text:
How digitisation is helping pave the way for climate neutrality
Digital: paving the way for climate neutrality
21 Oct 2020Planet news
By Joakim Reiter, External Affairs Director, Vodafone Group
With EU Green Week in full force, it's evident now more than ever that much must be done to achieve a carbon-neutral continent both in the immediate and longer-term. According to the Intergovernmental Panel on Climate Change, "scientific evidence for warming of the climate system is unequivocal".
The European Commission President Ursula von der Leyen has set the ambitious target tomake the EU the world's first climate neutral continent by 2050 and boldly increasing the 2030 target for emissions reduction from 40% to at least 55% in order to achieve this, we must put our best foot forward to transform this goal into reality.
Global warming poses an existential threat to both our people and planet – it is an issue our generation must engage with for the benefit of generations to come. According to new survey results released today from the Vodafone Institute, the European think-tank of Vodafone Group, nearly two-thirds of the EU citizens surveyed said that tackling climate change is an overarching issue for themselves, politics and businesses and more than half (54%) believe that digital technology can play a big role in solving the problems of climate change.
EU citizens feel so passionately about the imminent threat of climate change that the vast majority would accept stricter climate protection measures from the EU - even if this meant additional financial burdens. In fact, they expect Europe to take a leading role in the world.
The people have spoken, and it is now up to Governments and the business community to realise the change they seek in tangible ways. This is a key reason why Vodafone is committed to playing an active role in building a more efficient, circular economy. By helping to reduce and ultimately eliminate waste, we hope to achieve this, and have committed to an ambitious goal to reuse, sell or recycle 100% of our network waste by 2025.
What's more, our European network will be powered by 100% renewable electricity no later than July 2021, creating a Green Gigabit Net for customers across 11 markets that will grow sustainably using only power from wind, solar or hydro sources.
We're seeing many large companies commit themselves to climate neutrality and the range of resource-saving products is constantly evolving in almost all areas of life. Investors are exerting considerable pressure on listed companies to operate more sustainably and thus minimise long-term risks and consumers are becoming increasingly focused on how environmentally responsible brands are. In fact, this new research from the Vodafone Institute finds that 87% of EU citizens consider environmentally responsible behaviour when it comes to purchasing decisions.
Evidently, the issue of climate change is something we must all tackle together. It simply cannot be ignored, or de-prioritised, which is why Vodafone is committed to helping business customers who use its services reduce their own carbon emissions by a cumulative total of 350 million tonnes globally over 10 years between 2020 and 2030.
However, reducing CO2 emissions alone will not be enough. Smart solutions are needed to ensure global energy flows in the future. Digitisation plays a decisive role in this respect: whether in the efficient control of road traffic and international logistics flows, or in the reduction of energy consumption in private households. Digital power grids and smart meters, which are constantly being optimised by artificial intelligence, can lead to huge savings that would simply not be possible by purely human means.
As a new generation telecommunications company, delivering the next generation of networks, we are unrelenting in our commitment to driving this digitisation – not just within Europe, but globally.
Vodafone is the world's largest supplier of IoT services and we're pioneering new approaches to energy supply and delivery. Our smart meters and grids service improve efficiencies meeting the exact demands of customers at any given moment.
As the world responded to COVID-19, we collectively altered the way we interact with one another and our surroundings. So too did the Government. The European Commission has now asserted that all national recovery and resilience plans must focus strongly on both reforms and investments supporting the green transition. In fact, a minimum of 37% of the €672.5 billion recovery and resilience facility is earmarked to climate. By investing in digital infrastructures that drive efficiencies, reducing energy output and bringing people together when it's needed most, we can, collectively, tackle the issue of climate change. If a sustainable transformation of our economy is to succeed at the pace we need it to, digitisation is crucial. EU citizens recognise its potential. The time for Government and businesses to act is now.
Digital for Green


Nothing says comfort food like tacos and fried chicken. Milssam Tacorea brings another iteration of Korean-Mexican fusion to the city. This time in the heart of Yaletown at 1130 Mainland Street. The spot is cozy with a big fire pit up front, a selection of booth, regular table and communal table seats and a projector for hockey games.
Sister restaurant to Damso Korean Restaurant and Mr. Osmad Souffle Cheesecake in the West End, Milssam brings favorites from both, like the cheesecake that gives Uncle Tetsu's a run for their money, while creating new dishes for taco lovers.
My favorite taco of theirs, pictured in the middle above, is the cauliflower one. Taking inspiration from the classic Korean saucy noodle dish jjajangmyeon, the taco features warm and crunchy fried cauliflower, corn, avocado flavored with black bean mole sauce with pickled onions and greens. Unlike the usual chocolate mole, black bean mole has a salty richness that packs this taco with flavor.
The namesake taco, the Milssam Taco, was so popular at Damso that it sparked the creation of Milssam Tacorea itself. Beef belly, onion, lettuce and gochujang vin in a soft steam bun shell. The gochujang is mild with just enough of a sweet kick to linger delightfully on your lips.
Their seafood ceviche taco is a refreshing addition to the meal. Marinated shrimp and snapper, avocado, shallots and soy vin. The meat is tangy and sweet and the shell stayed crisp even though it was on the table for some time for pictures. The layering was well thought out to keep those conditions.
Milssam also has Korean-Mexican fusion drinks like a makuli and rum "majito" pictured above, Songria (soju and sangria) and Shikhae Horchata (horchata with malt, pictured below). Makuli, rum, mint, lime and soda are a fantastic combination. The idea of the soda threw me off when I read the menu but I highly recommend giving it a try.
This wouldn't be a taco place without Jarritos but in fusion cuisine fashion they've also imported the Korean 7-Up, Chilsung Cider.
Their KFC, Korean friend chicken is exactly what you would expect from a good Korean bar. Glad Yaletown finally has an option for this.
Seems a little confusing why a souffle cheesecake is on the menu at a Korean taco place but when you have a bite, you'll ask no questions and only be glad that it's there instead of halfway across town. I prefer it with the strawberry compote, but I have a sweet tooth. Light, fluffy, freshly made in 30 minutes. So, if you plan to have one, order it when you order your food. Just as good as Uncle Tetsu's without the lineup. I'm not sure this photo shows off the size of the cake, but I'll tell you that it can generously serve 4 after the fried chicken.


Day two began with dragging our bags down to the front desk, a small breakfast, picking up our bikes, and beginning our travels. We met some of our fellow travelers in the parking garage, but everyone seems very intent on staying with their groups and not really interested in too much inter socializing, which I appreciate. We started off with a trip through the park near the Praha train station, over the bridge, and on our way up the river.
Almost immediately upon crossing the river it felt like we were in a different land, it was amazing. We rode past a police dog training class and watched for a bit. The reward for the dogs attacking properly was getting to shake the dummy arm used to protect the officer's arm during the training. We ran into these dogs later at lunch time where I was almost mauled by one who still had his muzzle on. The police officers apparently were taking a short journey and just happened to be at the same spot we were having lunch at. One of the dogs was chained to the picnic bench by our bikes. The bench was not match as he lunged at me dragging the table behind him. I wasn't what I should do, I wasn't scared, but I was defiantly not comfortable. Finally the officer came over and signaled for the dog to stop and I was able to get my bike. I think this was quite amusing to the people at the restaurant.
Only a few times did we go further than we were supposed to on a trail, but it wasn't a very big mistake ever. We rode our bikes past palaces, through fields of corn, on the muddy small trail by the river, past little old men taking their small dogs for a Sunday bike ride in a basket on the back of the bike, and so many more amazing sights. We were able to carry our bikes up and down stairs, and even made a giant hike up to what we thought was an interesting museum, but turned out to be a butter making museum (we didn't go in).
At the end of the day we were exhausted, and rather damp from the semi constant rain. We were the last group in, and when we saw one of our fellow groups (from Austria) at dinner they asked if we had gotten lost. I told them no, we just stopped at places a lot, but was keenly aware that we were the youngest of the bunch and the slowest. Oh well I guess. Now to bed in preparation for another long day. Tomorrow we ride past a concentration camp, which should be very interesting/educational. The weather is supposed to be terrible, but I think we can manage.
It turned out to be a steep path to a town with a Butter Churning Museum.
Pot Bellied pigs along the bike path.


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Bringing Outside Artists to Light
They lived outside the arts world, but created works—now on view at Tufts—that are standing the test of time
Howard Finster, "Rhino," 1990, paint on cut board, paper on painted base, on view at the "Expressions Unbound" exhibition. Photo: Steve Briggs
By Rob Phelps
Provocatively posed and rendered in mud, Lady Liberty greets visitors near the entrance of Expressions Unbound: American Outsider Art from the Andrew and Linda Safran Collection, on display at the Tufts University Art Gallery through December 16. The earthen tones and curvaceous figure of Jimmy Lee Sudduth's Statue of Liberty lend a cheerfully brazen sensuality not typically associated with the national monument.
Another image of the iconic statue in the show, created by U.S. Marine Corps veteran and artist Benjamin Perkins, is more traditionally festooned, complete with stars on a blue background and patriotic phrases draping the statue's body in anthem-like visual rhythm.
"Both images prompt great questions about American iconography," said Dina Deitsch, director and chief curator for Tufts University Art Galleries. "How do symbols of our country manifest and move throughout different parts of the country and through people's visual imagination?"
Sudduth (1910-2007) lived in rural Alabama, where he worked as a farmhand. Entirely self-taught as an artist, he used mud, various grasses, berries, sugar, and molasses to create his works. "It was all there in the ground, every color of dirt and mud. I got twenty-three colors of dirt in my yard," he told art historian William Arnett, as recounted in Arnett's book Souls Grown Deep: African American Vernacular Art of the South.
"Sudduth's Statue of Liberty in mud can be construed however one sees it, which is what all great art does," Deitsch said. "It inspires you to see things in multiple viewpoints."
Born in 1904, Perkins had a career in the military and the government before becoming an ordained minister. He found his artistic calling in midlife after a divorce, when he moved from Virginia to his native Alabama and built his own church. While painting the structure red, white, and blue, he began adorning it with religious and patriotic images, and his work as an artist began.
In their own ways, Sudduth and Perkins exemplify outsider artists, both working "beyond the boundaries of accepted conventions and educational systems," according to the British scholar Roger Cardinal, who coined the term. Outsider artists invent their own means and methods to create their work with little or no knowledge of art history, technical training, or awareness of popular culture. Few have ever stepped into an art gallery or museum.
"Their expressions are fresh, literally unbound by artistic conventions that in some ways actually limit artistic expression within the discourses of the art world," said Deitsch. "Outsider artists don't work within these limits."
The exhibition includes all thirty-eight works from the Safran Collection of American outsider art, donated by Andrew Safran, A76, F77, and his wife, Linda, to Tufts. "This significant gift of works, largely by African-American artists, allows us to continue the project of opening the canon of art history, not only to a more inclusive global history, but also to a localized, national history that speaks to the divides in class, race, and educational opportunity that our field has unwittingly reinforced," Deitsch said.
The show and the donation of these works to the university's collection also provide opportunities for scholarship, from studying art and training artists to American history, sociology, and many other disciplines.
The nineteen artists in the show created their works from self-devised materials, ranging from Sudduth's mud and dirt to repurposed debris such as scrap wood and castoff corrugated tin roof panels, in addition to traditional tools like pencils, paints, and canvases. Many, like Perkins, came to their calling in midlife or later, expressing an accumulation of often bottled-up emotions and experiences.
Sanford Darling, who worked as a Hollywood stuntman, a commercial fisherman, and for twenty-five years as an engineer at a petroleum company, traveled in Europe and Asia in the early 1960s following his wife's death. Eager to document impressions and memories of his travels, he painted practically every surface—walls, beams, appliances, stairs—of his house, which became known as the "House of 1,000 Paintings." When he died in 1973, his house was disassembled, and his art works sold. The Tufts show includes a tropical scene on plywood from the exterior entrance.
After raising eleven children and enduring the abuses of an alcoholic husband, Bessie Harvey of Tennessee began creating sculptures, which she called her "dolls." She told Arnett that "they were my freedom from this world, that I could go into them and I could talk to God, and that the spirit would release me from all of the hurt, and I could hear him speak and talk to me." Her mixed-media sculpture The Spirit of Love, resembling an African totem, reflects her focus on African spirituality.
Howard Finster, who became a Baptist preacher at age sixteen in rural Alabama, started painting what he called his "sacred art" at age sixty. Similar to Sanford Darling's experience, he populated two and a half acres of his backyard with his sculpted constructions, calling the space "Paradise Garden." His whimsical paint on cut board sculptures Giraffe and Rhino are decorated with colorful motifs of humans in nature and are inscribed with Biblical messages stressing the distinction between humans and animals.
"These instructive and formative works broadly reflect the artists' own beliefs, fantasies and life experiences," writes Evie T. Joselow in the show's catalog. "Through their art, we, the viewers, bear witness to their keen perceptions from different environments and geographic locations. We also get a glimpse of their personal demons, demonstrating their perseverance in all kinds of situations to find unique, creative ways and means of expression."
Despite working almost entirely outside the established art world, many of the artists in Expressions Unbound have nonetheless made their mark on the art establishment. After two of his pieces appeared in the 1984 Venice Biennal, Finster was invited to design album covers for the pop bands R.E.M. and Talking Heads. In 1995, Harvey became the first black woman to present her work in the Whitney Biennial. Other artists in the show have had their work shown at the Corcoran Gallery in Washington, D.C., in New York's Museum of Modern Art and American Folk Art Museum and in documentary films.
"The idea," Deitsch said, "is that great art can come from anywhere."
The Tufts Art Gallery is located at 40 Talbot Avenue on the university's Medford/Somerville campus. Gallery hours are Tuesday through Sunday, 11 a.m. to 5 p.m., and Thursday until 8 p.m. For more information, visit http://artgallery.tufts.edu.
The University Art Galleries host the panel discussion "Does an Education Make an Artist?" on November 8, in which art historians specializing in both Outsider Art and major movements within the art history canon consider how artists have been trained and identified throughout history.
Rob Phelps is a freelance writer based in Quincy, Massachusetts.
American Outsider Art Comes to Tufts
Visions of the Future Present
We Are Not Your Subjects Now
Bringing Landmarks Like Notre-Dame Back to Life July 16, 2019

redpajama_set_name: RedPajamaCommonCrawl


Hello and welcome to My Gallery. It is my great pleasure to offer you my gathering of designs and needlework products. I know you will be able to navigate this Gallery quite smoothly, and find some wonderful treasures to thread your needles. Please do not hesitate to contact us if you have any concerns. I wish you endless hours of stitching pleasure.
This is a fun, quick, and simple miniature sampler, stitched in ONE Tudor Overdyed Silk, 12-ply..
I've created a series of bell pulls for your kitchen...... of fruits and flowers......to follow..
The first in the SOB series."SILKS ON BOARD"..... another new series of silk gauze creations. A wond..
Such a beautiful design and creatively constructed into a fine caddy for your favorite stitchin..


WAKADO: THREE WOMEN IN A SAUNA !
THREE WOMEN IN A SAUNA !
THREE WOMEN, TWO YOUNGER, AND ONE SENIOR CITIZEN, WERE SITTING IN A SAUNA.
SUDDENLY THERE WAS A BEEPING SOUND. ONE OF THE YOUNG WOMEN PRESSED HER FOREARM AND THE BEEP STOPPED.
THE OTHERS LOOKED AT HER QUESTIONINGLY. 'THAT WAS MY PAGER,' SHE SAID.
I HAVE A MICROCHIP UNDER THE SKIN OF MY ARM.
THE OLDER WOMAN FELT VERY LOW-TECH. NOT TO BE OUT DONE, SHE DECIDED SHE HAD TO DO SOMETHING JUST AS IMPRESSIVE. SHE STEPPED OUT OF THE SAUNA AND WENT TO THE BATHROOM.
THE OLDER WOMAN FINALLY SAID.........WELL, WILL YOU LOOK AT THAT... I'M GETTING A FAX!!


If you love peppermint bark candy, then you'll love these Candy Cane Brownies with chopped Hershey's kisses on top because they taste just like peppermint bark brownies!
The Holiday Food Bloggers are back, this time celebrating all things Christmas! How could we not when it's one of the biggest holidays in the world? When I think of Christmas desserts, candy canes, gingerbread, and eggnog come to mind. It's also that time of year to buy specialty Hershey's Kisses, like mint truffle, cherry cordial, and candy cane. I'm obsessed with seasonal candy for the limited edition flavors. Probably one of the best marketing tactics for companies, right? For today's recipe, I made Candy Cane Kiss Brownies, which are chocolaty brownies topped with a layer similar to peppermint bark. If you love candy canes and peppermint, you're not going to want to pass these up.
I saw a picture of brownies similar to these with crushed peppermints on top. However, I'm not a big fan of hard candy, so I used candy cane kisses instead. Much easier on my teeth without losing the candy cane flavor. It'd be an understatement to say I couldn't stop eating these brownies!
The brownie base recipe is from my favorite cookbook author Dorie Greenspan. I love that woman and can't wait to someday meet her. Seriously, if you are a baker, you need to buy Baking: From My Home to Yours for your collection (Amazon affiliate link). I used to be a part of Tuesdays With Dorie way back when, so I've baked a good bit of recipes from that book (since I couldn't keep up with the group).
The peppermint from the kisses is pretty strong, so I didn't add any peppermint extract to the brownies. I know several people who don't like peppermint (more for me), so feel free to substitute plain kisses.
Preheat oven to 350F. Line an 8x8-inch pan with parchment paper.
In a heatproof bowl over a saucepan of simmering water, melt the butter, bittersweet chocolate, and unsweetened chocolate. Remove the bowl from the heat and whisk in sugar (it'll be grainy). Beat in the eggs, one at a time, then beat in the vanilla. Add the espresso powder, salt, and flour.
Scrape into the prepared pan and bake 30-33 minutes or until a toothpick inserted in the center comes out clean. Cool completely.
In another heatproof bowl over a saucepan of simmering water, melt the white chocolate and shortening (white chocolate is very delicate to melt, so do not get any moisture in the chocolate or it'll seize up). Spread over the cooled brownies. Top with chopped kisses. Let the chocolate harden before cutting.
If you're looking for more Christmas recipes, check out the rest of the Holiday Food Bloggers! All you need to do is click on the thumbnail on the graphic above. Yes, each thumbnail is clickable and goes to each participating blog. Let me know if it's not working.
I think chocolate and peppermint were created to go together…what a fun idea!
i love peppermint bark. it sounds even better like this!
Oh, man, my family would inhale these luscious brownies! So yummy and festive!
I have to admit, I'm a sucker for white chocolate anything!
Oh my gosh… these are genius! I love peppermint-flavoured brownies, but those crunchy bits of candy cane always get stuck in my teeth and ruin the experience. Using candy cane kisses sounds perfect.
I have to admit I'm not a fan of peppermint (it doesn't do well in my system). Such a share because they are so pretty with the red and white kisses. I do see the possibilities with using different flavored kisses with these brownies. The white chocolate bark top would make any of the kisses stand out.
I like the peppermint, so you need to share with me! I spotted the candy cane and cherry cordial kisses at the market today and was so tempted to buy them, but I don't need to bake anymore stuff than I already am!
Those kisses are my favorite! And pairing them with brownies was a genius move!
We make peppermint bark every year because it's a Christmas thing. You've got to. I love that you used the peppermint Kisses on the brownies though, Carla. Much easier on the teeth!
Two great flavors together, these look pretty and YUMMY!


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Daniel Lloyd Little
Unheard, Unseen...
HomeUnheard, Unseen...Conflict in the NorthSubmarine Lost!Purchase PageAuthor GalleryDaniel's RecomendationsDisclaimersContactAnd finally...
Unheard, Unseen... by Daniel Lloyd Little is a riveting adventure thriller involving the Royal Canadian Navy submarine HMCS Corner Brook and US Navy SEALs in a dangerous covert operation that could save the world from future nuclear annihilation.
It is 2009 and North Korea has purchased four outdated submarine launched ballistic missile boosters from a French company tasked with destroying the rockets. Because they are not a weapon in and of themselves, international law governing the sale of strategic weapons can not prevent the acquisition. Both the US and Canadian governments however, know too well what the unstable North Korean leader intends for the rocket boosters in light of his failure to successfully launch a missile of his own.
When a carefully planned covert mission by US Navy SEALs to intercept the rockets unravels due to mechanical problems with the US Navy ultra quiet spy submarine USS Jimmy Carter, a replacement must be found and quickly! The surprising solution comes in the form of a Canadian submarine.
Commander Michael Simpson, nearing the end of his career with the Royal Canadian navy has few doubts concerning the capabilities of HMCS Corner Brook, and he knows that his crew is the best out there. The only answer to his orders is the same one that has echoed throughout the history of one of the world's best trained navies; 'ready, aye ready'. In Unheard Unseen, the author takes the reader on a thrilling adventure as the officers and crew of HMCS Corner Brook and a team of US Navy SEALs undertake a mission fraught with danger!
Here's a sneak preview...
Half an hour later he strolled into HMCS Corner Brook's junior mess to find a few of his crew spellbound as the SEALs checked over their equipment and weapons.
"Attention on deck!" Boris jumped up when he spotted Commander Simpson.
"We don't do that here lieutenant. People keep hitting their heads on things when they stand up too fast." Mike grinned and extended his hand to Boris.
"Thank you sir. Just being respectful first time meeting you, sir," Boris returned the smile and introduced Mike to his men.
"Good to have you aboard gentlemen," Mike announced after greeting each of the men in turn. "I hope you find the accommodations acceptable. If the crew become a bit too nosey, just let me or one of the other officers know and we'll shoo them out."
"No, they're just fine, sir," said Boris. "We don't mind educating them about our equipment and getting to know each other, and besides, when we're out there pounding on the hull to be let back in, we want to think of the whole boat being interested in picking us up," he finished with a grin.
"Okay lieutenant. But don't go recruiting any of them. I understand you met one of my petty officers and brought him back to us."
"Chief Cole? Yes sir. Good man…or snoop as we call him," Boris laughed, then narrowed his eyes. "Of course the Chief of Naval Operations might not see it that way," he added.
"Excuse me?" Mike asked, not sure if the lieutenant was joking or not.
"Oh, you'll have to ask Mr. Cole about that one, sir. I'm not even sure that it's not 'need to know'." The whole team laughed this time and Mike made a note to have a chat with his chief petty officer and find out what he had done. The CNO? Maybe it's time for young Mr. Cole to join the surface fleet, thought Mike, afraid to even imagine what might have happened.
To purchase Unheard, Unseen..., click here.
Unlikely? Click here to view an article from the Halifax Chronicle Herald.
Daniel donates copies of his novel to the Pointe-au-Pere Museum where the submarine
HMCS Onondaga is on display. Pictured here are Daniel, Anne-Marie Bourassa and Moe Allard.
Click the image above to visit the site.
In Unheard, Unseen, Daniel Little shows the Royal Canadian Navy at its best as the crew of the submarine HMCS Corner Brook is dispatched
to deliver a squad of US Navy SEALs on a mission that will see both of them forced to deal not only with the inherent suspense
of a special operation, but also with a deadly game of cat and mouse.
In the cutaway above, you can see the layout of the Victoria Class SSK's.
(Image used with permission of Canadian Government)
Visit my HalifaxAuthor Blog here. The Author can be reached at info@daniellittle.com
Unheard, Unseen... Reviews:
Dave Shirlaw
Editor of SeaWaves Magazine:
Kudos to Mr Little for having the ambition and drive to write a novel to this historically underserved market. There is enough potential visible to recommend this book as long as one remembers that this is a first effort. I look forward to seeing marked improvement in his second novel which I look forward to reading. Read The full review here.
Dwayne Hill
Just finished reading Unheard Unseen…good novel. Nice to see the canucks doing it right.
View Dwayne Hill's INCREDIBLE collection of radio control warships and submarines by clicking here.
Donald Courcy
While waiting for the winds to die down, I visited the Empress of Ireland display and saw your book Unheard, Unseen on the shelf. So I
bought it and read it the next day. What a book ! I really enjoyed it. Visit Donald's website by clicking here.
Chris Corso (Screen Writer)
Morning Mr. Little...I just finished Unheard, Unseen. Really a good read--dramatic and educational. A nice blend of fiction and the possible reality of...Your reverence for all things Navy shines through. A game of Chicken at seven-hundred feet ala "Rebel without a cause" that was fun.
Murray Austin
I spent 22 years in the Navy on the surface. I actually enjoyed the whole story although I don't have any background on the subs some of your general orders and such have more of an American slant than Canadian (A lot of our bridge orders and replies are throwbacks from the Royal Navy). I found it entirely believable overall. Just wanted to drop a line to say hi and look forward to rading your next endeavor.
Halifax, Nova Scotia, Canada - info@daniellittle.com - Support Your Public Library


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Small Produce Farm Environments Can Harbor Diverse Listeria monocytogenes and Listeria spp. Populations
ALEXANDRA BELIAS;
ALEXANDRA BELIAS
1Department of Food Science, Cornell University, 354 Stocking Hall, Ithaca, New York 14853
LAURA K. STRAWN;
LAURA K. STRAWN
2Department of Food Science and Technology, Eastern Shore Agricultural Research and Extension Center, Virginia Tech, 33446 Research Drive, Painter, Virginia 23420
MARTIN WIEDMANN;
MARTIN WIEDMANN
DANIEL WELLER
DANIEL WELLER *
(ORCID: https://orcid.org/0000-0002-6308-8025 [D.W.])
3Department of Statistics and Computational Biology, University of Rochester, Rochester, New York 14627
4Department of Environmental and Forestry Biology, SUNY College of Environmental Science and Forestry, Syracuse, New York, 13210, USA
*Author for correspondence. Present address: Department of Biostatistics and Computational Biology, University of Rochester, Rochester, NY 14620, USA. E-mail: wellerd2@gmail.com.
J Food Prot (2021) 84 (1): 113–121.
https://doi.org/10.4315/JFP-20-179
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ALEXANDRA BELIAS, LAURA K. STRAWN, MARTIN WIEDMANN, DANIEL WELLER; Small Produce Farm Environments Can Harbor Diverse Listeria monocytogenes and Listeria spp. Populations. J Food Prot 1 January 2021; 84 (1): 113–121. doi: https://doi.org/10.4315/JFP-20-179
A comprehensive understanding of foodborne pathogen diversity in preharvest environments is necessary to effectively track pathogens on farms and identify sources of produce contamination. As such, this study aimed to characterize Listeria diversity in wildlife feces and agricultural water collected from a New York state produce farm over a growing season. Water samples were collected from a pond (n = 80) and a stream (n = 52). Fecal samples (n = 77) were opportunistically collected from areas <5 m from the water sources; all samples were collected from a <0.5-km2 area. Overall, 86 (41%) and 50 (24%) of 209 samples were positive for Listeria monocytogenes and Listeria spp. (excluding L. monocytogenes), respectively. For each positive sample, one L. monocytogenes or Listeria spp. isolate was speciated by sequencing the sigB gene, thereby allowing for additional characterization based on the sigB allelic type. The 86 L. monocytogenes and 50 Listeria spp. isolates represented 8 and 23 different allelic types, respectively. A subset of L. monocytogenes isolates (n = 44) from pond water and pond-adjacent feces (representing an ∼5,000-m2 area) were further characterized by pulsed-field gel electrophoresis (PFGE); these 44 isolates represented 22 PFGE types, which is indicative of considerable diversity at a small spatial scale. Ten PFGE types were isolated more than once, suggesting persistence or reintroduction of PFGE types in this area. Given the small spatial scale, the prevalence of L. monocytogenes and Listeria spp., as well as the considerable diversity among isolates, suggests traceback investigations may be challenging. For example, traceback of finished product or processing facility contamination with specific subtypes to preharvest sources may require collection of large sample sets and characterization of a considerable number of isolates. Our data also support the adage "absence of evidence does not equal evidence of absence" as applies to L. monocytogenes traceback efforts at the preharvest level.
There is considerable Listeria diversity in the farm environment investigated.
Listeria subtypes were reintroduced or persisted over the growing season.
Four L. monocytogenes PFGE types were shared between feces and pond samples.
Agricultural water, Feces, Listeria, Pulsed-field gel electrophoresis, sigB
It is estimated that Listeria monocytogenes is responsible for ∼1,600 illnesses, 1,455 hospitalizations, and 255 deaths annually in the United States (31), and several high-profile outbreaks and recalls have been traced to L. monocytogenes contamination of fresh produce (8, 24, 44). For example, a 2011 outbreak was traced back to L. monocytogenes contamination of cantaloupes that resulted in 147 illnesses and 33 deaths (2). In addition, there have been >30 recalls since 2016 due to L. monocytogenes contamination of produce and ready-to-eat produce products (36). Although the majority of produce outbreaks and recalls due to L. monocytogenes contamination are traced back to the processing environment, L. monocytogenes can be introduced, or reintroduced, in the processing environment from the preharvest environment (e.g., on produce, in soil, on harvesting equipment, or on workers shoes (32)) or could be introduced onto product in the field and remain on product until consumption.
After detection of a Listeria-positive sample during routine monitoring or an outbreak investigation, processors or investigators often conduct extensive sampling to identify potential contamination sources (i.e., traceback analysis). Identifying how a Listeria isolate was introduced onto product or into the processing environment is a critical step in performing a root cause analysis and establishing effective corrective actions (to control potential contamination pathways). To determine whether an isolate originated in the preharvest environment, investigators and operators must collect environmental samples from farms that supply the product. However, the ubiquity of Listeria in the environment (1, 7, 11, 12) may make it challenging to prove a causal relationship between Listeria isolated from the preharvest environment and Listeria isolated from postharvest environments and/or product. Previous studies of preharvest environments isolated Listeria from soil (3, 25, 34, 35, 39, 40), wildlife feces (5, 19, 37, 40), agricultural water (1, 6, 17, 20, 33), and biological soil amendments (25). As such, a more comprehensive understanding of the prevalence, diversity, and distribution of Listeria on produce farms is needed to better understand on-farm sources of Listeria, understand transfer of Listeria between sample types on produce farms, and facilitate traceback analysis. Although some studies have investigated the diversity of Listeria in preharvest produce environments (3, 19, 20, 33–35), these studies have examined diversity over large geographic areas. For example, Chapin et al. (3) and Strawn et al. (34) investigated Listeria diversity on five New York state farms located up to 205 km from each other, whereas Stea et al. (33) examined Listeria diversity within an agricultural watershed in Nova Scotia, Canada, with a drainage area of 135 km2. Because there are limited data on Listeria diversity at small spatial scales (e.g., within individual farms), we aimed, in this study, to address this knowledge gap by examining the diversity of Listeria on a single produce farm.
Agricultural water and wildlife intrusion in produce fields have been identified as key sources and pathways for dispersal of Listeria contamination in preharvest environments (14, 26, 37, 39). Multiple studies have isolated L. monocytogenes from agricultural water and wildlife feces collected in produce-growing environments (6, 7, 19, 25, 34, 39). For example, Weller et al. (39) isolated L. monocytogenes from 45% (n = 11) of wildlife feces collected on 10 produce farms in New York state. Falardeau et al. (7) isolated L. monocytogenes from 10% (n = 223) of agricultural water samples collected from two watersheds in British Colombia, Canada. Furthermore, Cooley et al. (6) isolated L. monocytogenes from 43% (n = 1,405) of water samples collected from California's Central Coast agricultural region. As such, the primary objective of the study reported here was to characterize and compare the L. monocytogenes and Listeria spp., excluding L. monocytogenes, diversity in agricultural water and wildlife feces collected from a single produce farm in New York state.
Study design and sample collection
The study was conducted between May and July 2014 on a produce farm in the Finger Lakes region of New York state. Two separate irrigation water sources are located on the farm: (i) a pond and (ii) a stream. The water sources are within a rectangular 0.5-km2 (124-acre) area; the sampling sites in the pond and stream are ∼775 m (2,546 ft) apart.
Agricultural water and fecal samples were collected on 26 days. Fifty-two and 80 water samples were collected from the stream and pond, respectively; the Listeria prevalence and number of sigB allelic types (ATs) for the 52 stream samples and the 13 fecal samples collected near the stream were previously reported by Weller et al. (40). However, to better understand the Listeria diversity on the studied farm, these data were also included here. All water samples were collected as described in Weller et al. (40). Briefly, 250 mL of water was collected into a sampling cup (Nalgene, Rochester, NY) by using a sampling pole and was stored on ice until processing. Fecal samples (n = 77) located within 5 m of the pond (n = 64) or stream (n = 13) were collected opportunistically, as described previously (40); data for the fecal samples collected near the pond were not previously reported. A 5-m buffer around the pond and stream was used for practical reasons. All samples were stored at 4°C and processed within 3 h of collection.
Listeria enrichment and isolation
All samples were processed as described previously (40). Briefly, 10 g of the fecal samples was aliquoted and transferred to separate Whirl-Pak bags (Nasco, Fort Atkinson, WI). The water samples were each passed through separate 0.45-μm-pore-size filters, and each filter was transferred to a separate Whirl-Pak bag. Each sample was enriched in 90 mL of buffered Listeria enrichment broth (BD, Franklin Lakes, NJ) at 30°C. After 4 h, Listeria selective enrichment supplement (Oxoid, Cambridge, UK) was added to each sample. Each sample was incubated for a total of 48 h at 30°C. After 24 and 48 h of incubation, the enrichments were substreaked onto modified Oxford agar (MOX; BD) and L. monocytogenes plating medium (LMPM; Biosynth International, Itasca, IL). MOX plates were incubated at 30°C and LMPM plates were incubated at 35°C for 48 h. After incubation, characteristic Listeria colonies on 24- and 48-h MOX plates (i.e., gray, donut-shaped colonies) were streaked onto LMPM for isolation and incubated at 35°C for 48 h. One colony per presumptive Listeria-positive LMPM plate (up to four colonies total: one from the 24-h LMPM plate, one from the 48-h LMPM plate, one from the LMPM plate substreaked from the 24-h MOX plate, and one from the LMPM plate substreaked from the 48-h MOX plate) was selected, substreaked onto brain heart infusion agar (BD), and incubated at 37°C for 24 h. If blue colonies were present (characteristic of L. monocytogenes, L. ivanovii, hemolytic L. innocua due to phospholipase C action; (28)) on a given plate, they were preferentially selected to be substreaked on brain heart infusion agar. If a blue colony was not present, a round white colony was selected to be substreaked on brain heart infusion agar; this colony phenotype is characteristic of Listeria spp. other than L. monocytogenes, L. ivanovii, and hemolytic L. innocua. If no round white or blue colonies were present on any of the LMPM plates, the sample was considered negative for Listeria. If a round white or blue colony was present, sigB PCR was performed as a confirmation step and speciation of Listeria, as described below.
sigB sequencing and allelic typing
All isolates per presumptive-positive Listeria sample (i.e., up to four isolates per sample) were confirmed as Listeria via PCR amplification of sigB, as described by Nightingale et al. (22); all samples with presumptive-positive Listeria isolates in this study were confirmed as positive via sigB PCR. Sequencing of the sigB gene was then performed on one isolate per positive sample for further characterization: if the colony characterized was identified as L. monocytogenes, the sample was considered L. monocytogenes positive; if the colony characterized was identified as Listeria spp. other than L. monocytogenes, the sample was considered Listeria spp. positive. Although only subtyping one isolate per sample may lead to an underestimation of the true diversity present in the sample types investigated here, this approach allowed us to get a preliminary estimate of the diversity present. There were 22 samples with both blue and white colonies present on LMPM plates (of 209 total samples); for 21 of these samples, the blue colony was selected for characterization (2 of 3 fecal samples, 8 of 8 stream samples, and 11 of 11 pond samples). Because only L. monocytogenes was selected for pulsed-field gel electrophoresis (PFGE), preferentially selecting these blue colonies provided us with the larger sample size for further subtyping. A BLAST search was performed against an internal database (http://www.foodmicrobetracker.com/login/intro.aspx) to determine its sigB AT of each sequence. All isolates were archived as 15% glycerol stocks and stored at −80°C. A sigB AT phylogeny was constructed based on the 660-bp nucleotide sequence of the sigB gene by using the maximum likelihood method in MEGA7 version 7.0.26; the nucleotide substitution Tamura three-parameter model was used and 1,000 bootstraps were performed.
PFGE analysis
All confirmed L. monocytogenes isolates from the pond and the feces collected near the pond (n = 44) were characterized by PFGE. Only isolates from pond water and feces were characterized by PFGE as a subset analysis to investigate diversity over a small spatial scale (i.e., the ∼5,000 m2 around the pond). PFGE was performed using AscI and ApaI restriction enzymes, according to the Centers for Disease Control and Prevention PulseNet protocol (10).
All statistical analyses were performed using R version 3.5.1 (27). Chi-square tests were used to determine whether L. monocytogenes and Listeria spp. (excluding L. monocytogenes) prevalence differed between sample types. The Bonferroni correction was used to account for the three comparisons made for Listeria spp. (excluding L. monocytogenes) and for the three comparisons made for L. monocytogenes (pond versus fecal, pond versus stream, fecal versus stream; α = 0.05/3 = 0.0167). Although separate data for the feces collected near the pond and feces collected near the stream are provided in this study, these sample types were combined into a single fecal sample category for analysis due to the small number of fecal samples collected near the stream (n = 13 of 77 total fecal samples). To determine the number of unique ATs and PFGE types, AT richness and PFGE type richness were estimated using the breakaway package version 4.6.7 (43), which calculates the transformed species richness estimation with 95% confidence intervals (CIs) as outlined in Rocchetti et al. (29) and Willis and Bunge (42). To estimate AT and PFGE type diversity for each sample type, Simpson's index of diversity was calculated using the vegan package version 2.5-3 (23); 95% CIs were calculated for the Simpson's index of diversity, according to the methods described by Hunter and Gaston (13). The effective number of subtypes was also calculated to communicate Simpson's index of diversity in a more intuitive form (i.e., the estimated number of subtypes present in a sample type given a certain value of Simpson's index of diversity (15)).
If a reasonably large number of samples are collected from the preharvest environment in New York state and tested for Listeria ("seek"), it is likely Listeria will be "found." The overall prevalence of Listeria in the study reported here was 136 (65%) of 209; the prevalence of Listeria spp. (excluding L. monocytogenes) was 50 (24%) of 209, and the prevalence of L. monocytogenes was 86 (41%) of 209 (Table 1). Although the Listeria spp. (excluding L. monocytogenes) prevalence in the pond (25 of 80, 31%) and stream (14 of 52, 27%) samples was not significantly different (P = 0.735), the L. monocytogenes prevalence in the pond (31 of 80, 39%) and stream (33 of 52, 63%) samples was significantly different, even after the Bonferroni correction (P = 0.009; Table 1). The frequency of Listeria isolation from agricultural water has been reported in several previous studies conducted in New York state (3, 34, 35, 38, 39) and other produce-growing regions (1, 6, 7, 18, 33). In previous New York state studies, the L. monocytogenes prevalence ranged between 9% (39) and 59% (35) in pond samples, and between 10% (38) and 50% (39) in stream samples. As such, the frequency of L. monocytogenes isolation from pond samples in the current study falls within the range reported in the literature, whereas the frequency of L. monocytogenes isolation from the stream samples in the current study is higher than in previous studies. The higher L. monocytogenes prevalence in the stream sampled here, compared with those sampled in other studies, may be due to presence of two dairy operations immediately upstream of the sampling sites along the stream. In addition, run-off from the dairy operations could explain the significantly higher L. monocytogenes prevalence in the stream compared with the pond. Previous studies have indicated upstream livestock operations are associated with an increased prevalence of L. monocytogenes in stream water (7, 19, 41). Furthermore, another study suggested cattle may increase the prevalence and may distribute L. monocytogenes in the farm environment (22). Of note, the current study only tested 250 mL of water per sample, whereas other studies collected upward of 1 L per sample. As such, the prevalence reported for the pond and stream investigated in the current study may be an underestimation of the true prevalence. Regardless, although the current study supports agricultural water as a source of L. monocytogenes in the New York state preharvest produce environment, our findings also suggest future studies on agricultural water need to consider the impact of spatiotemporal factors (e.g., upstream land use) on downstream microbial water quality.
Frequency and prevalence of Listeria spp. and L. monocytogenes in fecal and agricultural water samples
The prevalence of Listeria spp. (excluding L. monocytogenes) and L. monocytogenes in the fecal samples collected here were 11 (14%) and 22 (29%) of 77, respectively (Table 1). Based on fecal shape and consistency, the majority of the fecal samples appeared to be produced by Canada geese (Branta canadenesis; n = 71). This is logical given the large flock of Canada geese that nested in the wetlands adjacent to the pond throughout the study. The remainder of the feces were produced by canids (n = 5) and an unknown animal (n = 1). The Listeria spp. (excluding L. monocytogenes) prevalence was 9 (13%) of 71 in the goose and 2 (40%) of 5 in the canid fecal samples. The L. monocytogenes prevalence was 20 (28%) of 71 in the goose and 2 (40%) of 5 in the canid fecal samples. Neither Listeria spp. (excluding L. monocytogenes) nor L. monocytogenes was isolated from the one unidentified fecal sample. Although multiple studies have isolated Listeria from wildlife feces (5, 19, 25, 34, 37, 39, 40), relatively few studies have examined the prevalence of L. monocytogenes in Canada goose feces specifically (5, 19). The L. monocytogenes prevalence in the goose fecal samples collected as part of the current study was greater than that of previous studies (5, 19). For example, a study conducted in Ontario, Canada, did not detect L. monocytogenes in any of the 18 goose fecal samples collected (19), whereas Converse et al. (5) isolated Listeria spp. from 47 (10%) of 495 of goose fecal samples. Although, the prevalence of L. monocytogenes in wildlife fecal samples varied between studies, our findings support previous studies' findings (33, 36, 37) that wildlife can act as source of L. monocytogenes in preharvest environments.
Absence of evidence does not equal evidence of absence
The composition of the Listeria population in the agricultural water and fecal samples was characterized using sigB allelic typing. Allelic typing was used to speciate the isolates and provide a preliminary estimate of diversity. In total, 136 isolates were selected for sigB allelic typing (i.e., one isolate per Listeria-positive sample); L. monocytogenes isolates were preferentially selected for subtyping based on colony morphology (blue colonies on LMPM). Of the 136 isolates, 20, 24, 6, and 86 isolates were identified as L. innocua, L. seeligeri, L. welshimeri, and L. monocytogenes, respectively. We identified 15 sigB ATs among the 50 Listeria spp. (excluding L. monocytogenes) isolates (∼1 unique AT per 3 isolates) and 8 ATs among the 86 L. monocytogenes isolates (∼1 unique AT per 11 isolates; Tables 2 and 3; Supplemental Table S1). In addition, we estimated two diversity parameters, richness and the Simpson's index, for Listeria spp. (excluding L. monocytogenes; Table 2) and L. monocytogenes (Table 3) in the pond, stream, and fecal samples. For example, the estimated L. monocytogenes AT richness for the pond water, stream water, and fecal samples was 8 (95% CI = 7 to 15), 9 (95% CI = 6 to 73), and 5 (95% CI = 5 to 8) ATs, respectively, indicating the presence of a considerable diversity on the farm investigated. However, it should be noted that this may be an underestimation of the true diversity present in these sample types on this produce farm, because only one isolate per sample type was characterized by allelic typing. Even with this limitation, our data indicate considerable Listeria diversity in the small area of the produce farm investigated. The proportion of ATs identified relative to the total number of isolates subtyped in the current study was similar to the proportion reported by previous studies conducted on New York state produce farms (3, 34). For example, applying the same analysis as in the current study, Chapin et al. (3) identified 50 sigB ATs among 186 Listeria spp. (excluding L. monocytogenes) isolates (∼1 AT per 4 isolates) and Strawn et al. (34) identified 12 sigB ATs among 107 L. monocytogenes isolates (∼1 AT per 8 isolates); both studies isolated Listeria from soil, drag swabs, water, and fecal samples collected from the five produce farms across New York state. Because all samples collected in the study reported here were from a <0.5-km2 area, compared with past studies that sampled larger areas (between 33 and 205 km apart), the similarity in the ratio of unique ATs relative to the total number of isolates subtyped in this and these previous studies (3, 34) suggests Listeria richness is similar across spatial scales. However, additional research is required to confirm this finding, especially in other geographical areas.
Diversity of Listeria spp. sigB allelic types (ATs) in agricultural water and fecal samples
Diversity of Listeria monocytogenes sigB allelic types and pulsotypes in agricultural water and fecal samples
Furthermore, multiple Listeria (including L. monocytogenes) sigB ATs were isolated from a single sample type on a given day on multiple occasions. In the pond, stream, and fecal samples, there was an average of 1.9, 1.5, and 1.7 Listeria (including L. monocytogenes) sigB ATs isolated per sampling day, respectively. There was also a maximum of three, two, and four Listeria (including L. monocytogenes) sigB ATs isolated per sampling day from the pond, stream, and fecal samples, respectively, indicating considerable Listeria diversity in the area sampled even on a given day. Six of the eight L. monocytogenes sigB ATs were isolated from at least one sample type on multiple dates (see Fig. 1, showing the isolation of L. monocytogenes sigB ATs over time; equivalent data for Listeria spp. [excluding L.monocytogenes] are shown in Supplemental Fig. S1). Notably, AT 57 was isolated from all three sample types (i.e., pond water, stream water, and feces) on multiple occasions and was isolated from at least one sample type on 23 of the 26 sampling dates. AT 57 represents 13 PFGE types (Fig. 2), consistent with previous reports that PFGE is considerably more discriminatory than sigB allelic typing (4). The high prevalence of a single AT (i.e., AT 57) makes it likely that less prevalent ATs are not isolated; for example, AT 57 may outcompete other ATs. In addition, the considerable AT diversity observed also makes it likely that samplings that do not include large sample numbers will not detect and isolate some Listeria sigB ATs. Hence, traceback investigations could mistakenly conclude that a given Listeria subtype is not present in a given preharvest environment, particularly in environments similar to the one investigated here.
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Date of L. monocytogenes sigB AT isolation from water and fecal samples. In total, 31, 33, and 22 isolates from pond, stream, and fecal samples were allelic typed, respectively. The colored squares indicate at least one isolate of the given AT was isolated on the given date from pond (blue), stream (yellow), or fecal (green) samples; dates when a sample was collected, but the specified AT was not isolated are represented by dark gray square, and dates when a sample was collected but was negative for Listeria are represented by light gray square. At each sampling event, three pond and three stream samples were collected. Fecal samples were collected opportunistically (i.e., when feces were present). If no fecal samples were collected, the square is white. The dates of Listeria spp. (excluding L. monocytogenes) sigB AT isolation is shown in Figure S1.
Date of L. monocytogenes PFGE type isolation from pond and fecal samples collected near the pond. In total, PFGE was performed on 31 and 13 isolates from pond and fecal samples, respectively. The colored squares indicate at least one isolate of the given PFGE type was isolated on the given date from pond (blue) or fecal (green) samples; dates when a sample was collected, but the specified PFGE type was not isolated are represented by dark gray squares, and dates when a sample was collected but was negative for L. monocytogenes are represented by light gray squares. At each sampling event, three pond samples were collected. Fecal samples were collected opportunistically (i.e., when feces were present). If no fecal samples were collected, the square is white.
In addition, a sigB AT phylogeny was constructed to investigate whether there was any relationship between sigB AT isolation and sample type (Fig. 3). The phylogeny suggests that neither specific sigB ATs nor specific sigB clades are associated with a given sample type. A Fisher's exact test also found no significant relationship between sigB AT and sample type (P = 0.30). Although our data could also be interpreted as suggesting that none of the ATs isolated in this study have an enhanced fitness in the different habitats sampled here, studies with larger data sets are needed to further probe niche adaptation of different Listeria species and clades.
Maximum likelihood phylogeny of Listeria isolated from pond, stream, and fecal samples from a single produce farm in New York state based on a 660-bp fragment of the sigB gene. Values on the branches are bootstrap values from 1,000 replicate trees. The scale displayed below the tree represents the genetic distance shown in the branch length.
In addition to characterization by sigB AT, we performed PFGE on the L. monocytogenes isolates from the pond (n = 31) and feces collected near the pond (n = 13) to better understand the diversity of the isolates at a very small spatial scale (∼5,000 m2). We identified 19 PFGE types among the 31 L. monocytogenes isolates obtained from pond water samples (∼1 PFGE type per 2 isolates) and 7 PFGE types among the 13 L. monocytogenes isolates obtained from fecal samples (∼1 PFGE type per 2 isolates). The estimated PFGE type richness for L. monocytogenes isolated from the pond water and fecal samples was 29 (95% CI = 19 to 207) and 9 (95% CI = 7 to 33) PFGE types, respectively (Table 3). Two concurrent studies (19, 20) conducted in a 200-km2 agricultural watershed in Ontario, Canada, also performed PFGE on L. monocytogenes isolates from agricultural water and wildlife feces; these studies used the same PFGE protocol as the current study (10). In the two Canadian studies, Lyautey et al. (19, 20) identified 21 PFGE types from 75 L. monocytogenes isolates from water samples (∼1 PFGE type per 3 isolates) and 18 PFGE types from 84 L. monocytogenes isolates from fecal samples (∼1 PFGE type per 5 isolates). As such, the ratio of PFGE types identified relative to the number of L. monocytogenes isolates subtyped in the current study, which was conducted in a 40,000-fold smaller area, was larger than the ratio found in the Canadian studies (19, 20) for both the fecal samples and water samples. Although this may suggest the L. monocytogenes population in the Finger Lakes region of New York is more diverse than Ontario, Canada, the greater diversity on the farm sampled in the study reported here may also be due to factors unique to the sampled farm. For example, because two dairy farms are located uphill of the sampled farm, continuous introduction of L. monocytogenes from the dairies may have resulted in a more diverse population of L. monocytogenes, compared with the other areas. To identify potential drivers of the greater PFGE type diversity observed in the study reported here, compared with Lyautey et al. (19, 20), a systematic survey of Listeria populations across and within regions is needed. Regardless, the diversity of Listeria identified in this study, and other studies, suggests it often may be difficult to determine whether an isolate found in a processing environment originated in a particular preharvest environment, as multiple subtypes may be present in a single environment. In fact, to either rule in or out any particular source in an environment with a considerable diversity, such as the source investigated in the current study, a large number of samples would have to be collected and a large number of isolates would have to be subtyped, including multiple isolates per sample, as suggested previously (9, 30). Consequently, the "absence of evidence" for presence of a given Listeria subtype (i.e., the Listeria subtype was not detected in any of the samples collected) in the preharvest environment in New York state and other preharvest environments with prevalent Listeria does not necessarily provide evidence that the given subtype is truly absent in this environment, particularly if sample size is small.
Should I stay or should I go
The L. monocytogenes PFGE data collected here indicate the survival ("stay") or continuous reintroduction ("go") of L. monocytogenes into the farm environment. Specifically, 8 of the 19 PFGE types isolated from the pond and 3 of the 7 PFGE types isolated from the feces collected near the pond were isolated on more than one date (Fig. 2). The repeated isolation of multiple PFGE types suggests these PFGE types were surviving or were continuously reintroduced in these sample types. For example, PFGE type CU-30,25 was isolated a total of eight times in this study, making it the most commonly isolated PFGE type (Fig. 2). This could indicate that the PFGE type CU-30,25 has an increased fitness in this environment or that there is a source of this PFGE type continuously contaminating the environment. In addition, 4 of the 22 PFGE types identified here were isolated from both the pond and feces collected near the pond (Fig. 2). Lyautey et al. (19, 20) found similar results in two of their studies: 5 of the 18 PFGE types from wildlife feces in the South Nation watershed in Ontario, Canada, were also isolated in water samples collected from the same watershed. Because the same PFGE types can be found in agricultural water and feces collected in proximity of the sampled water in both studies, this supports transfer of L. monocytogenes between feces and agricultural water and/or a common source leading to the contamination of both sample types in the investigated environment.
Allelic typing of the sigB gene has been used to speciate Listeria isolates and can provide initial characterization of Listeria strains present (16), but it is not of high enough discriminatory power to elucidate the actual diversity of a Listeria population in an environment. For example, in the current study AT 57 represents 13 different PFGE types, AT 61 represents 2 different PFGE types, and AT 67 represents 2 different PFGE types (Fig. 2). Strawn et al. (34) also performed PFGE on L. monocytogenes isolates with matching sigB ATs from a single sample type in a survey of soil, feces, drag swabs, and water samples from five farms in New York state (i.e., repeatedly isolated sigB ATs). For three of the four repeatedly isolated sigB ATs, they found multiple PFGE types corresponding to each individual AT. PFGE hence clearly provides improved discriminatory power (over sigB allelic typing), facilitating investigations of persistence, transfer between sample types, and continuous reintroduction (4). Interestingly, in our study, all isolates of the same PFGE type also had the same AT (Fig. 2). For example, all isolates of PFGE type CU-371,460 were AT 57, all isolates of PFGE type CU-107,177 were AT 57, all isolates of PFGE type CU-254,53 were AT 67, etc. This finding has not been directly reported in other studies; however, Liao et al. (16) have determined the sigB gene is genetically stable, because it is not a frequent site of homologous recombination or positive selection. As such, we expect this relationship between sigB ATs and PFGE types to hold true across different environments. Although sigB allelic typing is not as discriminatory as PFGE, it can still be useful in performing traceback analysis, as it can be used as an initial screen to determine which isolates should be selected for PFGE, or other fine resolution typing methods (e.g., whole genome sequencing) during a traceback investigation. As a result, to determine which isolates from an environment match an isolate from another environment (e.g., processing environment), allelic typing should first be performed ("work smarter"). Next, isolates from the preharvest environment that have the same AT as the isolate from the processing environment should be characterized by PFGE or whole genome sequencing ("work harder") to determine which are truly the same. This will save time and money, as PFGE and whole genome sequencing are considerably more expensive and time intensive to perform.
Overall, the data presented here and in previous studies (39) indicate Listeria is prevalent in the New York state preharvest produce environment; thus, it is likely for Listeria from the preharvest environment to contaminate produce and/or be introduced to the processing environment (from the farm). The considerable Listeria diversity observed in the small area within the produce farm studied, and the evidence of transfer between sample types or continuous reintroduction of Listeria, indicate that it often may be difficult to successfully trace isolates from finished products or the processing environment (e.g., environmental swab) back to farm sources (e.g., feces, water), unless a large number of samples are collected and a large number of isolates are subtyped. However, it should be noted that these conclusions may be limited to similar preharvest produce environments that are continuously contaminated with livestock runoff. As such, additional research is needed to determine whether the findings reported as part of this pilot study hold true for other produce farms and produce growing regions.
This work is supported by U.S. Department of Agriculture (USDA) Specialty Crop Research Initiative grant 2019-51181-30016 from the National Institute of Food and Agriculture. Manuscript preparation and revision were also partially supported by the National Institute of Environmental Health Sciences, National Institutes of Health (NIH), under award T32ES007271, and the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture, USDA. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the USDA or NIH.
Supplemental material associated with this article can be found online at: https://doi.org/10.4315/JFP-20-179.s1
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Spatial and temporal factors associated with an increased prevalence of Listeria monocytogenes in spinach fields in New York state
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Published 2021 by the International Association for Food Protection
This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/)
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Some say it's a jogging game that has become bloodsport. The player controls multiple characters and tries to run as far as possible with as many joggers alive as possible. It's an arcade-style survival game with in-game leaderboards and achievements. Grab your sneakers and experience the most brutal jogging game available!


Visual Index Waldorf•ish Online Courses that simplify your school year.
click on the text below the image to be taken directly to the lesson.
Oct - Week Four - dry/hard watercolor paints + paper, chalk or crayon (figure drawing).
November - Week One - Chalk/Pastel on black paper or chalkboard (Monet inspired).
November - Week Two - Dry/hard watercolor paint + paper. (Figures in a landscape).
April - Week Two - Watercolor (winter landscape) with watercolor paper + dry paints.
May - Week Two - Watercolor pencils + crayons, dry watercolor paints, paper, etc.


text:
| Filmography | Forum | Gallery
Dev Patel as Sonny in The Best Exotic Marigold Hotel (2012)
Source: The Official Trailer.
This photo, entitled "Dev Patel as Sonny in The Best Exotic Marigold Hotel (2012)", has been posted on Tuesday, September 27th, 2011 in the fan-made gallery dedicated to "Dev Patel".
Posted by G21635 - Tuesday, September 27th, 2011
"Dev Patel" ➤
Photo of Dev Patel, who portrays "Sonny" in "The Best Exotic Marigold Hotel" (2012).
"The Best Exotic Marigold Hotel" Copyright © Fox Searchlight Pictures
Photo of Dev Patel as "Sonny" in "The Best Exotic Marigold Hotel" (2012). As stated by the poster, this picture is from the video of the trailer available for "The Best Exotic Marigold Hotel".
Dev Patel is an actor.
As an actor, Dev Patel has been seen in movies such as The Last Airbender, released in 2010 portraying Zuko, The Second Best Exotic Marigold Hotel (2015) portraying Sonny Kapoor, Chappie (2015) (Deon), Slumdog Millionaire (2008) as Jamal Malik.
About "The Best Exotic Marigold Hotel":
Written by Ol Parker (adaptation), "The Best Exotic Marigold Hotel" is a Comedy/Drama film, released on March 09 of 2012 in the USA . Bill Nighy and Tom Wilkinson (Norman) are starring, alongside Judi Dench (Evelyn), Dev Patel (Sonny), Penelope Wilton (Jean) and Celia Imrie (Madge).
As an actress, Celia Imrie acted in movies such as Imagine Me & You, released in 2005 playing Tessa, Highlander (1986) (character: Kate MacLeod), The Borrowers (1997) in which she portrays Homily Clock, Frankenstein (1994) (character: Mrs. Moritz), The Second Best Exotic Marigold Hotel (2015) (character: Madge Hardcastle), and Nanny McPhee (2005) as Mrs. Quickly.
As an actor, Bill Nighy has been seen in movies such as Total Recall, released in 2012 playing Kuato, Page 8 (2011) as Johnny Worricker, The Magic Roundabout (2005) in which he portrays Dylan, Pirates of the Caribbean: At World's End (2007) (character: Davy Jones), Wrath of the Titans (2012) as Hephaestus, and Chalet Girl (2011) in which he plays Richard.
Part of a fan-made tribute gallery for "Dev Patel", this image ("Dev Patel as Sonny in The Best Exotic Marigold Hotel (2012)") has been uploaded on Tue, 27 Sep 11 11:12:30 +0000, considering that the poster has done it in a responsible and legal manner. This poster certainly thought doing something well by sharing this picture, however, if you consider that this image has not conformed to a fair-use or is inappropriate, please report this to Theiapolis.com immediately, and we will delete it as soon as possible. Thank you for your help.
This picture has been posted in the fan-made tribute gallery for "Dev Patel"
Check out the photo gallery for The Best Exotic Marigold Hotel
No message has yet been posted about Dev Patel.
Dev Patel To Be Cast in Neill Blomkamp's "Chappie"?
1 message - According to The Hollywood Reporter's Heat Vision Blog, Neill Blomkamp has Dev Patel in talks to take on the lead human character in his planned film...
1 message - bets best bestbest...
3 messages - IT COULDN'T BE BEAT! IT WAS AWESOME! THE BOOKS ARE GREAT. :)...
About Felipe Pazos Jr
6 messages - He's son of a great Cuban economist, Felipe Pazos....
Ellsworth 'Sonny' Wisecarver
An American actor
Jorge 'Sonny' Alarcón
A Mexican actor
Arsenio 'Sonny' Trinidad
Dev Malhotra
Sonny Greer
Resources: contributions from movie fans. Anyone can submit additionnal information and corrections, you can post a message into the forum, or you can contact me by e-mail. Information and materials are submitted by users and thus may not always contain up-to-date and correct information, so do not hesite to report mistakes, and submit corrections.Thank you!. Direct editing is no longer available, sorry.
theiapolis PEOPLE


text:
About West Vault Mining
Hasbrouck Overview
Hasbrouck Project Pre-feasibility Study
Interview Videos and Articles
Share Structure
2021 Annual Meeting Materials
ESTMA
West Vault Mining
Suite 838 - 1100 Melville Street
Canada V6E 4A6
TSXV : WVM
OTCQB : WVMDF
At Surface Gold
West Vault Closes Gold and Silver Stream Transaction With Sprott
VANCOUVER, BRITISH COLUMBIA, March 1, 2021 – West Vault Mining Inc. (WVM:TSXV, OTCQX:WVMDF) ("West Vault" or the "Company") announces the closing of a gold and silver Purchase and Sale Agreement (the "Stream Agreement") with Sprott Private Resource Streaming and Royalty Corp. ("Sprott") for 1.41% of all refined gold and silver (the "1.41% Stream") to be produced for the life of mine from the 100% owned and permitted Hasbrouck Gold Project, located southwest of Tonopah, Nevada. The Stream Agreement for the 1.41% Stream was first announced by the Company on February 22, 2021.
Under the terms of the Stream Agreement, Sprott has paid the Company an advance purchase deposit of US $6.0 million (the "Advance Deposit"). An additional purchase deposit of US $1.0 million (the "Additional Deposit") is to be paid to the Company within 10 days of the announcement of a Board-approved construction decision for the Hasbrouck Gold Project. Sprott may elect to add the Hill of Gold property to the Stream Agreement by paying a final purchase deposit of US $300,000 within 60 days of paying the Additional Deposit. In addition to the amortization of aggregate purchase deposits received by the Company, a cash transfer price payable upon delivery of refined gold and silver to Sprott has been set at 20% of the market value per ounce of metal, as quoted by the London Bullion Market Association on the date of delivery.
About West Vault Mining Inc.
Located and secure in Nevada, the Hasbrouck Gold Project is permitted for construction and operations. West Vault is focused on maximizing shareholder value for its 100% interest in the Hasbrouck Gold Project. After closing the Stream Agreement today and receipt of the Advance Deposit, the Company holds cash on hand in excess of US $7.0 million.
On behalf of West Vault Mining Inc.
"R. Michael Jones"
For further information please see the Company's website at www.westvaultmining.com or contact us by email at [email protected]
(604) 685 8311 / [email protected]
Disclaimer for Forward-Looking Information
This press release may contain forward-looking information or forward-looking statements (collectively "forward-looking information") within the meaning of applicable securities laws. Forward-looking information is typically identified by words such as: "believe", "expect", "anticipate", "intend", "estimate", "postulate" and similar expressions, or are those, which, by their nature, refer to future events. Forward-looking information in this news release includes, without limitation, the announcement of a Board-approved construction decision for the Hasbrouck Gold Project. Although West Vault believes that such information as set out in this press release is reasonable, it can give no assurance that such expectations and estimates will prove to be correct. The Company cautions investors that any forward-looking information provided by the Company is not a guarantee of future results or performance, and that actual results may differ materially from those in forward-looking information as a result of various factors, including the following: risks related to the novel coronavirus (COVID-19) global health pandemic, other global epidemics, pandemics or public health crises; the state of the financial markets for the Company's equity securities; the state of the market for gold or other minerals that may be produced generally; significant increases in any of the machinery, equipment or supplies required to develop and operate a mine; a significant change in the availability or cost of the labor force required to operate a mine; a significant increases in the cost of transportation for the Company's products; variations in the nature, quality and quantity of any mineral deposits that may be located; and the Company's ability to obtain any necessary permits, consents or authorizations required for its activities, to raise the necessary capital or to be fully able to implement its business strategies and other risks associated with the exploration and development of mineral properties. The reader is referred to the Company's public filings for a more complete discussion of such risk factors and their potential effects which may be accessed through the Company's profile on SEDAR at www.sedar.com.
Neither the TSX Venture Exchange nor its Regulation Services Provider (as that term is defined in the policies of the TSX Venture Exchange) accepts responsibility for the adequacy or accuracy of this release.
HASBROUCK DEVELOPMENT
Copyright 2020 West Vault Mining
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The look was inspired behind the beautiful cable-knitted cardigan from RW&CO's Fall/Winter collection. I chose to leave it by itself because it was a feature piece on its own, but to change it up I added a herringbone vest underneath. The sweater although is beautiful, on its own would have been to simple for a week like this. This week for me was a time for me to truly showcase my personal styling ability on myself and prove why I am called The Prep Guy. So underneath the vest I put on a light blue button down shirt, which had mini orange details on it. The base colour of the shirt was also blue so I knew I had leeway to introduce more colours. I wore a simple but very preppy burgundy and blue striped silk tie. Next I highlighted the orange detailing in the shirt by adding a orange cotton and silk scarf into the mix. It really made the orange pop and completed the upper look. The tricky part of the look was picking the right colour pants, I had two options, black or grey. The black slim fit jeans looked great but if the cardigan had been grey – it would have been perfect. So I decided to go with a great cotton trouser in a dark grey.


text:
Sureda ist der Familienname folgender Personen:

 Andrés Rigo Sureda (* 1943), spanischer Jurist
 Antonio Sureda (1904–1951), argentinischer Bandoneonist und Tangokomponist
 Jacobo Sureda (1901–1935), spanischer Maler und Dichter
 Jaume Santandreu Sureda (* 1938), spanischer Priester, Autor und Politiker
 Maria Pascuala Caro Sureda (1768–1827), spanische Mathematikerin und Priorin
 Pedro Caro y Sureda de la Romana (1761–1811), spanischer General

redpajama_set_name: RedPajamaWikipedia


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text:
Claudio Cobelli
Retired full professor. At the University of Padova as full professor.
Claudio Cobelli received the Doctoral degree (Laurea) in Electrical Engineering in 1970 from the University of Padova, Italy. From 1970 to 1980 he was a Research Fellow of the Institute of System Science and Biomedical Engineering, National Research Council, Padova, Italy. From 1973 to 1975 and 1975 to 1981 he was Associate Professor of Biological Systems at the University of Florence and Associate Professor of Biomedical Engineering at the University of Padova, respectively. In 1981 he become Full Professor of Biomedical Engineering at University of Padova. Since 2000 he is Affiliate Professor with Bioengineering at the University of Washington, Seattle, WA. From 2000 to 2009 he has been Chairman of the Graduate Program in Biomedical Engineering. From 2000 to 2011 he has been Chairman of the Ph.D. Program in Bioengineering at the University of Padova. His main research activity is in the field of modeling and identification of physiological systems, especially endocrine-metabolic systems. He has published 400 papers in internationally refereed journals. He is co-editor of Carbohydrate Metabolism: Quantitative Physiology and Mathematical Modeling (Chichester: Wiley, 1981), Modeling and Control of Biomedical Systems (Oxford: Pergamon Press, 1989) and Modeling Methodology for Physiology and Medicine (New York:Academic Press,2000).He is co-author of: The Mathematical Modeling of Metabolic and Endocrine Systems (New York: Wiley, 1983); Tracer Kinetics in Biomedical Research:from Data to Model (London:Kluwer Academic/Plenum Publishers, 2001) and Introduction to Modeling in Physiology and Medicine (San Diego: Academic Press, 2008). He is on the Editorial Board of Diabetes and Journal of Diabetes Science & Technology. In the past he has been Associate Editor of IEEE Transaction on Biomedical Engineering and of Mathematical Biosciencies. He has been on the Editorial Board of Control Engineering Practice, Diabetes, Nutrition & Metabolism, Diabetologia and American Journal of Physiology, Endocrinology & Metabolism. Dr.Cobelli has been Chairman (1999-2004) of the Italian Biomedical Engineering Group, Chairman (1990-1993 & 1993-1996) of IFAC TC on Modeling and Control of Biomedical Systems and IEEE EMBS AdCom Member (2008-9). He is a member of the Gruppo di Esperti della Valutazione (GEV), Area 09, of the Agenzia Nazionale per la Valutazione del Sistema Universitario e della Ricerca (ANVUR) for the period 2011-2013. He is President of the Organo di Indirizzo of the Azienda Ospedaliera Università di Trieste for the period 2011 – 2015. In 2010 he received the Diabetes Technology Artificial Pancreas Research Award. He is Fellow of IEEE, BMES and AIBME.
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