perspective. He/She shapes engaging learning environments and experiences that are supportive of and responsive to an emergent approach to playful inquiry and learning. The classroom teacher builds strong relationships with students, families and the team.
American United School (AUS) is located in the Sabah Al Salem Area of Kuwait at the Intersection of 6th Ring Road and Fahaheel Expressway. American United School is a modern, 12,000 square meter facility. The Founding Director came on board October 2012. Support staff which included a finance office, an admission/registration office, purchasing office, and administrative assistants began work in early 2013. The first phase of the facility, the elementary building opened September, 2013. Gaining approval/license from the Kuwait Ministry of Private Education required a name change and, therefore, American University School became American United School. The teaching staff of 24 teachers, 11 co-teachers and 39 staff members were on board to support grades Pre-K through grade 5 and the 68 students for SY 2013-2014. The end of year enrollment was 103 students and the student population represented 11 nationalities.
The second year (SY 2014-2015) saw 270% growth and the addition of the Middle School operating on the upper floor of the Elementary School Building. That second year, the teaching staff increased to 52 teachers, 23 co-teachers, 52 staff members and a student enrollment of 383 students. There were 28 nationalities represented in the student population. Additional personnel to assist and reinforce teaching and learning were added: Middle School Principal and Assistant Principal, Elementary Assistant Principal, Curriculum Specialist, Swimming Coaches, Information Management Officer, and other administrative support staff.
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AUS achieved International Accreditation by the Council of International Schools (CIS) on February 2018, and by the Middle States Association of Colleges & Schools (MSA-CESS) for the period May, 2018 to May 2018.
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This staggering war movie has left audiences with plenty to think about.
After watching 1917, many viewers are wondering if George MacKay has a girlfriend.
The cinematic year has gotten off to an impressive start with the release of such entertaining efforts as The Gentlemen, Jojo Rabbit and Uncut Gems. However, the film to truly make waves so far this year is 1917.
From director Sam Mendes (Skyfall) comes this World War I epic chronicling two soldiers as they attempt to deliver an imperative message across enemy lines to save the lives of 1600 men. It's an extraordinary piece of work and audiences have been quick to champion actor George MacKay in the central role of Lance Corporal Schofield.
The 27-year-old performer has previously starred in the likes of Captain Fantastic, Pride and Sunshine on Leith, but arguably turns in the best performance of his career so far in 1917. Fans are quickly becoming obsessed with him, leading to the inevitable question…
A REAL HERO: Who is Alfred Mendes?
Does George MacKay have a girlfriend?
No, at the moment George MacKay is single, as highlighted by Dating Celebs.
However, film fans will be keen to discover that he used to date Saoirse Ronan, who has delighted us all in films like Lady Bird and The Grand Budapest Hotel.
In a post-breakup interview with the Evening Standard back in 2016, the interviewer mentioned her and he simply said: "Saoirse is so brilliant and is doing amazing things."
Of course, he's not wrong. However, it's safe to say that they are both doing amazing things. Saoirse is actually nominated for Best Actress in a Leading Role at the 2020 Academy Awards for her performance as Jo March in Greta Gerwig's Little Women.
The earlier source notes that the pair met on the set of the 2013 film How I Live Now, in which George played Eddie.
According to the Irish Mirror, the film's director Kevin Macdonald said in a 2013 interview: "What happened during the course of film, and I'm hoping that I'm not speaking out of turn, is that they [George and Saoirse] fell in love and it was very easy… It was Saoirse's first proper boyfriend and, in a way, I think she was living through the same thing that the character is going through."
Since then, it's all been rather quiet.
THIS IS HOW I WIN! Uncut Gems ending explained
Twitter is in love with George MacKay!
Although the actor is single, there's certainly no shortage of people who would love to date him.
A number of people have taken to Twitter to praise the star. Check out a selection of tweets below:
THIS MEANS WAR! Wiley vs Stormzy explained
Is George MacKay on Instagram?
No, he's not.
As written by Emma Baty over at Cosmpolitan: "This man has literally no public social media. I have searched high and low, far and wide, and I cannot find even so much as an old, unused Twitter account. He might have personal profiles that are super locked down, but that doesn't help me…"
What a shame!
However, there is a fan account on Instagram at @gmackayonline. They have a bunch of content on there and boast over 4,000 followers. Be sure to take a look.
In other news, is Uncut Gems based on a true story?


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Susan from Canada sent an email to her loved ones and left her relatives and friends shocked.
The reason being that she needed to drop her wedding because her visitors wouldn't pay £900 for it EACH.
In a Facebook post shared to Mumsnet, details of Susan's requests rise, and, to say the least, it's each of the lots. The issue was an income issue, with visitors declining to stump up several pounds to bankroll her "exclusive" day.
At the point when her life partner proposed a modest and lively wedding in Vegas, she clearly "laughed at his face".
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This week, Kevin and Justin are joined by strength coach, speaker, and internationally published writer Lee Boyce.
Lee coaches clients and athletes for muscle, strength, and sport performance in Toronto, Canada. His work is regularly featured in many of the world's largest publications, like Men's Health, Men's Fitness, Oxygen, Esquire, The Huffington Post, Men's Journal, and T-Nation. He also ran track at the National level in university as a sprinter and long jumper.
On this episode, Lee describes his lengthy road to recovery from a devastating injury nearly two years ago. While playing basketball, Lee ruptured BOTH of his patellar tendons, requiring immediate major surgery, and leaving him unable to bend either of his legs. Lee explains how he coped with the injury, how he rehabbed back to full strength, and what he learned in the process.
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You can follow Lee here:
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Understanding the mechanisms of cross-species virus transmission is critical to anticipating emerging infectious diseases. Canine parvovirus type 2 (CPV-2) emerged as a variant of a feline parvovirus when it acquired mutations that allowed binding to the canine transferrin receptor type 1 (TfR). However, CPV-2 was soon replaced by a variant virus (CPV-2a) that differed in antigenicity and receptor binding. Here we show that the emergence of CPV involved an additional host range variant virus that has circulated undetected in raccoons for at least 24 years, with transfers to and from dogs. Raccoon virus capsids showed little binding to the canine TfR, showed little infection of canine cells, and had altered antigenic structures. Remarkably, in capsid protein (VP2) phylogenies, most raccoon viruses fell as evolutionary intermediates between the CPV-2 and CPV-2a strains, suggesting that passage through raccoons assisted in the evolution of CPV-2a. This highlights the potential role of alternative hosts in viral emergence.
The emergence of novel infectious diseases is often associated with pathogens jumping to new host species. However, the genetic and evolutionary mechanisms that determine how viruses cross species boundaries and adapt to new hosts are only partially understood (25). Well-documented examples of virus emergence in which both the donor and recipient species are known are rare but can reveal key aspects of this important evolutionary process.
Canine parvovirus (CPV) first emerged in the mid-1970s and then spread worldwide in 1978, causing a major disease pandemic in dogs (8). Feline panleukopenia virus (FPV) has long been known to infect many species within the order Carnivora, including large and small cats, mink, raccoons, and foxes (29, 33). CPV is clearly a variant of FPV that acquired the ability to infect canines through a small number of mutations in the capsid protein (VP2) gene that changed surface-exposed residues (1, 3, 19). These viruses use the transferrin receptor type 1 (TfR) as their primary receptor for attaching to and infecting cells. Canine cells resist infection by FPV because that virus cannot bind to the canine TfR, in particular because of a unique glycosylation site present in the canine TfR (3, 10, 16, 18). The first strain of CPV (named CPV type 2 [CPV-2]) was replaced globally during 1979 and 1980 by a genetic and antigenic variant named CPV type 2a (CPV-2a), which is the common ancestor of all the CPVs currently circulating in dogs worldwide (9, 24). The CPV-2a variant contained a number of important genetic changes, including replacements of four VP2 residues that alter capsid binding to the feline TfR and to antibodies and that likely increased the fitness of the virus in dogs and gave it the ability to infect cats (7, 16, 27, 30, 34). The CPV-2a strain has undergone additional evolution in dogs since 1979, generating genetic and antigenic variants known as CPV-2b and CPV-2c (9).
Here we further define the evolutionary basis of viral host range variation by refining our understanding of this classic example of a host-jumping virus—the emergence of CPV-2 and its variants in dogs—through the exploration of its evolution in alternative mammalian hosts.
Cells and cell cultures.CHO-derived TRVb cells, which lack an endogenous TfR (13), were grown in Ham's F-12 medium containing 10% fetal bovine serum. Crandell Rees feline kidney (CRFK) cells, Norden Laboratory feline kidney (NFLK) cells, and canine Cf2Th and A72 cells were grown in a 1:1 mixture of McCoy's 5A medium and Leibovitz L-15 medium with 5% fetal bovine serum.
Viruses.The new viruses examined here are listed in Table 1 and are highlighted in Tables S1 and S2 in the supplemental material. For virus isolation, homogenized tissue or fecal samples were filtered and were used to infect CRFK or NLFK cells. Viral DNA was PCR amplified either directly from tissue extracts or after a single passage in cell culture. Amplification was performed using Phusion Hot Start high-fidelity DNA polymerase (Finnzymes, Espoo, Finland). Amplicons were sequenced either directly or after cloning using a PCR cloning kit (Qiagen, Valencia, CA) and covered the complete VP2 protein sequence or, in some cases, the entire nonstructural and structural coding regions. Control viruses were derived from infectious plasmid clones of FPV (FPV-b), CPV-2 (CPV-d), and CPV-2b (CPV-39) (10, 20) and were purified as described previously (1). Viral inocula were prepared by passing viruses in NLFK cells, homogenizing the supernatant and cell fractions using a freeze-thaw cycle, and clarifying the inoculum through a Steriflip filter (Millipore, Billerica, MA).
Evolutionary analysis.A total of 433 VP2 sequences of different carnivore parvoviruses, 1 of which (GenBank accession number M24005) was sampled from a raccoon in 1978, were compiled from GenBank. These sequences were combined with 27 new sequences and with those determined here from other carnivore species, resulting in a final data set of 460 sequences of 1,755 bp (see Table S1 in the supplemental material). All VP2 sequences included in this analysis covered at least two-thirds of the VP2 gene (although similar phylogenetic results were obtained using 125 full-length VP2 sequences [data not shown]). An equivalent phylogenetic study was undertaken using 39 NS1 sequences collected from GenBank combined with 9 raccoon sequences determined here. This produced a total data set of 48 sequences of 2,007 bp (see Table S2 in the supplemental material). All sequence alignments were created using MUSCLE, version 3.7 (5).
Phylogenetic trees were inferred using the maximum-likelihood (ML) method implemented in PAUP*, version 4.0 (32), utilizing the general time-reversible model of nucleotide substitution incorporating invariant sites and a gamma distribution of among-site rate variation with four rate categories (model GTR+I+Γ4); this was the best-fit nucleotide substitution model determined by Modeltest, version 3.7 (28). Tree bisection-reconnection (TBR) branch swapping was employed in all cases. To assess the robustness of the phylogenetic groupings observed, a bootstrap resampling analysis was performed using 1,000 replicate neighbor-joining trees estimated under the ML substitution model described above, again utilizing PAUP*, version 4.0.
To assess the strength of temporal structure (i.e., clock-like behavior) in these sequence data, we performed a regression of root-to-tip genetic distances against the date (year) of sampling of each sequence by using the Path-O-Gen program (http://tree.bio.ed.ac.uk/software/pathogen/). This analysis was performed using the 48 NS1 and VP2 sequences that were available from the same viruses. Input genetic distances were obtained from the ML trees described above.
The time to the most recent common ancestor (TMRCA) of the raccoon sequences was estimated using the Bayesian Markov chain Monte Carlo (MCMC) method available in the BEAST package (4). This analysis incorporated the model of nucleotide substitution described above, in this case partitioned by codon positions, as well as a relaxed (uncorrelated lognormal) molecular clock and the conservative Bayesian skyline coalescent prior. Statistical support was depicted as values of the 95% highest probability density (HPD), and the MCMC analysis was run until convergence was achieved in all parameters. The BEAST analysis also allowed us to infer the maximum clade credibility (MCC) tree for these data, such that TMRCA values (i.e., divergence times) could be estimated for specific nodes. The MCC tree was topologically very similar to the tree produced in the ML analysis.
Finally, to determine whether recombination had occurred between the VP2 and NS1 genes, we employed the BOOTSCAN, GENECOV, and RDP methods available in the RDP3 package (12) using the default settings.
rTfR.The raccoon transferrin receptor (rTfR) was independently sequenced from a raccoon cell line (Pl 1Ut cells; ATCC, Manassas, VA) and also from heart tissue from a wild raccoon. For subsequent experiments involving binding to the rTfR (see below), mRNA from the wild raccoon was extracted using an RNeasy kit (Qiagen), amplified using the SuperScript III one-step reverse transcription-PCR (RT-PCR) system (Invitrogen, Carlsbad, CA), cloned into the pcDNA3.1(−) plasmid (Invitrogen), and sequenced as described previously (18) (GenBank accession number JN600499).
Antigenic analysis.Twenty-eight mouse or rat monoclonal antibodies (MAbs) against CPV-2, FPV, or CPV-2b have been described previously (21, 23, 27). For hemagglutination inhibition (HI) assays, antibodies were tested using standard methods, and titers were compared to the titer of the virus against which the MAb was raised (30).
Cell infection and binding assays.Infection of NLFK, A72, or Cf2Th cells was assessed using standard methods for 50% tissue culture infective dose (TCID50) assays (34). Cell binding was analyzed using purified virus capsids at 10 μg/ml incubated in a solution for 60 min at 37°C with NLFK or Cf2Th cells, or with TRVb cells transfected with a plasmid expressing the feline, canine, or raccoon TfR (6). Virus capsids were detected with MAb 2 and an Alexa 488-conjugated goat anti-mouse antibody (Sigma, St. Louis, MO), and binding was quantified using a Guava EasyCyte Plus flow cytometer (Millipore).
Raccoons (Procyon lotor) are native to North America but have become established in parts of Europe and Asia since the early 20th century (35). We sampled parvoviruses collected from raccoons that showed clinical disease during outbreaks in the United States between 2007 and 2010 (Table 1), and we compared these viruses to raccoon isolates collected between 1978 and 1990, as well as to the many FPV and CPV sequences from cats, dogs, and related hosts in GenBank (see Table S1 in the supplemental material). The infected raccoons exhibited disease symptoms characteristic of parvoviral enteritis, including lethargy, fever, diarrhea, and vomiting. Parvovirus infection was demonstrated by histopathology and immunohistochemistry of the intestine and lymphoid tissues (not shown).
Phylogenetic analysis of VP2 sequences showed that the raccoon virus isolates fell into several unrelated clusters across the phylogeny, indicating that there have been multiple cross-species transmission events involving raccoons (Fig. 1). Remarkably, 12 of the 17 raccoon virus sequences and the sequence of a virus isolated from a wild bobcat (Lynx rufus) fell at intermediate positions between the CPV-2 and CPV-2a sequences (Fig. 1). Due to the recovery of only a single intermediate virus from a bobcat, the significance of this species either as a long-term host for maintaining these viruses or simply as an incidental host due to raccoon predation is currently unknown. The raccoon VP2 sequences appear to be evolutionary intermediates in that they did not form a separate monophyletic group but instead occupied interleaved positions between the CPV-2 and CPV-2a sequences. The isolates in this intermediate group were collected between 2007 and 2010 from several areas of the United States, including states located between Florida and New York, as well as Wisconsin, while the bobcat isolate was collected in Kansas in 2010.
A molecular-clock analysis of the VP2 data set revealed that the 12 intermediate raccoon isolates and the bobcat isolate diverged from CPV between 24 and 31 years ago (95% HPD values) and hence have been circulating undetected for this period. The remaining five raccoon isolates fell outside the main cluster and were located either among the FPV (FPV/Raccoon/CA/208-A/10, FPV/Raccoon/NJ/RPV-6/90, FPV/Raccoon/TX/Rac1.2/78, FPV/Raccoon/TX/Rac2.2/78) or among the CPV-2a-derived (CPV/Raccoon/NJ/76836/10) sequences. These viruses may represent short-term spillover of FPV and CPV-2a into raccoon populations without sustained transmission, although addressing this question will require additional sampling. Molecular-clock estimates place the origin of these other raccoon viruses at various times during the past 45 years (full results are available on request).
Contrasting evolutionary patterns in the capsid and nonstructural genes.More-complex, and strikingly different, evolutionary patterns were observed in a phylogenetic analysis of 48 NS1 (nonstructural) gene sequences (Fig. 2). In this analysis, the raccoon viruses fell at three distinct phylogenetic positions, indicating multiple cross-species transmission events from canine viruses and giving no evidence that raccoons are intermediate hosts (Fig. 2). We used a Shimodaira-Hasegawa (SH) test to determine whether the NS1 and VP2 genes (the latter subsampled such that the same 48 viruses were used) gave different representations of the evolutionary history of carnivore parvoviruses. This confirmed that the NS1 and VP2 phylogenies were significantly different (P < 0.001) such that they have strongly contrasting evolutionary histories (which also means that they cannot be meaningfully combined into a complete genome phylogeny). Importantly, these dramatically contrasting phylogenetic patterns could not be explained by recombination, since we found no evidence for this process in our data (P > 0.1). Hence, the incongruent NS1 and VP2 trees are most likely the result of very different selection pressures acting on these genes.
There are good reasons to believe that the VP2 tree is the more accurate representation of evolutionary history. First, if the NS1 tree is correct, then both CPV-2 and CPV-2a, as well as the raccoon viruses, have evolved multiple times in parallel and do not form monophyletic groups, signifying a substantial amount of convergent evolution. Second, according to the NS1 tree, parvoviruses jumped from cats to dogs initially as the CPV-2a genotype and then evolved to CPV-2. Not only is this counterintuitive with respect to the known history of mutations in these viruses, but it runs counter to the time scale of parvovirus evolution. In particular, it is well documented that CPV-2 appeared in dogs and spread worldwide during 1978 and 1979, before CPV-2a emerged, as shown in the VP2 tree, and indeed CPV-2 isolates have earlier sampling times (1977 to 1980) than CPV-2a strains (late 1979 onward). Moreover, the correlation coefficients of root-to-tip genetic distance against sampling year were significant for the VP2 (r = 0.70; P < 1 × 10−5) but not the NS1 (r = 0.55; P = 0.171) data set. Because a strong temporal structure is present only in the VP2 phylogeny, we suggest that this is the correct representation of evolutionary history, although the sequencing of additional NS1 gene sequences will help resolve this issue.
Differences in capsid protein residues.After it emerged in 1979, the CPV-2a lineage replaced the CPV-2 strain worldwide within 2 years, indicating increased fitness in dogs. Importantly, the CPV-derived raccoon viruses also accumulated changes in several of the capsid codons that later helped define the CPV-2a lineage (VP2 residues 87, 101, 297, 300, and 305) (Fig. 1 and 3). Residue 300 appears to be a particularly important component of the host adaptive process. This residue is an Ala in FPV, CPV-2, and FPV-like raccoon viruses but is a Gly in all CPV-2a-derived viruses from dogs. However, 300Asp was seen in all raccoon isolates derived from CPV, irrespective of their positions on the phylogeny (Fig. 1 and 3), and hence must have arisen more than once in the raccoon viruses derived from CPV. The importance of the 300Asp change for CPV-derived viruses in hosts other than dogs is confirmed by the finding of this mutation in some viruses from leopard cats in Southeast Asia and in a CPV-2 isolate that had been passaged in cultured feline cells (11, 14, 22). The variants at residues 297 and 305 also have complex evolutionary patterns. Residue 297 was a Ser in CPV-2 and some older CPV-2a and CPV-2b sequences but was mutated to Ala in many of the raccoon viruses, reverted to Ser in some early canine isolates of the CPV-2a variant, and became fixed in dog viruses as an Ala throughout most of the world in the early 1990s (2, 9, 15). Residue 305 was an Asp in FPV and CPV-2, mutated to His in some of the raccoon viruses, and became a Tyr when the virus transferred to dogs during the emergence of CPV-2a.
Raccoon virus properties and host-specific receptor analysis.We confirmed that these mutations altered host range, TfR binding, and antigenic structure in the raccoon viruses. Tissue culture infectivity for dog cells reflects the canine host range of these viruses (34), and while the raccoon viruses tested infected feline cells similarly to the FPV, CPV-2, and CPV-2a prototypes, they did not infect canine cells (Fig. 4). Since a CPV-2 mutant with only the 300Asp change exhibited only 10-fold-lower infectivity for canine cells than for feline cells (Fig. 4), combinations of other residues in the raccoon viruses likely acted along with 300Asp to limit canine cell infection (Fig. 3 and 4).
The TfR is a key determinant of feline and canine cell susceptibility, and host-specific virus binding to TfR is controlled by capsid surface residues (10, 16). We therefore cloned the cDNA from a raccoon tissue sample and found that it exhibited 88% and 89% amino acid identity to the dog and cat TfRs, respectively, and that it lacked the glycosylation site that blocks the binding of FPV capsids to the canine TfR (16, 17) (see Fig. S1 in the supplemental material). When the raccoon, feline, and canine TfRs were expressed on hamster cells under identical conditions, the raccoon virus capsids bound the raccoon and feline TfRs to similar levels but did not bind to the canine TfR (Fig. 5).
The CPV-derived raccoon viruses were also antigenically distinct when tested with a large panel of MAbs prepared against FPV, CPV-2, or CPV-2b capsids (Fig. 6). All CPV-like raccoon virus isolates lost reactivity with several antibodies that recognized antigenic site "B," primarily due to the 300Asp mutation. The CPV/Raccoon/VA/118-A/07 isolate, which also contained the 224Arg mutation in the "A" site, lost reactivity with 26 of 28 MAbs tested (7, 30).
The most striking observation of our study was that a group of raccoon parvoviruses constituted a previously unidentified host range variant of CPV that has been circulating for at least 24 years and causing an epidemic of disease. In addition, the VP2 phylogeny revealed that most of these raccoon-associated viruses were evolutionary intermediates between CPV-2 and CPV-2a strains. Although virus surveillance has been conducted extensively in the domestic dog and cat populations since the emergence of CPV-2 in 1978, viruses intermediate between CPV-2 and CPV-2a have not been identified previously. In dogs, the CPV-2a strain rapidly replaced CPV-2 worldwide; this, coupled with the intermediate phylogenetic position occupied by most of the raccoon viruses, suggests that raccoons played a key role in the emergence of this fitter variant in dogs. The selective advantage of CPV-2a over CPV-2 may have involved both better adaptation to the canine host and partial evasion of the anti-CPV-2 antibodies that were widespread in the dog population by 1979 and early 1980 (24).
An unusual aspect of this adaptive process was that the evolutionary intermediates of CPV in raccoons could not themselves bind to the canine TfR or infect canine cells (Fig. 4 and 5). This system therefore illustrates the complexities of the evolutionary pathways and fitness landscapes that may occur during adaptation to new host species. While several raccoon-associated mutations (VP2 residues 87 and 101) were also found in the canine-adapted virus CPV-2a, the 300Asp mutation likely prevented binding of the capsids to the canine TfR in the background of the raccoon virus isolates (Fig. 5). The transfer of the raccoon virus back to dogs clearly required the crossing of a relatively low evolutionary barrier and involved transition mutations that converted VP2 codon 300 from Asp to Gly and VP2 codon 305 from His/Asp to Tyr and that resulted in the CPV-2a genotype, which infected and spread efficiently within dogs. However, it is possible that other carnivore hosts in addition to raccoons have played a role in the emergence of CPV-2a in dogs, such that more-comprehensive sampling of additional carnivore species would likely aid in determining those which are important in the evolution of these viruses.
Despite the clear phylogenetic links between the raccoon and canine viruses, the NS1 and VP2 phylogenies have strikingly different topologies, suggesting that NS1 and VP2 have been subject to very different selection pressures (especially given the apparent absence of recombination). Because our collection of canine isolates goes back to 1978, we can directly validate the inferred phylogenetic history through comparison with the known sampling dates of the viruses. In this case, the phylogenetic patterns seen for the VP2 gene fit well with the known history of the canine viruses. The raccoon virus sampling is less well distributed in time, but it is reasonable to expect that the VP2 gene analysis can similarly be used to more accurately infer the evolutionary history of these viruses. Future studies will be directed toward determining the selection pressures acting on NS1 and VP2 and how these might have shaped phylogenetic history.
At a more fundamental level, our study shows that the selection of small numbers of key host-adaptive mutations is readily accomplished in these single-stranded DNA viruses and can occur on multiple occasions; in this case, the transfers between a cat-like host, dogs, and raccoons were each associated with groups of such mutations. As a consequence, successful emergence is not always dependent on mutational availability (31). Similar observations have been made with respect to the adaptation of influenza viruses among mammalian species (25, 26) and further demonstrate that the strength of host phylogenetic relationships can act as a predictor of successful viral emergence (31).
Finally, and perhaps of most importance, this study highlights the potential role of alternative hosts (in this case raccoons) in providing indirect viral emergence pathways, even in cases where a fitter viral variant, such as the CPV-2a strain, appears to arise within the same host species as the original strain (CPV-2). While alternative and intermediate hosts are often proposed to provide a bridge for the transfer of viruses between widely separated host species, their role in facilitating increased adaptation to a single host has not been described previously. We therefore suggest that future attempts to survey virus biodiversity and to create theoretical models for predicting and forestalling viral emergence should pay close attention to the possible roles played by related hosts in providing favorable adaptive pathways.
A.B.A., J.D.B., M.G.R., and M.K.K. were supported by funds obtained through sponsorship by the fish and wildlife agencies of the member states of the Southeastern Cooperative Wildlife Disease Study (SCWDS). A.B.A. thanks Jeremiah Saliki at the University of Georgia for graciously allowing the use of his laboratory space. I.P. was supported by Marie Curie fellowship PIOF-GA-2009-236470. This work was also supported by NIH grants GM080533-05 (to E.C.H. and C.R.P.), AI028385, and AI092571 (to C.R.P.).
- Received 2 September 2011.
- Accepted 26 October 2011.
- Accepted manuscript posted online 9 November 2011.
Supplemental material for this article may be found at http://dx.doi.org/10.1128/JVI.06187-11.
- Copyright © 2012, American Society for Microbiology. All Rights Reserved.


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❶You get the whole day to explore. The Zoo is also en route to the Great Ocean Road.
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If you continue browsing, you agree to the use of cookies. More details can be found in our xay policy. It is a short Friends Bathurst ok drive from the city over the Westgate Bridge or via the Geelong Road. Public Geelong zoo free day Step 1.
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You can unsubscribe at any time. For more information, read our privacy statement. History 1 Wishlist 0 Log in. See gallery. Spend a day at Werribee Open Range Geelog and marvel at the variety of vay in natural settings. Enjoy a guided safari across open savannah, which is included with general admission.]Learn what goes into the creation and design of your own 3D virtual world and game!
During the Foundation program, Educators may wish to take the opportunity to co-learn freee the students. Co-Teach Feel confident with the program? Come team-teach with us!
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We have everything ready so have a freee how you would like to be part of the Foundation program. We will supply all the resources and let you shine. Imagine a new zoo will be built east of Geelong.
This zoo will be without any cages as it will use natural vegetation to protect and sustain the animals. The Geelong Zoo would like to promote the new zoo with a game that will educate, inspire and engage children to interact with the animals. Learn what goes into the creation and design of a 3D game and protect the animals!
This program can cater for up to 30 students. The Geelong Zoo would like to promote the new zoo with a game that will educate, Please let us know if you have 20 or more students so we can arrange free buses.
Please be advised that photos taken on the day of student work. Open 9am – zoo every day of the year (entry closes pm) Kids visit free! Zoos Victoria is a not-for-profit conservation organisation committed to fighting. Hi, may i know if the parking at werribee zoo is free? If not this way you can avoid the huge line that builds up when everyone starts to leave! enjoy your day. Find your booking with itinerary number.
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