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June screen printing workshop!
A one time workshop in a studio in Pardes Hanna, 138 Hadkalim st.
You will print your own unique designs (which you'll prepare beforehand) on your own fabric/paper/wood etc.
In the workshop we will learn:
- How to print with a screen on paper and fabric
- How to make a stencil out of a new screen
- To use inks and techniques for screen printing
In the studio you will have a screen with your image on it ready to use and all the equipment, inks and guidance you will need to creat gorgeous prints. The only thing you need to bring with you is items to print on: t-shirts, bags, pillowcases, paper, wood etc.
There will be only up to 4 people in every workshop.
Please wear clothes that you don’t mind getting splashed with ink!
Please send your artwork to [email protected] by the 7/6/19 as a PDF with 300 DPI in original size of 30x40 cm.
You can have max of 3 images/prints.
See you on the 14/6/19 at 9:00-12:30


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Los Angeles has found a solution to tackle a slice of the homeless issue in Los Angeles. More than 50 specially designed containers will be furnished and stacked in a little development next to the 110 and the 105 freeways. Mayor Eric Garcetti says that this form of permanent housing is faster to build.
"Let's figure out the models that work because we want to be able to take any piece of property anywhere in any amount of time with whatever funding we have and get something built."
Garcetti adds that 21 projects are now in the works because of voter approved measure HHH. The measure is a $1.2 billion bond to be allocated to the construction of 8,000-10,000 units of clean and safe affordable housing for the homeless, according to LA Chamber of Commerce.
"Nothing stops homelessness like actually getting to the root of the problem. Get people indoors first, but if you don't want them to go back outdoors, you have to treat them."
Garcetti goes on to say that the motivation for assisting the homeless is not tied to office accomplishments, but rather a need to assist fellow persons in need.
"This is not about a project. This is not about funds. This is about human beings. This is about flesh and blood," he says.
"A problem caused by the inaction of human beings can be solved by the actions of human beings here today."
The units will cost about $600,000 each, more than the median price of a condo in Los Angeles. The total cost of the units will be about 35 million dollars each. The one-bedroom units are expected to open next year.

domain: sociology


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The Gift that Keeps on Taking
I’m a sucker for holiday traditions. No matter how frenzied and unpredictable the year was, I know I can count on Chinese food on Christmas Eve and a new pair of gym shoes waiting for me under the tree. In keeping with that spirit, we’ll end the year of Explanation of Benefits the same way we did last year—with a page full of updates.
Employers rank the costliest
chronic medical conditions
As I’ve said before, I don’t mind being wrong, but I love being right. Last month, we warned employers to take diabetes more seriously. Well, guess what? Diabetes topped the list of the costliest chronic medical conditions for employers. That’s according to a survey of 530 employers by the International Foundation of Employee Benefit Plans for the IFEBP’s 2017 Workplace Wellness Trends report (bit.ly/1QX4LEc). Some 41 percent of the employers—431 from the U.S. and 99 from Canada—cited diabetes as the most expensive chronic medical condition their workers had (see chart). More than 90 percent said they offered wellness-related programs for their employees, but less than half—48.1 percent—said they included disease management services. Even less said they offered nutritional counseling or healthful food choices in cafeterias and vending machines, at 33.2 percent and 44 percent, respectively.
Doctors say much medical care
is unneeded, but blame patients for making them do it
In September, we talked about the use of unnecessary medical treatments and overspending on necessary medical treatments that could be provided for less (bit.ly/2hIqoB5). Both drive up health care costs for employers. As it turns out, most doctors agree with me. Researchers from the Harvard Medical School and the Johns Hopkins University School of Medicine asked 2,106 physicians about their views on wasted medical treatment, its causes and possible solutions. The consensus, published in the journal PLOS One, was that 25 percent of diagnostic lab tests are unnecessary, as are 22 percent of prescription medications, 21 percent of overall medical care and 11 percent of medical procedures (bit.ly/2xCIB6H). They cited fear of malpractice lawsuits (85 percent), pressure from patients (59 percent) and difficulty accessing patients’ previous medical records (38 percent) as the top three causes of overtreatment. Their top three solutions were: training

domain: news


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For travelers who want to take in the sights and sounds of Kumamoto, Kkr Hotel is the perfect choice...see more
For travelers who want to take in the sights and sounds of Kumamoto, Kkr Hotel is the perfect choice. From here, guests can enjoy easy access to all that the lively city has to offer. A haven of rest and relaxation, the hotel will offer total renewal just steps away from the city's numerous attractions such as Kumamoto Prefectural Traditional Craft Centre, Kumamoto Castle, Kumamoto Castle Park.
Offering hotel guests superior services and a broad range of amenities, Kkr Hotel is committed to ensuring that your stay is as comfortable as possible. To name a few of the hotel's facilities, there are restaurant, car park, laundry service, bar.
All guest accommodations feature thoughtful amenities to ensure an unparalleled sense of comfort. The hotel offers an excellent variety of recreational facilities, including massage. Superb facilities and an excellent location make the Kkr Hotel the perfect base from which to enjoy your stay in Kumamoto.

domain: travel


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Lottery is a form of gambling that involves the drawing of numbers at random. While some governments outlaw lotteries, others endorse them and organize state and national lotteries. It is a popular form of gambling in most countries, including the United States. Lottery numbers are drawn on a computer from millions of tickets sold each week.
Lotteries have long been used as a way to raise money for various causes. For example, the Old Testament instructs Moses to conduct a census of the people of Israel, and the ancient Romans reportedly used lotteries to give away property and slaves. In the United States, lottery games were first introduced by British colonists, who used them to fund public works. However, the practice was banned in ten states between 1844 and 1859.
The history of lotteries can be traced back to the fifteenth century in the Low Countries. Various towns held public lotteries in order to raise money for poor people and for city defenses. These lottery games were soon popular and were even hailed as a painless form of taxation. The first known lottery in France was the Loterie Royale of 1539, authorized by Francis I. Despite its early history, the first lottery was a flop. The ticket prices were too high and the social classes opposed the project. For two centuries, lotteries were largely banned in France but were tolerated in other countries.
Lotteries are considered a form of gambling, with a wide variety of games available. The most common lottery game is Lotto, where players select six numbers from a series of balls numbered from 1 to 50. While these games have no skill-based elements, they do offer the fantasy of becoming wealthy. But, in the end, it is just pure luck that determines whether or not you win.
A lot of modern lotteries use computerized methods for drawing winning numbers and symbols. The winning tickets are then divided between winners, or the money is transferred to the next drawing. This is called a jackpot or rollover, and it increases the top prize. However, some modern lotteries do not use computers.
Lotteries have a rich history in the United States. In colonial America, there were approximately 200 lotteries between 1744 and 1776. The money raised from these lotteries helped build roads, schools, canals, and bridges. A few colonies used lotteries during the French and Indian Wars, and the Commonwealth of Massachusetts organized a lottery to raise funds for an “Expedition against Canada” in 1758.
In the 1960s, lotteries and casinos re-emerged throughout the world. They were popular and profitable, and governments used them to generate revenue.

domain: finance


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For the month of May, the Coleman Book Club will be reading Confessions of a Sociopath: A Life by M. Thomas. Our meeting will take place on Friday May 8th at 10:30 AM - Book club members may pick their copies up at the library.
Join us for stories, songs, and rhymes as we promote early literacy skills and foster a love of books, Fridays in April and May @ 10:30AM. For children ages 2 to 5 and their caregivers. Siblings welcome!

domain: hobby


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Langhorne Farmers market a summer tradition
Some events are easily associated with summer: lounging on warm, sunny beaches; exciting nights at the carnival; eating half-melted, yet delicious, ice cream cones on a hot day.
For Langhorne, the borough farmers market has become a summertime and fall staple.
Dorothy Ortez, 73, said she's been coming for the past four years and doesn’t miss a week.
“It’s what they have ... There are things here you can’t find anywhere else,” she said.
The selection includes fruits, vegetables, breads, cookies, muffins, clothing, jewelry, bath and body products and even chili.
“We get stuff for everybody -- even our dog," said Langhorne resident Fern Thaler.
But it's not just the selection that keeps the residents coming-- it’s the fact that the locals know where it's coming from.
“They go the extra mile here.” said Ortez.
James and Jennifer Minnegan said they love the fact all the vegetables and fruits are locally grown.
Cindy McMahon, operator of Charlann Farms in Yardley and a vendor at the market, said people flock to the seasonal farmers market here and in other communities.
Although the knowledge of where the food is coming from is a big factor, some residents said they enjoy the sense of community that comes with the market.
“People in town love it. The locals can walk here and tell their neighbors about it (the market),” said Judy Sloane, a market organizer.
Jen Brozoski and her 3-year-old daughter walk to the market any chance they have.
“We’re sad when it’s over and can’t wait to come back.” said Brozoski, 33. Some of their favorite items at the market are the pumpkin bread and the cookies, she said.
Vincent Kopczenski, owner and vendor of 5 Flavors Chili, said he comes to the market to be with the people.
“You have a camaraderie here you don’t get at a local supermarket.” said Kopczenski, who's semiretired.
Terri Little, organizer and founder of the market, said she came up with the idea about five years ago. Little’s goal was to create a new local tradition in Langhorne.
“There are a lot of things that go on in this town, so why not a farmers market?” she recalled asking herself.
When the idea was presented to the local governing board, officials liked it and asked Little to organize the market. So, with the green light, Little asked Sloane to help and Langhorne’s Farmers Market was born.
Kelly Armstrong can be reached through editor Carl LaVO: 215-949-4227 or firstname.lastname@example.org


This work will cover to a standard method for generation of silver nanoparticles to be tested for inhalation toxicity on experimental animals. More detailed defining of the scope will be discussed in the following specific items. 1) Generator set-up: flat-plate ceramic heater was connected to an AC power supply and housed within a quartz case 2) Heater dimension: 50 × 5 × 1.5 mm3. 3) Surface temperature: about 1500? 4) Voltage : 85 V 5) Highest temperature on the local heating area, Tmax,: 1140? 6) air changes per hour in an exposure chamber: 12-15 times 7) loading mass of the silver lump : 160 mg 8) Nanoparticle range: 4.5-157 nm 9) Geometric mean diameters of silver nanoparticles: less than 100 nm in nonaggregated/ nonagglomerated form 10) Silver Nanoparticle measurement: SMPS (scanning mobility particle sizer) and CPC (Condensation particle counter), TEM (Transmission Electron microscopy).


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Let’s be honest, sometimes we just need simple, easy meals that don’t require a lot of thought. Yesterday was one of those nights. We had hiked for a little over an hour. I was tired and I just needed easy. Thankfully, I keep certain staples in the pantry. One of those staples includes a can or two of sloppy joe sauce. Of course, soy crumbles in the fridge or freezer are also important. You can use them for all kinds of things, so I keep them around.
I found some soft rolls at a quick stop, which happily were vegan, so Sloppy Joes were on the menu. I had to dress my crumbles up just a bit, because it makes dinner just a bit more special. I finely diced and sauteed some onions and then added garlic to the pan before I threw the crumbles in.
After that it was only a few minutes before I poured the can of Sloppy Joe mix over the top and let it simmer on low until heated through.
I served them on the soft rolls I had come across with corn on the cob as a side.
Bruce teases me about how I dress up my corn, but I don’t even care. It’s delicious with all theses spices!
Vegan Sloppy Joe Ingredients:
- Vegan Rolls
- Sloppy Joe Mix
- Soy Crumbles
- Onion, finely diced
Saute onion for 3-5 minutes, add garlic. Cook another min. Add Crumbles, saute 3-5 minutes. Add Sloppy Joe Sauce, heat through. Serve on Buns.

domain: food


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The new drama's costars party at their film's premiere. Plus: Jude Law, Bono, Lindsay Lohan and more
Updated: Monday Nov 22, 2004 | 06:00 AM EST
More from PEOPLE
See More News
9 of 15
Newly single Lindsay Lohan enjoys a day of shopping in Beverly Hills on Nov. 18. Lohan, who recently wrapped Herbie: Fully Loaded and set to release her debut CD, Speak, on Dec. 7, doesn't leave home without her red-string Kabbalah bracelet.
Copyright © 2013 Time Inc. All rights reserved. Reproduction in whole or in part without permission is prohibited.
In partnership with AOL

domain: entertainment


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Sunset View Cottage near Lake Michigan
Great Charlevoix location in this 4 bedroom, 2.5 bath cottage. Walking distance to Lake Michigan beach, the lighthouse and town! Amazing sunset views, sit on the porch and watch the boats coming and going through the channel. This home is full of character and is a great spot to spend with family and friends. The stone front porch was designed by Earl Young!
All rates are subject to change. Please check with your reservationist prior to booking.


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Studiovine is excited to launch the new Plymouth Raiders website.
Our Plymouth Studio has been at the forefront of the Raiders marketing for the past two seasons, and we recently secured a new 2-year partnership with the idea of re-marketing the club in a new direction.
Our Executive Director, Lee Nathan said “Here at Studiovine we have always valued our long and strong partnership with the Raiders. Having secured our next 2-year partnership, we wanted to work on a more dynamic feel for the brand and to enhance the fans experience when visiting the clubs website. Web design is not just about creating pretty layouts, it’s about understanding the needs of the users, and we believe we have done this.”
The fundamental task when building this new website was to put the user first, and we believe we have achieved this and more with the introduction of advertising space for sponsors which will help bring more revenue to the club. We researched various sports clubs across the world to take inspiration from and we used our knowledge of the sport industry to create the new website and marketing material to challenge some of the top teams.
The brief was clear, create a new, exciting identity with an online platform to compete against the best and to strengthen their position in the BBL for the 19/20 season.
Plymouth Raiders Digital Media Manager, Jack Hodgetts said “It was great to work with Studiovine on this project and I’m delighted to be able to showcase this to our fans now. The Studiovine team fully understood what we asked of them and brought our vision to life. I think the new branding showcases the Raiders brand and gives a huge step in the challenge for a place in the digital space in Basketball. The partnership with Studiovine is invaluable to us at Raiders, and we look forward to working with them for years to come.”
To view our work visit: plymouthraiders.com

domain: design


For most of us who read comprehensive news and science reports, we know that sea levels have been rising since the end of the last ice age, most rapidly about 12,000 years ago. In recent centuries, the average rate has been relatively uniform. The rate was higher during the first half of the 20th century than during the second. At around a couple of millimeters per year, it is a residual of much larger positive and negative changes locally. The risk from global warming is less than that from other factors (primarily geological).
Or so we thought.
Today, in glaring contrast of scientific research, we learned that drought, climate change and pollution are all because of our lack of faith in Allah.
That’s right folks, take my word for it, lest we forget, lest we get decapitated. Radical sheik Mohammed Omran said so, so it must be true. And he has proof too. What is his proof? He cites Mohammad:
What are the people now crying for? The prophet told you hundreds of years ago, ‘Look after the water’.
Oh, gee. What riveting, forensic proof. Al Gore, are you listening?


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Hail to you, enormous Earth, Mighty sea; program for humanityJust a phase for act and playYou are here but for a day; Hail for you, galaxiesYou don’t even notice me; You are nothing to beholdYou are getting older, getting old.
And it all comes down to thisThat the lady is firstThe one you miss. The fog blocks your mindIts wetness a deceiving type; You find your eyesScanning darkness on her behalf light. Tiny face, sheThe one you kissOnce you kissedWelcome home.
The fog blocks your mindIts wetness a deceiving sort; You find your eyesScanning darkness for her gentle; And it also all comes down to this -That the girl is firstThe one you missed, always miss; Tiny face, sheOne you kissedNow you missWelcome home.
Fate surrounds you like a doveCircling from aboveIn tongues the girl spokeOf the city associated with Love; You didn’t attain itNever even askedStill she warmed youAgainst her chest.
Nevertheless, England would later consider over the region. In some ways it can be stated that Hilton Head background properly begins in 1663 when William Hilton inadvertently discovered the island while wanting to locate Port Royal’s entrance. Typically the island was named Hilton Head in reference in order to the headlands his team observed as they discovered the area by marine. Both Spain and France started to occupy the Slot Royal Sound region in the 1520s, and right after some fighting the location went to Spain. He was pleased with the water plus air in the region, as well as the tall pine woods which implied that the soil was fertile. English colonization of the tropical isle began in 1698, but the island’s European human population still only consisted regarding 25 families by 1766.
Typically the Gullah slaves were separated, and within a couple of years of the end of the Civil Conflict they were the island’s just inhabitants-at least until 90.. The Civil War was another difficult time in Hilton Head history, with Union Troops quickly occupying the region after typically the Battle of Port Royal Sound in 1861.
Now i’m the doveI’m in Adore with the oneFrom above Fold my wings within the skyFallen, fallen, Don’t know whyGot it inside my heartAnd I know life is short; Life will be leanWhy can’t the thing is?.
Mountain sticks in the skyLifts me personally up, wonder why; I rise till the woods bury meBury meCan you hear my heart beating? All of nature, all of EarthDo you care of my condition? Did you notice of my birth?.
But now the air isAs of the dayHer sudden lossCrippled you awayAnd you wonderedAnd you wanderedTill now, The black fog’s upon you.
It times back to seasonal profession by the Woodland Indians in the fall plus winter of each 12 months till the area was colonized, continuing through the Us Revolution and the Civil Battle; yet the island lose interest little resemblance to modern day Hilton Head before the 1955s.. Hilton Head history is somewhat unusual inside that it dates back extremely far, yet the modern Hilton Head Island has not been incorporated as a city until 1983.
Soon our hopes will move in addition to breatheCarry me, carry me personally; Beat and beat in addition to beat the wheat; Kids and daughters Do behold us, nature’s conscienceDoes nevertheless scold usSeven cities, more effective seasAll the fruit regarding who we’ll be; More effective hearts with seven beatsSix of seven of these people sleepCrafty intellectuallyNow he goes up from the seaBrought to analyze humanityFor too long she’s analyzed meCalling, calling says typically the sea, “”Won’t you appear and swallow me? “”.
This particular development model has already been followed since, resulting within the popular resort vacation spot known today.. Whilst the land had generally been used for hunting and timber for several years, a group associated with investors led by Charles Fraser bought up the particular majority of the island’s land and developed this, beginning with Sea Pines plantation. His system of development consisted of respecting the environment while providing amenities such as golf courses and bike paths first, allowing development to follow around those amenities. Modern Hilton Head history began in 1951 with the introduction of electricity to the island.
It is believed by simply many historians that the particular unity and pride of Gullah culture can be traced to the job system of slave labour employed in the location. Instead of sets of slaves being pushed to operate, Gullah slaves were given a task to complete; upon finalization of the task, they were free to go home. Hilton Head farms thrived for many years after of which, and a unique servant culture which came to be able to be called Gullah produced on the island in addition to elsewhere in the location. This fostered greater self-reliance and also a sense of satisfaction inside their work not loved by most slaves, and it is this-along together with the distinctive Gullah language and traditions that developed-that helped Gullah culture to get strong enough that that survives to this day. There could be no doubt that the Gullah people had been essential to the growth of Hilton Head and everywhere else that they labored.

domain: history


NEW YORK (CNNfn) - Internet wunderkind Marc Andreessen, a co-founder of Netscape Communications and, until last month, chief technology officer of America Online Inc., has unveiled plans for his month-old Internet start-up, Loudcloud.com Inc.
While not disclosing many details, Andreessen said the new Menlo Park, Calif.-based firm will focus on accelerating Internet business creation and growth.
"We're being deliberately vague," Andreessen said Tuesday at a press conference. "We're going to come back in early 2000 and have more details."
Andreessen has teamed with several ex-Netscape employees, including Loudcloud's co-founder and CEO, Ben Horowitz, who also left AOL (AOL) in late September. Andreessen is listed as the company's co-founder and chairman. According to reports, Loudcloud is privately funded by Andreessen and other employees.
Andreessen co-founded Netscape with ex- Silicon Graphics CEO Jim Clark and served as CTO and executive vice president of products. He made the transition to AOL in the role of CTO following its acquisition of Netscape on Nov. 24, 1998.
"Internet sites themselves are becoming incredibly sophisticated and complex, and every company is under intense pressure to move as fast as possible to address increasing competitive challenges," said Andreessen.
"Loudcloud will capitalize on these trends by delivering technology and services high-growth Internet companies need to compete and win."
-- from staff and wire reports


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Personally, I think the traditional ones have a much stronger style than the digital one. But I’m sure that with enough practice you’ll get as comfortable using digital too. Thanks for sharing your work!
Best posts made by yeungtina
RE: Need some help with how to paint a character that is a memory (idk how else to word it)
@eric-castleman Love the coloring! I think the contrast between the two palettes works for the "memory" vibe you're going for.
Before you explained it, I thought that the kid in the car was doing the "remembering", because he's the one looking at the tree. Maybe he could be looking away?
Excellent work with such a tricky idea. Looking forward to seeing how it turns out.
Help a beginner with tone/lighting?
I’m new around here. Have been taking the fundamental curriculum of classes and it’s really helped me since I started drawing seriously just in April. I’ve been playing with some thumbnails to figure out tone for my latest drawing, but can’t decide which to go with. Would love to hear from y’all if you have any feedback?
RE: WIP March book cover composition. Critiques welcome.
@Aleksey This is adorable. I think the lily pad O is clever.


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Shiraz defines the Barossa Valley and varies enough between vineyards and the different approaches of winemakers to provide consumers with endless variations. Additional facets of the variety can be highlighted when Shiraz is blended with several other varieties. Mataro is one perfect partner and blends that Benjamin commenced ten years ago with Durif have highlighted another.
Indeed, we have formed the view that Durif has a strong future in the Barossa Valley as the variety blossoms in the additional warmth of the Barossa grape ripening period.
The variety came to Australia in the 1900s and found its way to Rutherglen in 1908 as part of new plantings to replace vineyards devastated by phylloxera. By the 1970s these Rutherglen reds had developed a cult following. Rutherglen is a warm climate so it was natural to see how the variety would adapt to the Barossa climate.
Like Shiraz from which it is derived Durif (Shiraz crossed with Peloursin) is noted for strong flavours and higher than average alcohol.

domain: agronomy


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While Hillsborough added 230,000 residents over ten years, Pinellas shrank.
The University of Florida released its latest estimates for changes to Florida's population between 2000 and 2010, and two adjacent Gulf Coast counties find themselves on opposite ends of the growth spectrum.
According to the university, Hillsborough County added 230,278 residents between 2000 and 2010, the third-largest numerical increase among all 67 counties in Florida.
Over that same time period, Pinellas County lost nearly 5,000 residents, and was one of just two Florida counties to see its population decline over the decade.
The data show that Tampa's growth is continuing to outpace that of St. Petersburg, while Pinellas' high population density is holding future growth back.
Overall, the state added roughly 2.8 million residents between 2000 and 2010, the university estimates. That's an increase of 17.6%, to 18.8 million residents.
Florida's population growth was the third-largest numerical increase in the country, and the eighth largest by percentage.


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This is the third in the series of posts on Disney’s newest photo package – Memory Maker. This post focuses on using the editing software after you’ve taken the photos.
If you’re planning to get a photo package at Disney or make Disney photo books, be sure to check out all my Disney themed posts on capturing and preserving those Disney memories. If you’re planning on buying a photo package, be sure to check out my two prior posts on getting a discount on Memory Maker and how to best utilize it while on your trip.
Buying the Memory Maker package
1) The best way to purchase a Memory Maker package is in advance of your trip. If you do that you only pay $149 instead of $199 – for $50 in savings. (Disneyland Memory Maker is only $69.00 purchased in advance.) If you purchase it online through your Disney account, it should be automatically linked. If not, here’s Disney’s Memory Maker help page which will prompt you on how to link it. If you purchase it in the parks, the staff will help you link it to your account;
2) Add friends and family to your account so they can contribute and download photos. If they are already linked to your MyDisneyExperience account, no need to do anything as they are already connected;
3) Take photos like crazy! Anyone in your group can get their photos taken whether or not you’re present. The PhotoPass photographers just scan their Magic Bands or PhotoPass cards. If they aren’t linked in your account, you can still add their PhotoPass cards later;
Viewing Your Photos
You can look at your photos in the photo centers as normal. If you’re traveling with a laptop, you can also immediately see your photos as your trip progresses. Remember though after you make your first download, you only have 30 more days to add photos to the package. Most of us will not be able to get more than one Disney visit into 30 days, but if you happen to live close to Disney World and make more than one trip in a month, you can easily use the package for more than one trip which is great.
1) Go to Disney PhotoPass site to pull up your photos. Pick your park and enter your login ID and password;
2) All your photos will be divided into the 4 parks under their respective tabs;
3) Click on the photo you want to edit;
4) You can zoom, change the orientation (portrait or landscape), convert to black and white, or change the photo size (if you plan to buy a print from Disney);
5) The fun part is adding borders and stickers to your photos. Select “All borders” or “All Stickers” to view all of the choices. There are currently about 40 stickers – about half of them being character signatures so you can make it look like Mickey, Anna, or Elsa personally autographed your photo. The others are mostly logos of the parks and character stickers. Currently there are about 60 borders, with various characters, attractions or messages;
6) Each photo expires 45 days after it is taken, so you need to be sure to view and download your pictures within 45 days. You’ll see an expiration date under the photo when you view it.
Tips and Thoughts on Memory Maker software
1) If you decide to add some embellishments, be sure to “Create a Copy” so your original photo stays untouched and then save it. You will want to have an unedited original to download;
2) It would be better if you weren’t limited as to which selection of borders or stickers you can use. For example we could only use Star Wars borders on the ones taken at the Star Wars themed character meal. I wasn’t able to use them on our other Hollywood Studios photos which would have been fun as well.
3) Unfortunately the Memory Maker software is very, very, very buggy. I would frequently lose my edited photos and have to do them over again. My intention was to make all my different versions and then download them all at once, but every time I would go into my account, I would find the photo count to be different and I’d have to figure out what went missing. It’s hard to explain. I wouldn’t actually lose any of the photos themselves, just various versions of them, sometimes the original unedited version, but there was always an edited version that I could revert back to its original form. But it was really annoying and I would often get error messages that something could not be saved etc.
Because of all these issues, I would recommend using the “download all” function to save all the photos to your computer from the start so you feel comfortable that you have all your originals secured. (You should go ahead and also make a backup of them for good measure.) Then go back and play with the software and add all your embellishments. By the way, as you’ll see from the screenshot at above, it says it will take 2 hours to complete the download, but it actually took only a few minutes. I have a pretty fast internet connection though, so it will likely vary depending on your service.
4) Buy prints with a vendor of your choice instead of buying them from Disney. The Disney prints are expensive. Two 4 x 6 prints or one 5 x 7 is $14.95 and one 8 x 10 is $16.95. When you download the photos a limited license document will also be saved to your computer. I doubt you’ll need to show it, but it’s good to have it just in case. The license pretty much states that the photos may be reproduced for personal use, so you may make prints, use them in your scrapbooks, photo books, photo mugs, t-shirts, calendars etc. I used mine from Disneyland for our Christmas cards.
Many of the photo book companies like Snapfish and Shutterfly also make prints and other custom photo gifts and sometimes offer free prints where you just pay shipping. Be sure to check out my photo deals page for big discounts!
5) Unfortunately, you can’t access your Memory Maker photos via mobile or an iPad, although hopefully that will change. I would be nice to easily access your photos from your phone especially while waiting in line for a ride. 🙂
Disney didn’t ask me to write this post nor do I receive any compensation if you buy Memory Maker. We all know that Disney doesn’t need any plugging from me, but I hope that you’ve found this post and all my other Disney posts helpful! This is stuff I wish I knew before my trip so I hope this info helps you plan your Disney vacation!
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The present study aimed to perform molecular identification of lepidopteran larvae from canopy fogging including gut-content analyses. A total of 130 lepidopteran larvae were selected from 37 fogging samples at the Panguana station, district Yuyapichis, province Puerto Inca, department Huánuco, Peru. Target trees were pre-identified and subsequently submitted to molecular confirmation of identity with three markers (rbcL, psbA and trnL-F). The COI gene of 119 lepidopteran larvae was successfully sequenced and found to belong to 92 species: Comparison of DNA barcodes with the reference database of adult moths resulted in 65 (55%) matches at species level, 32 (27%) at genus level, 19 (16%) at subfamily or family level, three just to order level. Three larvae could not be assigned to a family. For these larvae the fogged target tree now suggests a potential host-plant relationship. Molecular gut content analysis, based on High-Throughput-Sequencing was successfully tested for ten larvae corroborating feeding on the target plant in some cases but elucidating several other cases of potential ‘alternative feeding’. We propose a larger-scale approach using this rapid and efficient method including molecular gut-content analyses for comprehensively testing the ratio of ‘alternative feeders’ and pitfalls caused by collateral fogging of larvae from neighboring trees.
Citation: Hausmann A, Diller J, Moriniere J, Höcherl A, Floren A, Haszprunar G (2020) DNA barcoding of fogged caterpillars in Peru: A novel approach for unveiling host-plant relationships of tropical moths (Insecta, Lepidoptera). PLoS ONE 15(1): e0224188. https://doi.org/10.1371/journal.pone.0224188
Editor: Bernd Schierwater, Tierarztliche Hochschule Hannover, GERMANY
Received: October 7, 2019; Accepted: January 14, 2020; Published: January 30, 2020
Copyright: © 2020 Hausmann et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: All relevant data are within the manuscript and its Supporting Information files and in dx.doi.org/10.5883/DS-PANLARVA + dx.doi.org/10.5883/DS-PANPLANT Entered on GenBank, BioProject ID PRJNA593715 http://www.ncbi.nlm.nih.gov/bioproject/593715.
Funding: The authors GH, JD, AxH recieved financial support from the State of Bavaria in the framework of the SNSB Innovativ program (1/2017; Staatliche Bayerische Naturwissenschaftliche Sammlungen; https://www.snsb.de/index.php/de/) allowing to engage the author JM, as manager of AIM (Advanced Identification Methods GmbH, München, Germany), to perform part of the plant barcoding and metabarcoding analyses. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The funder (State of Bavaria) provided also support in the form of salaries for authors GH, JD, AF, AxH, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.
Competing interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: JM (manager of AIM, Advanced Identification Methods GmbH, MüCnchen, Germany) was paid from project grants for performing part of the plant barcoding and metabarcoding analyses of gut contents. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
Despite much valuable work on host-relationships of Neotropical moths, e.g. from Ecuador [1,2], or Costa Rica [3,4,5], the relevant literature is still scarce and patchy compared with the huge species diversity of Lepidoptera in Central and South America. Apart from the aforementioned programs only few original data are published for host-plant relationships of Lepidoptera and much of the work focused on caterpillars found on plants of economic importance (pests and potential pests) (e.g. [6,7,8]).
Exemplified from one of the most diverse moth families, Geometridae, the largest project in Costa Rica so far revealed the huge amount of 22,957 geometrid moth records, the barcoded reared adults clustering to 566 BINs, of which 162 currently having Linnean species names (D.J. Janzen & W. Hallwachs pers. comm.). Brehm  presented 48 neotropical geometrid species with host-plant records, with 11 records added by Dyer et al.  and 59 records by Bodner et al. . Thus, altogether for some 680 Neotropical geometrid species (about 270 of which with Linnean species names) host-plant relationships are known, covering approx. 8–10% (4% with Linnean names) of the described geometrid fauna of Central and South America (for estimations of total number of described geometrid species cf. Scoble et al. : 6433 species; Heppner : 7956 species).
Estimation for Neotropical species diversity is based on approx. 37,000 described Neotropical moth species (Heppner : 44,800 described Lepidoptera, including approx. 7800 Rhopalocera species ) and considering that (a) ‘Microlepidoptera’ are severely understudied and (b) the vast majority (>70%) of the Neotropical moth fauna is still undescribed as suggested by the ratio of undescribed species in some 380,000 Neotropical lepidopteran DNA barcodes on Barcode of Life Data Systems (‘BOLD’ ; accessed September 2019). Extrapolating the aforementioned data on species numbers and feeding records we estimate that for >98% of the putatively >100,000 Neotropical moth species authentic feeding records from nature are lacking.
Traditionally, most insect larvae are identified by rearing them to the adult stage and by analysing the morphology of the adult. Methodological constraints in this classic approach are (1) visual search and collecting on plant depending on the skills of the biologist, (2) the canopy region of trees hardly accessible, (3) nocturnal activity of many larvae requiring difficult search by night, (4) collecting without feeding observation may lead to misinterpretations ([14,15]: 20–50% „alternative feeders”on lichens, dead leaves, algae, etc), (5) beating, shaking, net-sweeping may obscure the real where-about of the larva, (6) feeding records in rearing may not reflect the natural host-plant association, (7) rearing to adult is time consuming, (8) rearing may fail (deseases, parasitoids), (9) identification and availability of host-plant (for rearing) often difficult.
Molecular identification of lepidopteran larvae and other insects through DNA barcoding (COI 5’) was repeatedly carried out successfully, e.g. [16,17,18,19,20], permitting an easy, cheap and rapid identification of larvae collected from their host-plants. Identification through DNA barcoding is possible even from dry skins after moulting and from empty pupal exuviae after hatching of the moths (own, unpublished data; ). Currently, there are large-scale projects devoted to the identification of larvae along with their host-plants in Papua New Guinea () and Costa Rica (). Both are based on an integrative approach combining morphology, rearing and molecular techniques for the identification of the reared adults and/or their parasitoids.
Miller et al.  and Matheson et al.  investigated and ascertained relationships between plants and caterpillars through a method based on the DNA identification of the larval gut content, an effective but (in earlier times) expensive and time-consuming approach, especially as a routine application in larger surveys. Later on, molecular gut content analysis was proposed for unveiling insect-host plant associations e.g. for beetles [23,24,25,26], and for soil insects .
The aim of this pilot paper was to establish methodology to infer host-plant relationships of caterpillars based on the identification of larvae collected by insecticidal knock-down (canopy-fogging) on their food-plants through DNA barcoding and to use gut HTS-based content analysis to estimate potential pitfalls due to ‘alternative feeding’ or due to collateral fogging from neighbouring plants, lianas etc. (cf. Discussion).
Material and methods
Collecting and canopy fogging
Canopy fogging was performed by AmH und AF from the ground with a Swingfog SN 50 fogger, using natural Pyrethrum, diluted in a highly raffinated white oil, as knock-down agent to prevent the introduction of persistent chemicals into the environment. For details of the fogging procedure see . In most cases, trees with dense foliage cover and little canopy overlap with neighboring trees were chosen. We made sure the fog reached the canopy and stood there for at least five minutes to affect the arthropods. In order to install the collecting sheets, larger saplings and other interferring vegetation elements were cleared below the tree projection area. All organisms dropping down from the trees were collected at least one hour after the fogging from expanded plastic sheets of 20 m2 size, covering an estimated minimum of 80% of the target tree canopy. All arthropods of each fogging event were pooled and then transferred into accurately labelled jars with 100% ethanol without pre-sorting. The following day the ethanol was renewed and excessive plant material with its high water content was removed. Samples were stored at room temperature for up to two weeks while in the research station Panguana. The ethanol was renewed again when the samples were added to the Zoologische Staatssammlung München in Bavaria, Germany (SNSB–ZSM).
The study site is located in the westernmost Amazonian Basin, eastern central Peru, department Huánuco, at the ACP Panguana station (-9.613393°N -74.935911°E; 222 m; see also ), the fogged target trees were all situated in a radius of less than 2000 meters around the station. Collecting was performed in the late afternoon, betwen 17 and 19 o’clock, from 24th of November to 8th of December 2017. For identification of the target trees see results.
Collection permits were released by Servicio Nacional Forestal y de Fauna Silvestre SERFOR: No. 007-2014-SERFOR-DGGSPFFS + No. 0406-2017-SERFOR-DGGSPFFS, export permits fauna: No. 003236-SERFOR; 003281-SERFOR; 003320-SERFOR; export permits flora: No. 003284-SERFOR + Resolution of General Direction No. 161-2018-MINAGRI-SERFOR-DGGSPFFS; No. 003333-SERFOR. Field site access was granted by SERFOR and Biological Research Station ACP Panguana. Further ethical approval was not required for the data analysis, since no in vivo experiments were performed.
Tissue sampling and identification of larvae (DNA barcoding, COI)
Out of 47 samples–each of them referring to a fogged tree (cf. Table 3 and S1 Table)–all lepidopteran larvae were separated, in total 130 specimens. The larvae were dried on paper, photographed and then separately stored in Eppendorf tubes. A list of all 130 larvae along with their fogging sample number is given in Table 1, examples are shown in Fig 1. Tissue sampling was carried out for all 130 larvae by using scissors and pincers, which were carefully cleaned after each tissue sampling in 100% alcohol followed by exposure to a burner to avoid contamination among samples. Tissues (one vertically cut segment, in very small larvae two segments) were transferred to a lysis plate, adding 0.5 ml of 100% alcohol to each well on the plate. On each plate one well was used for negative control.
(Upper) row 1: larvae nr. 5, 13, 26; row 2: nr. 28, 46, 52; row 3: nr. 62, 65, 71; row 4: nr. 81, 83, 85; row 5: nr. 88, 93, 100; row 6: nr. 109, 113, 115; (bottom) row 7: nr. 119, 122, 124 (numbers and identification of larvae see Table 2).
Tissue samples were submitted to the standard procedures of the Canadian Centre for DNA Barcoding (CCDB) for sequencing the mitochondrial 5’ cytochrome oxidase gene, subunit 1 (COI), the standard marker for the identification of most animals. LepF1 and LepR1 were the primers used for PCR and sequencing . Sequences were blasted against the complete sequence database of the Barcode of Life Data systems (BOLD, ) in order to infere the closest matches using the BOLD Identification Engine (http://www.boldsystems.org/index.php/ IDS_OpenIdEngine). Also morphology of larvae and related (genetically near) adult moths were considered to test the reliability of the results. Nomenclature of scientific taxon names follows the catalogue used on BOLD database, which in many families is in accordance with the currently available catalogues (e.g.  for Geometridae). Vouchers of larvae are stored at the Zoologische Staatssammlung München, Germany. Sequences, images and related metadata are available open access on BOLD under the dataset DS-PANLARVA (dx.doi.org/10.5883/DS-PANLARVA).
Tissue sampling and morphology-based identification of target trees
The 47 target trees have been pre-identified in the field based on morphology (shape of tree growth and shape of leaves, rarely blossoms or fruits) by the native caretaker of the Panguana Station, “Moro” Carlos Vásquez Módena, to Peruvian vernacular names (see Table 3 and S1 Table), which usually cannot be unequivocally referred to scientific plant names, however. For a tentative assignment of vernacular names to botanical taxa see S1 Table. For nomenclature of plant names we follow the “Plant List” (available online at www.theplantlist.org/1/). For most target trees a small branch was collected, pressed and kept in a herbarium for identification. Identification of a selection of sampled leaves was performed by Hamilton Paredes, Museo de Historia Natural, Lima. A small leaf piece was cut as tissue sample for DNA-Barcoding. In addition to that, sapwood/cambium tissue samples were taken of each target tree by using a leather punch to extract a core from the stem. Then, a thin slice of sapwood/cambium was cut and immediately dried over silica gel.
Identification of target trees through DNA barcoding (trnL-F, rbcL & psbA)
Because of the above mentioned uncertainties of target tree pre-identification, we have submitted plant tissue samples to DNA barcoding. For that purpose leaves were available for 37 out of the 47 target trees, pieces of cambium+sapwood for 46 trees. Plant tissues (leaves) were submitted to Sanger sequencing (AIM; Advanced Identification Methods GmbH– www.aimethods-lab.com) with two markers, rbcL and psbA using standardized protocols following [32,33]. An additional attempt was performed in CCDB (Guelph, Canada; primers: trnL-F, rbcL; standard Sanger sequencing procedure) using both leaves and sapwood samples, the latter supplementing those cases where no leaves were available for study. The third marker (trnL-F) was added for further resolving the identification to genus and species level in some cases. All resulting sequences were blasted against GenBank (NBCI) and BOLD data using standard blast functions. Sequences and related metadata are available open access on BOLD under the dataset DS-PANPLANT (dx.doi.org/10.5883/DS-PANPLANT).
Gut content analysis (rbcL, psbA)
For a subset of ten larvae, gut content analysis was tested for molecular identification of the larva’s ‘true’ diet. For that purpose, we performed a second vertical cut and submitted one segment of the larva to High-Throughput-Sequencing (HTS) with the two markers rbcL and psbA. Cut slices of the caterpillars were dried, homogenized and DNA extracted using the DNEasy Plant kit (Qiagen, Hilden, Germany). From each sample, 5 μL of extracted genomic DNA was used, along with plant TAQ (Bioline, Luckenwalde, Germany), and High Throughput Sequencing (HTS) adapted mini-barcode primers (trnH-psbA-f 5’-CGC GCA TGG TGG ATT CAC AAT CC-3’, trnH-psbA 5’-GTT ATG CAT GAA CGT AAT GCT-3’, [32,33], using the PCR conditions 95°C-4’– 35x 94°C-30”/55°C-30”/72°C-1’– 72°C-10’) were applied for PCR. Amplification success and fragment length were observed using gel electrophoresis. Amplified DNA was cleaned up and resuspended in 50 μL molecular water for each sample before proceeding. Successfully amplified products were used for a subsequent PCR reaction which adds Illumina Nextera XT indices to each PCR product, enabling a unique tagging of each sample. Illumina Nextera XT (Illumina Inc., San Diego, USA) indices were ligated to the samples in a second PCR reaction applying the same annealing temperature as for the first PCR reaction but with only seven cycles, and ligation success confirmed by gel electrophoresis (for detailed protocols see [34,35]). DNA concentrations were measured using a Qubit fluorometer (Life Technologies, Carlsbad, USA), and samples were combined into 40 μL pools containing equimolar concentrations of 100 ng each. Pools were loaded into a 1% agarose gel, run at 90 V for 45 minutes, bands of the target amplicon size were excised with sterilized razor blades, and purified with a GeneJet Gel Extraction kit (Life Technologies, Carlsbad, USA), following the manufacturer’s instructions. A final elution volume of 20 μL was used. High-Throughput Sequencing (HTS) was performed on an Illumina MiSeq using v2 (2*250 bp, 500 cycles, maximum of 20 mio. reads) chemistry. Negative controls for DNA metabarcoding analyses consisted of one negative-control-extraction (an empty DNEasy plant kit tube was extracted among the remaining ten caterpillar gut samples), one PCR negative control for each amplicon and one ligation negative control for each set of amplicons during library preparation with Nextera XT indices. Negative controls have been used to remove all OTUs with N(reads)< = 5 x sum of reads in negative controls (where sum of negative control reads is more than 20% of the number of reads in actual samples). All samples for each amplicon were separately pooled using equimolar amounts of 100 ng each. All samples were loaded on a single v2 2x250 bp MiSeq flow cell among other samples. Final DNA concentrations of amplicon pools were set to 380,000 total raw reads (190,000 paired end reads). The final concentration of the full library was 1.4 ng. Metabarcoding data are deposited and accessible on GenBank, BioProject ID PRJNA593715 (http://www.ncbi.nlm.nih.gov/bioproject/593715).
Identification of larvae
A total of 130 caterpillar specimens were collected from 37 of the 47 target plants. No lepidopteran larvae were found in the samples of ten target trees. COI sequencing (DNA barcoding) was successful for 119 larvae (91.5%). The larvae belong to 92 different COI clusters (BINs), which are a good proxy for different species .
When blasting the DNA barcodes of the larvae on BOLD database, 65 larvae (55%) belonging to 48 species showed ‘close genetic similarity’–here defined as lower than 2.5%–with adult reference vouchers. Such genetic similarity is interpreted here as ‘species (or sister species) level matches’ (Table 2). 27 species have Linnean names on BOLD database, 20 are listed under ‘interim names’ (name codes) which either refer to described but not-yet-identified taxa or to undescribed species.
For 32 larvae (27%) belonging to 27 species the blasting on BOLD database revealed genus level matches, in five cases with disputable reliability. For 19 larvae (16%) assignment to subfamily or family level was possible, the reliability of 12 of these assignments needs to be tested by further extension of the reference database, since long branch attraction effects may have influenced the results in a few single cases. In just three cases belonging to one single species no family suggestion could be given based on the COI barcode.
Identification of target plants
Pre-identification of target trees, as performed by the local administrator of the Panguana station (see S1 Table), was supplemented by molecular identification (sequencing of leaves and sapwood with the markers trnL-F, rbcL and psbA) of all but one of the target trees. For all but one of the target trees (98%) molecular identification through blasting on BOLD and GenBank brought a reliable identification to at least family level (see Table 3 and S3 Table). In four cases, however, the analysis of leaf and sapwood pointed to two different families which apparently is due to sampling errors, taking leaf and sapwood from different, neighbouring plants. In 37 target trees (79%) identification was possible to genus or species level (see Table 3 and S3 Table).
Gut content analysis
Gut content analysis was performed for ten larvae based on Next-Generation-Sequencing with two markers rbcL and psbA. The two highest numbers of HTS-reads for rbcL and psbA genes and their genetically most similar species as resulting from BLAST-search in GenBank is shown for each larva in Table 4.
When investigating host-plant relationships it is usually assumed that larvae feed on the plants from where they have been collected. This assumption is based on the behaviour of larvae usually resting on their feeding plant during their development. One needs to consider, however, that certain larvae abandon their host-plants searching for a hidden resting place during daytime and mature larvae often leave their food-plant in the last days before pupation looking for a suitable pupation site, sometimes far from their feeding plants. Moreover, in particular in rainwood forests “alternative feeders” may use epiphytes, lianas, lichens, algae, fungi or mosses [14,15], and in our fogging approach pitfalls are possible through collateral fogging of larvae from neighboring trees. Gut content analysis can shed light on true feeding biology.
Gut content matching identity of target tree
Only in one out of ten analysed larvae (see Table 4) the gut content revealed to match exactly the fogged target tree species: Physocleora AH02Pe (Geometridae; larva nr. 40) fogged from Mangifera indica. In a second case, Homoeopteryx near major (Saturniidae; larva nr. 98), the gut content revealed to be from the same plant family (Rubiaceae), genus Simira resulting from sequencing of plant sapwood and genus Faramea resulting from gut content analysis. The high percentage of eight out of ten larvae with a mismatch between target tree and gut content suggests that an a priori assignation of fogged larvae to the target trees usually is erroneous and that alternative feeding (epiphytes, algae, mosses etc.) or feeding on lianas and neighboring trees plays a major role. The rate of alternative feeding should be tested basing on a larger sample, ruling out a potentially biased ratio through external contamination of larvae by plant DNA (see below).
Gut content matching previously known host-plant but not the target tree
A larva of the genus Quentalia (Bombycidae, larva nr. 42, see Table 4) was fogged from a tree of the family Meliaceae, but the gut content pointed to feeding on Trophis racemosa (Moraceae). Since Quentalia larvae were previously recorded as feeding on Moraceae , Trophis racemosa is likely the true food-plant of the Quentinalia larva which may have been growing close to the target tree.
In a second case, Eudocima procus (Erebidae; larva nr. 65, see Table 4) was fogged from a tree of the genus Hirtella (Chrysobalanaceae) but the gut content pointed to feeding on Tinospora smilacina (Menispermaceae). Since species of the genus Eudocima are known to feed on Menispermaceae [4,37,38], Tinospora smilacina is likely the true food-plant of the Eudocima larva. Tinospora is a liana and likely was associated with the target tree.
A similar case is also referring to liana-feeding: larva nr. 83 (see Table 4) was fogged from a tree of Annonaceae or Malvaceae, but in its gut content we found the DNA of the neotropical liana Echites yucatanensis (Apocynaceae).
Hence in three out of ten cases (30%) feeding on lianas or on a neighboring tree was recorded. Although the rate of feeding on such associated or neighbouring plants should be tested basing on a larger sample, the results of this pilot study clearly show that an ad hoc correlation of target tree and feeding biology is often premature and incorrect.
Gut content not matching target tree but potentially pointing to alternative feeding
In four cases (larvae nr. 82, 102, 107, 108, see Table 4) the larvae were fogged down from trees (genus Paullinia, Sapindaceae; genus Annona, Annonaceae; genus Apeiba, Malvaceae), but the cut content was pointing to alternative feeding on mosses (Bryophyta). In the case of the Lascoria species (Erebidae; larva nr. 107) such alternative feeding is not excluded as larvae of this species were already observed when grazing on algae in Costa Rica . However, moss-feeding is very unusual in Lepidoptera, and this may also be caused by contamination since these fogging samples (under 80% alcohol) contained some leaves of Lejeunia mosses whose DNA may have invaded the larvae through their stigmata or contaminated them on their skin. Further research is needed to estimate the influence of contamination through the sample alcohol. For this purpose larvae should be de-contaminated by bleeching before sequencing. In addition, their gut content could be extracted carefully by cutting the larva longitudinally.
Inferring potential hostplant relationships (larvae without gut content analysis)
43 larvae with reliable identification to at least genus level, fogged from trees identified to at least genus level give first ‘hints’ on potential host-plants (Table 5). Almost all of them are new records, none of them was found in the ‘Hosts’ database  nor in Janzen & Hallwachs . Alternative feeding, however, is not excluded (see notes to larvae nr. 18, 38, 43, 74 and 84 in Table 5), hence all suggested host-plant relationships require confirmation.
Target tree confirming previously known host-plants (larvae without gut content analysis)
Among the 87 larvae successfully identified to genus or species and not subjected to gut content analysis, there are at least six cases where the fogged target trees match previously known host-plant relationships: larvae nr. 63 and 113 (Bombycidae, Anticla near antica) were knocked down from the trees nr. 14 and 34 (Moraceae); larvae nr. 117 (Geometridae; Semiothisa gambaria), 115 (Hesperiidae, Myscelus) and 144+126 (Erebidae, Sosxetra grata) from the trees nr. 35 and 41 (Meliaceae, Guarea), all confirming the relationships as previously recorded by Janzen & Hallwachs .
A powerful tool for future synecological research?
Our pilot study has revealed that (1) molecular identification of fogged, neotropical lepidopteran larvae works successfully in general and even down to species level (if already listed in BOLD), that (2) molecular identification of target trees usually works well at least to genus or family level and (3) molecular gut content analysis based on HTS techniques can be used for confirming or rejecting the feeding on the fogged target tree. With further completion of the DNA reference libraries in the future for (1) (Peruvian Lepidoptera; currently 12,746 sequences, 3532 BINs) and (2) (Peruvian plants) a better taxonomic resolution of identification will be achieved, whilst molecular gut content analysis (3) can be improved by de-contamination and/or isolated storage of the fogged larvae.
With that, the herewith presented approach has the potential for unveiling trophic interactions for primary consumers in tropical regions at a very large scale, which can be performed in a fast and cost-effective way considering the steadily dropping costs for DNA barcoding and HTS. The extremely high diversity of 92 species in 119 larvae in our study shows that canopy fogging and molecular analyses may improve synecological knowledge for a broad spectrum of arthropods. The availability of reliable data on trophic interactions is of great importance for forestry, agriculture, biodiversity and ecological research and–last but not least–for conservation purposes. Increasing such knowledge–particularly in megadiverse ecoregions–is an imperative in a world of unprecedented biodiversity losses. In this context, the proposed molecular approach of investigating host-plant relationships constitutes an important research tool, which fits well in the research plan of the recently launched BIOSCAN phase of the international Barcode of Life program (; see also https://ibol.org).
S1 Table. Morphology-based identification of target trees.
Morphology-based identification of target trees to Peruvian vernacular names (mostly provided by the administrator of the Panguana station, Moro Carlos Vásquez Modena) and attempt to assign scientific family / genus / or species names (partly provided by Hamilton Paredes (“HP”), Museum of Natural History, Lima, based on leaf samples). # = fogging sample from target tree without lepidopteran larva; * = no plant tissue available, so far (hence no molecular confirmation possible): 1 = same vernacular name with two different molecular identifications.
S2 Table. Target trees: Sequencing success and process identification numbers.
Data from BOLD, with fragment lengths in basepairs (bp). Sanger sequencing of rbcL, trnL-F and psbA genes, based on leaf (l) and cambium+sapwood (c) samples from the target trees.
S3 Table. Molecular identification of target trees.
Molecular identification of the target trees after Sanger sequencing (rbcL, trnL-F and psbA genes) of leaf (l) and cambium + sapwood (c) samples. Results from blasting on NCBI, BLAST matches (highest percent identity (‘Max ident’) of all query-subject alignments) usually >99.5%, otherwise indicated. Plant species/genera with blast matches sometimes not mentioned when plants are exclusively distributed on other continents. # = fogging sample from target tree without lepidopteran larva in the sample. Anac. = Anacardiaceae; Anno. = Annonaceae; Cann. = Cannabaceae; Chry. = Chrysobalanaceae; Clus. = Clusiaceae; Euph. = Euphorbiaceae; Faba. = Fabaceae; Malv. = Malvaceae; Malp. = Malpighiaceae; Meli. = Meliaceae; Mora. = Moraceae; Myri. = Myristicaceae; Nyct. = Nyctaginaceae; Rubi. = Rubiaceae; Sapi. = Sapindaceae; Sapo. = Sapotaceae; Viol. = Violaceae. 1 = same vernacular name with two different molecular identifications; 2 = potential sampling error (tissue sampling from neighboring tree or tube flip in the lab process); 3 = exclusively Indo-Pacific; 4 = exclusively Old World.
We thank “Moro” Carlos Vásquez Módena (Panguana) and Hamilton Paredes (Museum of Natural History, Lima) for identification of the target trees. We furthermore thank Julio Monzón (Lima / Freiburg) for providing the fogger and the chemicals. Víctor Meyer Ruiz Chota (Yuyapichis) was very helpful in taking the fogging samples, Johanna Brunner, Sarah Augustin and Stefanie Wothe (Munich) helped to take the cambium/sapwood samples. Dr. Andreas Fleischmann (SNSB, Botanische Staatssammlung, Munich) gave advice concerning the botanical aspects of the study. The genetic analyses have received considerable support from Paul D. N. Hebert and the Biodiversity Institute of Ontario (BIO) and the Canadian Centre for DNA Barcoding (CCDB University of Guelph). The data management and analysis system BOLD was provided by Sujeevan Ratnasingham. This study was part of the program ‘SNSB Innovativ’. We thank the Peruvian nature conservation authority and forestry office (Servicio Forestal y de Fauna Silvestre, SERFOR, Ministerio de Agricultura y Riego, MINAGRI) for collection permits (N° 007-2014-SERFOR-DGGSPFFS; N° 0406-2017-SERFOR-DGGSPFFS) and exportation permits (fauna: N° 003281-SERFOR, N° 003320-SERFOR; flora: N° 003284-SERFOR + N° 161-2018-MINAGRI-SERFOR-DGGSPFFS, N° 003333-SERFOR).
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Chief Prosecutor of U.S. Military Commissions Speaks About Reforms to War Crimes System
New York, December 3, 2012—Criticism of U.S. military commissions, while not unfounded, is outdated, Army Brigadier General Mark Martins told Columbia Law School students, faculty, and guests on Nov. 28, in a presentation on the legal protections afforded to accused war criminals under the Military Commissions Act of 2009.
|Brigadier General Mark Martins|
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|Brigadier General Mark Martins, left, and Professor Trevor W. Morrison '98|

domain: law


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domain: artistic


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(CMR) Former Governor Michael John Edward Gore, who served as the Governor of the Cayman Islands from 1992 until 1995, has died.
Mr. Gore was Governor when Her Majesty The Queen visited the Cayman Islands in 1994, meeting Her Majesty off Royal Yacht Britannia alongside his wife, Monica.
In expressing sadness at the death of Mr. Gore, Governor Martyn Roper said, “Governor Gore was passionate about the environment and biodiversity.”
“As a keen birder, he was instrumental in securing the land, which is now Governor Michael Gore's Bird Sanctuary in Spotts, which is owned by the National Trust for the Cayman Islands and serves as an urban oasis for waterfowl and other birds. He also opened the boardwalk at Tarpon Lake on Little Cayman in 1995,” he added.
“On behalf of everyone in the Cayman Islands, I send my deepest condolences to Governor Gore's wife Monica, family, and friends at this sad time,” Mr. Roper said.
Premier Wayne Panton also expressed condolences to Mr. Gore's family, stating that Mr. Gore was a friend of the Cayman Islands who served the islands well.
“It saddens me and the people of the Cayman Islands to learn of the passing of former Governor of the Cayman Islands Michael Edward John Gore,” he said.
Mr. Panton recalled that in his 1993 Throne Speech, Mr. Gore said, “Cayman is one of the more fortunate countries in the world with a standard of living and, perhaps even more important, a quality of life second to none. I should like to take this opportunity to pay tribute to the churches of all denominations who look after our spiritual well-being and serve the community in so many ways, to the voluntary organizations who look after those less able to look after themselves or concern themselves in areas which are outside the scope of Government, and to all residents who contribute to make the Cayman Islands such a wonderful place in which to live.”
The premier said, “The contributions of Gov. Gore to our country are immense and we remember him when we observe the Governor Michael Gore Bird Sanctuary, which contains the freshwater marshes of the Spotts-Newlands area which is a roosting and breeding habitat for several freshwater bird species.
As a matter of respect, the Union Flag and the Cayman Islands Flag are being flown at half-mast today and will fly at half-mast again on the day of his funeral or the day before if it falls on a weekend.


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By Michael Taylor,Nigel Thrift
This quantity charts the ways that multinational agencies contributed to the restructuring of the area economic climate, paying specific cognizance to the spatial outcomes of, and responses to, their operations at a couple of scales. The booklet takes as its subject the differential spatial results of the restructuring of alternative forms of multinational company.
By Mohammad N. Elahee,Mohammad Elahee,Farid Sadrich,Mike Wilman
By Michael J. Morley,Noreen Heraty,Snejina Michailova
Against the backdrop of historic cultures, a communist legacy and eventual institutional atrophy, a few of the societies of vital and japanese Europe have pursued competitive improvement trajectories because the early Nineteen Nineties. This a part of Europe is now characterised through a emerging fiscal heterogeneity and a swiftly altering socio-cultural context, underscored through waves of restructuring, privatization, expanding international direct funding and an rising individualism. whereas there was a transforming into curiosity within the transition economies some time past variety of years, the modern nature of human source administration in those societies isn't well-documented.
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By William H.A. Johnson
in growing new applied sciences and at one time was once the innovation king of the realm- yet that was once not less than seven centuries in the past! this day, the good oriental strength is making an attempt once more to take the throne of innovation for its personal. This wish to usurp the throne, which have been diligently taken by means of the West throughout the clinical Revolution, has put a virtually unrealistic emphasis on innovation. In Innovation in China: The Tail of the Dragon, the writer explores the problems and actors eager about making innovation the emphasis in China. He makes use of an easy platforms version of innovation and diverse perceptual lenses. The lenses are aimed toward the old, monetary, political, felony, academic and cultural parts of an innovation-based society. After interpreting the booklet the reader will comprehend extra approximately how innovation is going on in China and
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By Chekdar Bavli
By Vyuptakesh Sharan
While conserving the foremost contents of the sooner variants, the recent variation offers a transparent realizing of the most recent instruments and strategies utilized in overseas finance. The e-book bargains with modern monetary surroundings and methods, and the practical in addition to operational features within the foreign monetary area.
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Structural Revolution in foreign enterprise structure quantity 2 fills very important gaps within the latest literature of administration technological know-how by way of delivering new and superior equipment of optimum regulate method modeling. those study equipment are utilized in numerous difficulties of administration technological know-how and nationwide monetary administration. functions are on oil box improvement, strength method modeling, source modeling, time various regulate of dynamic procedure of nationwide economic system, and funding planning.
By L. Yuan
By Paul Glewwe
Surveying many elements of education—from administrative constructions to the provision of future health care to father or mother and pupil incentives—the individuals synthesize a magnificent variety of information, paying exact recognition to the gross imbalances in academic success that also exist among constructed and constructing nations. They draw out transparent implications for governmental coverage at numerous degrees, aware of monetary realities resembling finances constraints, and element to the most important components the place destiny examine is required. providing a wealth of insights into the most effective investments a kingdom can make, Education coverage in constructing Countries is a vital contribution to this so much pressing field.
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International NGOs are more and more vital gamers in the new relief structure yet their geographic offerings stay uncharted territory. This ebook specializes in styles of improvement tips, mapping, whereas analysing and assessing the rustic offerings of the biggest foreign NGOs. Koch's procedure is interdisciplinary and makes use of qualitative, quantitative and experimental ways to supply a transparent perception within the determinants of kingdom offerings of foreign NGOs.
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The publication is the 1st at the geographic offerings of overseas NGOs, and is as a result of substantial educational curiosity, in particular for these concentrating on improvement relief and 3rd area examine. in addition, the e-book presents particular coverage feedback for extra thought-out geographic judgements of foreign NGOs and their again donors.

domain: literature