Lamanda Case Study

Food prices are rising due to climate, peak oil and poor economy. Best time to start your city on Victory Gardens. Speech by Lamanda Joy to Great Lakes Bioneers tells how. With intro on food prices in Canada, UK, Australia, USA, clips from WWII garden propaganda. If you eat food, maybe you’ve noticed groceries costs more and more. Well stock up now, food inflation is just ramping up.

It’s not just the extra 200 million mouths to feed on the planet next year. Climate change is already re-arranging your food bill.

In North America, and around the world, one big driver is the record drought in the prime crop production areas of the United States this year. Many meat producers gave up, selling off their herds, temporarily keeping meat prices lower. The current cattle herd is the smallest since 1973. Once that sell-off goes through the supply chain, the high cost of corn and other grains will accelerate meat prices from 5 to 10% higher, according to one Canadian report.

Here is Peter Mansbridge of host of "The National" on Canadian Broadcasting Corporation on December 6, 2012.

"If you are buying extra groceries for the holidays, some experts suggest you also might think about stocking up for the coming year."

Next comes CBC Consumer Affairs reporter Aaron Saltzman [with a report from Canada’s premier agricultural university in Guelph, Ontario]:

"According to the University of Guelph’s annual food forecast, just about every basic staple will cost more next year. Dairy up as much as 3%; bread up as much as 4%; eggs up as much as 5%; but likely the biggest hike… ‘We would expect meat, particularly beef and pork to go up more significantly. We are saying probably as much as 4 and a half to 6%.’ The price of pork in particular expected to jump by as much as 10%.

According to the report the main driver behind most of these price increases is climate. The drought across North America this past year was one of the worst in recorded history. Among the hardest hit areas, the Great Plains states in the U.S. – America’s bread basket. That drove up grain prices and in turn, the cost of feed and livestock." …

…For those unwilling to go vegetarian, ‘Fill the freezer now, because it’s going to get tougher going forward.’

And, he says, if you are wondering how accurate the University’s predictions are, last year the forecast was bang on."

FOOD PRICES WAY UP SINCE 2002

Host Peter Mansbridge: "So some predictions there about food prices in the future. What about the prices we’ve already seen? Here are some numbers to consider. According to statistics Canada, the cost of meat has risen more than 30% in the past decade. Egg prices have risen by 50%. Bakery products are up by nearly 60%. By comparison, fresh vegetables cost about 1% less than 10 years ago."

Don’t be thrown off by comforting reports from the United Nations Food and Agricultural Organization. Their early December report says basic food prices fell by 1.5% in November. That was partly caused by a massive drop in the price of sugar.

But as the World Bank reports, food prices are currently "stable" but still very high. In fact, expensive food hovering near the record 2008 levels is the new normal. Unlike the UN, the World Bank food price index finds food prices are 7 percent higher than in 2011. Grains are 12 percent higher already from the previous year.

In the United Kingdom, the November Shop Price Index shows food prices up 4.6% from a year ago. Fresh fruits and vegetables are particularly high, causing what the Guardian newspaper calls "a nutrition recession" in Britain.

A series of reports in the Australian press say food prices there will hit a new record high in 2013. The cost of rice, wheat, pulses, edible oils, sugar and vegetables, are all rising in India.

Bloomberg business finds American meat prices are set to go much higher. Quote from Bloomberg: "The drought in the Midwest and Great Plains drove corn yields to a 17-year low and may last at least through February. U.S. consumers will pay 3 percent to 4 percent more for food next year, a half-percentage point above this year’s expected increase, according to the USDA."

It’s not just the drought. A weird bout of summer-like weather in March of 2012 caused many fruit trees to bloom early. The return of cold weather killed off the flowers, leading to a drop of the apple crop by as much as 80% in some regions. The world charity Oxfam has a special report on the impact of extreme weather events on the world food supply.

WHERE IS THE GOOD NEWS?

Wait a minute! Didn’t I promise you some good news this week?

Sure, if more people can’t afford red meat, their health will improve dramatically. Healthy vegetables are still the most affordable option in most places.

But it gets much better than that. We can quickly and cheaply convert most of our major cities into major food production centers. In this program, you will hear how it happened before. And how local urban food production is making a rapid come-back. The kicker is lots of folks are going for more than just the joy of healthy self-grown food. They come for the new sense of community as well.

Most of us can’t afford to just kick-off and head to the country. We need some income, at least to make the transition. So like Havana Cuba, after the Soviet empire died and stopped sending oil, we’ll have to feed ourselves where we are.

We are going to the City of Chicago, where the American Victory Garden movement was launched during World War Two. And where it is coming back strong. From the Great Lakes Bioneers conference, we have an excellent recording of Lamanda Joy, founder of the Peterson Garden Project. She’ll tell us about the Victory Garden movement, where MILLIONS of novice growers produced mountains of food for the War effort. How a whole nation can transform into local food production in just one year.

That’s not just something that happened in the past. Lamanda will tell us how Chicago is organizing once again, to bring back urban food production. Whether you are concerned about economic collapse, climate change, peak oil, or just healthy food – this speech is a message of hope for all of us.

This recording was made by Kelly Pierce of the Chicago Independent Media Center for Radio Ecoshock. Here is Lamanda Joy, speaking November 4th, 2012 to the Great Lakes Bioneers.

 

Transcript

My name is LaManda Joy and I am very happy to be with you today. Does everybody understand what the concept of a Victory Garden is? I see lots of nods so we will skip over that.


Why am I talking about them?


I know the conference these last couple of days has been a lot about inspiration and passion and what we can all do to make a difference. And I feel very attached to this Victory Garden story for a lot of reasons. The first of all is because my parents are Greatest Generation parents. They met in 1941 – love at first sight – at the Samuel Gompers Junior High School in Los Angeles. ( http://www.greatschools.org/california/los-angeles/2109-Samuel-Gompers-Middle-School/) (now called Samuel Gompers Middle School). Something else significant happened that year. We went to war. So my parents as young adults lived that whole experience or what you call The Greatest Generation. That’s my handsome father. He was drafted and served in Occupied Forces just after the end of the war with occupied forces in Japan. So I grew up with a family that was talking about WWII a lot. It was the biggest thing in their lives and it continues to have an impact today.

After the war was over, my parents moved to Oregon, where I was born, just outside of Portland. And my dad learned how to garden. Notice in the back ground here. This is my great-grandfathers garden. During the war it was a Victory Garden. After the war it was just a garden again. My dad learned to garden there and then when I came along in the late 60’s he taught me how to garden as I was growing up. So that’s a little bit about that ethos. You know I have this sort of unbroken chain of what happened during those times that I think is really valuable. So I am very attached to this story. And how it can make a difference for us today.


Who here likes to garden? I hope that there’s a few people. Ok. Good.


Now. Who likes to eat? Everybody. Ok good. Well if you an eater than you are a good gardener. Or at least, you have good potential to be a good gardener.


Those of you who are gardeners know that once you get that bug, it’s something that is kind of with you for a life time, right?


So, I got that bug very early growing up in Oregon helping my dad garden. Then I moved to Chicago in ’94 and I lived for a few years in a Condo there. Which was lovely. It was a vintage condo, it overlooked the lake which was very nice. But there was one problem. There was no where to garden. So every spring I got a little obnoxious. You know. Where are going to garden? How are we going to garden? You know. We can find neighbors with backyards. There was no community garden so it was a little bit of a problem.


So, it was in February of 2006 that my husband woke up one morning and he said (timidly) “Should we go look for a house?” “Should we go look for a yard?”


So, yes “Let’s go look for a yard” – go ahead and hit it. So we bought a yard with a house attached to it. And we called it ‘The Yarden’ (theyarden.com). That is what it looked like when we got. A four foot chain link fence. Lots of sun. We didn’t have any old trees that were casting shade. We didn’t have any apartments looking down on it. It was nice. So we bought this yard and there happened to be a house attached to it. Then the next year we put in this. So this is our ‘yarden’. It’s seventeen hundred square feet. It is all organic. We do all open pollinated.


We have espaliered fruit trees. This was my happy place. Right?


I think many people are like this. Or maybe I’m just like this. But I think that everybody is like me. Like maybe everybody has had the same experience. So I assumed that everybody knew how to garden and can and bake and all these things that my parents had taught me and I quickly learned that that wasn’t the case. I started blogging about and learned that ‘Whoa’. People don’t really know how to do this stuff and they really want to do this stuff.


So these are two of the factors that brought me into this story. But there’s one more. And that’s community. I moved into this neighborhood. I had lived in as many as ten different places since I moved here in nineteen ninety four (1994). Then I moved to this neighborhood and we were going to stay put so I really wanted to support the community. And I was at my local butcher shop waiting for my order when I looked at the wall and I saw this photo. And I was like “What is that photo?” So, Reuben and Irv told me the story that this photo was taken at the building where their butcher shop was in 1943 actually. The woman who took it lived in the neighborhood her entire life and when she passed away her family came to the butcher shop and said “Hey we found this photo. Do you want it?” They collected old photos. They said “Sure”. And they stuck it on the wall.


I came in and saw it many years later and it was, like, “Hah”.


So all of these things sort of came together. Once again, like “Hah. I really want to understand the Victory Garden Movement. And I had heard a lot of statistics. Maybe you have heard them too. 240 million people gardened. 40% of the produce consumed was home grown. And I a little OCD (Obsessive Compulsive) so I said “I am going to figure out how they did this.”


That’s a lot of people learning to garden. And that’s a lot of people growing food. How do you teach an entire city to grow their own food? So that became my research question. I really dove into what Chicago did to grow their own food during WWII.


So we are going to talk a little bit about the world at war. You know the war started in 1941 and ended in 1945. And in between we had four summers where people were Victory Gardening. 42, 43, 44, and 45. Most of my information is about 1942 and 1943. So, I don’t have the full spectrum but that’s ok, we’ll talk about those two years.


So war on the home front. During the last part of 1941 when the country was actively preparing for war, for national defense there were already signs of food shortages and rising prices and especially in the category of fresh vegetables. Great industries were being transformed into munitions plants. And this quickly absorbed much of the country’s pool of unemployed from the depression. One of the first industries to feel the pinch of the labor shortage was that of ‘fresh vegetable growers’ as lower paid field workers found higher paying jobs in munitions and other war industries. So the farm workers were going to find better jobs. So there weren’t people to grow the food. Another issue was transportation. Transportation lines were quickly conscripted to move troops and munitions, which also impacted the delivery of fresh vegetables from farms to urban areas.


As one of the higher ups in the Chicago Victory Garden movement phrased it “Uncle Sam needs trains for munitions so we will have to grow our own carrots.”


So, from the smallest family groupings, and I hope the mother was taking this picture and not pulling the plow, but we don’t know because people were desperate in these times. So, from the smallest family groupings to the largest organizations people used whatever they could to prepare their Victory Gardens.


Here you see the Mayor of Boston plowing up the Boston Commons. Everyone was drafted into the Victory Garden effort. Religious Orders. Movie stars. This is Veronica Lake. She was a very famous movie star at the time and she was known for this beautiful hair style that she would wear. She was sort like the ‘peek-a-boo’ girl, she looked from behind her hair. Well. People were emulating this hair style so much that is was a problem because they were all becoming ‘Rosie the Riveters’ and their hair was getting caught in machinery. So, they talked to Veronica Lake about doing a campaign called ‘Hair Wins The War’. If you google that you will find a very interesting video about how ‘Hair Wins The War’ (a 1 minute and 49 second clip). (http://www.youtube.com/watch?v=mgpvKXLTwr8) And how you need to change your hairstyle. For the duration of the war, she wore her hair like this to encourage women to wear their hair up. I usually wear my hair like this when I am gardening and presenting but I was just too busy this morning.


Movie stars were involved. Cartoon characters. Super heroes. And of course, popular culture. You know you got really thirsty doing your Victory Garden so you might need to drink a Dr. Pepper. Or, if you got a little more thirsty, maybe you needed to drink your Victory Garden beer. Or if you were really thirsty, maybe you just drank a whiskey and let someone else to the gardening.


But, here I am just joking. I showing you these just to show you how much this effort was a part of popular culture. It wasn’t something that was in the shadows. It was part of everything that people were doing.


So back to the serious note.


Industries. Steel mills. Railway stations. Sears. Sears started 24 thousand Victory in the Los Angeles area. Newspapers. And, back to popular culture, this is Marshall Fields. This is the window from Marshall Fields. And that sign says “To make your Victory Garden grow, wear gay and chic garden apparel located in the sport department on floor 6.”


So, of course, industries, especially garden industries, jumped in full force. International Harvester, which is a Chicago based manufacturer of plows, they are still here, they are called Navistar, was known to be a big supporter of the Victory Garden movement.


And, I’m just going to take a little detour. You know, you gotta eat year round but you can only garden for a part of it. Canning was huge during WWII. So, hand in hand with food production came food preservation. Modern freezing techniques were not widely used and tin and other materials for store bought canned foods were being used for munitions. So women were encouraged to can, dehydrate, brine, and otherwise preserve the harvest. And salute while you are doing it.


So, you can see that industrious preservation was a serious business. It was a real war job. It wasn’t just ‘Oh, we’ll make some jam to give away as Christmas gifts.’ There would be no food if you did not can.


One of the research pieces that I ran across said that five billion pints were preserved during WWII – every summer.


<>Has anyone done any canning this summer? Yeah? You know it’s a lot of hot work, right? 5 billion pints nationwide.

Pressure cookers and canning supplies were in such demand that their production and distribution were overseen by the government. Serious business.


So, at the height of WWII, 24 million meals per day, on the 50 plus battle fronts and fronts that the U.S. participated in. So you can see that the Victory Garden movement wasn’t just a morale building luxury, it was a dire necessity.


So, here are some words from those times:

Americans started gardening. Gardens began sprouting behind sign posts, at the bases of railroad embankments, in school yards and church yards, and in window boxes. They were tended by housewives who left babies in sandboxes while they knelt to scoop and sift the soil. And by little girls who wheeled their trowels and rakes into their plots in doll buggies. And by war workers who were back to the good earth for the first time since they waded barefoot in mud puddles as children. And so, the historic gardening campaign of WWII began.

So that’s an overview of the nation. But let’s talk specifically about Chicago. You know I mentioned a bit earlier that I am OCD. You all know what that means. If you notice the acronym of this organization, they are OCD as well. So they did keep things very organized. Let’s talk about that Offices of Civilian Defense. Fiorello LaGuardia (http://en.wikipedia.org/wiki/Fiorello_La_Guardia) was appointed as the person in charge of the Office of Civilian Defense. He was not happy about this. He had other things he wanted to do. And they did not include working in the Office of Civilian Defense. But, the original Civilian Defense organization was centered around preventative measures against air attacks, sabotage and the like. You know there was a fear that our enemies would actually make it to our shores. So people were preparing for air raids and stuff like that.


It was recognized that that same organization might well function in the Victory Garden field. And the OCD Administrator in DC issued a memorandum in January 1942 urging local coordinators to set up machinery for promoting Victory Gardens. Now, I have to tell you this from my extensive research not all cities used the Office of Civilian Defense as the mechanism for that. Chicago did heavily. Other cities did different things but it seemed to work for us.


So the activities of the Office of Civilian Defense were air raid surveillance, attack training, scrap and fat collection, war bond sales, morale committees and victory gardens. Rumor has the Office of Civilian Defense was the brain child of Eleanor Roosevelt. She understood the impact of the First World War on the nation and she wanted people to feel involved, feel like they were active and not just sitting here passively while all the horribleness was going on overseas. So, it was her idea. However, you know, the wife of the President at that time couldn’t be the brain child of something like that or run something like that. So, for 18 months she went to New York where she was Fiorello La Guardia’s secretary. He didn’t want to do it so she became his secretary and sort of ran things from the shadows.


So, Chicago. Here are some words from the time:



  • Victory gardening in WWII was radically different in the character and extent from the corresponding activity in WWI
  • Lack of sustained promotion, absence of any organized scheme of promoting gardening and general haphazard and wasteful methods destroyed much of the usefulness and productivity of the so-called war garden movement of WWI.
  • In WWII, at least, we learned from our mistakes, at least in Chicago, and most of these faults were recognized before actual fighting took place.

  • A thorough and complete scheme of organization was set up largely due to the aid of the Chicago Park District
  • In the local Office of Civilian Defense there was a continuous program of promotion, stimulation, guidance and active help

This was our mayor at the time, Mayor Kelly, and he quickly appointed some Park District big-wigs to run the program. You know, we were lucky at the time, the Park District was probably the strongest it has ever been. Less than ten years before 1941 – during the mid-thirties we had three Park Districts that were all run separately. Then these were all conglomerated into one Park District. As a result they were very well organized. They had gone through this process of making their communications good. And we also were in the depression, so Chicago could afford really high-class, ivy-league horticulturalists and business people to help with the Park District. So, we were in a really lucky spot at that time to have a really strong Park District to lead the way.


The goals of the Victory Garden movement were to stimulate as many people as possible to apply themselves during their leisure hours – they were not expecting people to quit their jobs or do whatever – this was a leisure hour pursuit. They wanted to provide novice gardeners and those not familiar with growing vegetables, the information needed to make their efforts most productive. And you know this was written by a gardener because the final one was to lay a foundation for the expansion of the program in peace time.


Maybe if that had happened we would be in a different world today. But we are not.


The resources to accomplish this were a lot of vacant lots, backyards, strong park systems, school districts, libraries, corporate support, and a devoted citizenry of which 90% had never gardened before.


It wasn’t that they had landscape gardened before but didn’t know how to grow food. They had never gardened before.

To accomplish this and the other mandates of the Office of Civilian Defense, the city was divided into seven divisions. And there were more divisions in the suburbs. Division 7 doesn’t show on here because it wasn’t deemed to be important or possible to grow food. And all 77 Chicago neighborhoods were segregated into one of these 7 Divisions. Each Division had a Division Chairman that reported to the OCD. And each neighborhood had a Community Chairman that reported to the Division Chairman. And each block had a Block Captain that reported to the Community Chairman. And then there also other committees. The Advisory Committee, the Committee on Demonstration Gardens, the General Committee, the Committee on Food Preservation, the Committee on Planning and the Committee on Kinds of Vegetables.


There was lots of organization going on. But that’s a good thing because that meant that we are able to have the information that we have today. Because they kept track of everything. There was this level of bureaucracy that people had to work through. And it was very successful.


Block Captains are the ones who actually got most of the work done. That was their duty. To find land three or four blocks from homes of interested parties, prepare the site, arrange for plowing, participate in the Victory Garden Council, maintain enthusiasm and plan Harvest Festivals.


Let’s take a little detour and talk about Harvest Festivals. This was a festival that happened in Streeterville, which is not far from Michigan Avenue. I like to show this photo because we have been interviewing senior citizens that remember victory gardening. And one woman lived in this neighborhood and she said that down by the lake, although this was technically in Division 7, she said that down by the lake there were victory gardens as far as the eye could see. So people took whatever space they could ‘as far as the eye could see’ to grow their victory gardens.



This image shows the Soldier Field Harvest Festival. It was held in the exhibition hall underneath the seats at the south end of the old Soldier Field. That’s the old old Soldier Field. It was sponsored by the Chicago Sun and practically every community in the Chicago Metropolitan Area participated. Hundreds of exhibits of garden produce, flowers and canned goods were featured. The Park District and commercial growers also participated. Thousands of visitors participated and pledged to grow victory gardens in 1944.


So Chicago, as you can see was really serious and very organized about their victory garden effort to the point where they passed an ordinance that if someone were to damage or pilfer or steal from a victory garden they would be fined. This was made effective in July of 1944, and to me this is like some big-wig’s wife got her tomato stolen so they decided that was the time to pass the ordinance. The fines range from $50 to $200 levied on people who were caught, busted, you know, pilfering victory gardens. In todays currency that would be $650 to $2600. A lot of money.


The question is: How would you know if something was a Victory Garden or just a ‘garden’? How would it apply?


What is believed to be the first system of reporting or recording Victory Garden efforts for statistical purposes of any part of United States, was established in Chicago in 1942. A simple distribution system of decals in exchange for information was used and a great deal of energy on the part of the Block Captains was required. They called it ‘decal-a-mania’. I think 6 color printing was a big deal back then. But 75,000 were distributed in 1942 and another 100,000 were distributed in 1943. And in return you gave your name and home address and the location and size of the ‘garden’. Downright boring, sleep inducing statistics. But, the reason we that we had 20,000,000 Victory Gardens and 40% of the produce was grown is because this idea took off. It was Chicago’s idea. It took off to the rest of the country and other metropolitan locations across the nation adopted this statistical method. So we thank Chicago for those sound bites that we now hear.


Then you started seeing these photos start showing up in all sorts of press photos. This is a demo garden at Lincoln Park. This is Little Village. This is another one. Now this tall gentleman here is the man responsible for the decals – he worked for the Park District, he was on the Victory Garden committee. He was the one who was responsible for the decal. He was very proud of that and was pointing one out.


I just want to point that 90% of these people had never gardened before, right. You can tell by the clothing that they are wearing. So, of course, it was very important to educate people because people didn’t know what they were doing. An extensive program of lectures in six different locations throughout the city were given. Horticultural staff and Park District led those lectures. Leaders in the seed trade, commercial growers and, in addition, individual lectures were provided by garden clubs and other organizations. In 1943, 162 sanctioned garden lectures happened across these six locations. And then countless garden club lectures, which I assume were unsanctioned happened but they are not in the statistics. And then there were mass meetings showing movies or slide shows.


If you go to YouTube and search for ‘Victory Garden Movie’, it will pull up one of these movies with a lot of sweeping orchestration and very serious narrator talking about how the family is growing their own food (in hushed tones). What they are doing. It’s interesting. I like to think that all these people got together and watched these movies.


(Movie excerpt:) You can have vegetables. Lots of them. On your table next winter. You can have your own fresh vegetables on your table this summer. If you have your own Victory Garden. Yes, there is no restriction on home canning and home processing of vegetables and garden fruits and berries. Plan your Victory Garden now! Get your garden plot lined up. Get the advice of a garden expert if you need it. And be prepared to ‘grow your own’ for victory. Join a garden club, or community garden movement. Or share a garden with your neighbor. You can help win the battle of food production. You can help our fighting men get the food they need. You can help save the vital metals used in commercial canning if you grow your own Victory Garden in 1943.


Newspapers were also involved. They passed out a lot of pamphlets that talked about Victory Gardens. Now this wasn’t ornamental, make it look pretty. This was just the down and dirty of growing food. At time we were just coming out of the Depression so about 30% of the people, the men, that may have been drafted could not be drafted because of malnutrition. So vitamins and nutrition played a big part in this information. Much like we need to day. Very similar situation.


There were also plans that people would hand out. You know if you have this size of garden here is a plan for a family of four. And here’s another one for a family of five. And these are very specific. They have a little bit of succession planting information. They recommend crops that are good for Chicago. This was all very specific to Chicago. There were articles in the papers. Special interest articles and weekly columns from garden experts.


And I love this. This is a radio program called ‘Know your Onions. On WGN’. It happened on Monday through Friday. And what happened is. You know – the Block Captains would be out in their gardens and you would be hearing about what the problem is. This year we had a lot of problems with squash vine borers, and a few other things. So, they would listen to the problems and this would trickle up through the OCD Bureaucracy and they would be very quickly able to address these problems and teach people ‘Ok. You are having a problem with squash pine borers. Here is how you deal with it.’


But there are also some public interest stories that happened on this show and I would like to share with you some of the headlines that I found in my research.


Rejoice kids.Spinach seems to be scarce.”


Did anybody read that book a couple of years ago? The 25 Dollar Victory Garden. No? It was a pretty good book. Anyway. That was a big profit on 64 cents. Back then it was a 64 cent Victory Garden.


Joe Green busy in his Victory Garden forgets taxes.


Then we get to the ‘fishing stories’.


The four foot radish?


Maybe. I don’t know. Maybe.


41” bean. Maybe.

12 pound cabbage. I can believe that.


But here is my favorite headline from the times.


Man who traded Victory Garden truck for beer. Wife divorces him.


Social Media.


How can that be. They didn’t have twitter back then. But they actually did have something that was social. They had these little kiosks that were at every garden and every corner. And so if there was a newspaper article, or ‘Hey we are all going to get together and weed.’ Or ‘Hey. Go see “Hair wins the war” at the library this weekend.’ All the information that you might need to know about your garden or about your community, what was happening, would be displayed in these little kiosks. And I have seen so many photos of these with little girls standing in front of them. I have seen them all over at the Library of Congress. It is just very interesting. That was a key point of how people got their information.


Now I have to tell you a funny story. The first time I did this lecture years ago, I got to this section – it was at the Chicago Flower and Garden Show – and a family walked in, saw this slide and turned around and walked out.


Plowing doesn’t sound like a very important or interesting topic but it was very important. And now I will proceed to show my collection of what I like to call ‘Plow Porn.” And it usually involves some proletariat looking guy sitting on a tractor shaking hands with someone in a fedora. And I mentioned International Harvester earlier. Early in 1942, International Harvester was approached and they very patriotically and promptly turned over three new plows to plow in the vacant properties within the city limits. The Park District supplied operators, fuel and supplies for these plows. Demand for plowing was so intense that rules were set up to regulate it.


These were some of the rules:



  • The plot had to be certain size with a certain opening
  • you had to have written permission from the land owner
  • it had to be inspected to ensure the soil was good, level, free of debris, it had to be sunny and fertile,
  • four families were required to participate

So, those were the rules for the Victory Gardens. At least as for where the plowing was concerned. And this wasn’t just community gardens. This also could be – well, families had to be involved but this could be small gardens or huge gardens. The plows were busy.


Let me read what the back of this photo says.


“This shows the tractor plow provided by the Victory Garden Department – Office of Civilian Defense through the cooperation of the Chicago Park District and the International Harvester Company. This Victory Garden is being prepared at the vacant lot at 5400 Ferdinand Street. The man with the rake is Frank L. Bennett, head of the Concert Bureau, who resided in the building shown in the background and was the moving spirit in developing this Victory Garden. This shows the same area included in photo A, except now Mr Bennett and his neighbors have something to show for their labors. The garden is located in the North Austin community where Mr. Bennett is Victory Garden Community Chairman. Notice the shrinkage in his waistline as a result of his gardening activity.”


This photo was taken at the inception of the community Victory Garden at 50th and Ingleside, a congested apartment house district of Chicago. The tractor plow provided by the International Harvester Company for the Victory Garden Committee is preparing the soil at an unsightly vacant lot that later will be transformed into a neat and productive garden. A local Civilian Defense group is studying the soil.


This next photo doesn’t say anything on the back of it but it is at Howard and Bell, which is the dividing line between Chicago and Evanston. And I love this photo because you see the plow, you see the little police man checking it out.


And this is another photo. Another ‘Plow Porn’ photo.


It sprung originally from the green prairie acres of the mid-West. “Chicago goes back to the soil as their citizens begin a tremendous Victory Garden campaign. One of the more than three hundred projected Victory Gardens on the North Side,(Division 6) is this portion of the Emerald Turf back campus of the Presbyterian Theological Seminary.”


So I was doing this lecture at De Paul, and when I was done the instructor said “,ou know that’s right around the corner.” And I said “Oh sure.” And he said “No. I mean literally.” So I gathered my stuff and went around the corner and it was literally ‘right around the corner’. So that shows where they were actually plowing. And this area past there is a giant soccer field. So, between the shrub and the soccer field there is about 30 feet by about 150 feet. So I’ve been picking on them and lobbying them about putting in another Victory Garden. Telling them it would be a good educational thing for the students to understand they have all that empty space. And I did learn that they put a Victory Garden in, a small garden. It is a little farther this way. But it is on that place. So it does make me happy that the land is being used for that again.


And this is the tally for plowing in 1942. This doesn’t include people who were gardening in their homes. It just includes community gardens that were plowed by the city. 508 separate community plots, some as large as 6 to 8 acres. An additional 1500 community plots, including Division 5, the largest Victory Garden in the nation, which is located at St. Louis and Foster – Northeastern University. It was 32 acres and 800 families gardened there.


I mentioned earlier that I had been talking with the senior citizens about their remembrances and one of the guys that I talked to, loved him, Ron. He gardened at that garden. And he used to be the Historian for the Tribune. So he said he’s like “LaManda, I believed it then, I believe it now, I believe that my Victory Garden is what won the war.” And that was the point of the Victory Garden movement. It was so people could feel like their effort made a difference. He told me once that he had to go speak there because he was a historian. He pulled into the parking lot and it was very close to where their garden was. He said he couldn’t get out of the car. He said he sat for ten or fifteen minutes just remembering the effort, 32 acres, 800 people and how he felt participating in something like that. It was a really beautiful story.


KIDs. Kids were not safe from the OCD. Recognizing the rising enthusiasm for Victory Gardening by adults might demand much of the lawn space of Chicago parks, the Victory Garden Department suggested in the latter part of 1942 that school children’s plots be set aside on park property during the 1943 season. All city parks were surveyed to determine suitable spaces, where good soil and sunlight were available and good recreational uses would not be interfered with. Working with public schools and youth leaders 5th through 8th graders were provided with plots on park property. Poor weather conditions in 1943 interfered with this plan. By the time growing could start many of the children had lost interest so instead of 30,000 plots being assigned only 14,000 were assigned.


I mentioned earlier that parades and hoo-haa was important and here is the children’s parade. This is what this photo said:


Thousands of children in the upper grammar grades are being assigned plots 5 by 12 in size in 65 of Chicago’s parks this week and next week. The typical planting plan for these plots includes a row of petunias at one end and zinnias at the other, with rows of lettuce, radish, snap beans, kohlrabi, carrots and beets in between. Complete seed packets containing just the right amount of seeds for each of these rows has been prepared and provided for the children with the compliments of Marshall Field and Company.


Here are some the children marching to their plots in parade form from Garfield park.


Did you ever see those Frankenstein movies where the villages are going after Frankenstein with the torches and .. this is like the daylight children’s version. These kids are so into it. Like that little girl. I wish I had my pointer and I could point some of these things out. That little girl with her digger. She is serious. There is some serious business happening here.


I also like it because you see some of these posters which we now think of as ‘retro’ but that was them, modern, carrying them around. The symbols of the war. This little guy in the front holding a sign that shows what he is going to garden. What he is going to grow. His different seed packets and his little plan.


I am going to veer off a little bit. We are talking about urban gardening but there was a big effort with rural areas too. You know 4H is a big program in rural areas for farm kids to get to show their livestock, etc, etc. Well there was a big program sponsored by Sears, nationwide, where they found young women garden leaders that were working in their 4H groups to lead people into developing victory gardens and establishing that. And this was an event that happened in 1943 where those young ladies were all pulled together from across the nation and rewarded for their efforts. I really love their little hats. And their little 4H aprons.


But back to Chicago.


This was a Boy Scout troop that found that they liked to march in formation to their garden because it prepared them for when they were going to be drafted.


Here are some ladies who are ready to garden for victory. They are little more cheerful than the would be soldiers in the previous picture.


This is a Chicago Park District photo.


This is one of the many successful Victory Garden Projects conducted through Chicago Public Schools. And this one was in the charge of a woman by the name of Miss Flursham, Principal of the Brandon School, and a nearby vacant lot was turned into a very fine flower and vegetable growing plot. A special ceremony and party was held the day the garden was started and plots were assigned. Right through the summer including the hot vacation period these youngsters continued their cultivation and insect control. Because of Miss Flursham’s excellent leadership a fine crop of vegetables resulted.


This was the photo that appeared in the newspaper. Here is the other photo that didn’t get shown. And I think this really illustrates what people who work with children go through. This guy in the front, I know he got busted for something. I like to joke that maybe he had a ‘sugar high’ but since sugar was rationed maybe it was molasses high. But if you look at the pandemonium of what those teachers had to put up with to make something like this happen. I really like that photo.


And of course they all had to report up to the OCD. And these were their report cards which showed their plot number, their row number, the gardeners name and address. Had the weeds been removed. Had the insects or disease been taken care of. Etc. Etc. Then all this information was collected and tallied on up through the ranks. The kids were not immune to the OCD of the OCD.


Then to the harvest.


We were leading the nation in 1942. We had 12,000 family gardens on 509 community plots. 290 acres. And in 1943 53,000 family gardens on 1500 community plots. 14,000 childrens gardens. And all this on 908 acres. Now this is just community gardens. This doesn’t include people that have home gardens or people that weren’t joiners that didn’t want to fill out the form to get the sticker. This is just based on the information that we have. And a lot happened in the suburbs but we are not talking about that right now.


So, how did Chicago do it?


We had government support. There were over-arching organizational structures. There was a donation of space and equipment. There was mass education. Promotion. Corporate and individual commitment. And recognition.


So let’s move on to today.


We have talked about 1942, 1943. And people often ask “Well, did the Victory Gardens just end at the end of the war?” And technically they did and they stopped being called Victory Gardens. But people continued to garden. Immediately after the war ended they became Freedom Gardens. Because the ‘Cold War’ was starting to percolate. So you would see information like ‘Food fights the red menace.’


You know, it was still a disaster over in Europe and people still had to get food. And whoever was giving them food probably was the side they were swinging toward. So if it was Russia, they could potentially become part of the Cold War Soviet Union. If it was America .. so food was a real tug-of-war in Europe.


After that, around the late 40’s early 50’s and the G.I.’s started coming back, inflation started happening. So, the gardens were called ‘Thrift Gardens’. People continued gardening to save money.


After that, around the mid-fifties, people were real (real) tired of gardening. You know, the world was changing. We were becoming the American Dream. People were moving to the suburbs and getting big cars. And they wanted to “keep up with the Jones’” in different ways. And that was sort of the end of the names to the Victory Garden movement. Of course people continued gardening through that but from a movement perspective, it really sort of moved on to landscape gardening and my house being better looking than my neighbors house. You know. My roses are better. My clematis is better. Whatever that is.


So, let’s talk about today. We have a new motivation for victory gardening. We don’t have a universal effort towards defeating an enemy but we all have our own individual enemies or our own little worries that we are fighting against. And they are all valid. It could be the economy. Your need to save money. It could be the transportation costs. Your carbon footprint. Food safety or other environmental concerns. GMO’s, GE, loss of seed diversity. Self sufficiency. And if you have grown your own food you know that home grown food tastes better. This is like preaching to the choir in this audience but you should know that this is the first administration since WWII and Eleanor Roosevelt to have a food garden at the White House. It tool a long time to get that garden there.

And just a little aside. The USDA did not want Eleanor Roosevelt to have a food garden at the White House. They did not want her to have a Victory Garden. They were worried that those precious resources, those precious seeds would be squandered by those city slickers trying to grow their own food and failing. Because that is what happened during WWI. After the first season and cities like Chicago did such a good job, the USDA was more into it.

I have found only one photo of Eleanor Roosevelt’s Victory Garden and she wasn’t even tending it. It was the Vice-President liked a photo op with his little girl. So it was interesting that the USDA wasn’t interested. Which leads me to my next point.

We can all collectively boo Tom Vilsack (http://en.wikipedia.org/wiki/Tom_Vilsack) but one thing that he has done which is interesting. Tom Vilsack is the USDA Secretary of Agriculture and one of his first mandates when he got his position was to start the People’s Garden program. So the USDA is considered the people’s department. It was started by Abraham Lincoln, and on the anniversary of his birth Vilsack put out this memorandum to all the USDA facilities around the world (there is 30,000 of them) that they should start a garden or they should participate in a garden. So now every State and every Province has a People’s Garden. They are all over Europe and they have given almost a million pounds of produce to local food banks.


I point this out because I feel it is the first time we have had the political environment to make a movement like this – a Victory Garden like concerted effort in cities to have these huge food gardens happen.


Let’s also talk a little about communities today. You know people lament that nobody goes to church any more or there are not social organizations. People don’t go outside because they have air conditioning. You know. Whatever those things that we don’t have now that we think people had back then, and we are missing something. But I like to point that people are forming communities in new ways today. And they want to be together. But they are also using social media in a lot of ways.

If you study statistics about how people learn to garden the number one statistic of all time is from a neighbor or a friend. That’s the best way. The number two statistic up until 2009 has been from books. In 2009 it became the Internet. So, your friends or neighbors are your first option. And your second option is the Web. I show you this because it kind of combines the best of both worlds. This book was written using Social Media. The authors reached out to people who had never gardened before and asked them how they would like to learn and what they would like to learn. Then they wrote this book using different private groups on Facebook and Ning and different social networks. They wrote the book in ninety days and published it. It is a very good gardening book. A basic gardening book. Good stuff. But the difference is that if you have a question on page 72 about the asparagus pea you can send them a ‘tweet’. You can send them an email. You can go to their Facebook page. So you still are able to have that community relationship with people despite the fact that you are reading a book.

The book is titled “Grocery Gardening” by Jean Ann Van Krevelen.


This is a garden at 77th and the lake. This is Rainbow Beach. It is a community garden that was an original WWII Victory Garden. The one existing community garden in Chicago that has lived its whole life from WWII until today.


I heard when I was speaking a couple of weeks ago that there are three gardens in Evanston that have continued to be gardens since WWII which is fascinating.


There is little funny story about this garden. It’s on the bus line. You know the Park District doesn’t like fencing gardens because they want their property to be available to everybody. That is what the Park District is all about. Well, this garden was on the bus line, and the bus driver would stop in front of the garden and let people go to the garden and pick food who weren’t gardeners, and then get back on the bus and then drive off. So the Park District agreed that maybe in these circumstances it would be OK to have a fence. And a lock on the gate. So the gardeners didn’t grow all of their food for the people on the bus.


So. I have been doing all of this research and kind of got excited about the idea of Victory Gardens and what sort of impact it could have today. And they seemed to do a really good job during WWII. They had this nice plan. So, I thought “Hmmm. What would it be like if we just followed those steps and just recreated a Victory Garden?”


That photo on the far side, circled, in my neighborhood. That was an empty lot. Everything else had been built up. But that one piece was an empty lot. So, I was driving down Peterson with my husband. I had been studying all of this and we had been talking to parents about it. You we had just bought our house a few years earlier and like .. “That’s the lot from the picture.” My husband said “What are you talking about?” I said “The lot from the picture. The lot from the picture.” He exclamed “You’re going to kill us!” Because we were driving. And I had just had my epiphany and said “That’s the lot from the picture.” And what? Maybe we should try and put in another Victory Garden.


So, I went to my Alderman and said “Hey. Did you know that this was a Victory Garden. I am a Master Gardener. I have been studying WWII history. Why don’t we try to put a Victory Garden in?” He said: “Go for it.” So we got a lot of support.


So here was my game plan, right. Government support. Donations of space and equipment. Over-arching organizational structures. Mass education. Corporate and individual commitment. And recognition. Not re-inventing the wheel. Just doing exactly what they did during WWII.


So what happened is we were trying to repeat history. History repeating itself. Or, as I like to say History re-eating itself.


So we started the Peterson Garden Project. This was our plan. As I mentioned, I am a Master Gardener. I have been a food gardener all of my life. So I tried to think through… Ok. We wanted to invite people who had never gardened before. We want to keep it as simple as possible. We want to remove as much complexity as possible. So people can go in here and really succeed. So that’s how we designed the garden.


So I was the Block Captain. And remember one of the Block Captain’s duties is morale boosting. So this was after the ground-breaking. And getting everyone to do a ‘sing-a-long’. I am not going to make you do a ‘sing-a-long’ today, so don’t worry. That’s me doing the ‘sing-a-long’ and that’s our alderman laughing at me.


We used social media to launch this garden. We launched it on April 27th of 2010 and within 5 days we had 50 people in the community that were meeting at a restaurant to talk about this garden. I thought if 20 people wanted to do it, it would be fun. And it ended up between those people and the people who joined after that becoming the largest edible garden in the city at that time. We used ‘Twitter’. And these are some of the people that started with us at that first spot. And we had to educate people. 90% of Victory Gardeners of WWII didn’t know how to grow their food. So, we invited people who had never grown their food. We did it very simply. All of our beds are raised beds. I am not advocating one way or another. There are a million ways to garden. And if your food is growing you are doing it right. But we did it specifically using square foot gardening techniques. And there is a tool on Gardener Supply called The Kitchen Garden Planner. So, we had homework for people. What do you want to eat? Here’s the size of the beds. Go to this website, pick what you want to eat, it has a ‘drag and drop’ tool where you can grab peppers and it will show you how many per square, and then people would bring their homework and we would teach them from there. This is an actual video. And I will talk through this because someone always asks “What kind of soil did you use?” “Where did you get the water?” So, I’ll answer some of those questions during this video.


Now this is a time-lapse of when we put the garden in 2010 and I also like to point out that like 1943 it was an incredibly wet, rainy, spring. So every time I planned a volunteer activity we got rained out. This was the bed. We put in 157 raised beds. It had been a foundation so there was a building but it was gone. As a result there was no top soil. I personally like raised bed gardening for many reasons. If you want to know what those are, we can talk afterward. But we put down 250 cubic yards of mulch. That is a lot. We built the beds. You can see how they are lining them all out now. We put a weed barrier on the bottom. And then we got 110 cubic yards of mushroom compost organic garden mix. We do all organic gardening. We teach everybody organic gardening.


Then people became members of The Peterson Garden Project. They paid a small fee to join. And then they got assigned their beds. Then you see the individuals starting to put in the soil. Everybody had to shovel their own soil. And then if you look carefully you will start to notice how square foot markings start to develop. So you can that those are some of the new gardeners. And this was done in the space of about 5 days. We condensed it down into this three minute thing. And we had some rain in there. But once we got the beds put in it went pretty fast. It was a bit of a ‘hair pulling’ exercise getting things delivered. Like our gate never seemed to be wide enough for the dump trucks coming so we would have to take the fence down and put the fence back up. And if the guy didn’t show up when he said he would – then what? It was a lot of drama at first. We sure learned a lot though, that first year.


But sometimes you stumble into some really wonderful things unsuspecting. I never really thought anything more than to satisfy my own curiosity of ‘What if we did this?’ But we learned. You know there is a famous permaculturalist that said “All of the problems of the world can be solved in the garden.”


We learned that very quickly in this garden. People were there for a lot of reasons. And mostly it was for the community. So the food growing was a good thing. But the community that formed out of it was a big deal.


So we decided this worked pretty good. Since we have done it before, what could we do to do more of this? Share this model and make this happen. We decided that we needed to be a ‘not for profit’ and we decided that we had a motto that was to recruit, educate and inspire a new generation dedicated to making urban food production the norm not the exception.


That was 2010. 2011 we started getting very serious about doing this on a bigger scale. As a result we spent a lot of time in 2011 building interest in the press. We had a lot of visits. A lot of people really like how the model worked. How the allotment cell garden worked for people who had a sense of community but they were still responsible for their individual piece. It wasn’t here is a huge space and let’s all agree on what we are doing. Which is a good thing. A good model. But we wanted to have a big garden and we wanted to have a lot of people learning. So that model seemed to work. This year was the seventieth anniversary of the first Victory Garden season of WWII. So, before the ‘Occupy Wall Street’ movement happened we had a program we were calling ‘1% for Victory’. We couldn’t call it that after ‘Occupy Wall Street’ but we essentially wanted to do 1% of what they had done in 1942.


Our goal was to put in 5 community gardens. Teach 7500 people how to garden. And have 5% of the produce donated to food agencies.


We were starting a course up in Division 6 which is where I live and it is very close to our original garden. These were the gardens that we worked on this year. The Land on Lincoln, we did not get our lease on time but we are building that one right now. Vegewaters on Edgewater (we like to have a sense of humor), Montrose Green is right is right by the CTA Brown Line stop so if you are on that platform you can look down on that garden. Global Garden is at Lawrence and Sacramento and is a 1.5 acre property that we share with an urban refugee training farm. And Stars Garden is on the property of an old Notell Motel across from the drive-thru Starbucks on Lincoln and Jersey which is very interesting because it still has the Stars sign up there. We were also very fortunate in that we put in a community garden at the Field Museum. This is Diane Whealy of Seed Savers Exchange (www.seedsavers.org) or (en.wikipedia.org/wiki/Seed_Savers_Exchange). They have really influenced a lot of my personal choices about food and gardening. It was a great pleasure to get to work with Diane. She is a very funny lady. She said “You know, I always like it when people have a little bit of sculpture in their garden.” This was in reference to the Brachiosaurus in the background. We ended up calling it the ‘Brocolli-asaurus’.


In all we were able to put in 4 of those gardens. And we were able to put in the Field Museum garden in this year. And for next year… who knows. We ended up having about 2600 people gardening with us on 690 plots. We gave 5% of the food from each of those gardens to local food pantries around the gardens. These are the statistics. But what this doesn’t show is the community. You don’t see how taking these spaces and giving people a skill that they can share and that applies to everybody. Because everybody eats. That’s the part that you don’t really see.


I get to talk about this stuff a lot. But we have a community of people that stepped up from the very beginning. You know, when I am out standing in the middle of that big empty lot and thinking to myself “Am I insane? What was I thinking of? How can I possibly do this.” Then people started coming. And saying “I could this” “I am in marketing.” “I am in fund raising.” And this amazing community formed to make this project happen. As I said, I stumbled on this idea. I had these stories from my parents. I love gardening. I was curious. And,.. I like to show this next photo. These are my parents today.


And this is a quote from JFK – “One person can make a difference. And everyone should try.”


During WWII everybody had to try. Everybody had to do it. And today we live in a nicer world in a way. You know we are not in conflicts like that. We are in conflicts. Don’t get me wrong. You know we are living in a world that WWII created. But I still feel that it is really important that we try. It is good to ‘not let the perfect get in the way of the good.’ Just get out there. You know if I had thought more about that project, I might not have embarked on it. I was traveling 80% of the time for work. And I felt that this was really compelling to me, so surely it would be compelling to other people. And it has certainly worked out.


I also like to point out sometimes with certain audiences that this is one lifetime and my mother says that when she was little everybody had chickens. We knew how to cut the head off a chicken. We got your eggs. We had a neighbor that had milk. Even in urban areas you had access to that stuff. And the food system has changed so dramatically in one lifetime. I sometimes visualize that I had my mother who taught me to can, and cook, and bake, and my dad who taught me gardening and many other really useful skills. And they are old. They are 85 years old. And I sometimes feel like that stuff is slipping away. Those skills are slipping away. And it is very important not to let that happen.


I feel very committed to the fact that we need to take care of ourselves a little bit more. So, that I do, in honor of my my mom, my ‘Rosie the Riveter’ mom.


And that is what I have to say today about Victory Gardens.

Citation: Lamanda A, Cheaib Z, Turgut MD, Lussi A (2007) Protein Buffering in Model Systems and in Whole Human Saliva. PLoS ONE 2(2): e263. https://doi.org/10.1371/journal.pone.0000263

Academic Editor: Axel Imhof, University of Munich, Germany

Received: November 21, 2006; Accepted: February 9, 2007; Published: February 28, 2007

Copyright: © 2007 Lamanda et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This study was supported by the University of Bern, Switzerland. None of the authors have any financial arrangements with any commercial organisation that could benefit from this study.

Competing interests: The authors have declared that no competing interests exist.

Introduction

When food enters our alimentary canal through the mouth, it first comes into contact with saliva, mainly excreted by the three major salivary glands. Acid containing beverages and foods as well as acids originating from the stomach are menaces to the teeth as these agents contribute to the erosion of tooth surfaces [1], [2]. Enamel and dentin are composed primarily of a carbonate substituted calcium deficient hydroxyapatite. When hydroxyapatite is in contact with water (saliva), hydroxyl ions (OH) can remove from the tooth surface during an erosive challenge like drinking an apple juice [3], vomiting [4] or gastro-oesophageal reflux [5]. If this process is repeated frequently, a loss of tooth substance, also known as erosion, may be the consequence [6], [7]. Dissolution ends and remineralization of the dental hard tissues occurs when the pH in close proximity to the tooth begins to rise [2]. This rise in pH is caused by saliva that permanently covers the structures forming the oral cavity. The salivary components responsible for the increase in pH are the three buffer systems, carbonate, phosphate and protein buffer system [8], [9]. The carbonate and phosphate systems have been well characterized [8], [10], [11], [12], [13]. With the exception of the knowledge regarding that the total protein concentration varies from 0.15 to 0.65% [14] and that 940 different protein species are present in saliva [15], [16], [17], [18], [19], [20], [21], [22], the information about the protein buffer system is scarce [23], [24], [25], [26], [27].

Over the last 40 years, the prevalence of dental erosion increased continuously [28], [29]. As the buffer characteristics of saliva can influence the erosion process [3], the aim of the present study was to quantify the buffer characteristics of a model system for salivary protein buffering and proteins prepared from whole saliva.

Materials and Methods

Acid/base titrations

Ten milliliters (ml) of the analytes (saliva samples or solutions) were placed in a vessel in a water bath and stirred at 37°C. First, 5 ml of NaOH 0.01 mol/l were added in steps of 200 µl to enclose the buffer range of di-hydrogenphosphate (pH 6.1–8.1) and then 25 ml of HCl 0.01 mol/l were added in steps of 200 µl. The pH was measured with a micro glass pH electrode 3 mm in diameter (DG 101-SC, Mettler Toledo, Schwerzenbach, Switzerland) and recorded after each addition step. Data points were fitted with Sigmaplot V9.0. Buffer values β, in [mol/(l×pH) [30]], were calculated as β = −ΔC/ΔpH [31] where ΔC is the amount of the titrator used (acid/base) and ΔpH is the change in pH caused by the addition of the titrator. The buffer value was used to quantify the buffer capacity. The buffer optimum was determined at the pH with highest buffer value within the buffer range. The buffer range, in pH units, was used to describe the pH interval where the buffering reaction of one or a mixture of compounds took place. The buffer power B, in µmol (H+) and µmol (OH), was used to quantify the amount of acid and base that can be buffered by a substance or a mixture of substances. The first derivative of the normalized titration curve (ΔC/ΔpH) was plotted against the pH and B was determined at the point with maximum slope. The experimentally measured values were compared to those that were calculated. The buffer power B [mol/l] was calculated according to the formula B = c2/2c, where c is the concentration, in mol/l, of the buffer component(s) [31]. Purified human salivary protein was analyzed by an automated titration system (Mettler-Toledo DL53, and the Software Lab X pro V 2.10.000) with the same titration parameters except that only 50 µl of acid were added per step.

Control group (human saliva samples)

The saliva was collected using a widely accepted procedure [32] under resting conditions, between 9:00 am and 10:00 am, from unmedicated volunteers who refrained from eating, drinking, smoking and performing oral hygiene measures for 2 hours (hr) before collection. Prior to saliva collection, the procedures were explained to the patients and an informed consent was taken from each of them. After collection, the buffer capacity of the saliva samples was determined using the CRT®buffer test (Ivoclar Vivadent, Schaan, Lichtenstein) as follows: the entire reaction pad was wetted with saliva. The saliva excess was dropped off from the test strip. After 5 minutes (min) of reaction time, the final color of the reaction pad was compared to the color of the standard color code chart. The samples were subjected to titration immediately after collection to prevent discrepancies caused by protease activity and the formation of ammonium by urease. For protein precipitation saliva samples were collected following the same procedure as described above at 9:00 am, 13:00 and 17:00.

Averaging saliva titration curves

Unstimulated saliva samples of 5 male subjects aged 35 to 45 with buffer capacities ranging from low to high according the CRT® buffer test were subjected to acid base titration as described above. The pH measurements were recorded with constant increments due to the monotone nature of the titration. As all pH measurement points in the saliva titration curves corresponded to each other, they were averaged and the standard deviation was calculated.

Search for model proteins

First, the availability (>1 g) of high-pure water soluble proteins at reasonable costs (<100 €/g) was checked. In this regard, about 100 proteins were selected. Then, the isoelectric point (theoretical best buffering point) of the selected proteins was calculated with the ProtParam analysis tool [33] at www.expasy.org. The proteins with buffer optima beyond the buffer range of hydrogencarbonate and di-hydrogenphosphate (pI within the range pH 3 to 5 or pH 8 to 10) were selected (group A).

Secondly, a list containing all known human salivary proteins was created from the literature. Their isoelectric points were calculated with the ProtParam analysis tool [33]. The proteins with buffer optima beyond the buffer range of hydrogencarbonate and di-hydrogenphosphate (pI within the range pH 3 to 5 or pH 8 to 10) were selected (group B). The amino acid sequences of the proteins in group A were aligned against the amino acid sequences of the proteins in group B with BLAST [34] or LALIGN [35]. A sequence in group A was selected if more than 30% of its amino acid sequence was identical to the amino acid sequence of a protein in group B.

Solutions

Inorganic buffer compounds.

Water: 10 ml deionized water was used. Di-hydrogenphosphate solution: 0.68 g (5 mM) KH2PO4 (Merck, Dietikon, Switzerland, for analysis, M = 136.09 g/mol, pKa = 7.1) was dissolved in 1000 ml deionized water. Hydrogencarbonate solution: 0.84 g (10 mM) NaHCO3 (Merck, for analysis, M = 84.01 g/mol, pKa = 6.1) was dissolved in 1000 ml deionized water. Hydrogencarbonate and di-hydrogenphosphate solution: 0.68 g (5 mM) KH2PO4 and 0.84 g (10 mM) NaHCO3 were dissolved in 1000 ml deionized water.

Organic buffer compounds.

Amyloglucosidase solutions: 10 µM (0.1%), 20 µM (0.2%), 50 µM (0.5%) amyloglucosidase from Aspergillus niger (Fluka BioChemika, Buchs, Switzerland Swissprot P69328, 640 amino acids, Mr = 98 kDa, pI = 4.35) was used as a model for human α-amylase (Swissprot P04745, 511 amino acids, Mr = 57.8 kDa, pI = 6.4). 0.01, 0.02 or 0.05 g amyloglucosidase were dissolved in 10 ml deionized water. Lysozyme solution: 340 µM (0.5%) lysozyme from hen egg white (Fluka BioChemika, Swissprot P00698, 147 amino acids, Mr = 14.6 kDa, pI = 9.4) was used as a model for human salivary lysozyme (Swissprot P61626, 148 amino acids, Mr = 16.5 kDa, pI = 9.4). 0.05 g lysozyme was dissolved in 10 ml deionized water. α-amylase solutions: 20 µM (0.1%), (40 µM) (0.2%) and 100 µM (0.5%) α-amylase from hog pancreas (Fluka BioChemika 10080, Swissprot P00690, 511 amino acids, Mr = 57 kDa, pI = 6.5) was used as a model for human salivary α-amylase. 0.01 g, 0.02 g and 0.05 g were dissolved in 10 ml deionized water. Amyloglucosidase and lysozyme solution: 10 mg (0.1% 10 µM) amyloglucosidase and 50 mg (0.5%, 340 µM) lysozyme were dissolved in 10 ml deionized water. α-amylase and amyloglucosidase solutions: 0.01 g (0.1% 10 µM) amyloglucosidase and 0.02 g (0.2%, 40 µM) α-amylase from hog pancreas were dissolved in 10 ml deionized water. α-amylase and amyloglucosidase solution: 0.01 g (0.1% 10 µM) amyloglucosidase and 0.02 g (0.2%, 40 µM) α-amylase from hog pancreas were dissolved in 10 ml deionized water.

Combined organic and inorganic buffer compounds.

Amyloglucosidase, lysozyme hydrogencarbonate and di-hydrogenphosphate solution: 0.01 g (0.1%, 10 µM) amyloglucosidase and 0.05 g (0.5%, 340 µM) lysozyme were dissolved in 10 ml of a solution containing 0.68 g (5 mM) KH2PO4 and 0.84 g (10 mM) NaHCO3 per 1000 ml deionised water. α-amylase, amyloglucosidase hydrogencarbonate and di-hydrogenphosphate solution: (0.1% 10 µM) amyloglucosidase and 0.05 g (0.5%, 100 µM) α-amylase from hog pancreas were dissolved in 10 ml of a solution containing 0.68 g (5 mM) KH2PO4 and 0.84 g (10 mM) NaHCO3 per 1000 ml deionized water. α-amylase, amyloglucosidase hydrogencarbonate and di-hydrogenphosphate solution: (0.1% 10 µM) amyloglucosidase and 0.02 g (0.2%, 40 µM) α-amylase from hog pancreas were dissolved in 10 ml of a solution containing 0.68 g (5 mM) KH2PO4 and 0.84 g (10 mM) NaHCO3 per 1000 ml deionized water. Salivary protein solution: The fresh prepared salivary proteins from 10 ml of stimulated saliva were dissolved in 10 ml deionized water. After adjustment of the pH to 7, all solutions were stored in gas-proof closed vessels.

Precipitation and dialysis of salivary proteins

Ammoniumsulphate was added to 10 ml of fresh collected resting saliva under constant stirring at 0°C. When 75% of ammoniumsulphate saturation was reached the mixture was stirred for additional 30 min. After centrifugation at 14000 rpm on a Hicen 21 centrifuge (Jepson Bolton, Watford, England) for 30 min at 4°C, the supernatant was removed and the obtained precipitate was dissolved in 5 ml deionised water. To remove all inorganic ions, the solution was dialyzed (Sigma dialysis sacks D6191-25EA, Sigma, Buchs, Switzerland) overnight at 4°C against deionized water. After dialysis, the volume of the dialyzed solution was adjusted to 10 ml.

Electrophoresis

Electrophoretic separation (SDS Page) was performed on a Mini-PROTEAN® 3 cell (BioRad, Rheinach, Switzerland) using a 17.5% polyacrylamide gel as previously described [36].

Fast RuBPS polyacrylamide gel staining

The polyacrylamide gels used for protein separation were visualized with Ruthenium II tris-bathophenantroline disulfonate (RuBPS). RuBPS was synthesized according to Rabilloud [37] and the staining was done as previously described [36] with modifications. In brief: the gel was placed in 50 ml of 40% Ethanol/10% acetic acid containing 1 µM RuBPS for 1 hr. After 20 min of destaining in 40% Ethanol/10% acetic acid, the gel was washed for 10 min in water and then scanned with an Amersham Storm 860 scanner (Amersham Bioscience, Freiburg, Germany). Images were processed with the advanced image data analyzer software (AIDA, v4.10, Raytest, Straubenhardt, Germany).

Determination of the protein concentration

The protein concentration was determined with the colorimetric method according to Bradford [38].

Results

Search for human salivary α-amylase and lysozyme substitutes

15 proteins fitted to the selection criteria of group A and 346 proteins of group B. Three proteins were chosen to serve as model proteins: lysozyme from hen egg which has an isoelectric point of 9.4 and 57% sequence similarity to human salivary lysozyme, amyloglucosidase from Aspergillus niger which has an isoelectric point of 4.35 and 35% sequence similarity to human α-amylase and hog pancreatic α-amylase which has an isoelectric point of 6.5 and 86% sequence similarity to human α-amylase. Among the α-amylases of all species, hog pancreatic α-amylase has the closest affinity to human salivary α-amylase. Therefore only the high sequence similarity was taken in account.

Inorganic buffer compounds

Water.

Water (Fig. 1,f) was found to have no measurable buffer power or buffer value.

Figure 1.

Titration curves with 150 pH measurements per curve of (a) 5 mM di-hydrogenphosphate, (b) 10 mM hydrogencarbonate, (c) 10 mM hydrogencarbonate plus 5 mM di-hydrogenphosphate, (d) 10 µM (0.1%) amyloglucosidase, 340 µM (0.5%) lysozyme, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (model system I), (e) 10 µM (0.1%) amyloglucosidase, 40 µM (0.2%) α-amylase, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (model system II) and (f) deionized water. The calculated buffer power is indicated in µmol per 10 ml of the analytes, in the internal scale.

https://doi.org/10.1371/journal.pone.0000263.g001

5 mM di-hydrogenphosphate.

A solution of 5 mM di-hydrogenphosphate (Fig. 1,a) was found to have buffer power of 30 µmol acid (hydrogen ions, H+) and 24 µmol base (hydroxyl ions, OH). Optimal buffering was measured at pH 6.7 with 0.003 mol/(l×pH). The calculated buffer power was 25 µmol acid and base.

10 mM hydrogencarbonate.

A solution of 10 mM hydrogencarbonate (Fig. 1,b) was found to have a buffer power of 74 µmol (H+) and 8 µmol (OH). Optimal buffering was measured at pH 6.2 with 0.005 mol/(l×pH). The calculated buffer power was 50 µmol acid and base.

10 mM hydrogencarbonate plus 5 mM di-hydrogenphosphate.

A solution of 10 mM hydrogencarbonate plus 5 mM di-hydrogenphosphate (Fig. 1,c), was found to have a buffer power of 110 µmol (H+), and 22 µσmol (OH). The distances between inflections Ia and Ib as well as between Ib and Ic were larger than the calculated 25 µmol (H+). Optimal buffering was measured at pH 6.5 with 0.008 mol/(l×pH). The calculated buffer power was 75 µmol acid and base.

Organic buffer compounds

10 µM (0.1%), 20 µM (0.2%), 50 µM (0.5%) amyloglucosidase.

A solution of 10 µM (0.1%) amyloglucosidase in water (Fig. 2,b, Fig 3,a) was found to have a buffer power of 68 µmol (H+) and 0 µmol (OH). The buffer range spanned from pH 3.3 to 5.3 with optimal buffering at pH 4.35 and a buffer value of 0.004 mol/(l×pH). A solution of 20 µM (0.2%) amyloglucosidase in water (Fig. 3,b) was found to have a buffer power of 140 µmol (H+) and 0 µmol (OH) and a buffer value of 0.02 mol/(l×pH) at pH 4.35, and a solution of 50 µM (0.5%) amyloglucosidase (Fig. 3,c) was found to have a buffer power (H+) that was out of the scale of this experiment (>200 µmol (H+)) and 0 µmol (OH) with a buffer value of 0.02 mol/(l×pH) at pH 4.35. The increase in the amyloglucosidase concentration from 0.1% to 0.2% increased the buffer value 7 times at pH 4.5.The pH difference (ΔpH) between the 0.1% and 0.2% titration curves (Fig. 3 curves a and b) was 1.9 pH units after addition of 100 µmol acid and 0.2 pH units after addition of 50 µmol base.

Figure 2.

Titration curves with 86 pH measurements per curve of (a) 340 µM (0.5%) lysozyme in water, (b) 10 µM (0.1%) amyloglucosidase in water, (c) 10 µM (0.1%) amyloglucosidase plus 340 µM (0.5%) lysozyme in water, (d) 40 µM (0.2%) α-amylase and (e) 10 µM (0.1%) amyloglucosidase, 40 µM (0.2%) α-amylase.

https://doi.org/10.1371/journal.pone.0000263.g002

Figure 3.

Titration curves with 80 pH measurements per curve of amyloglucosidase in concentrations (a) 10 µM (0.1%), (b) 20 µM (0.2%) and (c) 50 µM and α-amylase in concentrations (d) 20 µM (0.1%), (e) 40 µM (0.2%) and (f) 100 µM (0.5%).

https://doi.org/10.1371/journal.pone.0000263.g003

340 µM (0.5%) lysozyme.

A solution of 340 µM (0.5%) lysozyme in water (Fig. 2,a) was found to have no measurable buffer attributes, although the protein had 32 titrable groups [39].

20 µM (0.1%), 40 µM (0.2%), 100 µM (0.5%) α-amylase.

A solution of 20 µM (0.1%) α-amylase in water (Fig 3,d) was found to have a buffer power of 54 µmol (H+) and 4 µmol (OH). The buffer range spanned from pH 5 to 7 with optimal buffering at pH 6.3 and a buffer value of 0.0005 mol/(l×pH) and from 7.2 to 9 with optimal buffering at pH 8.4 and a buffer value of 0.0006 mol/(l×pH). A solution of 20 µM (0.2%) α-amylase in water (Fig. 2,d and 3,e) was found to have a buffer power of 66 µmol (H+) and 22 µmol (OH) with a buffer value of 0.003 mol/(l×pH) at pH 6.3 and a buffer value of 0.01 mol/(l×pH). A solution of 50 µM (0.5%) α-amylase (Fig. 3,f) had a buffer of 80 µmol (H+) and 24 µmol (OH), and a buffer value of 0.004 mol/(l×pH) at pH 6.3. In the 0.1, 0.2 and 0.5% α-amylase solution buffering was measurable from pH 5 to 8.4. The increase in the α-amylase concentration from 0.1% to 0.2% increased the buffer value 6 times at pH 6.3 and 15 times at pH 8.5. The pH difference (ΔpH) between the 0.1% and 0.2% α-amylase titration curves (Fig. 3, d and e) was 0.1 pH units after addition of 100 µmol acid and 2.1 pH units after addition of 50 µmol base.

10 µM (0.1%) amyloglucosidase and 340 µM (0.5%) lysozyme.

A solution of 10 µM (0.1%) amyloglucosidase and 340 µM (0.5%) lysozyme in water (Fig. 2,c) was found to have a buffer power of 84 µmol (H+) and 0 µmol (OH). The buffer range spanned from pH 3.3 to 5.3 with an optimal buffering at pH 4.5 and a buffer value of 0.007 mol/(l×pH).

10 µM (0.1%) amyloglucosidase and 40 µM (0.2%) α-amylase.

A solution of 10 µM (0.1%) amyloglucosidase and 40 µM (0.2%) α-amylase (Fig. 2,e) was found to have an acidic buffer power of 114 (H+) and 8 µmol (OH) with a buffer optima at pH 4.5 and a buffer value of 0.007 mol/(l×pH). Buffering was measurable between pH 3.5 to 5.5.

Buffering of purified salivary proteins.

10 ml of a solution containing the purified proteins from 10 ml whole stimulated human saliva collected at 09:00 am (Fig. 4,a) was found to have a buffer power of 11 µmol (H+, 100%) and 5 µmol of (OH, 100%). 10 ml of a solution containing the purified proteins from 10 ml whole stimulated human saliva collected at 13:00 (Fig. 4,b), was found to have a buffer power of 9 µmol (H+, 82%) and 3 µmol of (OH, 60%). 10 ml of a solution containing the purified proteins from 10 ml whole stimulated human saliva collected at 17:00 (Fig. 4,c), was found to have a buffer power of 8 µmol (H+, 73%) and 6 µmol of (OH, 120%). The buffer range reached from pH 5 to 8 with a buffer optimum in all three samples at pH 6.7. The buffer value was 0.0008 mol/(l×pH, 100%) in the 09:00 sample, 0.0005 mol/(l×pH, 63%) in the 13:00 sample and 0.0008 mol/(l×pH, 100%) in the 17:00 sample. The protein concentration was 1.83 g/l in the 09:00 am and 17:00 sample and 1.76 in the 13:00 sample. The difference between the 9:00 am and 17:00 sample and the 13:00 sample was 0.16%.

Figure 4.

Titration curves with 80 pH measurements per curve of purified salivary protein from 10 ml saliva. Saliva samples were taken at (a) 9:00 am, (b) 13:00 and (c) 17:00. Next to the titration curves the corresponding electropherograms sections containing proteins from 50 to 110 kDa are shown. Proteins were visualized by modified ruthenium (ii) tris bathophenantroline staining.

https://doi.org/10.1371/journal.pone.0000263.g004

Combined inorganic and organic buffer compounds

Model system I, a solution of 10 µM (0.1%) amyloglucosidase and 340 µM (0.5%) lysozyme, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (Fig. 1 curve d, 5A curve b, 5B curve b) was found to have a buffer power of 158 µmol (H+) and 38 µmol of base. There were two discrete buffer optima within the buffer range between pH 3.4 to 7.5. The first was at pH 4.3 with a buffer value of 0.005 mol/(l×pH), whereas the second was at pH 6.5 with a buffer value of 0.01 mol/(l×pH).

Model system II, a solution of 10 µM (0.1%) amyloglucosidase and 40 µM (0.2%) α-amylase, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (Fig. 1 curve e, 5A curve c, 5B curve c) was found to have a buffer power of 132 µmol (H+) and 45 µmol (OH). The solution had a buffer zone from pH 3.5 to 8 with buffer values starting from 0.004 mol/(l×pH) at pH 3.5 ascending to 0.008 mol/(l×pH) at pH 6.4 and descending to 0.003 mol/(l×pH) until pH 8.

Human resting whole saliva (Fig. 5A,a) was found to have a buffer power of 168 µmol (H+) and 42 µmol (OH). Human resting whole saliva had a buffer zone from pH 3.4 to 8 with buffer values starting from 0.005 mol/(l×pH) at pH 3.4 ascending to 0.01 mol/(l×pH) at pH 6.5 and descending to 0.004 mol/(l×pH) until pH 8.

Figure 5.

Panel A: Titration curves with 150 pH measurements per curve of (a) human saliva, (b) 10 µM (0.1%) amyloglucosidase, 340 µM (0.5%) lysozyme, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (model system I) and (c) 10 µM (0.1%) amyloglucosidase, 40 µM (0.2%) α-amylase, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (model system II). Panel B: Titration curve with 150 pH measurements per curve of (a) titration curve with 150 averaged pH measurements (5 per pH measurement point) of 5 male subjects with standard deviations indicated by grey bars. (b) 10 µM (0.1%) amyloglucosidase, 340 µM (0.5%) lysozyme, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate, (c) 10 µM (0.1%) amyloglucosidase, 40 µM (0.2%) α-amylase, 10 mM hydrogencarbonate and 5 mM di-hydrogenphosphate (model system II).

https://doi.org/10.1371/journal.pone.0000263.g005

The average of the human resting whole saliva collected from 5 individuals (Fig. 5B,a) was found to have a buffer power of 154 µmol (H+) and 36 µmol (OH). Average human resting whole saliva had a buffer zone from 3.5 to 8 with buffer values starting with 0.004 mol/(l×pH) at pH 4 ascending to 0.008 mol/(l×pH) at pH 6.5 and descending to 0.003 mol/(l×pH) until pH 8.

Electrophoresis

The electropherograms of purified salivary proteins obtained from the 3 whole saliva samples collected at 9:00, 13:00 and 17:00 showed 32 protein bands of molecular weights from 14 kDa to 250 kDa (Fig. 4). 31 protein bands had unchanged band intensity whereas one 50 kDa protein band showed changed intensity. After subtraction of the background the numerically integrated band areas of the 50 kDa protein (Fig 4a,c) were 10876 LAU (linear arbitrary units, 100%) at 9:00 am to 4476 LAU (40%) at 13:00 and 9989 LAU (90%) at 17:00.

Discussion

In the present study, the buffer attributes (value, power, range and optimum) of two model systems for human saliva, purified salivary proteins and single proteins were quantified by acid base titration. In the first step, the procedure was done by dissolving each of the model compounds in water separately. In the second step, the same compounds having the concentration as in the human saliva were mixed. In the third step, the procedure was done with human saliva and purified human salivary proteins. Then, the data obtained from the model systems, human saliva, purified salivary protein and single protein were compared. Amyloglucosidase from A. niger, lysozyme from hen egg and α-amylase from hog pancreas were used as model proteins because purified genuine or recombinant expressed salivary proteins were not available in the desired purity, quantity or at reasonable costs. α-amylase from hog pancreas has almost the same amino acid sequence as human salivary α-amylase and is its closest relative. Amyloglucosidase and lysozyme have the ideal physicochemical properties to demonstrate buffering beyond the buffer ranges of di-hydrogenphosphate and hydrogencarbonate. Amyloglucosidase and lysozyme have a high amino acid sequence similarity to their human counterparts whereas lysozyme from hen egg has almost the same molecular mass and the same isoelectric point as human salivary lysozyme. Moreover, this approach was feasible as the buffer function of a protein is dependent on its isoelectric point but independent from its catalytic properties or the species where it originates from. The total amyloglucosidase and lysozyme concentration in model system I (0.6%) as well as the total amyloglucosidase and α-amylase concentration in model system II (0.3%), did not exceed the total protein concentration found in human saliva [14].

346 human salivary proteins had their buffer optima beyond the buffer range of hydrogencarbonate and di-hydrogenphosphate (pH 5.1 to 8.1) what pointed out the plausibility that buffering beyond pH 5.1 to 8.1 could be based on proteins. Finally, buffering in saliva is likely to occur from proteins as in the rest of the human body where proteins are the most potent buffer substances [40].

In the present study, the experimentally determined buffer attributes of 5 mM di-hydrogenphosphate and 10 mM hydrogencarbonate were in agreement with the published data [8], [31] except for the observation that the carbonate system buffered 48% more acid than expected by calculation. The reason for this finding was attributed to the open system and is in agreement with the published data [30], [41].

Human whole saliva had a buffer zone spanning from pH 3.4 to 8 compassing the buffer ranges of hydrogencarbonate (pH 5.1 to 7.1) and di-hydrogenphosphate (pH 6.1 to 8.1). However, buffering in the range of pH 3.4 to 5 was not attributed to the buffering of hydrogencarbonate or di-hydrogenphosphate. It is known that at pH 4.3, hydrogencarbonate and di-hydrogenphosphate exhibit a maximum of 3% of their optimal buffer values [31]. The buffer values of the saliva samples measured in this study were in agreement with those published by Bardow [8] and even high concentrations of di-hydrogenphosphate and/or hydrogencarbonate can exhibit little buffer effect at pH 4.3 [31]. Therefore, it would be reasonable to conclude that we had the evidence that salivary buffering at pH 4.3 could be derived from the proteins.

The results of this study showed that the buffer value at pH 4.3 of model system I was 20 times higher than expected from 5 mM di-hydrogenphosphate and 10 mM hydrogencarbonate. However, at pH 4.3 model system I had exactly the same buffer value as human saliva and a buffer power that varied very little compared to the human saliva. The buffer value of model system II at pH 4.3 was 18 times higher than expected from 5 mM di-hydrogenphosphate and 10 mM hydrogencarbonate and almost identical to model system I and human saliva. This study showed that the purified salivary protein from 10 ml of whole saliva had the same buffer value at pH 5 as 5 mM di-hydrogenphosphate and 50% of the buffer value measured for 10 mM hydrogencarbonate. At pH 4.5, the buffer value measured for the salivary proteins was two times higher than for 5 mM di-hydrogenphosphate and 10 mM hydrogencarbonate. At pH 4, the buffer value measured for the salivary proteins was 6 times higher than for di-hydrogenphosphate and hydrogencarbonate where as at pH 3.5 the buffer value measured for the salivary protein was 6 times higher than for di-hydrogenphosphate and 4.5 times higher than for hydrogencarbonate.

For the combination of hydrogencarbonate and di-hydrogenphosphate with amyloglucosidase and lysozyme, 75% of the buffer value at pH 6.5 derived from hydrogencarbonate and di-hydrogenphosphate. The remaining 25% derived from amyloglucosidase and lysozyme. These results were unexpected as the fraction of the buffer value derived from proteins were responsible for only 3% of the buffer value at pH 6.5 [31]. Therefore, these findings were concluded as the evidence of the contribution of proteins to a larger fraction of the buffer value at pH 6.5 than hitherto assumed. These results, therefore, both support the hypothesis of Sellmann [42] regarding that proteins buffer at low pH values and the assumption of Freidin [25] who proposed protein buffer activity in a zone from pH 5.5 to 7.8.

The results of this study showed that a change in protein concentration (e.g. α-amylase) as small as 0.1% may change the buffer power up to two times and the buffer value up to 15 times. This change was within the same range as measured for the total protein content of the saliva samples taken at 9:00 am and 17:00 that had a 0.16% higher concentration than the samples taken at 13:00. As only one 50 kDa protein band of a total of 32 protein bands showed a lower intensity in the samples taken at 13:00 it was reasonable to conclude that this changed band caused the chances in basic buffer power and the buffer value at pH 6.7. Two time repetition of the experiments confirmed these results.

The 50 kDa protein band was subjected to protein identification which was performed by mass spectrometry and peptide mass fingerprinting (results not shown). Two proteins, α-amylase and serum albumin were identified. Although there are isoenzymes of α-amylase known with a masses around 50 kDa [43], the identification did not reach significance level. This was also the case for serum albumin. As SDS Page probable cannot provide high enough resolution to separate the different protein species that may be present in single protein band, further studies applying 2D electrophoresis will be necessary for unambiguous protein identification. In this study, the role of carbonic anhydrases and urease was neglected because both enzymes are found mainly in the enamel pellicle [9], [44] which was not included in the experiments.

The present study demonstrated that salivary buffering between pH 3.4 and 5 was not based on hydrogencarbonate and di-hydrogenphosphate but rather on proteins. Buffering between pH 5.1 and 8 was found to be based mostly on hydrogencarbonate and di-hydrogenphosphate but also seemed to be dependent on a larger fraction of proteins than thought before [8], [23]. There is some evidence that α-amylase could be one of the protein buffers in human saliva. In this context, it is worth mentioning the recently discovered human salivary α-amylase subproteom which consists of 67 amylase subspecies with isoelectric points ranging from pH 3.5 to 7.6 [43]. These α-amylase variants may provide like zwitterionic buffers [45], [46], a buffer system operational between pH 3.5 and 5 and auxiliary buffering through anionic and cationic sites present as non-interacting carboxylate and ammonium side chains between pH 5 and 8. However, further studies have to be undertaken to identify the protein buffer components in the human salivary proteome. The “Bufferomic” approach, as demonstrated in this article, is maybe only the first step in this direction.

Author Contributions

Conceived and designed the experiments: AL. Performed the experiments: AL ZC. Analyzed the data: AL ZC. Contributed reagents/materials/analysis tools: AL. Wrote the paper: AL AL MT.

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