|Publication number||US8100169 B2|
|Application number||US 11/929,578|
|Publication date||Jan 24, 2012|
|Filing date||Oct 30, 2007|
|Priority date||Feb 1, 2002|
|Also published as||US8104533, US20030155434, US20080048052, US20090014562|
|Publication number||11929578, 929578, US 8100169 B2, US 8100169B2, US-B2-8100169, US8100169 B2, US8100169B2|
|Inventors||Daniel P. Rini, H. Randolph Anderson, Guochang Zhao|
|Original Assignee||Rini Technologies, Inc.|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (10), Referenced by (2), Classifications (36), Legal Events (2)|
|External Links: USPTO, USPTO Assignment, Espacenet|
The present application is a continuation of U.S. patent application Ser. No. 10/342,669, filed on Jan. 14, 2003, now abandoned which claims the benefit of U.S. Provisional Patent Application Ser. No. 60/398,244, filed Jul. 24, 2002 and U.S. Provisional Patent Application Ser. No. 60/353,291, filed Feb. 1, 2002, all of which are hereby incorporated by reference in their entirety.
In recent years, attention has been focused on methods of high heat flux removal at low surface temperatures. This is due to the advancing requirements of the electronics industry that prevent high temperature heat transfer due to the operating conditions of electronics. Though the heat transfer process is very complex and still not completely understood, many evaporative spray cooling experiments have been performed which indicated the high heat removal capability of this cooling technique. The spray technique generally works in the following way; a spray nozzle is use to atomize a pressurized liquid, and the resulting droplets are impinged onto the heated surface. A thin film of liquid is formed on the heat transfer surface in which nucleate boiling takes place. The droplet impingement simultaneously causes intense convection and free surface evaporation. When a liquid with high latent heat of vaporization (such as water) is used, over 1 kW/cm2 of heat removal capability has been demonstrated.
The temperature of the cooled surface is determined by the boiling point of the liquid. Since the resulting heat transfer coefficient is very large (50,000 to 500,000 W/m2C) the surface temperature will be only a few ° C. above the boiling point of the liquid.
This type of cooling technique is most appropriately implemented when used to cool high heat flux devices such as power electronics, microwave and radio frequency generators, and diode laser arrays.
As stated above, diode laser arrays and microwave generators are devices that can be cooled with this type of impinging spray technology. Current market forces are driving these devices to increased power and size requirements. As a result, high heat flux devices are now being designed with surface areas much larger than 2 cm2. New high heat flux devices may be 10 cm2 to 1000 cm2, or larger. Spray cooling techniques offer the ability to provide superior cooling if atomizer nozzle and nozzle arrays can be developed that provide optimal cooling characteristics. Typical atomizer nozzles for painting, fuel atomization, and humidification do not meet the cooling need of these next generation high power electronic devices.
The subject invention pertains to a spray nozzle apparatus and method of use. The subject spray nozzle can be used to spray atomized fluid. The atomized fluid can then be incident on a heated surface such that heat is transferred from the heated surface to the atomized fluid. The subject spray nozzle can incorporate a means to initially destabilize a primary fluid inputted into the spray nozzle. A secondary fluid can then interact with the initial destabilized primary fluid so as to further destabilize the primary fluid such a combined flow of the primary fluid and the secondary fluid exits the spray nozzle and is atomized upon exiting the spray nozzle.
The subject invention also relates to a spray-cooling system and method of use. The subject spray-cooling system can incorporate a spray nozzle and a heat transfer plate. A device to be cooled, such as a laser diode, microwave amplifier, or other high power electrical device, can be placed in direct thermal contact with the heat transfer plate. The spray from the spray nozzle can then be sprayed onto the heat transfer plate. In a specific embodiment, a cellular transfer plate can be used and the spray from the spray nozzle can be sprayed into a cellular cavity or compartment, within the heat transfer plate. Heat is transferred from the heat source to the sprayed liquid via the wall of the heat transfer plate. The heat transfer plate can be attached to the spray nozzle housing, such that as the sprayed liquid exits the spray nozzle housing, the sprayed liquid enters a cellular cavity of the cellular heat transfer plate. Removing the cellular heat transfer plate from the spray-cooling assembly can allow the apparatus to be used as a conventional spray nozzle.
In a specific embodiment, the subject spray nozzle housing can house an inner nozzle portion and an outer nozzle portion. In a specific embodiment, the inner nozzle portion can incorporate swirl-developing channels. The inner spray nozzle can be placed within the outer nozzle portion. In a specific embodiment, the outer nozzle portion can be integral with the nozzle housing. A primary fluid can enter the inner nozzle via a connection to a primary fluid manifold or other fluid delivery means, flow through the inner nozzle, and exit the inner nozzle via, for example, one or more openings at, or near, the tip of the inner nozzle. In a specific embodiment, the primary fluid can flow through the inner nozzle portion via one or more fluid channels, such as holes drilled through the inner nozzle portion. The primary fluid exiting the inner nozzle portion can be initial destabilized as the primary fluid is inputted to a mixing chamber.
A secondary fluid can interact with the primary fluid as the primary fluid exits the tip, or near the tip, of the inner nozzle and enters the mixing chamber such that the primary fluid is further destabilized by the interaction with the secondary fluid. In a specific embodiment, secondary fluid can enter an outer cavity, or mixing chamber, surrounding the outside of the inner nozzle via a secondary fluid inlet port. The secondary fluid can flow from the outer cavity in channels cut on the outside of the inner nozzle such that the secondary fluid flows between the outside nozzle portion and the inner nozzle within the channels. Near the tip of the inner nozzle, the channels can spiral around the outside of the inner nozzle as the channels approach the tip of the inner nozzle. The channels can terminate near the tip of the inner nozzle near where the outer nozzle portion separates from the outside of the inner nozzle to form a small mixing chamber proximate to the tip of the inner nozzle portion where the primary fluid exits the inner nozzle portion. The secondary fluid can interact with the primary fluid exiting the tip of inner nozzle portion such that the primary fluid and secondary fluid leave the housing through a discharge port as a combined fluid flow. The combined fluid flow can be atomized upon exiting the discharge port.
Various alternative embodiments can be utilized to enable the interaction of a primary fluid and a secondary fluid creating a swirl in a mixing chamber so as to produce an atomized spray. A specific embodiment can have secondary fluid channels drilled into an inner nozzle portion from the proximal end of the inner nozzle portion, matching up with holes drilled from the distal end of the inner nozzle, such that the fluid exiting the secondary fluid channels exit the channels with a velocity having a radical component and a component tangent to the outer wall of the inner nozzle portion such that the secondary fluid exits in a spiral fashion. Additional alternative embodiments can deliver the secondary fluid via one or more tubes positioned to project spiraling secondary fluid into the mixing chamber. Other embodiments can incorporate alternative means for delivery of the secondary fluid in accordance with means known to those skilled in the art.
The subject spray nozzle can incorporate a primary fluid manifold which can function as an inlet port for the primary fluid to enter the one or more fluid channels of the inner nozzle. In alternative embodiments, each fluid channel of the inner nozzle portion can be individually served with primary fluid. In a specific embodiment, the subject invention can relate to a vapor atomizer which incorporates a pressure atomizer to initially destabilize a primary fluid wherein the initially destabilized primary fluid then enter a mixing chamber into which a secondary fluid is introduced which further destabilizes the primary fluid such that a combined flow of the primary and secondary fluids is atomized upon exiting the mixing chamber
The subject invention pertains to a spray nozzle apparatus and method of use. The subject invention also relates to a spray-cooling system and method of use. The subject spray-cooling system can incorporate a spray nozzle and a heat transfer plate. A device to be cooled, such as a laser diode, microwave amplifier, or other high power electrical device, can be placed in direct thermal contact with the heat transfer plate. The transfer of heat between the coolant fluids and the heating device takes place through the heat transfer plate. In a specific embodiment, a cellular heat transfer plate 1, as shown in
The assembly can be fastened together with a fastening device, such as a bolt, which can be held together with fastener holder 5, such as a threaded bore. Fastener holder 5 can be a simple through hole and another fastener holding mechanism can be used. In an alternative embodiment, the fastening system can be completely external to the assembly in which case fastener holder 5 would not be required. Other methods of fastening may also be utilized, such as brazing, welding, or other means known in the art. To prevent leaking between the parts in the assembly an o-ring 10 or other gasket mechanism can be used.
A heat source 20 can attach to the heat transfer plate 1. The nozzle assembly can then spray coolant fluid into cavity 25. The spray interacts with the rear wall of cavity 25 of the heat transfer plate 1, and heat is transferred from the heat source 20 into the coolant fluid being sprayed.
The spray enters the cavity 25 after exiting the discharge port 40 of the outer nozzle portion 38 as shown in
The inner nozzle portion housing 55, shown in
In alternative embodiments, more or fewer exits ports 125 can be used. Exit ports can also be positioned at other positions, for example near tip 75, in order to introduce initially destabilized primary fluid into mixing chamber 42 such that the secondary fluid introduced into mixing chamber 42 further destabilizes the primary.
The secondary fluid can enter the secondary fluid cavity 85 via inlet port 80. The secondary fluid then flows into one or more channels 70 cut into the outside of the inner nozzle. In a specific embodiment, as the secondary fluid flows in the channels toward the tip of the inner nozzle, the channels can turn from an axial direction, to a more radial direction as shown in
The secondary fluid exiting the end section of channel 115 can further destabilize the primary fluid so as to further increase the surface to volume ratio of the primary fluid. Initial destabilization of the primary fluid can happen when, for example, the primary fluid is separated into a plurality of streams, or otherwise physically destabilized. This initial destabilization allows the secondary fluid to further destabilize the primary fluid with less energy. The physical geometry of the subject mixing chamber allows the further destabilization of the primary fluid with much less energy, supplied by the secondary fluid, than prior techniques. In a specific embodiment, channel 115 makes an angle of between about 0° and about 90° with respect to the axis of atomization as the channel terminates into the mixing chamber 42 such that the secondary fluid exiting the channel enters the mixing chamber at about the same angle. This angle is labeled θ in
The primary fluid can enter the inner nozzle portion 56 via a primary fluid manifold 90, shown in
The assembly can incorporate a single nozzle and a single spray chamber 25, or multiple nozzle housings and inner nozzles, for example if the heat transfer plate incorporates multiple spray chambers 25. In alternative embodiments, the secondary fluid manifold can be replaced with a tube connection. In a specific embodiment, the secondary fluid is delivered via a tube for a single nozzle.
As the secondary fluid nears the distal end of inner nozzle portion, the outer wall of the inner nozzle portion tapers in toward tip 75 creating mixing chamber 42 between the inside wall of outer nozzle portion 38 and the tapered or cone-shaped portion and tip of inner nozzle portion 56. In the embodiments shown in
Although a variety of droplet sizes and velocities can be produced in accordance with the subject invention, in a specific embodiment an atomizing spray nozzle can produce droplets having mean diameters in a range from about 10 microns to about 200 microns and provides the droplets a velocity in a range from about 5 meters per second to about 50 meters per second. Preferably, the size and velocity of the particles are such that the effects of gravity are negligible. Producing small droplets at high velocity can allow the method and apparatus of the subject invention to be used with heated surfaces 2 oriented in a variety of directions (e.g. vertical or horizontal) and can make it easier to provide coverage of the surface 2 with the spray coolant.
Although the nozzle array assembly shown in
This atomized fluid then enters a corresponding compartment 16 of cellular heat transfer plate 15. The atomized fluid then interacts with the walls of compartment 16 and heat is transferred to the atomized fluid. In this way, heat from heat source 20 can be transferred through a wall of heat transfer plate 15 to the atomized fluid. The heated fluid is then discharged via one or more discharge ports 50. In alternative embodiments, the heated fluid can flow to the atmosphere such that discharge ports 50 are not needed. In addition, the atomized spray can be sprayed onto other types of surfaces from which heat is to be removed or can just be sprayed into the air.
With respect to the subject invention, for example in the embodiment shown in
In a specific embodiment of the subject invention, a pressure-vapor atomizer can incorporate two separate nozzles, or nozzle stages where a first nozzle, such as a pressure atomizer, can initial destabilize a primary fluid. The output from this first nozzle stage can then be inputted to the second nozzle stage for further destabilization. Referring to
In a specific embodiment, the subject nozzle sprays evenly onto an essentially circular area having a diameter of about 10 mm at a distance of about 13 mm from the tip of the nozzle. Table I provides comparison data for two of the best commercially available vapor atomizers two nozzles and a specific embodiment (RTI) of the subject invention. The subject nozzle (RTI) provides a much higher quality spray at much lower energy costs. The energy required for producing a spray comes from the pressures and flow rate of the vapor and liquid entering the nozzle. Therefore, the lower the pressure and flow rate the more efficient the nozzle operates, providing that the performance of the atomization is similar.
Hole size (mm)
Overall size H × D (mm)
25.4 × 22.8
25.4 × 22.8
13.1 × 6.2
Pressure Drop (psi)
Vapor Flow rate at 10 psi (scfh)
Mass flux (g/sec)
Nozzle performance can be quantified by the distribution of the sprayed liquid on a particular target. The three nozzles characterized in Table I were tested for spray distribution. Ideally, the distribution of the sprayed liquid would be perfectly even over the target area.
The SU22B nozzle produces an uneven distribution, with the highest mass flux rate (˜2.2 g/cm2 sec) in the center of the spray pattern. The spray pattern has decreasing mass flux moving away from the center of the spray toward the edge of the spray pattern and approaching essentially zero at the edge of the spray pattern. In contrast, the specific embodiment of the subject nozzle (RTI) referenced in Table I produces a more even spray pattern. Although, the mass flux is higher in the center of the spray pattern, which is cone-shaped, the variation of mass flux is less than about 40% across the spray pattern. The results of spray cooling with the specific embodiment and a prior art nozzle are shown in
The subject invention also relates to a heat transfer apparatus having an enhanced surface which can increase the rate of heat transfer from the surface to an impinging fluid. The subject enhanced surface can be incorporated with any of the heat transferred surfaces disclosed in the subject patent application or incorporated with other heat transfer surfaces. The subject invention also pertain to heat transfer apparatus, such as heat transfer plates, which incorporate the subject enhanced surfaces. The subject enhanced surfaces can also be utilized for heat desorbtion from a surface. In a specific embodiment, the subject system can comprise: a housing, a fluid pump or compressor, a nozzle array consisting of one or more nozzles, and a high heat flux source interface plate. The process begins with the housing. The housing contains the working fluid. The process as shown in
Evaporative spray cooling is enhanced by maintaining the thinnest liquid layer possible on the heat transfer surface. Pressure atomizer nozzles use high pressure liquid and vapor atomizer nozzles use compressed vapor to atomize the liquid coolant. Both types of nozzles can be used to produce a high velocity and lower droplet density spray. The result is a spray of liquid coolant onto the extended surface area which takes advantage of the additional surface area.
The pump 346 draws in the liquid coolant and pressurizes it to the desired pressure. The pressurized liquid goes to the liquid inlet port of spray nozzle 353. Compressor 350 draws in coolant vapor and pressurizes it to the desired pressure. The pressurized coolant vapor is sent to the vapor inlet port on spray nozzle 353. The compressed vapor and the pressurized liquid coolant combine in nozzle 353 to form small liquid droplets with a high velocity.
The spray nozzle 353 can be a vapor atomizer nozzle as shown using both compressed vapor and liquid coolant or a pressure atomizer nozzle, not shown, which uses only pressurized liquid.
The droplets impinge on cooling plate 360. Multiple surface area enhancements 370 are connected to cooling plate 360 as shown in
In a specific embodiment, protrusions, and/or indentations, having a height and/or depth, to diameter ratio of between about 0 to about 10 can be utilized. In further specific embodiments, a height, and/or depth, to diameter ratio of between about 1 and about 5 can be utilized. In another embodiment, protrusions, and/or indentations, having a height to spacing between adjacent protrusions, and/or indentations, ratio of between about 2 and 4 can be utilized. In a further embodiment, a height, and/or depth, to diameter ratio of about 3 can be utilized. In a specific embodiment, the number of protrusions, and/or indentations, density/spray cooling area is between about 1 and about 100 per square centimeter. In a further specific embodiment, the number of protrusions, and/or indentations, density/spray cooling area is between about 10 and about 20 per square centimeter. In a specific embodiment, the subject surface enhancements can increase the surface area, as compared to a smooth surface, by about 1 to about 5 times. In a further specific embodiment, the subject surface enhancements can increase the surface area by about 1.1 to about 2. In a specific embodiment, the center to center spacing of the subject protrusions, and/or indentations is between about (0.1) d and about 10d, where d is the diameter (or mean diameter) of the protrusions, and/or indentations. In a further specific embodiment, the center to center spacing is about d. In a specific embodiment, the roughness of the subject enhanced surface can have a RMS of between about optically smooth and about 100 micrometers.
The vapor coolant can then flow to a condenser, such as coil 342. The vapor condenses on the condenser coil 342 and forms liquid. The liquid then flows into reservoir 345. A heat extractors 341, removes the heat from the condenser 342 via thermal connection 340. The heat extraction can be a refrigeration cycle or an ambient heat exchanger.
A series of control devices including thermocouples, flow meters and level indicators are used to control the process in order to maintain the desired operating conditions.
Sample and embodiments described herein are for illustrative purposes only and various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and the scope of the appended claims.
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US5220804||Dec 9, 1991||Jun 22, 1993||Isothermal Systems Research, Inc||High heat flux evaporative spray cooling|
|US5353865||Mar 30, 1992||Oct 11, 1994||General Electric Company||Enhanced impingement cooled components|
|US5522452 *||Nov 22, 1993||Jun 4, 1996||Nec Corporation||Liquid cooling system for LSI packages|
|US5606201||May 24, 1993||Feb 25, 1997||Mannesmann Aktiengesellschaft||Fluid-cooled power transistor arrangement|
|US5718117||Apr 10, 1996||Feb 17, 1998||Motorola, Inc.||Apparatus and method for spray-cooling an electronic module|
|US5854092||Nov 12, 1997||Dec 29, 1998||Motorola, Inc.||Method for spray-cooling a tunable semiconductor device|
|US5934555 *||Mar 4, 1997||Aug 10, 1999||Abb Research Ltd.||Pressure atomizer nozzle|
|US20030155434||Jan 14, 2003||Aug 21, 2003||Rini Daniel P.||Spray nozzle apparatus and method of use|
|GB1356114A||Title not available|
|JPS62119947A||Title not available|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|US20100323312 *||Jun 9, 2010||Dec 23, 2010||Oztiry Akiler Madeni Esya Sanayi Ve Ticaret Anonim Sirketi||Electrically operated gas/oil burner|
|WO2014138263A1 *||Mar 5, 2014||Sep 12, 2014||Frto-Lay North America, Inc.||Shelf-stable snack with textured sauce|
|U.S. Classification||165/80.4, 239/398, 165/104.33|
|International Classification||A61P17/02, F25B39/02, F25B19/02, C12Q1/68, C12N5/10, F28F7/00, C07K14/52, A61K38/00, F25B1/00, G01N33/574, C07K16/18, B05B7/04, F28F13/02, C12P21/08, H01L23/473, B05B7/00, C07K16/24, C07K14/78, C12N15/12, C12N15/09|
|Cooperative Classification||B05B7/0475, F25B39/02, F25B1/00, H01L23/4735, F25B2339/021, F25B2341/0014, F28F13/02, F25B19/02, B05B7/0012, H01L2924/0002|
|European Classification||H01L23/473J, F28F13/02, B05B7/04C3D|
|May 8, 2012||CC||Certificate of correction|
|Jul 8, 2015||FPAY||Fee payment|
Year of fee payment: 4


text:
Never thought of attaching gutters to a wall to grow lettuce. Check out these vertical vegetable garden ideas that will solve your garden space problems so you can start growing your own veggies today!
Shanty to Chic always does amazing things. I think her kids must self sustain themselves with all she can done and then make it miraculously easy for the rest of us. Thanks for making this DIY Farmhouse table so frugal! More DIY Fixer Upper Farmhouse Styl
What to Plant in your Winter Vegetable Garden in February (or August!)
Growing a winter vegetable garden? Here is what to plant in February (or August in southern hemisphere). Garden, gardening, permaculture, vegetable gardening, Veggie gardens, permaculture design, mulching, mulch, self sufficient, Potager garden Landscaping, Backyard ideas, Permaculture, Compost, get started, start vegetable garden, tips, skills, frugal, survivalism, homesteading ideas, simple living, self sufficient small farm hacks, urban garden, #gardening #gardens

domain: agronomy


text:
The chelmer and blackwater navigation is the canalisation of the rivers chelmer and blackwater in essex in the east of england the navigation runs for 13 75 miles 22 13 km from springfield basin in chelmsford to the sea lock at heybridge basin near maldon it was opened in 1797 and remained under the control of the original company until 2003. It s a lovely walk into maldon and there.
Hoe mill lock. Motorhome pitches no. The site is a camping and caravanning club certificated site and we are able to accept up to five caravans motorhomes or trailer tents. 9 10 from 6 reviews.
Hoe mill lock is slightly more tucked away on the chelmer and blackwater navigation and hence is usually much less busy than paper mill lock east of here malden and the river estuary aren t far away just a few locks but it s generally a less busy stretch of water through some lovely farmland and countryside. These privately owned sites allow up to 5 motorhomes or caravans and 10 tents at any given time. Sup hoe mill lock river chelmer essex.
Hoe mill lock in essex. Hoe mill lock and the sugar mill. Hoe mill lock the causeway ulting maldon essex browse area cm9 6ra tel.
Hoe mill lock is a less frequented and well sheltered stretch of the chelmer blackwater navigation that s great for busy times of year and or stronger winds. Hoe mill lock has the largest drop on the navigation at 8 feet and 3 inches. Hoe mill lock the causeway ulting maldon essex browse area cm9 6ra tel.
9 10 from 6 reviews. 01245 382156 visit their website check digital tv coverage see weather forecast. As a result there is a danger of swamping the boats when filling the lock and so the lock paddles on the upper gates are located in underground side culverts.
Hoe mill lock cm9 6ra maldon uk rated 4 8 based on 4 reviews a beautiful place to stay and very friendly. Description this is a club member only site you may be able to join on site please check direct with the site. Hoe mill lock will remain accessible to vehicles via the causeway crouchmans farm road ulting road b1019 maldon road hatfield road langford road b1018 heybridge approach a414 maldon bypass spital road a414 maldon road oak corner a414 chelmsford road herbage park road church hill the street rectory road hoe mill road and vice versa.
Small member exclusive campsites each with their own individual charm. Paddleboarding hoe mill lock river chelmer. There is a mown grass area suitable for camping alongside the navigation at hoe mill adjacent to the car park.
01245 382156 visit their website check digital tv coverage see weather forecast rating. Maldon hoe mill lock maldon hoe mill lock certificated sites. It is now run by essex waterways ltd a wholly.
Please note that photos are not always of the actual campsite. Open all year rating.

domain: geography


text:
Ms Lim obtained a Bachelor of Science degree with Honours in Human Nutrition from the University of Otago, New Zealand, where she was awarded the prestigious Muriel Bell Memorial Prize. She gained a fellowship to pursue a Master degree in Community Nutrition at the University of Queensland.
Ms Lim has extensive experience in nutrition research at an international level as well as having pioneered intervention and educational programs here in Brisbane. She has been the nutritional consultant for the Heart Beat Centre, the Clear Mountain Health Lodge, and the Aboriginal and Islander Community Health Service. She is a board member of the Oceania College of Clinic Nutrition. She is author of the nutrition course run by the Open Learning Institute of TAFE.

domain: medical


PennDOT Offers Tips on how to Maintain Driveways During Winter Weather
With the arrival of the winter season in the northwest region, the Pennsylvania Department of Transportation (PennDOT) is reminding residents of the steps they can take to properly maintain driveway entry points.
Additionally, PennDOT is offering some tips for securing mailboxes during the winter:
- Ensure the mailbox has a strong support
- Construct a cantilever mailbox support that will "swing" a mailbox out of harm's way
- Clear snow from in front and on the sides of the mailbox, disposing of it properly, not on the roadway
- Use reflective tape or other material to make the mailbox easier to see during storms or dark hours
PennDOT also urges residents to not shovel snow onto state roads.
The Department of Transportation recommends moving snow to the right of the driveway and to clear a section prior to the driveway to lessen the amount of snow a plow might push onto the driveway.
For more information, visit PennDOT's website


text:
Sarah Jessica Parker and Matthew Broderick's two Manhattan townhouses both feature classic brick fa?ades and bath rooms any woman would like to own. Among their restrooms features a big white tub with local floating wood shelves and a newly installed bathtub with glass entrances.
If your New Year's resolution is to finally clean the chaos from your home, this year's minimalistic trend may inspire you to stick to it! Get rid of old accessories and cover appliances you don't use every day. Choose your kitchen counter d?cor properly and keep these elements small and sleek. Plan your kitchen this year for operation to keep chaos away.
Although most of us will never experience this lavish way of life, it's fun to see where well known celebs spend their time and find out about their flavour in home design. Join me as I take you inside the restrooms of five of the very most admired celebrities of our time.
Think about the design of your home and bathroom and that means you can filter down a few appears that'll go with your home design. Updating your lighting is a superb way to up grade your bathroom without having to spend lots of time and money home improvement the complete room. First, get a concept of which kind of lighting exists. Let's review a few types and that means you have a much better idea.
As the heart of the house, a kitchen can be an area that needs to be welcoming and useful. If you have an out-of-date kitchen, you may be considering a kitchen remodel this season. While it has much larger costs in advance, it can have a great impact on the ROI for your home. The common cost to redesign a kitchen is $18,727, with most homeowners spending between $12,986 and $21,306. Materials tend to be the costliest items when it comes to a kitchen remodel.

domain: design


text:
A little bit of history
In 1916, Mister Theodor Lauber bought the "Sonne" pension after a bankruptcy. He runned it successfully for several years until the age of 63 before selling it to Mr. Ulrich Imboden, from St. Niklaus (VS), as nobody wanted to take it over in his own family.
Mr Imboden was one of the biggest businessmen in the canton (state) of Valais at that time and was also a renowned politician (member of the state Grand Council). He sold the property to Mr. Stanislaus Kronig in 1948 in order to pursue other projects.
First steps for the Kronig family
Zermatt native and long time single, Mr.Stanislaus Kronig firmly believed that money should always be invested in real estate. That's the reason why he bought two years later (1950) the adjacent property from the Zermatter “bourgeoisie” for the sum of CHF 1'200.00. This deal also included the rights to use the Triftbach gorge, located next to the building.
After his death, the legacy was passed on to his close family: his sister Victoria and her brother Wilhelm, who had four daughters: Martha, Pia, Monika and Maria. Maria, married with Mr. Alban Lingg, received the modest pension "Sonne" in heritage.
Shortly before these events, Mr. and Mrs. Lingg had begun the construction of the luxurious "Albana" hotel, located in the Schluhmattstrasse. They saw in the pension "Sonne" an interesting complement for their business and renovated the property adding dormitories. This was the ideal solution for groups and for people with fewer resources. On the following picture, you can see the main building and the annex (both built in 1860) and further a third renovated building. You can notice that the chalets built on the hill above the hotel are still there nowadays.
The Forster-Lingg family
Following those renovations, Ms. Käthy Lingg, the daughter of Alban Lingg and Maria Kronig, took over the management of the hotel "Sonne". Ensuring the commercial part at the beginning, she became the Executive Director following the death of his father in 1974. She married Mr. Leo Forster a year later. Together, they invested substantial sums of money in the company to reach, a few decades later, the 4 star superior category.
The last transition comes in 2012 with the arrival of their two son, Alexander (born in1983) and Fabian (1985) as the fourth generation of the family to take control of the hotel.

domain: finance


text:
Roger Snoble announced today that he will leave soon after seven years as CEO of the Los Angeles County Metropolitan Transportation Authority. Release after the jump.
Los Angeles County Metropolitan Transportation Authority (Metro) CEO Roger Snoble today announced he would retire once his successor is on board. Snoble, 63, has headed Metro for seven years and, under his leadership, the agency garnered top national awards for excellence. He also played a key role in securing last fall’s passage of Measure R, a new half cent sales tax that will fund a comprehensive package of new rail and bus and street and highway improvements in the county over the next 30 years.
“After 43 years of public service in the transit industry, I think this is an opportune time to move on and pass the baton to another executive who will insure the implementation of Measure R,” Snoble said. “I did a lot of soul-searching and decided it’s in the best interest of the agency to have an executive on board soon who can commit the next four to five years in guiding the development of dozens of critical transit and highway projects that will be funded by the new sales tax, which will be collected starting next July.”
Snoble was praised by loal elected officials including Mayor Antonio Villaraigosa and Los Angeles County Supervisor Zev Yaroslavsky, who both serve on the Metro Board of Directors.
“Roger Snoble is a dedicated public servant who has provided solid leadership and a steady hand in restoring the credibility of the MTA while improving public transportation for millions of Los Angeles County residents and commuters,” said Mayor Villaraigosa.
Supervisor Yaroslavsky added, “Roger Snoble has led Metro to new heights in the delivery of expanded, quality public transportation to the citizens of Los Angeles County. What was a troubled and moribund agency more than 10 years ago is now recognized as one of the highest performing bus systems in the nation. On his watch the Orange Line, Eastside Gold Line and Exposition light rail line have been or are being built. With the passage of Measure R last month, Metro is poised to build the integrated public transit network that this region so desperately needs.”
Metro will recruit for a new CEO to head one of the nation’s largest public transportation agencies that is responsible not only for bus and rail operations that transport more than 1.5 million boarding passengers on an average weekday but also construction of new transportation projects such as the six-mile extension of the Metro Gold Line to East Los Angeles that is nearing completion. Metro also is Los Angeles County’s lead transportation planning agency and programs local, state and federal funds for transportation for the 88 cities, county, Caltrans and transit operators.
Snoble plans to attend next summer’s opening of the Metro Gold Line Eastside Extension, a project that is ahead of schedule, within budget and has maintained the best construction safety record of any public works project in modern history – nearly 4 million construction work hours without a lost time injury.
This project’s record is just one of the reasons that Metro earned the 2006 Outstanding Public Transportation System award for large transit properties given annually by the American Public Transportation Association (APTA).
Snoble guided the Eastside light rail project from inception. He also oversaw development of the highly successful Metro Orange Line busway in the San Fernando Valley and expansion of the Metro Rapid system that now has 28 lines crisscrossing the county. Since Snoble joined Metro Oct. 1, 2001, the agency also has taken a lead role in expanding its green power initiatives. Today Metro operates more than 2,500 buses powered by compressed natural gas. Its El Monte transit facility earned a gold LEED (Leadership in Energy and Environmental Design) rating for sustainability, and installation of solar panels on the roofs of Metro bus divisions annually saves Metro hundreds of thousands of dollars in energy costs.
Moreover, extensive surveys show customer satisfaction for Metro continues to improve. One sign of that is the fact that Metro ridership remains high despite plunging gas prices.
Snoble’s career in public transportation spans four decades. Besides heading Metro, he also was the chief executive at Dallas Area Rapid Transit (DART) for seven years (1994-2001), and he served as president and general manager of the San Diego Transit Corporation where he worked for 20 years before DART. Snoble started his career in 1965 as a planner for the TriCounty Regional Planning Commission in Akron, Ohio. He also worked for Akron Metro Transit.
Snoble has earned numerous awards for his leadership. APTA named him Transit Manager of the Year in 1998. Under Snoble’s leadership, DART was cited by APTA as the Transit Agency of the Year in 1997.

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Crustaceans that initially colonize a freshwater temporary pond can strongly bias the subsequent genetic composition of the population, causing nearby populations to be genetically distinct. In addition, these crustaceans have various reproductive modes that can influence genetic differentiation and diversity within and between populations. We report on two species of tadpole shrimp, Triops newberryi and Triops longicaudatus “short”, with different reproductive modes. Reproduction in the tadpole shrimp can occur clonally (parthenogenesis), with self fertilization (hermaphroditism), or through outcrossing of hermaphrodites with males (androdioecy). For all these reproductive modes, population genetic theory predicts decreased genetic diversity and increased population differentiation. Here we use mitochondrial control region (mtCR) sequences and nuclear microsatellite loci to determine if the difference in reproductive mode affects the high genetic structure typical of persistent founder effects. Previous authors indicated that T. newberryi is androdioecious because populations are composed of hermaphrodites and males, and T. longicaudatus “short” is hermaphroditic or parthenogenetic because males are absent. In our data, T. newberryi and T. longicaudatus “short” populations were highly structured genetically over short geographic distances for mtCR sequences and microsatellite loci (T. newberryi: ΦST = 0.644, FST = 0.252, respectively; T. l. “short”: invariant mtCR sequences, FST = 0.600). Differences between the two Triops species in a number of diversity measures were generally consistent with expectations from population genetic theory regarding reproductive mode; however, three of four comparisons were not statistically significant. We conclude the high genetic differentiation between populations is likely due to founder effects and results suggest both species are composed of selfing hermaphrodites with some level of outcrossing; the presence of males in T. newberryi does not appreciably reduce inbreeding. We cannot exclude the possibility that males in T. newberryi are non-reproductive individuals and the two species have the same mating system.
Citation: Horn RL, Kuehn R, Drechsel V, Cowley DE (2014) Discriminating between the Effects of Founding Events and Reproductive Mode on the Genetic Structure of Triops Populations (Branchiopoda: Notostraca). PLoS ONE 9(5): e97473. https://doi.org/10.1371/journal.pone.0097473
Editor: Norman Johnson, University of Massachusetts, United States of America
Received: January 2, 2014; Accepted: April 19, 2014; Published: May 13, 2014
Copyright: © 2014 Horn 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: The research was supported by the New Mexico Agricultural Experiment Station and by support to DEC from the Cooperative State Research, Education and Extension Service, US Department of Agriculture under Agreement numbers 2008-34461-19061 and 2008-40549-04328 in cooperation with New Mexico State University and the Texas A & M University System. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Aquatic invertebrates that disperse passively via an encysted embryo use a variety of transport methods to colonize new habitats. Abiotic factors, such as water and wind, , , , ,  and biotic vectors, such as birds, mammals, insects, amphibians and human activity can disperse invertebrates large distances , , , , , , , , , . Colonization of new habitats by a combination of these factors can be relatively quick, especially if ponds are located in close proximity , . The potential for dispersal, however, does not always equate to the actual immigration into ponds that is occurring by aquatic invertebrates . It is commonly observed that populations of many aquatic invertebrates can have a high degree of genetic differentiation despite being located in close proximity , , , a result not expected if contemporary dispersal is frequently occurring between populations.
In cyclically parthenogenetic zooplankton, De Meester et al.  emphasized the importance of local adaptation for monopolizing resources, thereby creating genetic differentiation between ponds in close proximity. Boileau et al.  concluded that founder events, not contemporary gene flow, have a pronounced effect on the population genetic structure of aquatic invertebrates that produce resting eggs. To demonstrate that genetic “barriers” are formed to inhibit immigration into populations, Boileau et al.  used simulations to show that FST does not decay for at least 2000 generations in a large population established by a few founders and subsequently experiencing migrant influx.
In addition to founder events, the mode of reproduction can also influence the amount of genetic structure and diversity in large Branchiopods . A population with individuals that reproduce via selfing experience a heterozygote deficit and decreased diversity due to small effective population sizes , . In addition, compared to species that outcross, populations of selfing individuals are genetically more isolated because of limited gene flow and often experience demographic fluctuations , .
The tadpole shrimp (Triops sp.) is a passively dispersing aquatic crustacean that has been said to use several forms of reproductive modes including parthenogenesis, hermaphroditism, androdioecy (a mix between outcrossing and hermaphrodites) and gonochorism (males and females that outcross) , , . Within Triops populations, low genetic diversity, deviations from Hardy-Weinberg equilibrium, large inbreeding values (FIS) and large population differentiation have been observed and has been attributed to founder events and the degree of outcrossing between individuals , , , , , .
Many of the previous studies have focused on Triops populations that are separated by distances of hundreds or thousands of kilometers between sampled ponds , , . Large geographic distances between populations makes it difficult to determine if it is the mating system influencing the genetic structure and diversity of Triops populations or if dispersal of encysted embryos is simply limited over long distances. The current study is designed to aid in differentiating between the influence of founding events, dispersal and mating systems by using nine Triops populations located within 30 km and encompassing two putative species with different presumed reproductive modes. Two of the species of Triops in the northern Chihuahuan Desert are T. longicaudatus “short” and T. newberryi , . Different reproductive modes are presumed for T. l. “short” and T. newberryi based on the male (absence of a brood pouch) to female (presence of a brood pouch) ratio within populations; T. l. “short” is comprised of all females and is assumed to reproduce via parthenogenesis or hermaphroditism whereas T. newberryi is thought to be androdioecious, with populations comprised of hermaphrodites that outcross with males . A recent phylogeny of Triops showed that T. l. “short” and T. newberryi are not monophyletic, calling into question whether species status is warranted .
The first objective of this study is to assess the genetic structure of each Triops species and determine what factors (founding events or contemporary dispersal) influence population differentiation. Secondly, we compare the effect of different presumed reproductive modes and the degree of inbreeding to the genetic diversity and structure of the Triops populations. We hypothesize, based on population genetic theory, that the androdioecious species will have more alleles, higher allelic richness, fewer private alleles, higher observed heterozygosity, lower FIS and FST, and relatively greater genetic variance within as opposed to between populations. The last objective is to evaluate whether the two putative species of Triops in southern New Mexico are reproductively isolated in the ponds in which they co-occur.
Materials and Methods
Sampling Methods and Study Sites
There were no special permits required to sample Triops as they are not listed as a protected species. Triops newberryi and T. l. “short” samples were collected in 2008–2011 at nine temporary ponds in southern New Mexico on public land, the Chihuahuan Desert Rangeland Research Center (CDRRC) and the Jornada Experimental Range (JER), both owned by New Mexico State University: two natural playa lakes (PL-07, PL-09), six modified playa lakes (PL-03, PL-05, PL-08, PL-11, PL-33, PL-36), and one man-made flood retention pond (FP-03) (GPS coordinates: Table S1 in File S1). All ponds were located near Las Cruces, New Mexico, USA within the Chihuahuan Desert (Fig. 1). The two species co-occurred at three of the playa lakes (PL-03, PL-05 and PL-07) (Fig. 1). Live samples were collected using 3 mm mesh seines and immediately placed in 95% ethanol for preservation. A description of field sampling methods, sample locations, and morphological verification of species, are given by Macdonald et al. . Each specimen was evaluated for the presence or absence of a brood pouch to determine sex of the individual. Samples were stored at −20°C until DNA isolation.
Mitochondrial Control Region Sequencing
Extraction of DNA followed a modified version of the HotShot method described by Montero-Pau et al. . Aliquots of 75 µl of the lysis buffer and neutralizing solution were added and samples were incubated at 95°C for 45 minutes. Amplification of the mitochondrial control region (mtCR) was performed by polymerase chain reaction (PCR) with newly developed primers (dloopF 5′GCACGAGTTAAGCCGATCTT; dloopR 5′CCACATGATTTACCCTATCAAGG) for T. newberryi (n = 160) and T. l. “short” (n = 66). Reaction volumes of 25 µl consisted of 10 µl GoTAQ Green Master Mix (Promega Corp., Madison, WI), 400 pM of each forward and reverse primer and 1 ng/µl of genomic DNA. PCR reactions were run in a Fisher thermocycler (Fisher Scientific Inc., Pittsburgh, PA) with the following conditions: 94°C for 2 minutes, followed by 35 cycles of 94°C for one minute denaturation, 50°C for one minute annealing, 72°C for one minute elongation and a final extension of 72°C for 15 minutes. PCR products were checked for strength of amplification on a 1% agarose gel. Purification of PCR products was performed with ExoSAP-IT (USB Corporation, Clevland, OH) following manufacturer’s protocol. Purified PCR products were then sequenced by NMSU’s MOLBIO Molecular Analysis Service in both forward and reverse directions (http://mmas.research.nmsu.edu).
Sequences were aligned using the assembly function in the program Geneious Pro v5.4.6 . Summary statistics, which included number of haplotypes, number of substitutions, number of transitions/transversions, number of polymorphic sites, nucleotide diversity (π), and haplotype diversity (h), for each playa and all samples collectively were obtained by Arlequin v3.5 . Relationships between haplotypes (maternal lineages) were resolved using the program TCS  to construct a haplotype network at the 95% confidence level.
The appropriate substitution model (TrN+I) for the data was selected based on the Akaike information criterion (AIC) using Modeltest v3.7 . The genetic distance between populations and between haplotypes using the p-distance and the Tamura-Nei model were calculated in the program MEGA v5.05 with uniform rates among sites and gaps treated as missing data .
The program Arlequin v3.5  was used to calculate an AMOVA and also generate pairwise ΦST values. A sequential Bonferroni analysis  was utilized to correct for multiple, nominal tests in the pairwise analysis. Pairwise genetic distances [ΦST/(1−ΦST)] were directly compared to log transformed pairwise geographic distance between the playas to test an isolation by distance hypothesis with the IBDWS v3.21 utility on the web that uses a Mantel test for the analysis , .
Genomic DNA for the microsatellite analysis was extracted by the phenol-chloroform protocol  and DNA was stored at −20°C. A total of 163 T. newberryi from six ponds and 156 T. l. “short” samples from six ponds were genotyped for eight loci developed specifically for Triops species found in southern New Mexico (TL-L-1, TL-S-5, TL-S-9, TL-S-13, TN-6, TN-7, TN-13, TN-14 ) and one microsatellite designed for T. cancriformis (TCB-99 ). To guarantee the loci are informative for both species , loci were chosen based on the primers’ ability to PCR-amplify DNA in both species and secondly to contain more than one allele per locus in order to avoid ascertainment bias as described in Ellegren et al. . PCR reactions were done in an UNO II cycler (Biometra, Göttingen) in a 15 µl reaction volume containing 0.3 pmol/µl forward and reverse primers (biomers.net, Ulm), 0.1 pmol/µl Cy5-labled M13 , 1X PCR-Buffer (10x reaction buffer without detergent or MgCl2; BD Solis Biodyne, Tartu, Estonia), 2.5 mM MgCl2, 0.2 mM DNTPs, 0.04 U/µl Taq-Polymerase (Fire Pol DNA polymerase, Solis Biodyne, Tartu, Estonia) and 2 ng/µl genomic DNA. Microsatellite PCR reaction conditions were 95°C for 3 minutes, followed by 35 cycles of 94°C for 30 seconds denaturation, primer specific annealing temperature for 60 seconds, and 72°C for 60 seconds elongation, before a final extension at 72°C for 3 minutes. Genotyping of all samples was performed on an Automated Laser Flourescence (ALF) II express (Amersham Pharmacia Biotech, Nürnbrecht). Internal and external standards as well as one reference sample (previously sequenced sample) were included on each gel to facilitate consistent scoring across gels. Alleles were scored with the AlleleLinks 1.02 software (Amersham Parmacia Biotech).
The expected and observed heterozygosity of each population and linkage disequilibrium were calculated in Genepop on the Web v4.0.10  and Bonferroni corrections were applied . The dataset was checked for null alleles using the program FreeNA  and applying the EM algorithm  to compare amplification success of non-specific microsatellite markers on the different species. Values are not reported due to the difficulty of assessing heterozygote deficit vs. null alleles in organisms with high inbreeding  and are instead used only as a proxy for successful marker amplification. GenAlEx v6.41  was used for an AMOVA to determine amount of variation between and within populations. The overall and pairwise FST, their significance values, FIS (the inbreeding coefficient), and allelic richness was calculated in FSTAT v18.104.22.168 as well as a two-sided statistical test to compare allelic richness, observed heterozygosity, FIS and FST between species . The proportion of selfing (S) in each population was calculated, based on the estimated FIS values, using the equation S = 2FIS/(1+FIS) . It is noted that there can be a bias when estimating S if the FIS values do not accurately represent inbreeding, and are instead from genotyping error or population substructure . Estimates of inbreeding and selfing rate were compared to the percentage of males in T. newberryi populations, as it would be expected that an increased proportion of males would cause an increase in the rate of outcrossing, therefore lowering selfing estimates. The migration rate (Nm)  between populations of Triops was calculated in GenAlEx v6.41 . Similar to the mitochondrial data, genetic distance, defined as FST/(1−FST), was compared to log geographic straight line distance between playas to detect the presence of isolation by distance using the IBDWS v3.21 utility on the web that uses a Mantel test for the analysis .
To visually determine the genetic structuring between populations within a species, a discriminant analysis of principal components (DAPC ) was performed using the adegenet v1.3–5  package in the R platform v2.15.2 . DAPC overlooks the within group variation and summarizes the amount of between group variation, making this method superior for assessing relationships between populations . A factorial correspondence analysis (FCA) of both species together was performed in GENETIX v4.05.2  to visually assess species designation and if hybrid individuals are present in the sample set .
Population Structure - mitochondrial Control Region
The T. newberryi mtCR ranged in length from 548 to 551 base pairs long with an overall total of 24 polymorphic sites, 21 substitutions and a transition to transversion ratio of 20∶4 (Genbank accession numbers KJ627793–KJ627799). Summary statistics for each playa and for all playas combined are in Table 1.
A TCS statistical parsimony network of T. newberryi mtCR sequences revealed that most individuals are of two haplotypes; 55 individuals had Haplotype 1 and 46 individuals had Haplotype 2 (Fig. 2). The placement of Haplotype 2 and 3 was not certain, but Haplotype 2 and 3 will connect to Haplotype 1 through an additional 12 and 16 mutational steps respectively, at 92% (data not shown). Two singleton haplotypes were observed from PL-07. The average distance between haplotypes was 6.6 mutational steps, excluding Haplotypes 2 and 3. In populations with more than one haplotype, the haplotypes were strongly divergent, with the average number of mutational steps between haplotypes greater than 6.6 steps, except for the singleton haplotypes in PL-07. For example, the 30 samples from PL-11 had two highly divergent haplotypes (with no connection at the 95% confidence level) indicating two genetically different maternal lineages within a population (12 mutational steps between Haplotypes 1 and 2 at 92%).
Each circle of the network represents a haplotype (Hap), the size of the circle is proportional to the number of individuals sequenced with that haplotype, each color represents a different sampled location, each line equates to one mutational step, and the small black circles are hypothetical haplotypes.
The overall ΦST value was highly significant (ΦST = 0.644, P<0.0033) indicating strong genetic differentiation across all populations of T. newberryi. Estimates of pairwise ΦST values indicated significant structure between sampled playas (P<0.0033, sequential Bonferroni correction), except for the comparison between PL-05 and PL-07 (ΦST = 0.059, P = 0.009) (Table 1). AMOVA indicated most of the variation was among populations (64.5%) versus within populations (35.5%) (Table S2 in File S1). Divergence between populations was not significantly associated with geographic distance (Mantel test, P = 0.65).
Of the 66 T. l. “short” samples sequenced, in contrast to T. newberryi, there was only one mtCR haplotype 549 bp in length (Genbank accession number KJ627792). The closest T. newberryi haplotypes to the T. l. “short” haplotype were Haplotypes 5 and 6, which differed by 18 pairwise differences. There were 18 polymorphic sites between the T. l. “short” haplotype and T. newberryi Haplotypes 5 and 6 consisting of 17 substitutions and 1 indel. No connection was found between the T. l. “short” haplotype and the T. newberryi haplotypes in the TCS network at any confidence level.
Population Structure - microsatellites
Results of the AMOVA for T. newberryi indicated that 70.4% of the variation was within populations compared to 29.6% of the variation among populations (Table S2 in File S1). In contrast, the AMOVA for T. l. “short” indicated that 35.2% of the variation was within populations compared to 64.8% of the variation among populations (Table S2 in File S1). There was a significant difference (P = 0.001) when comparing the species overall FST values; T. l. “short” has a significantly greater degree of genetic structure and reduced gene flow than T. newberryi. The overall FST value for T. newberryi was 0.252 and every pairwise FST comparison was significant after Bonferroni correction (Table 2). The FST values ranged from 0.080 between FP-03 and PL-36 to 0.491 between PL-03 and PL-36. Pairwise comparisons between PL-03 and every other sampled playa consistently had the largest FST values in the dataset (FST≥0.276; Table 2). The overall FST value for T. l. “short” was 0.600 and pairwise FST comparisons were significant after Bonferroni correction (Table 3) except for the comparison between PL-03 and PL-07. The FST values ranged from 0.024 between PL-07 and PL-08 to 0.794 between PL-03 and PL-33.
The shape of the DAPC scatterplot for T. newberryi was similar to that representing an island model of population structure, with four population clusters that overlapped (Fig. 3A ). The first axis of the DAPC separated the population ellipses consisting of individuals from PL-03 and FP-03 from the other populations (Fig. 3A). There were three distinct clusters in the DAPC scatterplot of T. l. “short” populations that were genetically more similar to each other than to the remaining populations [a (PL-09); b (PL-03, PL-07, PL-08); c (PL-05, PL-33); Fig. 3B]. Along the first axis in the DAPC for T. l. “short”, PL-05 and PL-33 were separated from the other populations (Fig. 3B). The second axis of the DAPC separated PL-09 from the remaining populations. The shape of the T. l. “short” scatterplot for the DAPC was similar to the population structure observed in a hierarchical island model , different from the island model seen in T. newberryi.
The three clusters of the DAPC for T. l. “short” are designated as (a) PL-09, (b) PL-03, PL-07, PL-08 and (c) PL-05, PL-33. The insert graph displays the discriminant analysis eigenvalues with the largest two values in dark gray: T. newberryi, first eigenvalue was 287.2; the second eigenvalue was 157.8; T. l. “short”, first eigenvalue was 6167; the second eigenvalue was 219.9.
The Nm values between T. newberryi populations were generally below one, indicative of gene flow that is below the threshold for mitigating the effects of genetic drift  (Table 2). Despite assumptions that are likely violated in natural populations when calculating Nm , comparison of the relative degree of migration between species can still be made. Three pairwise comparisons had values that were slightly above one (Nm = 1.097 to 2.152): FP-03 vs. PL-07; FP-03 vs. PL-36; PL-07 vs. PL-36 (Table 2). The Nm values between T. l. “short” populations were below one, except for three pairwise comparisons (Nm = 1.660 to 7.168): PL-03 vs. PL-07; PL-03 vs. PL-08; PL-07 vs. PL-08 (Table 3).
Genetic Diversity & Reproductive Mode
In T. newberryi populations, there were in total seven haplotypes, but the number of haplotypes in a population varied from one in PL-36 to five in PL-07. Although FP-03 had the least amount of sequenced individuals (n = 10), it had four haplotypes. The amount of nucleotide and haplotype diversity also varied from the lowest nucleotide and haplotype diversity in PL-36 at zero to the highest nucleotide diversity of 0.014 in FP-03 and the highest haplotype diversity in PL-07 at 0.625 (Table 1). For all populations combined, the nucleotide diversity was 0.015 and the haplotype diversity was 0.747. Modeltest results utilizing the AIC selected the TrN+I model as the best fit for the T. newberryi mtCR data. The smallest Tamura-Nei distance was between PL-05 and PL-07 (0.71%) and the largest occurred between PL-36 and FP-03 (2.34%) (Table S3 in File S1). The average genetic distance between playas was 1.60% and 1.79% based on p-distance and the Tamura-Nei model, respectively (Table S3 in File S1).
The number of alleles in T. newberryi microsatellite loci ranged from one to 14 with an average of 3.15 alleles across all loci (Table 1). The microsatellite TN-13, developed specifically for T. newberryi, was the most variable marker (21 alleles), while T. l. “short” specific markers were less variable (TLS-13, 2 alleles; TLS-5, 3 alleles; Table 1). The average allelic richness across all loci and populations was 4.12 (Table 1). The largest allelic richness was in PL-07 (4.36) and the smallest was in PL-11 (2.01). There was no evidence of linkage disequilibrium. Significant heterozygote deficiency after Bonferroni correction was observed for all loci in each population (Table 1).
For T. l. “short” the number of alleles ranged from one to seven with an average of 2.37 alleles across all loci (Table 1). Markers developed specifically for T. l. “short” (TLS-9 and TLS-5 ) were the most and least variable with nine and three total alleles, respectively. The average allelic richness across all loci and populations was 3.34 (Table 1) and was not significantly different to the allelic richness observed in T. newberryi populations (P>0.05). The largest allelic richness was in PL-07 (2.97) and the smallest was in PL-33 (1.54). Evidence of linkage disequilibrium was found in one playa lake (PL-09) only between loci TCB-99 and TLS-9. Observed heterozygosity was smaller than expected heterozygosity in each population and a significant departure (after Bonferroni correction) from Hardy-Weinberg expectations was observed in all populations (Table 1). The difference in observed heterozygosity between T. l. “short” and T. newberryi was not statistically significant (P>0.05).
In each population of T. l. “short” and T. newberryi, the individuals were examined for the presence or absence of a brood pouch, indicative of female or male. Every T. l. “short” individual had a brood pouch, congruent with the literature and the proposed parthenogenetic or hermaphroditic reproductive mode . Also consistent with the prediction of T. newberryi as an androdioecious species , there were males observed within every T. newberryi population. The amount of males per population ranged from one male out of 30 samples (3.3% males) in PL-07 to eight males out of 28 (28.6%) in FP-03 (Table 1).
Inbreeding analysis indicated a significantly large overall FIS value in T. newberryi: 0.601 (Table 1). All populations had large FIS values, ranging from 0.394 in PL-11 to 0.889 in PL-03 (Table 1). The estimated proportion of selfing per population was smallest in PL-11 (S = 0.565), largest in PL-03 (S = 0.941) and a (geometric) mean for all T. newberryi populations was S = 0.740 (Table 1). There was no correlation between the values of FIS or S in T. newberryi and the percentage of males per population. For example, PL-03 and PL-11 had the same percentage of males (13.3%), but the FIS and S estimates for these populations were the largest and smallest observed, respectively. Like T. newberryi, inbreeding analysis in T. l. “short” indicated a significantly large FIS value over all populations: 0.547 (Table 1). The population FIS values for T. l. “short” ranged from 0.277 in PL-08 to 0.658 in PL-05. A statistical comparison of the overall FIS values for the two species of Triops was non-significant (P>0.05). The proportion of overall selfing in T. l. “short” populations (geometric mean S = 0.689) was slightly lower than T. newberryi, with the largest proportion of selfing in PL-05 (S = 0.794) and the smallest in PL-08 (S = 0.433) (Table 1).
Visual Assessment of Species Designation
The FCA separated T. newberryi populations from T. l. “short” populations, with the exception of one T. l. “short” individual that did not cluster with either species group (Fig. 4). There was one of the eight loci (TN-14) in this individual (PL07-42; Fig. 4) where the allele combination 157/161 was observed, but was not present in any other T. l. “short” at the same locus. The first axis of the FCA accounted for 11.8% of the variation within the data set and the second axis accounted for an additional 7.4% of the variation. Within T. l. “short”, there was a clustering pattern in the FCA similar to the DAPC, in which individuals from PL-05 and PL-33 separated along the second axis from the remaining four T. l. “short” populations (Fig. 4).
The first axis of the FCA represents 11.8% of the variation within the data and the second axis represents an additional 7.4%. Each colored square represents a different population and the population clusters are labeled by species on the graph.
The results indicated that populations of Triops were highly structured genetically, even across short geographical distances. There was no evidence of a positive relationship between FST and geographic distance in either species. The genetic diversity varied across Triops populations, with slightly greater, but non-significant, overall genetic diversity in T. newberryi than in T. l. “short” for microsatellites and substantially higher haplotype diversity in the former species. Different models of population structure were observed for T. newberryi (island model) and T. l. “short” (hierarchical island) and both species of Triops had low estimated migration (Nm) between playa lake populations. Populations of T. newberryi and T. l. “short” were characterized by low diversity, a large degree of inbreeding and high proportions of selfing in all populations. There was no statistically significant difference between any of these measures (FIS, observed heterozygosity, allelic richness) between the species, but all interspecific differences except FIS and the proportion of selfing (S) were consistent with population genetic theory. The presumed androdioecious species (T. newberryi) had higher diversity, fewer private alleles, higher heterozygosity, lower FST, and greater within-population genetic variance. Lastly, there appears to be a clear genetic distinction between the putative species in southern New Mexico and no evidence of hybridization despite co-occurrence in some playa lakes.
Population Genetic Structure
Despite the potential for dispersal across habitats located in close proximity, playa lake populations of Triops in this study have a high degree of genetic differentiation. For example, PL-03 and PL-05 are separated by a distance of only 1.45 km, however the microsatellite results indicate T. newberryi and T. l. “short” are each genetically distinct across the two ponds. There is no evidence of isolation by distance in either species with mtDNA or microsatellites, indicating factors beyond immigration and contemporary dispersal are influential in structuring Triops populations in southern New Mexico.
A survey of T. cancriformis in Europe  and of Lepidurus packardi in California  attributed the high genetic structure observed between populations to founder events and high selfing rates. Similarly in the current study, there is evidence that founder events and genetic drift, not contemporary gene flow, is responsible for the genetic structure of T. newberryi and T. l. “short” in southern New Mexico populations. The migration estimates between most populations are too small (<1)  to counteract the effects of genetic drift, suggesting that isolation and drift after the initial founding event is responsible for the genetic structure. There is an excess of private alleles (3–29%) in each species that can be due to populations that have been separated over time and have experienced little to no gene flow .
Despite both species being highly structured genetically, there was a significantly higher degree of differentiation (FST values) among the T. l. “short” populations. The difference is illustrated in the different shapes of the DAPC plots; the hierarchical island model as seen for T. l. “short” populations is congruent with increased differentiation as compared to the island model for T. newberryi. The differences in the FST values between species could be due to a variety of factors including influence from the mating system (discussed below), different population sizes, a genetic bottleneck, and/or distinct colonization events. For example, the T. l. “short” PL-05 and PL-33 populations cluster together in the DAPC scatterplot, possibly indicative of the same genetic lineage colonizing and successfully reproducing in both of these locations. In addition, T. l. “short” PL-03, PL-07, and PL-08 populations cluster together and have slightly increased Nm values (>1), which may indicate founding lineages and historical connectivity. Zierold et al.  demonstrated the possibility of long distance dispersal and range expansions within European populations of T. cancriformis after glacial maximum, but significant differentiation was still observed. This trend observed in populations of Triops is more consistent with founder events being relatively more influential for the genetic structure than contemporary dispersal.
Comparison of Reproductive Mode
The highly inbred mating systems proposed for Triops species in southern New Mexico can also affect the genetic structure of populations. Populations of T. l. “short” in this study consist of all females (individuals have a brood pouch) that are thought to reproduce either through parthenogenesis or hermaphroditism , . Androdioecy is the presumed reproductive mode for T. newberryi because there can be some proportion of “males” (individuals lacking a brood pouch) in a population and individuals with a brood pouch can reproduce in isolation , , . In the current dataset, there was a ratio of “males” in T. newberryi populations ranging from 3.3%–28.6%. In comparing the results of the two Triops species with different presumed mating systems, T. l. “short” had fewer alleles, decreased allelic richness, and smaller HE and HO than T. newberryi (Table 1), however the comparison between species for these values were not statistically significant. Despite an absence of statistical significance, a selfing species, such as T. l. “short”, should have a lower number of alleles, reduced allelic richness and low observed heterozygosity . Also consistent with the effect of mating systems between species is the increase in the greater number of private alleles in T. l. “short”, the hierarchical island model of population structure and larger between species variation in T. l. “short” when compared to T. newberryi.
Inconsistent with the theory for a selfing species is the inbreeding coefficient and proportion of selfing, as T. newberryi has slightly higher values than T. l. “short”, which should be lower if T. newberryi populations experience some benefit of outcrossing with androdioecious matings between hermaphrodites and males. The reduced microsatellite variation, greater population differentiation and the single mtCR haplotype for T. l. “short” that we observed could also be evidence for demographic differences between species including more population bottlenecks in T. l. “short” , that T. l. “short” is an evolutionarily more derived species than T. newberryi  or that the persistence of founder events is stronger within T. l. “short” populations. The differences in diversity between species could also be due to genotyping error or amplification success of microsatellite markers within a species, however, there is no indication that allele dropout with cross-species microsatellites is more prevalent in T. newberryi or T. l. “short” (results not shown). With mitochondrial genes, Vanschoenwinkel et al.  suggested close genetic affinity between T. newberryi and T. l. “short” and Macdonald et al.  reported only modest differences between Triops species. Despite some males within T. newberryi populations, the FIS values were large for both species of Triops in southern New Mexico indicating an overall high degree of inbreeding. There was also no correlation between the fraction of males in each population and the FIS values of T. newberryi, as would be expected if populations with more males experience more outcrossing, and therefore, a smaller inbreeding coefficient.
Using allozyme data, Sassaman et al.  concluded that there was a difference in genetic variation between self fertilizing T. longicaudatus populations and androdioecious populations of T. newberryi in the southwestern United States, but inbreeding values were not given. Zierold et al.  did not observe significant differences of FIS values, heterozygosity, or allelic richness between T. cancriformis populations that had some males compared to populations with no males. Velonà et al. , however, did observe greater diversity in populations of T. cancriformis that contained males and are presumed to be outcrossing. While the genetic diversity may have differed between androdioecious and selfing populations, the inbreeding coefficient, as in this current study, was not able to distinguish between those populations that have some outcrossing compared to parthenogenetic populations .
Based on population genetic theory, there should be strong differences in the diversity and structure of a self fertilizing species compared to a species that experiences some outcrossing . Our results suggest that there are genetic differences between the two species with different presumed mating systems, consistent with the predictions of population genetic theory, however, there was a lack of statistical significance when comparing T. l. “short” and T. newberryi. The aforementioned evolutionary history and/or demographic differences between the species may explain why T. l. “short” has decreased genetic diversity compared to T. newberryi despite both species experiencing the same level of inbreeding. The inbreeding coefficients and selfing rate estimates are inconsistent with what would be predicted if T. newberryi populations were composed of hermaphrodites that can outcross with males (androdioecy). This might also suggest that although T. newberryi has slightly greater genetic diversity, the role of androdioecy does not alleviate the effects of inbreeding with regard to loss of alleles with genetic drift. In a confirmed androdioecious clam shrimp , there was a linear decline in inbreeding as the proportion of males increased, but we did not observe this in our data. Our results call into questions the role of males in T. newberryi. If there is an insufficient level of outcrossing to decrease the estimate of inbreeding, then do males have a reproductive role within a population? We cannot exclude the possibility that individuals lacking a brood pouch are non-reproductive and do not contribute genetic material to subsequent generations. If this is true, then T. newberryi and T. l. “short” populations could be composed of hermaphrodites that can outcross with other hermaphrodites and both species could have the same mating system. Outcrossing in both species is supported by the large values of FIS and selfing rates that are not near unity in value, which would indicate complete self fertilization as expected in a pure hermaphroditic scenario. Further clarification of the mating system will require a more direct method, such as a progeny array, to determine the reproductive mode of the Triops species within southern New Mexico. Work is also needed to clarify how adaptable the species are to their respective environments and if these differences contribute to the genetic diversity and structure of T. newberryi and T. l. “short” populations.
Putative Species Designation
We presently have analyzed three ponds in which T. l. “short” and T. newberryi co-occur in Southern New Mexico in this study. In a global phylogeny of the Notostraca, Vanschoenwinkel et al.  concluded that T. longicaudatus “short” and T. newberryi were not monophyletic and should be considered conspecifics. It is of interest to know if the species hybridize when they co-occur or if they are reproductively isolated , . The two species share a majority of their microsatellite alleles, although some alleles are more prevalent in one species.
The FCA of both species showed one individual from PL-07 that was ordinated in an intermediate position between the T. l. “short” and T. newberryi population clusters (Fig. 4); both species occur in PL-07. The morphology of this individual was consistent with T. l. “short” . The alleles of one microsatellite (TN-14 locus, alleles 157/161) make this sample different. Lacking pedigree information, we are unable to determine if this condition is identical in state or identical by descent and thus a hybrid origin.
The population genetic structure of T. newberryi and T. l. “short” in southern New Mexico appears to be strongly influenced by founder events and genetic drift. The high degree of genetic structure between populations at a local scale suggests that contemporary gene flow is not rapidly eroding persistent founder effects. There is evidence that Triops cysts may have the potential for dispersal via a variety of methods , , , , , , , but the results here suggest that the potential for dispersal is not realized either because immigration is too low to homogenize the genetic diversity or that migrants have a decreased ability to hatch and/or reproduce in ponds that are non-native. Despite slight differences in the amount of genetic diversity between the presumed selfing (T. l. “short”) and androdioecious species (T. newberryi), the results indicate similar levels of inbreeding occurring in both species. We conclude both species could be composed of hermaphrodites that can outcross with other hermaphrodites and androdioecy for T. newberryi remains unconfirmed. There are clear differences genetically between T. l. “short” and T. newberryi and little evidence to suggest hybridization between the species, which supports T. l. “short” and T. newberryi being distinct species as suggested by Macdonald et al. . Additional work could further clarify the mating systems and determine adaptability of migrants to new habitats.
We thank R. Sallenave, K. Macdonald, J. Alleman, N. Harings, M. Schiavon, M. Serena and B. Leinauer for help with sample collection.
Conceived and designed the experiments: RLH DEC RK. Performed the experiments: RLH VD. Analyzed the data: RLH. Contributed reagents/materials/analysis tools: DEC RK. Wrote the paper: RLH DEC RK.
- 1. Cáceres CE, Soluk DA (2002) Blowing in the wind: a field test of overland dispersal and colonization by aquatic invertebrates. Oecologia 131: 402–408.
- 2. Cohen GM, Shurin JB (2003) Scale-dependence and mechanisms of dispersal in freshwater zooplankton. Oikos 103: 603–617.
- 3. Vanschoenwinkel B, Gielen S, Vandewaerde H, Seaman M, Brendonck L (2008a) Relative importance of different dispersal vectors for small aquatic invertebrates in a rock pool metacommunity. Ecography 31: 567–577.
- 4. Vanschoenwinkel B, Gielen S, Seaman M, Brendonck L (2008b) Any way the wind blows - frequent wind dispersal drives species sorting in ephemeral aquatic communities. Oikos 117: 125–134.
- 5. Vanschoenwinkel B, Gielen S, Seaman M, Brendonck L (2009) Wind mediated dispersal of freshwater invertebrates in a rock pool metacommunity: differences in dispersal capacities and modes. Hydrobiologia 635: 363–372.
- 6. Proctor VW (1964) Viability of crustacean eggs recovered from ducks. Ecology 45: 656–658.
- 7. Green AJ, Figuerola J (2005) Recent advances in the study of long-distance dispersal of aquatic invertebrates via birds. Diversity Distrib 11: 149–156.
- 8. Frisch D, Green AJ, Figuerola J (2007) High dispersal capacity of a broad spectrum of aquatic invertebrates via waterbirds. Aquat Sci 69: 568–574.
- 9. Green AJ, Jenkins KM, Bell D, Morris PJ, Kingsford RT (2008) The potential role of waterbirds in dispersing invertebrates and plants in arid Australia. Freshw Biol 53: 380–392.
- 10. Vanschoenwinkel B, Waterkeyn A, Vandecaetsbeek T, Pineau O, Grillas P, et al. (2008c) Dispersal of freshwater invertebrates by large terrestrial mammals: a case study with wild boar (Sus scrofa) in Mediterranean wetlands. Freshw Biol 53: 2264–2273.
- 11. Vanschoenwinkel B, Waterkeyn A, Nhiwatiwa T, Pinceel T, Spooren E, et al. (2011) Passive external transport of freshwater invertebrates by elephant and other mud-wallowing mammals in an African savannah habitat. Freshw Biol 56: 1606–1619.
- 12. Beladjal L, Mertens J (2009) Diaspore dispersal of Anostraca by flying insects. J Crustacean Biol 29: 266–268.
- 13. Waterkeyn A, Vanschoenwinkel B, Elsen S, Anton-Pardo M, Grillas P, et al. (2010) Unintentional dispersal of aquatic invertebrates via footwear and motor vehicles in a Mediterranean wetland area. Aquatic Conserv: Mar Freshw Ecosyst 20: 580–587.
- 14. van Leeuwen CHA, van der Velde G, van Groenendael JM, Klassen M (2012) Gut travellers: internal dispersal of aquatic organisms by waterfowl. J Biogeogr 39: 2031–2040.
- 15. Bohonak AJ, Jenkins DG (2003) Ecological and evolutionary significance of dispersal by freshwater invertebrates. Ecol Lett 6: 783–796.
- 16. Hebert PDN, Finston TL (1996) Genetic differentiation in Daphnia obtusa: a continental perspective. Freshw Biol 35: 311–321.
- 17. Pálsson S (2000) Microsatellite variation in Daphnia pulex from both sides of the Baltic Sea. Mol Ecol 9: 1075–1088.
- 18. Zierold T, Montero-Pau J, Hänfling B, Gomez A (2009) Sex ratio, reproductive mode and genetic diversity in Triops cancriformis. Freshw Biol 54: 1392–1405.
- 19. De Meester L, Gomez A, Okamura B, Schwenk K (2002) The monopolization hypothesis and the dispersal-gene flow paradox in aquatic organisms. Acta Oecologica 23: 121–135.
- 20. Boileau MG, Hebert PDN, Schwartz SS (1992) Non-equilibrium gene frequency divergence: persistent founder effects in natural populations. J Evol Biol 5: 25–39.
- 21. Velonà A, Luchetti A, Scanabissi F, Mantovani B (2009) Genetic variability and reproductive modalities in European populations of Triops cancriformis (Crustacea, Branchiopoda, Notostraca). Ital J Zool 76: 366–375.
- 22. Holsinger KE (2000) Reproductive systems and evolution in vascular plants. PNAS 97: 7037–7042.
- 23. Charlesworth D, Wright SI (2001) Breeding systems and genome evolution. Curr Opin Genet Dev 11: 685–690.
- 24. Longhurst AR (1954) Reproduction in Notostraca (Crustacea). Nature 173: 781–782.
- 25. Zaffagnini F, Trentini M (1980) The distribution and reproduction of Triops cancriformis (Bosc) in Europe (Crustacea Notostraca). Monitore Zool Ital 14: 1–8.
- 26. Sassaman C (1991) Sex ratio variation in female-biased populations of Notostracans. Hydrobiologia 212: 169–179.
- 27. Sassaman C, Simovich MA, Fugate M (1997) Reproductive isolation and genetic differentiation in North American species of Triops (Crustacea: Branchiopoda: Notostraca). Hydrobiologia 359: 125–147.
- 28. Mantovani B, Cesari M, Luchetti A, Scanabissi F (2008) Mitochondrial and nuclear DNA variability in the living fossil Triops cancriformis (Basc, 1801) (Crustacea, Branchiododa, Notostraca). Heredity 100: 496–505.
- 29. Korn M, Green AJ, Machado M, Garcia-de-Lomas J, Cristo M, et al. (2010) Phylogeny, molecular ecology and taxonomy of southern Iberian lineages of Triops mauritanicus (Crustacea: Notostraca). Org Divers Evol 10: 409–440.
- 30. Stoeckle BC, Cowley DE, Schaack Y, Macdonald III KS, Sallenave R, et al. (2013) Microsatellites for North American species of Triops (Branchiopoda: Notostraca). J Crustacean Biol 33: 48–55.
- 31. Zierold T, Hanfling B, Gomez A (2007) Recent evolution of alternative reproductive modes in the ‘living fossil’ Triops cancriformis. BMC Evol Biol 7: 161
- 32. Macdonald III KS, Sallenave R, Cowley DE (2011) Morphologic and genetic variation in Triops (Branchiopoda: Notostraca) from ephemeral waters of the northern Chihuahuan desert of North America. J Crustacean Biol 31: 468–484.
- 33. Vanschoenwinkel B, Pinceel T, Vanhove MPM, Denis C, Jocque M, et al. (2012) Toward a global phylogeny of the “living fossil” crustacean order of the Notostraca. PLOS One 7: 4 e34998 doi:10.1371.
- 34. Montero-Pau J, Gómez A, Muñoz J (2008) Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs. Limnol Oceanogr Methods 6: 218–222.
- 35. Hogan BLM, Costantini F, Lacy E (1986) Manipulating the Mouse Embryo. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Woodbury, New York, USA.
- 36. Drummond AJ, Ashton B, Buxton S, Cheung M, Cooper A, et al. (2011) Geneious v5.4, Available: http://www.geneious.com/.
- 37. Excoffier L, Lischer HEL (2010) Arlequin suite ver 3.5: A new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Resour 10: 564–567.
- 38. Clement M, Posada D, Crandall KA (2000) TCS: A computer program to estimate gene genealogies. Mol Ecol 9: 1657–1660.
- 39. Posada D, Crandall KA (1998) Modeltest: testing the model of DNA substitution. Bioinformatics 14: 817–818.
- 40. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, et al. (2011) MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance and maximum parsimony methods. Mol Biol Evol 28: 2731–2739.
- 41. Rice WR (1989) Analyzing tables of statistical tests. Evolution 43: 223–225.
- 42. Jensen JL, Bohonak AJ, Kelley ST (2005) Isolation by distance, web service v.3.23. BMC Genet 6: 13 Available: http://ibdws.sdsu.edu/.
- 43. Ngan EC (2006) Isolation by distance web service with incorporation of DNA data sets. Master’s Thesis, San Diego State University.
- 44. Weber JL (1990) Informativeness of human (dC-dA)n.(dG-dT)n polymorphisms. Genomics 7: 524–530.
- 45. Ellegren H, Primmer CR, Sheldon BC (1995) Microsatellite ‘evolution’: directionality or bias? Nature Genetics 11: 360–362.
- 46. Schuelke M (2000) An economic method for the flourescent labeling of PCR fragments. Nat Biotechnol 18: 233–234.
- 47. Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86: 248–249.
- 48. Chapuis M-P, Estoup A (2007) Microsatellite null alleles and estimation of population differentiation. Mol Biol Evol 24: 621–631.
- 49. Dempster AP, Laird NM, Rubin DB (1977) Maximum likelihood from incomplete data via the EM algorithm. J R Stat Soc Series B Stat Methodol 39: 1–38.
- 50. David P, Pujol B, Viard F, Castella V, Goudet J (2007) Reliable selfing rate estimates from imperfect population genetic data. Mol Ecol 16: 2474–2487.
- 51. Peakall R, Smouse PE (2006) GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol Ecol Notes 6: 288–295.
- 52. Goudet J (1995) Fstat version 1.2: A computer program to calculate Fstatistics. J Hered 86; 485–486.
- 53. Hedrick PW (2011) Genetics of Populations. Massachusetts: Jones and Bartlett Publishers. 461 p.
- 54. Wright S (1951) The genetical structure of populations. Ann Eugen 15: 323–354.
- 55. Jombart T, Devillard S, Balloux F (2010) Discriminant analysis of principal components: a new method for the analysis of genetically structured populations. BMC Genet 11: 94.
- 56. Jombart T, Ahmed I (2011) adegenet 1.3-1: new tools for the analysis of genome-wide SNP data. Bioinformatics 27: 3070–3071.
- 57. R Core Team (2012) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available: http://www.R-project.org/.
- 58. Belkhir K, Borsa P, Chikhi L, Raufaste N, Bonhomme F (2004) GENETIX 4.05, logiciel sous Windows TM pour la génétique des populations. Laboratoire Génome, Populations, Interactions, Université de Montpellier II, Montpellier, France.
- 59. Pritchard VL, Jones K, Cowley DE (2007) Estimation of introgression in cutthroat trout populations using microsatellites. Conserv Genet 8: 1311–1329.
- 60. Slatkin M, Barton NH (1989) A comparison of three indirect methods for estimating average levels of gene flow. Evolution 43: 1349–1368.
- 61. Whitlock MC, McCauley DE (1999) Indirect measure of gene flow and migration: FST≠1/(4Nm+1). Heredity 82: 117–125.
- 62. Aguilar A (2012) Range-wide and local drivers of genetic structure in an endangered California vernal pool endemic crustacean. Conserv Genet 13: 1577–1588.
- 63. Ishida S, Taylor DJ (2007) Mature habitats associated with genetic divergence despite strong dispersal ability in an arthropod. BMC Evol Biol 7: 52
- 64. Galtier N, Depaulis F, Barton NH (2000) Detecting bottlenecks and selective sweeps from DNA sequence polymorphism. Genetics 155: 981–987.
- 65. Moritz C (1994) Applications of mitochondrial DNA analysis in conservation: a critical review. Mol Ecol 3: 401–411.
- 66. Pannell JR (2002) The evolution and maintenance of androdioecy. Annu Rev Ecol Syst 33: 397–425.

domain: literature


text:
WASHINGTON — Hillary Clinton is calling for a 4 percent “fair-share” income-tax surcharge on people making more than $5 million a year — an idea immediately denounced by rival Bernie Sanders as “too little, too late.”
The surcharge on the .02 percent of taxpayers who make more than $5 million a year is expected to raise $150 billion over a decade.
“I want to implement the Buffett Rule: Make sure there’s a minimum tax they have to pay,” Clinton said while campaigning Monday in Iowa.
The reference was to billionaire Warren Buffett, who has demanded higher taxes on the rich.
Clinton said she would later announce plans to go even further than the “Buffett Rule,” which would establish a minimum tax rate of 30 percent on those earning more than $1 million a year.
“I want to impose what I call a fair-share surcharge on multimillionaires because right now, we’re behind, and we need to get the wealthy and the corporations to pay more of their fair share,” said Clinton.
Sanders spokesman Mike Briggs, said the plan was inadequate.
“At a time of grotesque income and wealth inequality . . . Secretary Clinton’s proposal is too little too late,” Briggs said.
With Post Wires


text:
I remember the lunches that Mom packed for my sister and me every day when we went to St. Rita’s Elementary School. The school did not have a cafeteria that served lunch, so we had to bring our own every day.
We carried our lunches in metal lunch boxes with a hinged lid. They even had a matching thermos with its own cup attached to the top of it. The outside of the box might feature the image of a superhero, like Bat Girl, or maybe an image of a favorite TV show, like “Lost in Space.” Since they were metal, they were pretty sturdy and would last for several years.
Mom would pack us sandwiches wrapped in aluminum foil. They could be baloney, peanut butter and jelly, tuna, cheese, and even once in a while ham and cheese on white bread with a little mustard or mayo. Mom filled our insulated thermoses with juice or chocolate milk.
These were the foods and drinks that we were used to having and it was fun to carry our colorful lunch boxes to school.
A lot of kids today bring their own lunches to school, and I think that is a great way to have a sustainable school lunch. Students can carry reusable boxes or sacks as well as thermoses or reusable drink bottles. Aluminum foil wrap can be brought home to be washed and reused or recycled.
And today, there are so many more sustainable options for packing school lunches. Beeswax wrap and reusable sandwich bags made of organic cotton are just a few of the possibilities. There’s really no reason to create more waste or spend money on disposable plastic sandwich bags for school lunches.
Another bonus to a home-packed lunch is that parents can control the contents to ensure the food is healthy and eco-friendly!
Sometimes, the things that we had to do in the past out of necessity, like bringing our lunches to school, wind up being the best things that we can do for ourselves and for the Earth.

domain: food


text:
|Now it squirts loose, and the Ice Wolves have it.||Сада се тргну, а Ледени вукови га имају.|
|The ice, too, makes sounds.||И лед ствара звукове.|
|My feet were like ice, my head was on fire.||Стопала су ми била попут леда, а глава у ватри.|
|You're gonna scare away the Ice.||Уплашићеш лед.|
|What do they construct on the ice?||Шта они граде на леду?|
|Then, oddly, the hailstorm of ice bullets abruptly stopped.||Тада је, зачудо, туча ледених метака нагло престала.|
|I've got a guy. He wants it packed in ice and on the next plane.||Имам момка. Жели да је спакована у лед и на следећем авиону.|
- "Ice, damn you! Send to the nearest pub. And telephone at once for a doctor."
- W-we're half way up this ice wall.
- A road surface not covered by water, snow, ice or other substances. B.VII-22.
- It makes it seem almost as if we're looking through the ice to the ocean beneath.
- It's the same thing, except I don't have to see you sitting there like a pile of melted ice cream.
- The weight of the ice shelf on top of the ocean must be pushing plankton-rich sea-water up into the hole."
- What do you do if you wake up and your finger's in a bowl of melted ice?
- The ice cracked behind us and was driven with force towards the north; a breeze sprang from the west, and on the 11th the passage towards the south became perfectly free.
- Then two weeks before the Russians attacked us on the ice, I called Sergei, and it was Ruskov on the end of the line.
- Top it all off with two scoops of pistachio ice cream, and finish it with another sesame-seed bun.
- The wooded banks, with their base-lines of flashing ice, were slipping by.
- The wall of ice was five feet above the ground on which the boat rested.
- So anyone who wants to be president, line up by the ice cream machine.
- The CO2 emissions from fires help accelerate global warming, which in turn leads to more fire, while soot emitted from fires is often deposited on ice leading to accelerated melting.
- Rachel had plunged through the ice on a frozen pond while ice-skating when she was seven.
- Well, ice is melting, oil spilt, black president...
- Rachel gazed out at the silvery path of ice before them and tried to imagine what the hell the administrator of NASA was doing up here.
- For instance, as the Arctic ice cap melts, less sunlight is reflected.
- I feel like I want to giggle behind my hand at a lot of things and hold, but not eat, a big ice cream cone.
- By contrast, observe the lava's affect on this ice swan.
- I told Andy that he was on very thin ice when he snuck away for three months to the Caribbean.
- The champagne, and those girls and their dancing... those beautiful silver things, and the ice, and the caviar. . . .`
- Looks like they hit the ice at full impulse.
- I'd like, uh dozen bagels, a pound of gravlax and what kind of ice cream do you have?
- But with decreases in seasonal sea ice, not only is the Arctic now open to this wave noise, but the number of storms and the intensity of storms in the Arctic has been increasing.
- Just ice water in his veins, ticker tape coming from his heart.
- Greenland's ice sheet suffers from greenhouse gases... emitted elsewhere on Earth.
- Interstices, however, form not as pockets of saltwater but rather as highly branched networks of brine ice whose tendrils are as wide as a human hair.
- Just like water can be ice, steam and water.
- In the winter, it's incredibly cold, so you get these huge stepping stones, put them in a line across the ice, and as the ice melts, they drop and form a line.
- Decreasing sea ice is also causing increased erosion along coastal villages, and changing prey availability for marine birds and mammals.
- This decrease in sea ice is sometimes referred to as an increase in the open water season.
- Do you realize how thin the ice is you're skating on?
- A two-months' beard covered his face; and the beard, in turn, was matted with the ice of his breathing through the long seventy-mile run.
- But when the ice is frozen solid, and there are no big temperature shifts or current changes, the underwater Arctic has some of the lowest ambient noise levels of the world's oceans.
- We bring the body to him, he removes the kidney, packs it in ice, and we're good to go.
- Ice skating and hot chocolate at Kafe Leopold.
- Remember, with the ice and skating?
- He, um, took a break from the drive to do some ice fishing.
- A four-year-old boy fell through the ice of a frozen lake in Hanover, Germany.
- To look at a vast expanse of open water at the top of our world that used to be covered by ice.
- I want this ice fishing operation up and running by 0800 hours.
- Why aren't they here fetching you ice chips and morphine?
- "Bring some ice with you," I called after her. She nodded and ran.
- Dom Perignon's on ice and Bloomberg's closing Broadway for the ticker-tape parade.
- Ice fishing..that's all anybody..
- Another photograph was taken of the renowned American bandit nestled in his bed of ice.
- He wants to go ice fishing in October.
- You tell ice about Nick's lust for killing his brother?
- You ever come across a civilization with cookie dough ice cream?
- The horns of the crescent were blue-white from the polar ice caps.
- Making the raging river of air even more unnerving to Tolland was the slight downwind grade of the ice shelf.