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Myanmar - People and Life
SOCIETY, RICH CRONIES AND POVERTY IN MYANMAR
SOCIETY IN MYANMAR
Burma-Myanmar has feudal traditions, Buddhist traditions, democratic traditions but no caste system. According to the Joshua Project: "The single most important social institution in the village is the temple. It symbolizes unity among the villagers, and provides a wide variety of activities for the people. The Burmese do not recognize clans or lineages. Marriages are monogamous, and rarely arranged by the parents
According to Countries and Their Cultures: "Not only is poverty widespread, there is marked inequality. Essentially, the society is divided into a tiny elite, a fairly small middle class, and a large number of very poor people. While there are traditional elites within most of the ethnic groups and new elites in some groups whose wealth comes from smuggling, the national elite is overwhelmingly Burmese. In recent years income from the narcotics trade has been an important source of wealth for members of the elite. Although some segments of the middle class have prospered from the economic reforms of the late 1980s, most have not done well and remain poor. [Source: Countries and Their Cultures everyculture.com ]
The middle class is increasingly becoming more political involved. "In Burma, the middle class is very thin," said a 38-year-old graphic designer who in 2004 helped found an undercover nonprofit group that recruits potential political leaders, told the Washington Post. "We need to grow, strengthen that. Most democratic countries have a broader middle class. It is the only way to go forward."
See Repression
Rich in Myanmar
Many of those who are considered rich in Myanmar are either connected with the military regime or have connections to the drug trade or gem and timber smuggling enterprises. The most visible are ethnic Chinese and the sons and daughters of the military elite. Some or Myanmar's elite live in white concrete and glass, neo-Thai mansions outside of Mandalay. The Rangoon elite have traditionally lived around the lake Aung San Suu Kyi lives on central Yangon.
A diplomat told Newsweek, "It's tale of two nations. The elite live in the First World of Rolexes, Land Rovers and gold rings. The rest live in the 'Third World.'"
Life of Myanmar's Generals
The military leaders live in isolated compounds with schools, hospitals and housing that are much better than those of ordinary Burmese. The expensive villas of the top officers have satellite dishes, huge lawns, polished teak floors, high ceilings and gorgeous gardens. These officers drive fancy cars, eat sandwiches with the crust cut off delivered by servants, and spend their time playing golf. Before the their new capital in Naypyidaw they like to play at the Yangon City Golf Resort, where the membership in the 1990s was $3,000 (20 times Myanmar's per capita income at that time).
The generals enjoy things—such as cell phones, computers, air conditioners, Rolexes and Land Rovers—that not every one else in Myanmar has access to. They always have electricity and water unlike the rest of the country which often receives these services only a few hours a day. In 1997, the military paid $.15 a gallon for gasoline while ordinary people paid $1.75.
The children of the generals have become Myanmar's spoiled elite. They wear expensive hip-hop fashions, dance to rap and techno music, drink $3 beers at the Yangon's Galaxy disco and watch bootleg DVDs of "Titanic" or "Star Wars: the Phantom Menace" . The irony of all this is that ordinary Burmese are denied these things because they are regarded as too Western and decadent.
Many of the homes and offices of the ruling generals in Myanmar have teak-paneled reception rooms, teak-paneled bedrooms, and teak-paneled hideaway bars.
The generals in the military regime love to play golf. They play on courses built when the British controlled the country and on a few new ones built by businessmen with connections to the generals. Brook Larimer wrote in Newsweek: "Every mourning at the City Golf Course n Rangoon, dozens of military officers in creased khakis and saddle shoes traips off the first tee, followed by platoons of young female caddies. The girls, who wear bright red lipstick and easy smiles, perform different jobs for their 35-cent fee: one hauls the clubs, another holds the parasol, one lines up the putts—and all applaud politely after each successful shot."
See Separate Article MILITARY RULE AND MYANMAR'S SUPERSTITIOUS, HIGH-LIVING AND PARANOID GENERALS
Hill Station for the Generals
The two military academies in Pin U Lain, a new town built from scratch near the British hill station Maymo, have new buildings and young cadets walk about in sharp uniforms. The golf course has a helipad.
Describing Pin U Lain, Bertil Lintner wrote in the Washington Post, "Built in the lush hills northeast of Mandalay, the new town is a kind of refuge — but for the Burmese military. Instead of the British Victorian-style mansions of the old Maymyo, you'll find gaudy luxury villas in the new one. The town is also home to the Defense Services Academy, Burma's West Point, which trained many of the generals. [Source: Bertil Lintner, Washington Post, September 30, 2007 \/]
"When construction on the officers' town began in late 2005, the Irrawaddy, a magazine published by Burmese exiles in Thailand, reported that "no expense has been spared to allow the generals to live in what basically is a resort, complete with an artificial beach and a man-made stretch of water to lap onto it." The theme-park retreat will also include replicas of a famous pagoda in Rangoon, the old royal palace in Mandalay and a popular beach resort — which, the magazine dryly noted, "is probably where the fake beach comes in." \/
Thanks to a newly upgraded airport, the retreat is a quick plane ride to Burma's new capital, Naypyidaw, built in the wasteland and jungle 200 miles north of the old capital, Rangoon. Naypyidaw means "Abode of Kings," and kings are precisely what the Burmese generals see themselves as. On the capital's parade ground stand newly erected, larger-than-life statues of three famous pre-colonial warrior kings whom the junta's leader, Senior Gen. Than Shwe, sees as his role models. \/
See Separate Article on Naypyidaw
Than Shwe's Daughter's Lavish Wedding
One of Than Shwe's few trips outside the capital was to his daughter's wedding in Yangon, in 2006. The event angered many Burmese because it cost $300,000 and the couple received wedding gifts worth $50 million, according to The Irrawaddy. Michael Casey of Associated Press wrote: Than Shwe's "government suffered a rare paparazzi-style scandal, when a 10-minute video clip, filmed at the wedding in the old capital Yangon, surfaced on the Internet purporting to show the bride, Thandar Shwe, swathed in sumptuous jewels - revealing the utter disparity in wealth between the military elite and the impoverished general population. The champagne, five-star comforts and other opulence became a sore point among exile-based dissidents and the butt of jokes mocking Than Shwe and the junta's insistence that his military regime is not corrupt." The same week the video appeared Transparency International ranked Myanmar, along with Somalia, as the most corrupt country in the world. [Source: Michael Casey, Associated Press, October 1, 2007]
Jonathan Watts wrote in The Guardian: "Strings of diamonds, cascades of champagne and tens of millions of dollars worth of gifts would be considered ostentatious at any wedding. But in Burma - one of the poorest countries in Asia - people are said to be up in arms at the luxury on display in a video of the wedding laid on by the head of the junta, General Than Shwe, for his daughter...Opponents of the military regime claim that spending on the couple's marriage in July was more than three times the state health budget. [Source: Jonathan Watts, The Guardian, November 2, 2006 /]
"In the most opulent sequence, the camera zooms in on glittering jewelled clusters in the hair of the bride, Thandar Shwe, then pans down from her diamond ear-studs to at least six thick strings of what appear to be diamonds. At a lavish reception, the groom - Major Zw Phyo Win, an army officer and deputy director at the ministry of commerce - pours champagne over a cascade of glasses and helps his bride slice into a five-tiered cake. What is not seen are the gifts, which reportedly include luxury cars and houses worth a total of $50m (£26m). According to south-east Asian newspapers, the rush to buy jewels as presents and decorations pushed up the price of precious stones in the run-up to the wedding. The wedding video appears to have been filmed with the approval of the the married couple and guests. It is unclear how it was leaked on to the internet or how widely it can be seen in Burma. /\
"Such mindless indulgence - smiling, well-fed guests wrapped in their finest clothing and most expensive jewels - is an affront to the millions of Burmese suffering under the incompetence and brutality of the country's military leadership, and the millions of Burmese migrants trying to scratch out a living on foreign soil because no proper employment is available at home," wrote editor Aung Zaw in Irrawaddy. "Than Shwe was the one who accused other top leaders of corruption whenever he wanted to remove them. It's the pot calling the kettle black." /\
"The minutiae of the wedding arrangements provided material for observers of the secretive regime who believe Than Shwe may be preparing to step back from the day-to-day running of the country. "In the seating arrangement, Than Shwe and his deputy were on one table and all the other junta members were on a very distant table. That tells you a lot about the hierarchy," Soe Aung of the Bangkok-based National Council for the Union of Burma was quoted as saying by Reuters. In footage of the ceremony at a state hall in Yangon, Than Shwe walks beside his daughter in white shirt and a traditional orange wrap called a longgyi, a rare sight of a general almost always seen in military uniform. Many other guests were in uniform. /\
Wikileaks Cables: Myanmar General Considered Buying Manchester United
Former Myanmar leader Than Shwe once considered spending a billion dollars to buy Manchester United as a gift to his grandson, a soccer fan.
Robert Booth wrote in The Guardian: "The leader of Burma's military junta considered making a $1 billion (£634m) bid to buy Manchester United football club around the time it was facing rising anger from the United Nations over its "unacceptably slow" response to cyclone Nargis.Than Shwe, commander in chief of the armed forces and a fan of United, was urged to mount a takeover bid by his grandson, according to a cable from the US embassy in Rangoon. It details how the regime was thought to be using football to distract its population from ongoing political and economic problems. The proposal was made prior to January 2009; only months earlier, in May 2008, the Burmese junta had been accused of blocking vital international aid supplies after Nargis struck, killing 140,000 people. [Source: Robert Booth, The Guardian, December 6, 2010 ^]
"Than Shwe reportedly concluded that making a bid for United might "look bad" at the time, but the revelation that the proposal was even considered is likely to fuel criticism of the regime's cruelty. The senior general instead ordered the creation of a new multimillion dollar national football league at the same time as aid agencies were reporting that one year on, many survivors of the cyclone still lacked permanent housing, access to clean water, and tools for fishing and agriculture. ^
"The mooted price tag for Manchester United was exactly the same as the aid bill to cover the most urgent food, agriculture and housing for the three years after the cyclone, as estimated by international agencies including the UN. The proposal revealed that the regime, which is increasingly exploiting its oil and gas reserves, felt confident of finding such a sum. According to Forbes magazine's valuation of the club at the time, $1 billion would have been enough to acquire a 56 percent controlling stake. ^
"One well-connected source reports that the grandson wanted Than Shwe to offer $1 billion for Manchester United," said the June 2009 cable to Washington. "The senior general thought that sort of expenditure could look bad, so he opted to create for Burma a league of its own." Than Shwe then reportedly coerced and bribed eight leading business and political figures to establish teams and ordered them to spend large sums on imported players and new stadiums. ^
"The cable revealed that in January 2009, selected Burmese business people were told "that Than Shwe had 'chosen' them to be the owners of the new professional soccer teams. [The informant, a top executive at one of the sponsor companies] said the owners are responsible for paying all costs, including team salaries, housing and transportation, uniform costs, and advertising for the new league. In addition, owners must build new stadiums in their respective regions by 2011, at an estimated cost of $1 million per stadium." ^
"The Magway team was spending $155,000 a month on salaries while the Kanbawza team, linked to a bank, had budgeted $2 million for the 2009 season. Rangoon United hired five players from Africa and Delta United recruited several Argentinians. "When asked why the owners would participate in such an expensive endeavour, [an executive with one company sponsor] observed that they had little choice," the embassy reported. "'When the senior general asks someone to do something, you do it with no complaints,' he stated." ^
"He added that several of the business people expected to receive incentives from the regime, such as construction contracts, new gem and jade mines, and import permits, which would more than offset their costs. The owners of the clubs in the Burma national football league, which launched on 16 May 2009, include "regime crony" Zaw Zaw, who also chairs Burma's football federation and drew up plans for the league with the senior general's grandson. "Zaw Zaw hired Senior General Than Shwe's grandson to play on the team," a separate cable adds. But according to the dispatch, "many Burmese businessmen speculate the regime is using it as a way to distract the populace from ongoing political and economic problems or to divert their attention from criticism of the upcoming 2010 elections". ^
Cronies in Myanmar
Simon Denyer wrote in the Washington Post, "A new English word has entered colloquial Burmese, a word that could not even be uttered in public until recently. The word is "crony," and it describes the business elite who exploited their closeness to the country's military rulers to amass vast wealth in the past two decades. These well-connected elite made their money in industries such as construction, rubber and logging, as well as in arms dealing and drug smuggling. Their gains have only increased in the past two years, a result of changes that have privatized many state-owned assets and enterprises — and allowed the rich to buy them up at bargain prices. [Source: Simon Denyer, Washington Post, March 26, 2013]
"Burma's business elite have been investing some of their wealth by erecting hotels and office buildings in Rangoon and other cities. But outside these gleaming new buildings, cycle rickshaws still ply the streets, and there are few signs of a more general boom. Small-business owners and shopkeepers say consumer demand remains tepid. The business elite, say critics such as Zaw Aung, a former political prisoner who is a research fellow at Thailand's Chulalongkorn University, have the power to crush potential competitors, corner the benefits of Burma's reform process and prevent a new, more diverse middle class from emerging.
Aung Zaw wrote in The Irrawaddy, "Indeed, there are many questions that need to be asked. Can cronies become builders of industry and national economic power? How can they contribute back to society by building philanthropic foundations and provide life-long assistance to society? Many showy tycoons and cronies in Burma are not interested in helping society. In fact, critics have charged that contributions from cronies are tiny compared to the money they spend on their posh Italian sports cars." At same time some "cronies have quietly supported Suu Kyi and the opposition movement and donated to the Burmese community. [Source: Aung Zaw, The Irrawaddy, January 28, 2013 ////]
"Sean Turnell, an expert on Burma's economy at Macquarie University in Sydney, Australia, thinks the cronies are destructive and resistant to reform. "I think the majority are cronies of the destructive sort—but some might turn out for the better." "They are rent-seekers pure and simple rather than builders of genuine enterprise," he added. "[They are] living off government regulatory largesse, the recipients of monopoly and quasi-monopoly profits and so on. As such, they are political animals as much as economic ones. But certainly there are some too who may emerge as something else. On this front, I guess we have to hope so, since they are amongst the few with sufficient capital to do transformative things, if this is what their desire is." ////
Jill Drew wrote in the Washington Post, ""All they know is stealing," seethed one taxi driver as he took a passenger on a circuitous route to the airport, slowing in front of the house of Tay Za, the owner of a local airline who is close to Senior Gen. Than Shwe, leader of the junta. The villa had an open garage, with two Ferraris inside, one red and one yellow. "They want money, money, money. And we have nothing," he said. The driver keeps a notebook hidden under newspapers on his dashboard. In it he writes, in Japanese characters, how the government controls gasoline sales to siphon money for themselves. He wants to smuggle the notebook out of the country so foreign media can report on the system. The government limits official gas sales to two gallons a day. To buy more, drivers must purchase black-market gasoline — obtained by sellers who pay kickbacks to government-appointed filling station managers — at nearly double the official rate.[Source: Jill Drew, Washington Post, October 24, 2007 \]
See Separate Article LOCAL GOVERNMENT, BUREAUCRACY, CRONIES AND CORRUPTION IN MYANMAR
Former Imprisoned Crony Still Looking for Ways to Make Money
The Washington Post reported in 2008: "The climate of nepotism and capricious junta policies means that uncertainty pervades even among the most seemingly successful. In his sparsely furnished living room, an avowed former "crony" of senior generals recounted how he grew a small logging firm that traded rosewood and teak to China into a sprawling foreign investment firm that eventually bankrolled three ministers and a mayor, all of them senior military officers. In return for supplying licenses and contracts, the four received large deposits in private Singapore bank accounts, he said. [Source: Washington Post, August 16, 2008 /|]
"Profits, however, one day started to slip, the deposits to those bank accounts slimmed, and the businessman was thrown in jail, charged with the very thing that swelled the officers' accounts, he said — using a local company as a front for illicit foreign dealings. But nearly eight years behind bars hasn't dissuaded him from attempting another trek down Burma's twisted path to prosperity. Only six months since he was released, gray-haired and frail, from Insein prison, he says he searches the Internet daily for information on how to tap the booming emigrant industry — funneling unskilled Burmese workers to jobs outside the country. "This is not a legal way. It is a form of trafficking," he said. For help, he said, he would be turning to old friends in the Home Ministry. As for his clients, he added, they don't really know what they're getting into. But "if they have a chance to go abroad, they can make money." /|\
Zaw Zaw: One of Myanmar's Premier Cronies
Simon Denyer wrote in the Washington Post, "Zaw Zaw, the head of the giant Max Myanmar conglomerate, is one of Burma's richest men, having made his fortune through lucrative government contracts, helping to build the country's new capital, Naypyidaw, in the past decade, and constructing roads and extracting tolls. He profited handsomely when state-owned assets were sold off in 2011 in the largest privatization push in Burma's history, picking up a banking license and cement factory. He runs roadside gas stations, controls the auto import trade, owns a jade mine and rubber plantations, and is fast expanding into luxury resorts. Zaw Zaw insists that he pays taxes and creates jobs, and he says Burma will not prosper if everyone in the business community is vilified as being cronies. "At this time, we all have to cooperate together to build the country," he said. [Source: Simon Denyer, Washington Post, March 26, 2013]
Aung Zaw wrote in The Irrawaddy, "Zaw Zaw—who is still in his mid-forties— is media savvy and friendly. He will proudly tell visitors and the media that he once washed dishes in Japan before coming back to Burma to run his own business selling used cars and later getting involved in the jade mining business in Kachin State. "I have nothing to hide," he told me. He was a university student during the 1988 uprising in Rangoon and he witnessed the crackdown and his fellow students being gunned down. [Source: Aung Zaw, The Irrawaddy, January 28, 2013 ////]
"Since his early days, Zaw Zaw's business empire has expanded considerably. In addition to his mining interests, he now has his own bank (Ayeyarwady Bank, one of the largest in Burma), a cement factory, gas stations and a major construction company. The latter company was awarded numerous lucrative contracts in Naypyidaw, the new capital, including a stadium for the 2013 Southeast Asian Games. ////
"Zaw Zaw may be rich, but he also know that he needs to contribute to society. In 2010, he set up the Ayeyarwady Foundation, a charitable organization. Since then, he has been building schools across Mon and Karen states and Irrawaddy and Mandalay divisions. Recently, the chairman of the Max Myanmar also attended the wedding of a former student leader and member of the 88 Generation Students group.////
Tay Za: Another One of Myanmar's Premier Cronies
Makoto Ota wrote in the Yomiuri Shimbun: "A mysterious tycoon lurks behind Myanmar's military government. Tay Za, said to be 43 years old, is known for his close relationship with the junta leadership. The United States earlier this month imposed fresh sanctions on Myanmar, freezing bank accounts of businesses close to the junta, apparently targeting Tay Za. But observers have said the sanctions will not sway the tycoon, who is said to be cunning enough to help the junta in skirting most U.S. and EU sanctions. [Source: Makoto Ota, Yomiuri Shimbun, October 30, 2007 ||||]
"Tay Za built his fortune since 1990 through logging, teak log exports, hotels and tourism. The exact size of his fortune and personal details are not known, but his close proximity to the junta and its help in his success is well reported. He is the owner of Air Pagan — in which the wife of the junta's leader, Senior Gen. Than Shwe, is an investor. Tay Za also is the only authorized import agent for the Russian military industry. He, together with Vice Senior Gen. Maung Aye, the junta's No. 2, clinched a deal to purchase MiG-29 fighter-bombers from Russia in 2002. It also is rumored that Tay Za helped, through Air Pagan, Than Shwe's wife and others leave the country for locations such as Vientiane and Dubai during the peak of September's antigovernment demonstrations. ||||
"Tay Za is said to have numerous subsidiaries, bank accounts and luxurious condominiums in Singapore, and he has "no trouble lending his money out," according to a source in Yangon. This diversification means the U.S. sanctions, which only freeze assets in the United States, probably will have little impact on the tycoon. ||||
Aung Zaw wrote in The Irrawaddy, "Tay Za is known to be close to Burma's senior military leaders, including ex-dictator Snr-Gen Than Shwe....However, he told me that he never met the reclusive former strongman until after his helicopter crashed on a snow-capped mountain in the far north of Kachin State in February 2011. Than Shwe—who refused for a full month to allow foreign aid workers into Myanmar after Cyclone Nargis claimed more than 140,000 lives—immediately ordered hundreds of troops to conduct a search-and-rescue mission for Tay Za and his crew, all of whom survived. Tay Za told me he later went to the residence of the recently retired junta supremo to express his heartfelt gratitude. Tay Za also quietly met Suu Kyi soon after her release in November 2010. He had reportedly offered to assist the NLD. Party sources told me Suu Kyi did not reject his offer. [Source: Aung Zaw, The Irrawaddy, January 28, 2013 ////]
Myanmar of Cronies Try to Improve Their Image
Simon Denyer wrote in the Washington Post, "Faced with a lot of bad publicity, the cronies have been fighting back. Some have warned of legal action to try to silence critics; Khin Shwe has threatened to attack the farmers who have occupied his land. But most have been trying to improve their images — to make themselves more acceptable business partners in the new Burma, because they fear retaliation and losing their wealth, or simply because they don't like criticism. "It gives me so much pain, I often cannot sleep at night," Zaw Zaw, the head of the giant Max Myanmar conglomerate, said in a rare interview last month while watching a soccer match in Rangoon, the former capital, also known as Yangon. [Source: Simon Denyer, Washington Post, March 26, 2013]
Aung Zaw wrote in The Irrawaddy, "One Rangoon–based observer said that Zaw Zaw and other cronies need to show not only that they support current political reforms, but also that they are willing to make a long-term commitment to the development of civil society. They should also return land that they acquired under the former regime, he said. "The cronies must show that they are part of the solution, not part of the problem," the observer added. [Source: Aung Zaw, The Irrawaddy, January 28, 2013 ////]
Providing cash and building schools and hospitals here and there isn't enough, one Rangoon-based diplomat said firmly. Some cronies now realize that strong recommendations from Suu Kyi and prominent activists such as Min Ko Naing and Ko Ko Gyi and other actors in the civil society movement are important, as the US is closely monitoring them.
Burmese Nouveau Riche
Foreign investment in the 1990s created a class of instant millionaires in Yangon, opened a yacht club, three new golf driving ranges and nightclubs that sold lobster for $100 a plate. At that time there were many new German, Korean and Japanese cars even though as of 1996 Myanmar didn't have a single new car dealership.
Myanmar greatest entrepreneur in the 1990s was Thien Tun, a man who went from being bean exporter in 1990 to an owner of hotels, factories, and a bank and the exclusive importer of a number of items, including Safeguard soap and Chivas Regal Scotch. His first major venture, a Pepsi bottling factory churned out 600,000 bottles of soft drink a day in 2000. At that time there were plans to boost the figure to 5 million bottles after a new plant is opened in Mandalay. "I wouldn't say it's a gold mine, but it's a good investment" Theien Tun told the Los Angeles Times. "I already do gold mining, and up to now, I haven't made money there yet." he said his motto is: "Low profile, move fast, make money, live peacefully."
When Thein Tun was contacted by Pepsi-Cola in 1990 about forming a joint venture, he had never heard of the soft drink company but his son Than Zin Tun—a handicraft dealer in Thailand—told him, "Daddy trust me, this business can't fail." Among Tun's planned joint ventures were a brewery with Carlsberg, a whiskey distillery with Seagrams, an "energy" drink factory with Japan's Marubeni, a plastics factory with South Korea, a Mitsubishi car distributorship and a gas turbine power plant with Hong Kong's China Light and Power Co. [Source: Michael Hirsh and Ron Moreau, Newsweek, June 19, 1995]
Khin Shwe, the owner of hotel and construction company that posted revenues of $60 million in 1994, at that time owned a large home, three new cars and 14 thoroughbred horses.
Poverty in Myanmar
Myanmar is one of the world's poorest countries. Population below poverty line: 32.7 percent (2007 est.) The United Nations ranked Myanmar 138 out of 166 countries in its 2009 Human Development Report and ranked it as one of the twenty poorest countries in the world, with an estimated annual per-capita income of five hundred dollars. The per capita income in Myanmar is less than half that of India and below Bangladesh and Nigeria.
In the 1990s a lack of foreign exchange was offered as an excuse why there was nothing—no brooms, no soap, no toilet paper. Many people could only afford to have meat once a week and water generally had to be boiled before it could be consumed. The government spent a lot of money on supporting the army and buying new military hardware while most of the people lived in poverty.
Thomas Fuller wrote in the New York Times, "Many if not most of Myanmar's people are either too poor or too isolated to raise their voices. Their main preoccupation is survival. The World Food Program has quadrupled the amount of food it distributes in Myanmar over the past four years, but still feeds only 300,000 people, a fraction of the 5 million who it says do not have enough food. Tony Banbury, the regional director of the U.N. World Food Program, says poverty and malnutrition in Myanmar are "one of the sad, forgotten stories of Southeast Asia." "There were a lot of wars and conflicts in Southeast Asia, but most countries have moved on," he said. "There are millions of people in Burma that the world has simply forgotten about." [Source: Thomas Fuller, New York Times, October 25, 2007]
Thousands of people have died of AIDS because of the near-absence of retroviral drug treatment outside Rangoon and Mandalay. In 2008, the government failed to provide aid to millions of people affected by a devastating cyclone in the Irrawaddy River Delta. In early 2012, Myanmar President Thein Sein said The government aimed to cut the poverty rate from 26 percent of the resource-rich country's 60 million people to 16 percent by 2015 and use "all means possible" to fight graft, which he said was one of the biggest threats to the country's progress. [Source: Aung Hla Tun, Reuters, March 1, 2012]
"The World Food Programme [WFP] provides food aid to 500,000 people across Myanmar [Burma] but that really only represents the poorest of the poor," Paul Risley of the WFP told the BBC. "What we've found is that over the last decade, opposite to virtually every other country in Asia where slowly poverty is being gnawed away at and food security is becoming more commonplace, in Myanmar there are more people living below the poverty line and more people facing food insecurity," he said.
Poor People and Poverty in Myanmar
In the Irrawaddy delta, where many Burmese villagers live, the poor live meal-to-meal in flimsy thatch huts on bamboo stilts along coffee-brown rivers and rice paddies, according to AFP. After being isolated from the rest of the world for so long, many are used to expecting very little in a country where running water and electricity are still considered luxuries in many areas.
In 2007, Andrew Marshall wrote in Time, "Burma has a grave and worsening humanitarian crisis. Half of Asia's malaria deaths occur here; a third of the children under 5 years old are malnourished; most of its people live on less than a dollar a day. "People have been successfully intimidated into keeping their heads down — maybe," says Shari Villarosa, chargé d'affaires at the U.S. embassy in Rangoon. "But it's still a struggle for them to survive — to feed and educate their families, to get health care. So there could be another eruption." [Source: Andrew Marshall, Time, October 11, 2007]
In the 1990s, the United Nations estimated that one in three children under five are malnourished, a condition that makes children who 15 and 16 look nine or ten. To earn money to pay off loans for food some families sent their daughters to Thailand, where they often were forced to work as prostitutes and returned home with the HIV virus. When trains stopped at bridges people sometimes approached the windows and moaned "We are hungry. Have pity." The passengers on the train usually toss them food. One woman told Swerdlow, "It wasn't like this when I was young." [Source: Joel Swerdlow, National Geographic, July 1995]
On the streets of Yangon you can see children sleeping on pieces of cardboard, groups sleeping on bundles, women with malnourished children, and vendors selling worthless objects. In Mandalay, people ties bags with leftovers to lampposts to give to the poor and beggars of all ages are a common sight at Maha Myat pagoda, "We're not hungry or dying," a Rangoon shopowner told the Los Angeles Times. "But there's a lots of hardship. People with contacts in the ruling class get richer, but the average person can barely make ends meet."
Myanmar's Poor Eek Out a Living with Leftover Gold and Jade
Reporting from Kharbar, Myanmar, Paul Watson wrote in the Los Angeles Times, "Squatting along the rocky banks of the Nmai Hka River, villagers labor from dawn till dusk over large wooden pans, scrounging for crumbs from the junta's table. Children barely big enough to swirl the heavy slurry toil alongside men and women, doing backbreaking work that exposes them to toxic mercury. Every few minutes, they pause and tilt their dripping pans to catch the sunlight, hoping for the glint from a few golden flecks that haven't been scooped up with the rest of Myanmar's vast mineral wealth by the ruling generals and their cronies. [Source: Paul Watson, Los Angeles Times, December 24, 2007 |]
"On a recent day by the river, Ja Bu, 46, strained to lift shovel loads of slurry as a 10-year-old boy, ankle-deep in the cold, muddy water, worked a pan big enough for him to bathe in. Sixty miles west, Ja Bu's younger brother was searching for jade in the drainage ditch of a mine exhausted years ago by the junta. The few dollars that Ja Bu and her brother manage to scratch together each day from what the generals didn't take buys food, clothes and shelter for 10 people. |
"The junta tightly controls access to its large gem and jade mines, but remote places such as Kharbar" are beyond their reach. "But the junta doesn't let much trickle down to places like Kharbar, a remote northern stretch near the Himalayan foothills, close to the Chinese border. It's a spectacularly beautiful, unforgiving place where villagers live in thatched huts with walls woven from bamboo. Thin as cardboard, they are flimsy shelter against frigid winter winds. And as the cost of food and fuel rises, so does the villagers' resentment, which roils like the rapids of the Nmai Hka that taunts them with tiny gifts of gold. |
"Dong Shi, a wiry man in a green sweater splitting at the seams, has been working the brown slough and bamboo sluices here for three years. On a good day, he finds $8 worth of gold flakes, the biggest about the size of a pinhead. Like other prospectors, he must pay $250, or more than half an average person's annual income here, to the owner of the land for permission to pan just 10 square feet of riverbank.After Dong Shi pays his stake's owner, his share of the diesel to run a generator and sluice pumps, school fees for his four kids and other mounting expenses, he has little left. "We eat all that I earn," he said. "I have nothing left in my pocket. Tomorrow I go back to work on the river, just to have some more food." |
"It is grueling, risky work. To separate gold particles from the slurry, miners squeeze drops of mercury from strips of cloth soaked in quicksilver, exposing them and the river fish they eat to dangerous levels of the heavy metal, which can damage kidneys and the nervous system. For all the prospectors' pain and risk, most pans come up bust. So they dig deeper, push the limits harder.Desperate to hit pay dirt, dreaming of finding a rare nugget instead of just flecks, some villagers rig up hand pumps onshore to homemade breathing hoses, and wade to the middle of the river. They work up to three hours at a time underwater. |
"Child labor is an essential part of production at the bottom end of outdoor factories that surround Mandalay's jade market. Children huddle on their haunches around glowing embers in metal braziers, melting doping wax on the end of dop sticks, plucking small pieces of jade from a cup, and carefully placing them on the wax blobs. They blow gently to harden the seals and then hand the sticks up the line to other children. |
"On a recent day, one boy sat on the edge of a stool, stretching his leg to reach a wooden pedal that he pumped to spin a bamboo cylinder, wrapped in sandpaper, as he ground pieces of jade to a refined sheen. Once they'd done their best with small hands scraped by the grinders, the boys passed the jade along to men. It takes an experienced hand to get the shimmering polish that will bring the best price, a small piece of the profits that keep Myanmar's military in power. That can't be left in the shaky hands of children. |
Economic Gap Between Thailand and Myanmar
On the economic and social disparities between Myanmar and Thailand, Thomas Fuller wrote in the New York Times, "On one side, millions of Myanmar's people suffer from chronic malnutrition. On the other, Thais enjoy a much more affluent society, where people are generally so well fed that obesity among children is a big concern. Children die in Myanmar of diseases so easily preventable that most people in Thailand have never heard of them. Burma was considered one of the most promising economies in Asia during the immediate postwar years. Today, the comparison with Thailand highlights Myanmar's missed opportunities under the grip of its military government and the breadth of the country's problems. [Source: Thomas Fuller, New York Times, October 25, 2007 ]
"There are significant differences in the salaries of construction workers: a daily wage the equivalent of 100 baht, or about US$3, on the Myanmar side, compared with double that in Thailand. A teacher on the Myanmar side earns the equivalent of US$47.50 a month, residents say, compared with upward of US$158 in Thailand. Crossing into Myanmar means stepping back in time. Bicycle rickshaws are a major mode of public transportation on the Burmese side — not because they are good for the environment, but because many people are too poor to be able to afford a car or a smuggled motorcycle from Thailand, which earns US$10 billion a year shipping cars and pickup trucks around the world.
"In both Thailand and Myanmar, the military has been deeply involved in politics in recent decades. Thailand has had more than a dozen coups since the 1930s and, after the overthrow last year of a democratically elected government, power remains in the military's hands. The salient difference, says Sean Turnell, an expert on the Burmese economy at Macquarie University in Sydney, Australia, is that Thailand's leaders have allowed businesses to thrive.
"During 45 years of misrule, Myanmar's generals have almost entirely dismantled the economy, he said. There are no effective property rights and contract enforcement is nonexistent. "If in other countries ruling regimes behave occasionally as mafioso in skimming a cut from prosperous business, then Burma's is more like a looter — destroying what it can neither create nor understand," Turnell said. The economic dysfunction means there is no financial underpinning for better healthcare or more widespread distribution of medicines.
"According to UNICEF, 10 out of 100 children in Myanmar die before reaching the age of five. In Thailand, two out of 100 die. A woman has a one in 75 chance of dying in childbirth in Myanmar, compared with one in 900 in Thailand. Because children in Myanmar are malnourished, 32 percent are significantly below the expected height for their age compared with 13 percent in Thailand. Cynthia Maung, a Burmese who runs a nonprofit health clinic on the Thai side of the 1,800km border, has become accustomed to detecting malnourished children. "The skin peels easily," she said. "The hair becomes brittle. The eyes look drowsy. There's muscle wasting — you can't see the muscles, just bone and skin." "
"Terrence Smith, a US gynecologist and obstetrician at the clinic, says most of his patients are unnaturally lean. "We do ultrasounds and the transducer goes straight to the organs," he said. In one corner of Smith's ward were two tiny, malnourished newborns dropped off and abandoned by their mothers. Maung's clinic was set up to treat sick patients from Myanmar who cannot afford health care inside their country. The clinic treated about 2,000 patients in 1989 when it opened. Last year, 100,000 Myanmar people came for treatment."
Image Sources:
Text Sources: New York Times, Washington Post, Los Angeles Times, Times of London, Lonely Planet Guides, The Irrawaddy, Myanmar Travel Information Compton's Encyclopedia, The Guardian, National Geographic, Smithsonian magazine, The New Yorker, Time, Newsweek, Reuters, AP, AFP, Wall Street Journal, The Atlantic Monthly, The Economist, Global Viewpoint (Christian Science Monitor), Foreign Policy, burmalibrary.org, burmanet.org, Wikipedia, BBC, CNN, NBC News, Fox News and various books and other publications.
Last updated May 2014


text:
Кубок Словаччини з футболу 2020–2021 — 28-й розіграш кубкового футбольного турніру в Словаччині. Титул вдруге поспіль здобув Слован (Братислава).

Календар

1/8 фіналу

| colspan="3" style="background:#fcc;"|6 квітня 2021

|-
| colspan="3" style="background:#fcc;"|7 квітня 2021

|}

1/4 фіналу

| colspan="3" style="background:#fcc;"|13 квітня 2021

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| colspan="3" style="background:#fcc;"|14 квітня 2021

|}

1/2 фіналу

| colspan="5" style="background:#fcc;"|28 квітня/4 травня 2021

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| colspan="5" style="background:#fcc;"|29 квітня/5 травня 2021

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Фінал

Посилання
 Кубок Словаччини на futbalnet
 Кубок Словаччини на soccerway

2020—2021
Словаччина
Словаччина
2020 у словацькому спорті
2021 у словацькому спорті

redpajama_set_name: RedPajamaWikipedia


The Eyeo Festival brings together an exciting community of people doing fascinating things with technology. Artists, data designers, creative coders, AI explorers, storytellers, tool & platform developers, and researchers all cross paths at Eyeo.
Join us for 4 days of enthralling talks, unique workshops, a code summit, and thought provoking conversations with passionate thinkers and makers. You'll love it. If you're looking for a creative refresh, to find collaborators, to learn about the state-of-the-art in creative technology, or just to find out what the future could look like, join us this June for Eyeo. Come hang with and hear from this exciting community. Share ideas. Connect with people IRL. Converge to Inspire.
If you're drawn to the intersection of these worlds you should definitely check out who'll be there this year. It's probably a place you want to be too. See the full lineup.
Come hear from over 30 speakers leading the way in a range of arenas, from creative coders like Daniel Shiffman and Sarah Groff Hennigh-Palermo, or data designers like Moritz Stefaner, Nathan Yau and Catherine D'Ignazio, or AI explorers like Helena Sarin, Mario Klingemann and Hannah Davis, or interactive creators like Mike Tucker, Christina "Phazero" Curlee and Meow Wolf, plus artists, astronomers, activists, and some people who are just too tricky to categorize.
Mike Tucker, Moritz Stefaner, Stephanie Dinkins, Onyx Ashanti, Mario Klingemann, Nicole Aptekar.
Get hands on and take a workshop.


text:
Tomcsányi Gusztáv (Ósva, 1852. február 13. – Máramarossziget, 1911. október 3.) erdőmérnök, erdőtanácsos, szakíró.

Életpályája
A középiskolát Kassán járta ki. 1871–1874 között a selmecbányai erdészeti akadémián végezte el tanulmányait. 1874-ben lépett állami szolgálatba: Ungváron, Besztercebányán (1881), Budapesten (1883–1890), Bustyaházán (1890), majd Máramarosszigeten (1899) dolgozott haláláig mint az erdőigazgatóság vezetője. 1876-ban erdészeti államvizsgát tett. 1877–1878 között az erdészeti akadémia növény- és erdőműveléstani tanszék tanársegéde volt. 1883-ban a földművelésügyi miniszterium erdészeti osztálya az erdőmérnöki állás ellátásával bízta meg, így 1884-től erdőmérnök lett. 1894-ben főerdőmester lett, 1895–1898 között pedig a lugosi erdőigazgatóság vezetője volt. 1899-től a máramarosszigeti erdőgazdaság igazgatója volt. 1910-ben (valóságos) miniszteri tanácsos lett.

Irodalmi munkái az Erdészeti Lapokban jelentek meg, főleg erdőműveléssel foglalkozott. Ő volt a légfűtésre berendezett magpergető és a tölgymakk-telelő tervezője.

Művei
 Az erdei facsemeték neveléséről (Budapest, 1889)

Díjai
 a vaskorona-rend III. osztálya (1904)

Jegyzetek

Források

 Szinnyei József: Magyar írók élete és munkái. Budapest, 1891-1914. Hornyánszky Viktor
 http://mek.oszk.hu/01800/01841/01841.pdf
 Tomcsányi Gusztáv, a tölgyesek megmentője

További információk
 Tomcsányi Gusztáv (Erdészeti Lapok, 1911)
 A magyar legújabb kor lexikona. Szerkesztette: Kerkápoly M. Emil. Budapest, 1930.

1852-ben született személyek
1911-ben elhunyt személyek
Magyar erdőmérnökök


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text:
Hurricane Electric and Digital Realty expand partnership to APAC
Digital Realty confirms that Hurricane Electric has deployed points of presence (PoPs) at its data centres in Singapore, Hong Kong and Sydney.
These new PoPs will give customers access to Hurricane Electric's IP transit network and broadens Digital Realty's global relationship with Hurricane Electric to the Asia Pacific (APAC) region.
Driven by rising demand for on-demand video, mobile gaming, and online content, APAC is now the fastest developing region for data centre colocation globally. This in turn gives rise to Data Gravity, the challenge of moving data at scale, slowing enterprise workflow performance, heightening security & regulatory concerns, and increasing costs.
"With businesses everywhere facing an explosion in the volume and complexity of the data they are managing, Hurricane Electric's expansion to our facilities in Singapore, Hong Kong and Sydney will help our customers access a variety of new connectivity options to overcome data gravity barriers and scale their digital businesses," said Govind Choudhary, head of service provider and market strategy, Asia Pacific, Digital Realty.
"This extension of PlatformDIGITAL across Asia Pacific through Hurricane Electric's global network connectivity will further enhance Digital Realty's position as an industry leading interconnection platform."
Through this partnership with Hurricane Electric, Digital Realty's customers can connect directly to Hurricane Electric's IPv4 and IPv6 network using 100GE, 10GE and GigE ports with access to over 4,000 customers in 53 metros across 27 countries on six continents.
Additionally, customers will be able to exchange IP traffic with Hurricane Electric's global network, which offers over 30,000 Border Gateway Protocol (BGP) sessions with more than 10,000 different networks via more than 280 major exchange points and thousands of customers and private peering ports.
"Hurricane Electric is delighted to expand our relationship with Digital Realty by extending our global network to its fast-growing customer base at key locations in the APAC region," said Walt Wollny, director of interconnection strategy at Hurricane Electric.
"This network expansion builds on Hurricane Electric's commitment to ensure that we provide connectivity to as many places as possible across the globe."
News NewsInfrastructure and NetworksData CentresFibre

redpajama_set_name: RedPajamaCommonCrawl


and help you make fitness an integral part of your life.
Every session is a full body workout that demands conscious and continuous engagement of the abdominals and core musculature throughout the whole 55-minutes of the class and promote long lasting effects that will extend beyond your training into your daily living and support your life style.
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Whether a group or a private session, each workout is specifically designed to the needs of the bodies in front of my eyes with personalized cuing that leads to noticeable results.


Data Availability: All data files are deposited in Dryad, doi:10.5061/dryad.4tn48.
Funding: This work was funded by the Institute of General Medical Sciences, National Institutes of Health (R01GM105244) and by UK Biotechnology and Biological Sciences Research Council as part of the joint NSF-NIH-USDA Ecology and Evolution of Infectious Diseases program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Infectious agents can rapidly evolve in response to health interventions . Here, we ask whether pathogen adaptation to vaccinated hosts can result in the evolution of more virulent pathogens (defined here to mean those that cause more or faster mortality in unvaccinated hosts).
Vaccination could prompt the evolution of more virulent pathogens in the following way. It is usually assumed that the primary force preventing the evolutionary emergence of more virulent strains is that they kill their hosts and, therefore, truncate their own infectious periods. If so, keeping hosts alive with vaccines that reduce disease but do not prevent infection, replication, and transmission (so-called "imperfect" vaccines) could allow more virulent strains to circulate. Natural selection will even favour their circulation if virulent strains have a higher transmission in the absence of host death or are better able to overcome host immunity. Thus, life-saving vaccines have the potential to increase mean disease virulence of a pathogen population (as assayed in unvaccinated hosts) [2–4].
The plausibility of this idea (hereafter called the "imperfect-vaccine hypothesis") has been confirmed with mathematical models [2,5–9]. Efficacy and mode of action are key. If the vaccine is sterilizing, so that transmission is stopped, no evolution can occur. But if it is non-sterilizing, so that naturally acquired pathogens can transmit from immunized individuals (what we hereafter call a "leaky" vaccine), virulent strains will be able to circulate in situations in which natural selection would have once removed them . Thus, anti-disease vaccines (those reducing in-host replication or pathogenicity) have the potential to generate evolution harmful to human and animal well-being; infection- or transmission-blocking vaccines do not [2–9]. Note that the possibility of vaccine-driven virulence evolution is conceptually distinct from vaccine-driven epitope evolution (antigenic escape), in which variants of target antigens evolve because they enable pathogens that are otherwise less fit to evade vaccine-induced immunity. The evolution of escape variants has been frequently observed [4,10].
The imperfect-vaccine hypothesis attracted controversy [11–14], not least because human vaccines have apparently not caused an increase in the virulence of their target pathogens. But most human vaccines are sterilizing (transmission-blocking) or not in widespread use or only recently introduced . Moreover, unambiguous comparisons of strain virulence and the impact of vaccination on transmission require experimental infections in the natural host—clearly impossible for human diseases. The situation is different for veterinary infections. Here, we report experiments with Marek's disease virus (MDV), a highly contagious oncogenic herpesvirus that costs the global poultry industry more than $US2 billion annually . We test a key prediction of the imperfect-vaccine hypothesis: that vaccination will elevate the fitness of highly virulent strains above that of less virulent strains.
Chickens become infected with MDV by inhalation of dust contaminated with virus shed from the feather follicles of infected birds. In a contaminated poultry house, chicks are infected soon after hatching and remain infectious for life . The virus can remain infectious in poultry dust for many months [17,18]. As originally described, Marek's disease (MD) was a paralysis of older birds, but by the 1950s, "acute MD" characterised by lymphomas in multiple organs in younger birds was occurring. This became the dominant form of MD, with increasing virulence, characterised by more severe lymphomas and mortality at increasingly early ages and, under some circumstances, paralysis and death in the first weeks of life, well before lymphoma formation [15,19].
MDV has been evolving in poultry immunized with leaky anti-disease vaccines since the introduction of the first vaccines in 1970 [15,19–24]. All MD vaccines are live viruses administered to 18-day-old embryos or immediately after hatch, and vaccinated birds can become infected and shed wild-type virus [25–28]. Wild-type MD viruses are so-called serotype 1 viruses. First-generation vaccines include a serotype 3 herpesvirus of turkeys called HVT; second-generation vaccines are a combination of HVT and SB-1, a serotype 2 isolate. Third-generation vaccines are based on an attenuated serotype-1 virus isolate CVI998, the so-called Rispens vaccine [15,19–24].
Our first three experiments involved Rhode Island Red (RIR) chickens, a breed that has not been subject to the intensive selective breeding and outcrossing that characterizes modern commercial chicken strains. Specific pathogen-free (SPF) parent birds were unvaccinated, and so offspring used in our first two experiments were free from maternally derived antibodies. In our first experiment, we infected 8-d-old chicks with five strains of MDV chosen to span the virulence spectrum defined by Witter and colleagues [21,29]. The viral strains varied from the less virulent HPRS-B14, which killed 60% of unvaccinated birds over 2 mo, to the highly lethal Md5 and 675A, which killed all unvaccinated birds in 10 d (Fig 1, top panels). When age-matched birds were vaccinated 8 d earlier with HVT, the first MDV vaccine to go into commercial use, survival improved dramatically, with a few deaths occurring only late in the experiment, and then only in birds infected with the most virulent strains (Fig 1, top panels).
Fig 1. Impact of vaccination on mortality and viral shedding of five strains of MDV.
Experiment 1. Groups of 20 Rhode Island Red chickens were unvaccinated (dotted lines, light shading) or HVT-vaccinated (solid lines, dark shading) at 1 d of age and challenged with viral strains HPRS-B14 (black), 571 (purple), 595 (green), Md5 (blue), or 675A (red) 8 d later. Viral strains vary in virulence in unvaccinated hosts, and vaccination protects against death (top panels, with strains arranged in order of increasing virulence from left to right.). Vaccination suppresses the concentration of virus in dust, but by keeping hosts alive, prolongs the infectious periods of hyperpathogenic MDV (middle panels). This means that cumulative number of virus genome copies (VCN) shed per bird is suppressed by vaccination for the least virulent strain and enhanced by several orders of magnitude for the most virulent (bottom panels). Error bars and shaded regions indicate 95% confidence interval (c.i.) Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
We collected dust from the isolators containing infected birds and measured the concentration of virus genomes in the dust using real-time PCR. At contemporaneous time points, vaccinated birds shed fewer virus genome copies than unvaccinated birds infected with the same viral strain (Fig 1, middle panels). Those patterns reflected viral loads in the feather follicles (S1 Fig). Critically, the infectious period of unvaccinated birds infected with our two most virulent strains was less than a week because hosts died so rapidly. During that week, barely any virus was shed (Fig 1, middle panels). In contrast, the infectious period of the least virulent strains continued for the entire experiment (almost 2 mo). Thus, the least virulent strain shed several orders of magnitude more virus from unvaccinated birds than did the virulent strains (Fig 1, bottom panels). By preventing death, vaccination greatly increased the infectious period of the most virulent strains, increasing the total amount of virus shed by several orders of magnitude, and increasing it above that of the least virulent strain (Fig 1, bottom panels). Thus, the net effect of vaccination on both host survival rates and daily shedding rates was to vastly increase the amount of virus shed by virulent strains into the environment.
To confirm that virus shed into the environment was a robust proxy for overall bird-to-bird transmission potential, we co-housed birds infected with our three most virulent strains with immunologically-naïve sentinel birds (Experiment 2). When unvaccinated birds were infected with the two most lethal strains (Md5 and 675A), they were all dead within 10 days (Fig 2A), before substantial viral shedding had begun (S2 Fig). Consequently, no sentinel birds in those isolators became infected (Fig 2B) and none died (Fig 2C). In contrast, when HVT-vaccinated birds were infected with either of those hyperpathogenic strains, they survived for 30 days or more (Fig 2A), allowing substantial viral shedding (S2 Fig). All co-housed sentinels consequently became infected (Fig 2B) and went on to die as a result of MDV infection (Fig 2C). Thus, in accordance with the imperfect-vaccine hypothesis, vaccination enabled the onward transmission of viruses otherwise too lethal to transmit, putting unvaccinated individuals at great risk of severe disease and death.
Fig 2. Vaccination enhances transmission of hyperpathogenic MDV.
Experiment 2. Groups of ten birds were HVT-vaccinated (solid lines) or not (dotted lines) and experimentally infected with one of our three most virulent MDV strains, 595 (green), Md5 (blue) and 675A (red), and co-housed with ten unvaccinated sentinel birds. Vaccination prolonged the survival of experimentally infected birds (A), ensuring that sentinel birds became infected (B) and, hence, died (C). In B and C, solid lines denote sentinels cohoused with vaccinated experimentally infected birds and dotted lines denote sentinels cohoused with unvaccinated experimentally infected birds. Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
Interestingly, the viral strain 595 was slightly less virulent than the other two viruses (taking a day longer to kill half of the unvaccinated birds, and 6 d longer to kill them all) (Fig 2A). This slightly reduced mortality rate prolonged the viral shedding from unvaccinated birds, so that about 100-fold more virus was shed into the environment by the 595-infected cohort than from the cohorts infected by the two more lethal strains (S2 Fig). This was evidently sufficient for transmission, because all co-housed sentinels eventually became infected (Fig 2B) and went on to die (Fig 2C). Thus, slight reductions in lethality can be sufficient to allow onward transmission. Nonetheless, even for strain 595, vaccination led to more rapid infection of sentinels (Fig 2B; median time to positivity 9 d earlier than in unvaccinated birds, p < 0.05), thus increasing the rate at which secondary cases were generated, a critical determinant of both viral fitness and case incidence in a rising epidemic.
The high mortality rates we observed in unvaccinated chickens infected with our most virulent strains are due to early mortality syndrome, which involves the rapid onset of paralysis, disorientation and an inability to feed and move, followed by death [30–33]. In today's modern industry, parental birds are almost always vaccinated against MDV, which results in the transfer of maternal antibody to chicks. These antibodies appear to be protective against the early mortality syndrome [30–33]. This raises the prospect that vaccination of laying hens could also permit onward transmission of viral strains that would be too lethal to otherwise transmit from offspring birds. We tested this possibility with further experiments using our most (675A) and least (HPRS-B14) virulent virus strains, again in Rhode Island Red birds, but this time including chicks derived from hens vaccinated 4–5 wk prior to egg lay with a standard commercial Rispens vaccine (Experiment 3).
Vaccination of hens enhanced the survival of offspring experimentally infected with HRPS-B14 (Fig 3A, p < 0.05). Maternally derived antibody had no detectable effect on the replication of that viral strain in the feather tips (S3 Fig panel A, p > 0.05) and, while it somewhat suppressed the amount of infectious virus shed into the environment early in infections (Figs 3B and S3B), it did not affect the rate at which sentinel birds became infected with HRPS-B14 (Fig 3C, p > 0.05) and few sentinels died (Fig 3D). Thus maternal protection had little impact on the transmission success of our least virulent strain.
Fig 3. Maternal vaccination enhances viral shedding and onward transmission of hyperpathogenic MDV.
Experiment 3. Groups of ten unvaccinated chicks produced by hens that were Rispens-vaccinated (solid lines) or not (dotted lines) were infected with viral strains HPRS-B14 (black) or 675A (red) and cohoused with sentinels of the same maternal antibody status. Maternally derived antibodies prolonged the survival of experimentally infected birds (A) and enhanced the amount of virus shed into the environment by the hyperpathogenic strain (B), making possible the infection of sentinels with the most virulent strain (C), which led to their death (D). Shaded regions represent 95% c.i. for unvaccinated (light) and vaccinated (dark). Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
However, presence of maternal antibody greatly impacted the transmission success of the most virulent strain (675A). As expected, the offspring of vaccinated hens survived for longer following infection with 675A virus than did maternally derived antibody-negative chicks (Fig 3A, p < 0.05). As we found in our first two experiments, very little of the highly virulent strain was shed from birds with no immune protection before they died (Figs 3B and S3). Consequently, no sentinels became infected (Fig 3C). But birds with maternal protection survived longer to shed more virus (Figs 3A, 3B, and S3B), so that all sentinel birds became infected (Fig 3C) and died (Fig 3D). Maternal vaccination was not as protective as direct vaccination of offspring (cf. Fig 3A with Fig 2A and the top panels of Fig 1). Nonetheless, vaccination of laying hens, like the vaccination of offspring, enabled the onward transmission of the hyperpathogenic strain from offspring (Fig 3C). Again, these data are consistent with the imperfect-vaccine hypothesis.
Our experiments above show that direct vaccination of birds or vaccination of parent hens makes possible the onward transmission of viral strains otherwise too lethal to transmit, and thus that unvaccinated individuals are put at increased risk of severe disease and death. However, in a modern commercial broiler setting, all birds in a flock would originate from vaccinated hens (and so would be positive for maternally derived antibody), and also be vaccinated. We thus set out to determine whether our most virulent strain could transmit to vaccinated sentinels, a necessary condition for persistence of hyperpathogenic strains in the modern industry (Experiment 4). To mimic the current commercial broiler situation, we obtained modern commercial broiler birds derived from Rispens-vaccinated hens and, at 1 d of age, we HVT-vaccinated all the birds we would experimentally infect. Those birds were then infected with our most virulent viral strain (675A) at 8 d of age. We cohoused those experimentally infected birds with sentinel birds, which were either HVT vaccinated or not. We performed this experiment twice. To accommodate changing regulatory requirements (see Methods), we did the first replicate with birds housed in isolators until 35 d of age, after which they were moved to floor until they were 11 wk old (Experiment 4a), and the second replicate with birds maintained in floor pens from 1 d of age until 7 wk of age (Experiment 4b).
All sentinels became infected, irrespective of vaccine status (Fig 4A). Thus, vaccinated maternal antibody positive commercial birds shed wild-type virus that caused infections in both vaccinated and unvaccinated maternal antibody positive birds. Vaccination only slightly suppressed viral replication in the infections acquired by the sentinel birds (Fig 4B). Importantly, all sentinels, vaccinated and unvaccinated, became virus positive in the feather follicles, meaning that they themselves started shedding. Vaccination protected sentinel birds from death (Fig 4C), prolonging infectious periods by about 2 wk (Fig 4D; standard error of the difference ±3.2 d, F1,36 = 19.9, p < 0.0001). Thus, not only does our most virulent strain transmit between modern commercial broilers when they are vaccinated, the duration of shedding in the next step in the transmission chain is also increased by vaccination.
Fig 4. Transmission of hypervirulent MDV to modern commercial birds.
Experiment 4. Groups of ten modern commercial broiler chicks derived from Rispens-vaccinated hens were HVT-vaccinated at 1 d of age and experimentally infected with the hypervirulent MDV strain 675A. Those experimentally infected birds were co-housed with groups of ten sentinel birds from the same commercial stock (and thus also derived from Rispens-vaccinated hens) which were HVT-vaccinated (solid lines) or not (dotted lines). The experiment was performed twice (experiment 4a, red; experiment 4b, blue). Independent of their vaccine status, all sentinel birds became infected (A), with high levels of virus replication in feather follicles (B). Vaccination prolonged the survival of sentinel birds (C), and consequently their infectious period (D). In panel D, "x" denotes death because of MDV. Error bars, 95% c.i. Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
MDV became increasingly virulent over the second half of the 20th century [19,21–24]. Until the 1950s, strains of MDV circulating on poultry farms caused a mildly paralytic disease, with lesions largely restricted to peripheral nervous tissue. Death was relatively rare. Today, hyperpathogenic strains are present worldwide. These strains induce lymphomas in a wide range of organs and mortality rates of up to 100% in unvaccinated birds. So far as we are aware, no one has been able to isolate non-lethal MDV strains from today's commercial (vaccinated) poultry operations [19,23]. Quite what promoted this viral evolution is unclear. The observation that successively more efficacious vaccines have been overcome by successively more virulent viral strains has prompted many MDV specialists to suggest that vaccination might be a key driver [19–24,34–37], though identifying the evolutionary pressures involved has proved challenging. There is no evidence in Marek's disease that vaccine breakthrough by more virulent strains has anything to do with overcoming strain-specific immunity (e.g., epitope evolution); genetic and immunological comparisons of strains varying in virulence suggest that candidate virulence determinants are associated with host–cell interactions and viral replication, not antigens . The imperfect-vaccine hypothesis was suggested as an evolutionary mechanism by which immunization might drive MDV virulence evolution , but there has been no experimental confirmation. Our data provide that: by enhancing host survival but not preventing viral shedding, MDV vaccination of hens or offspring greatly prolongs the infectious periods of hyperpathogenic strains, and hence the amount of virus they shed into the environment.
Our data do not demonstrate that vaccination was responsible for the evolution of hyperpathogenic strains of MDV, and we may never know for sure why they evolved in the first place. Clearly, many potentially relevant ecological pressures on virulence have changed with the intensification of the poultry industry. For instance, as the industry has expanded, broilers have become a much larger part of the industry, and broiler lifespans have halved with advances in animal genetics and husbandry; all else being equal, this would favour more virulent strains , so too might greater genetic homogeneity in flocks  or high-density rearing conditions , or indeed increased frequencies of maternally derived antibody if natural MDV infections became more common as the industry intensified in the pre-vaccine era (Fig 3) . But whatever was responsible for the evolution of more virulent strains in the first place (and there may be many causes), our data show that vaccination is sufficient to maintain hyperpathogenic strains in poultry flocks today. By keeping infected birds alive, vaccination substantially enhances the transmission success and hence spread of virus strains too lethal to persist in unvaccinated populations, which would therefore have been removed by natural selection in the pre-vaccine era.
The relaxation of natural selection against hyperpathogenic strains revealed by our experiments arises because vaccination enhances host survival. In serial passage experiments with a rodent malaria, immunity induced by whole parasite immunization  or vaccination with a recombinant antigen  also promoted the evolution of virulence. However, by design, those experiments did not allow host death to impact pathogen fitness, and so the evolution towards increased virulence was driven in a different way. Evidently, immunity in that system is disproportionately efficacious against less virulent strains. Our MDV experiments were not designed to test for within-host selection, but there is some suggestion that vaccine-induced immunity better controlled the replication of the least virulent strain (Figs 1 and S1). In principle, these two evolutionary pressures (within-host selection favouring virulent variants for their ability to evade immunity and vaccine-induced relaxation of between-host selection against virulence) could together generate very potent selection for more virulent strains . Within-host competition between strains could add further selection for higher virulence [41,42].
Vaccine failure in the face of virulent pathogens has been documented for at least two viruses other than MDV: feline calicivirus  and infectious bursal disease virus in poultry . Both cases are also associated with long-term use of leaky anti-disease vaccines. Our data are also consistent with hypotheses purporting to explain virulence increases in two well-studied wildlife systems. First, strains of the poultry pathogen Mycoplasma gallisepticum in North American house finches have become increasingly virulent, probably due to the increasing incidence of partially immune birds after the bacterium emerged in finch populations in the 1990s . Second, after well-documented declines in virulence following its release as a biocontrol agent in Australia, myxoma virus became increasingly virulent; that virulence evolution was most likely a consequence of increases in the genetic resistance and hence survival of wild rabbits in response to natural selection imposed by the virus . In both cases, anti-disease protection induced by natural immunization (finches) or by genetic resistance (rabbits) prolonged the infectious periods of otherwise highly lethal strains. These cases and our data raise the prospect that a variety of disease mitigation technologies have the potential to drive virulence evolution, including disease-ameliorating drugs [7,47] or genetic enhancements of host resistance . If these technologies prolong infectious periods of hyperpathogenic strains, as we have shown vaccination can, they too could create conditions favouring the emergence of highly lethal strains. This does not mean that such technologies should be avoided, particularly when alternative options are limited. Vaccination has massively reduced yield losses due to MD, despite the evolution . However, when protecting all individuals is impossible, or evolution is ongoing, the use of additional transmission-blocking interventions such as improved hygiene might be essential.
We suggest that the risk of outbreaks of hyperpathogenic strains be considered wherever disease interventions improve host survival without preventing pathogen transmission. Such situations might include vaccination against Newcastle disease  and avian influenza in poultry [51–53] and vaccination against Brucella in domesticated mammals , as well as genetic enhancement of agricultural animals including fish and poultry. Whether leaky human vaccines could also create the conditions in which more virulent strains can thrive will depend, among other things, on the selective factors currently preventing the emergence of hyperpathogenic strains in human populations. Our data emphasize that a comprehensive understanding of the impact of vaccines on pathogens cannot end with Phase III clinical trials or post-implementation studies of antigenic or serotype frequencies [2,4,10,55].
The HVT vaccine virus strain FC126 was second chick embryo fibroblast (CEF) passage stock from commercial HVT vaccine (Poulvac, Fort Dodge Animal Health). Commercial CVI988/Rispens vaccine virus (Nobilis Rismavac) was from Intervet. The challenge virus strains (seventh duck embryo fibroblast passage stocks) were a gift from Dr. A. M. Fadly (Avian Disease and Oncology Laboratory, United States). In the MDV literature, virulence (pathotype) is defined in terms of vaccine break-through [21,27,29], with virus strains categorized into pathotypes denoted as mild, virulent, very virulent, or very virulent plus (mMDV, vMDV, vvMDV, vv+MDV). In our experiments, we used up to five strains chosen to cover this spectrum. The strains were HPRS-B14, 571, 595, Md5, and 675A. HPRS-B14 has not been formally pathotyped, but would likely be categorised at the lower end of vMDV. The remaining four of these strains have been pathotyped as vMDV, vvMDV, vvMDV, and vv+MDV respectively . Note, however, that for the purposes of the present paper, defining virulence in terms of vaccine resistance introduces semantic circularity. Consequently, in the main text and what follows here, we instead explicitly define (measure) virulence as lethality in immunologically naïve birds.
For amplification of virus stocks, and to ensure there was no variation in virus passage history between experiments, 5-d-old Rhode Island Red (RIR) chickens were inoculated with 1,000 plaque forming units (pfu) of virus, via the intra-abdominal route. Lymphocytes isolated from spleens harvested at 14 d post infection (dpi) were cultured with primary CEF cells for 7 d, when cytopathic effect was clearly visible. The cells were passed two further times in CEF to produce virus stocks. The cell-associated virus stocks were titrated and stored in liquid nitrogen. CVI988/Rispens and HVT vaccines were administered via the subcutaneous route (neck), and challenge virus via the intra-abdominal route.
All studies and procedures involving animals were in strict accordance with the European and United Kingdom Home Office regulations and the Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012, under the authority of the Project Licenses PPL 30/2621 and PPL 30/3169. Birds were individually identifiable with wing bands and had access to water and a vegetable-based feed ad libitum. Any bird deemed to have reached the humane endpoint was culled. In the main text, the humane endpoint was taken as the time at which "infection-induced death" occurred. Chickens which reached the humane endpoint from 5–10 dpi (early mortality phase) showed a rapid onset of paralysis, disorientation, reluctance to feed, reluctance to move. and reduced weight gain. In our experience, this endpoint precedes natural death by less than two hours. Chickens which reached the humane endpoint from 15 dpi onwards showed a gradual onset of reluctance to feed, lethargy, and reduced weight gain. In our experience, this endpoint precedes death by up to 24 h. These endpoints, and our estimates of their timing with respect to viral-induced death, were arrived at from small-scale pilot experiments that determined the necessity for close monitoring because of the rapid onset of virus-induced death. Any bird that was found dead was reported to the UK Home Office. The majority of culled chickens showed enlarged spleen with gross lymphoid lesions. The prevalence of visceral lesions was broadly in line with those described by Witter (1997, his Table 4) for strains of corresponding pathotypes, despite differences in the breed and maternal antibody status of test chickens, and slight differences in the passage number of viral stocks.
For Experiments 1–3, chickens of the outbred Rhode Island Red (RIR) breed were hatched from the eggs of specified-pathogen-free flocks maintained at The Pirbright Institute. Chicks hatched from the eggs of unvaccinated hens were considered free from maternally derived antibody against MDV, and are hereafter referred to as MtAb-neg chicks. Chicks having maternally derived antibody against MDV (MtAb-pos) were hatched from eggs collected from RIR hens 4–5 wk after these hens were vaccinated with one commercial dose of Nobilis Rismavac CVI988/Rispens MDV vaccine (Intervet).
In Experiments 1–3, chicks were housed in positive pressure, high efficiency particulate air (HEPA)-filtered avian isolators (Controlled isolation Systems, US) within rooms in the Experimental Animal House at The Pirbright Institute, Compton. Chickens were monitored up to four times daily, and any chicken considered to have reached the humane endpoint was culled by cervical dislocation. When experiments were terminated, any surviving birds were culled. Post mortem examination was performed on all culled chickens and the presence or absence of gross Marek's disease lesions recorded.
The isolators are designed to house 20 1-d-old chickens or five adult chickens. In groups in which mortality following infection was low, it was necessary to reduce crowding in an isolator at intervals, by culling some birds. In these cases, birds to be culled were randomly selected, and the number of infected and sentinel birds culled was arranged to maintain the appropriate infected:sentinel ratio. Any birds culled for the purposes of reducing crowding were not included in survival data calculations.
For experiments 4a and b, commercial broiler breed chicks of the "Aviagen slow growing broiler line" were hatched from eggs supplied by Aviagen. Eggs were from CVI988/Rispens vaccinated hens, and therefore, all birds used were MtAb-pos as confirmed by ELISA (see below). We used different housing protocols for Experiment 4 from those adopted in Experiments 1–3 because regulatory requirements changed over the course of our studies, with work with adult birds in isolators becoming strongly discouraged. This meant we moved to floor-housing birds for at least part of Experiment 4, a condition that, anyway, more closely resembles housing conditions in the poultry industry. In Experiment 4a, birds were housed in isolators (as described above) until they were 35 d of age when groups were moved into floor pens within separate experimental rooms. Floor pens were constructed from metal barred caging panels that could have sections added to the layout to increase the pen area as birds became larger. Compressed straw pellets were used as bedding. In Experiment 4b, birds were housed within floor pens from 1 d of age in separate experimental rooms. To restrict the dissipation of dust and dander in first few weeks, the initial floor pen (measuring 1 m × 1 m) was partially contained using Perspex sheets attached to the wire caging to form a housing cube with open edges. As birds increased in size, additional non-Perspex covered cage sections were added to the initial cube to increase the pen area.
Two hundred (n = 200) MtAb-neg 1-d-old chicks were randomly allocated to ten groups, each group of 20 chicks being housed in a separate isolator, with two isolators (A and B) per room (S1 Table). In each room, chicks in one isolator (A) were vaccinated with HVT at 1 d of age, while chicks in the second isolator (B) were not vaccinated. At 8 d post vaccination (dpv), all chicks were challenged with one of five strains of MDV, each strain being used to infect a group of unvaccinated chicks, and a group of HVT-vaccinated chicks (S1 Table). Doses of vaccine and challenge viruses were approximately 1000 pfu and 300–600 pfu per chicken, respectively.
From each group, ten pre-selected chicks were feather-sampled [56,57] twice weekly until 55 dpi or until they reached the humane endpoint. Dust samples were also collected from each isolator twice weekly or until no chickens remained. Each time dust samples were taken, the pre-filter on the isolator air exhaust was removed and replaced with a new, clean filter. Within the isolator, the removed filters were shaken into a polythene bag to collect poultry "dust," which was transferred to tubes and stored at −20°C.
One hundred and twenty (n = 120) MtAb-neg 1-d-old chicks were randomly allocated to six groups, each group of 20 chicks being housed in a separate isolator, and there being two isolators (A and B) per room (S2 Table). In each group, ten chickens were randomly selected to be the "shedder" birds (i.e., experimentally infected), while the remaining ten chickens were selected to be "in-contact sentinel birds". In one isolator from each room (A isolators), the shedder birds were vaccinated with HVT at one day of age. In B isolators, the shedder birds were not vaccinated. At 8 dpv all shedder birds were challenged with one of three strains of MDV. Sentinels were neither vaccinated nor challenged. Doses of vaccine and challenge viruses were approximately 2,750 pfu and 1,000–1,500 pfu per chicken, respectively.
Sentinel birds are necessary to directly measure natural transmission rates, but once sentinels themselves become infectious, they make it difficult to determine how much virus is being shed by experimentally infected birds. For our studies of 675A, we therefore added two additional treatment groups (Groups 4A and 4B, S2 Table) by randomly allocating 20 (n = 20) additional MtAb-neg 1-d-old chicks to two additional isolators, with ten birds in one isolator being vaccinated as above (A isolator), and the ten in the other isolator not being vaccinated (B isolator). All 20 of those additional birds were then experimentally infected at 8 dpv, as above. The existence of these two extra groups allowed us to estimate the viral shedding rates for birds experimentally infected with 675A without any issue of viral contamination from sentinels. We did not, however, have the resources to run analogous extra groups for Md5 or 595, and so the dust data for those strains (S2 Fig) includes dust shed from sentinel birds, which may contain virus after about 20 d post-experimental infection, when the first sentinels began to become infectious (Fig 2, S2 Fig).
Feather samples were collected from every shedder bird cohoused with sentinels twice weekly until 52 dpi or until they reached the humane endpoint. From 3 d onwards, at the same time points unless there were none alive, 150 μL blood samples were collected from every sentinel into 3% sodium citrate. At the same times, dust was collected and stored as described above. For the analysis of the 675A data, we used the survival data for all 20 experimentally infected birds of identical vaccine status (i.e., pooling the relevant data from the Group 1 and Group 4 isolators), the dust data (S2 Fig panel B) from the Group 4 isolators and the feather data (S2 Fig panel A) from the Group 1 isolators (groups defined in S2 Table).
Sixty (n = 60) MtAb-neg 1-d-old chicks and 60 MtAb-pos 1-d-old chicks were randomly allocated to groups, each group being housed in a separate isolator, and there being two isolators (A, containing 20 chicks, and B, containing ten chicks) per room (S3 Table). In A isolators, ten chickens were randomly selected to be shedder birds (i.e., experimentally infected), while the remaining ten chickens were selected to be in-contact sentinels. At 9 d of age, all shedder birds were challenged with one of two strains of MDV; sentinels were not challenged. In each B isolator, all ten chickens were challenged with one of two strains of MDV, and these isolators were used for collection of dust so that shedding of MDV from ten infected chickens could be accurately determined without "dilution" by dust from non-infected sentinels. Doses of challenge viruses were 100–500 pfu per chicken.
Blood (150 μL) was collected from five pre-selected chickens from each of the B isolators prior to challenge. Serum was stored at −20°C. Feather samples were collected from every shedder bird in the A isolators twice weekly until 52 dpi or until they reached the humane endpoint. From 3 dpi onwards, blood samples were collected from every sentinel bird at these same time-points (or until such time that no chickens remained). At each of the above time-points (or until such time that no chickens remained), dust was collected from B isolators and stored as described above.
All tested chickens from the MtAb-neg group were negative for anti-MDV antibody (assayed by ELISA; see below), while all tested chickens from the MtAb-pos group were positive for maternal antibody.
In both Experiments 4a and 4b, 40 (n = 40) commercial broiler breed chicks were hatched from eggs produced by CVI988/Rispens vaccinated hens and therefore all were MtAb positive. The basic experimental design for Experiments 4a and b was the same; four test groups of age-matched chicks; two groups acting as MtAb-pos, HVT vaccinated, experimentally infected shedder birds housed independently with one of two groups of MtAb-pos sentinel birds that differed in their vaccination status, i.e., either HVT vaccinated or not (see S4 Table).
In both Experiments 4a and 4b, 40 1-d-old chicks were randomly placed into four groups across four isolators (S4 Table). In each experiment, all birds except the ten within each unvaccinated sentinel group were vaccinated with approximately 1,500 pfu HVT FC126 via the subcutaneous route at one day of age. At 8 d of age, the two groups serving as shedders were challenged with approximately 725 pfu of vv+ MDV 675A via the intra-abdominal route. Blood (150 μL) and feather samples were collected twice weekly from all birds until 21 dpi and thereafter weekly. Dust samples were collected weekly from the housing air extract filters, as above.
Viral titres were assayed indirectly by PCR as follows. Peripheral blood lymphocytes (PBL) and feather tip samples were prepared as previously described [56,57]. Each dust sample was measured into triplicate 5 mg aliquots. Total DNA was prepared from each PBL, feather and dust sample using a DNeasy-96 kit (Qiagen), according to the manufacturers' instructions for extraction of DNA from cells (PBL) or from tissues (feather tips and dust). Real-time quantitative duplex PCR (q-PCR) to amplify the MDV-1 meq gene and the chicken ovotransferrin (ovo) reference gene was used for absolute quantification of MDV genomes as previously described . This assay does not detect HVT vaccine virus. All reactions using feather tips or dust samples as target DNA contained 10 μg bovine serum albumin to overcome the inhibitory effect of melanin pigment . Standard curves, prepared using 10-fold serial dilutions of DNA from MDV1-infected CEF (for meq reaction) and non-infected CEF (for ovo reaction) and accurately calibrated against plasmid constructs of known target gene copy number, were used to quantify MDV genomes per 104 cells or per μg dust.
Enzyme-linked immunosorbent assay (ELISA) was performed to measure maternally derived antibody against MDV in serum samples from hatched chicks . Serum samples were tested in duplicate at 1:100 dilution on ELISA plates coated with MDV-infected or non-infected cell lysates. Serum from a non-infected chicken was used as a negative-control, and serum from an MDV1-infected chicken as a positive control.
All data and the R code used to create all the figures are deposited in the Dryad repository: http://dx.doi.org/10.5061/dryad.4tn48 .
For groups of chickens, mean values for virus genome copy number for PBL, feather tips or dust, were determined using the log10 transformed copy number for each individual sample. For feather tip data, 95% c.i. of the means were calculated using the t-distribution. For dust data, 95% c.i. of the mean data were approximated as ±2 standard errors. Plotted values of virus concentration in dust are for samples based on the cumulative dust shed since the previous plotted sample (when filters were last changed). For each sentinel chicken, the time at which MDV was first detected in PBL by qPCR (time to positivity) was recorded. Time to positivity was taken as the first sampling time at which the q-PCR Meq Ct value was <40 for successive time-points. Statistical comparisons of survival or time to positivity were made using the Mantel-Cox test applied to Kaplan-Meier survival curves plotted using GraphPad Prism v5.
To assess transmission potential, a key component of viral fitness, we calculated the cumulative virus genome copy number (VCN) shed rate over the lifetime of an infection. Cumulative VCN shed is a good proxy for transmission potential because virus shed from feather follicles is the only source of infectious MDV, and because MDV-contaminated dust remains infectious for many months [16–18]. Cumulative VCN shed can be uniquely determined from three components; the dust shed from a bird over time, the concentration of VCN per unit shed dust over time, and the lifespan of an infection. We directly measured the latter two values in Experiments 1–3, and the former value has been previously estimated . Details of how we used these measures to calculate cumulative VCN are given in S1 Protocol.
In Experiment 4, we estimated viral genome concentration in feather follicles of sentinel birds. Because these birds were co-housed with experimentally infected birds, virus-negative feather shafts can become contaminated with dust from infected birds. We therefore set quantitative thresholds for virus positivity in feather pulp, as described in S2 Protocol. Duration of infectious period (Fig 4D) was taken as the time from when viral titres first exceeded this threshold until bird death.
S1 Fig. Vaccination suppresses but prolongs viral replication in the feather tips of experimentally infected birds.
Experiment 1. Groups of 20 Rhode Island Red chickens that were unvaccinated (dotted lines) or HVT-vaccinated (solid lines) at 1 d of age and challenged with viral strains HPRS-B14 (black), 571 (purple), 595 (green), Md5 (blue), or 675A (red) 8 d later. Virus genome copy numbers were estimated by qPCR from the pulp of feathers plucked from individual birds. Error bars are 95% c.i. of the mean. Large error bars are from time points where few birds remained alive. Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
S2 Fig. Vaccination suppresses but prolongs viral replication in the feather tips of experimentally infected birds, increasing total viral genomes shed into the environment.
Experiment 2. Groups of 20 Rhode Island Red chickens were unvaccinated (dotted lines, light shading) or HVT-vaccinated (solid lines, dark shading) at 1 d of age and challenged with one of our three most virulent viral strains 595 (green), Md5 (blue), or 675A (red) 8 d later. Top panel shows virus replication in the feather follicles, middle panel shows virus concentration in dust collected from isolator filters, and lower panel shows estimates of cumulative viral genomes shed from an experimentally infected bird. Error bars and shaded areas are 95% c.i. of the mean. Note that estimates of cumulative viral genomes shed from vaccinated 595- and Md5-infected birds are biased upwards after around day 20, when sentinels began to shed virus (see Methods and S2 Protocol for discussion). Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
S3 Fig. Maternal vaccination prolongs the replication of the most virulent strain of MDV in feather tips of infected chicks and hence shedding.
Experiment 3. Groups of ten unvaccinated chicks produced by hens that were Rispens-vaccinated (solid lines) or not (dotted lines) were infected with viral strains HPRS-B14 (black) or 675A (red). Viral genome concentration in feather follicles (top panel) and in dust (bottom panel). Error bars are 95% c.i. of the mean. Large error bars in top panel are from time points where only two birds remained alive; after day 41, only one unvaccinated HPRS-B14-infected bird remained alive and so there are no error bars. Raw data can be found at http://dx.doi.org/10.5061/dryad.4tn48.
S1 Protocol. Calculation of cumulative virus genome copy number of lifetime of an infection (Fig 1, lower panels, and Figs 3B and S2) [28,60,61].
S2 Protocol. Controlling for background viral contamination of feather pulp (Experiment 4, Fig 4B and 4D).
S1 Table. Design of Experiment 1: Effect of HVT-vaccination on shedding of five strains of MDV.
S2 Table. Design of Experiment 2: Effect of HVT-vaccination on transmission of three strains of MDV.
S3 Table. Design of Experiment 3: Effect of maternally-derived antibody on shedding and transmission of two strains of MDV.
S4 Table. Design of Experiments 4a and 4b: Transmission of MDV strain 675A in commercial maternal-antibody–positive HVT-vaccinated birds.
We thank A. Fadly (ADOL-USDA) for virus strains; Aviagen for eggs; staff of the Animal Services Department of The Pirbright Institute for provision of chicks and subsequent husbandry; A. Morrow (DEFRA) for triggering all this; C. Morrow (previously of Aviagen) for introducing AFR to the poultry industry; K. Crouch for initial experimental work; K. Atkins, T. Day, P. Dunn, S. Gandon, M. Mackinnon, S. Nee, and members of the Research and Policy in Infectious Disease Dynamics program of the Science and Technology Directorate, Department of Homeland Security, and the Fogarty International Center, National Institutes of Health, for discussion; and K. Atkins, C. Jackson, P. Kerr, C. Morrow, and R. Woods for comments on the manuscript.
Conceived and designed the experiments: AFR SJB CP LPS SWWB VKN. Performed the experiments: SJB CP LBK LB LPS. Analyzed the data: AFR SJB CP DAK. Wrote the paper: AFR SJB CP DAK. Conceived the study and raised the funding: AFR VKN.
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