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Arlington (18)
Vegetarian Restaurants: (16 Listings)
Amsterdam Falafelshop
3024 Wilson Blvd ~ PH: 703-243-2110
Inexpensive-- Vegan-friendly, Middle Eastern, Salad bar, Fast food, Take-out--- Additional Info: Franchise falafel shop offering top-it-yourself falafel and Dutch-style fries with dipping sauces. Gluten-free suitable except the pita. This location opened Oct 2014. Mon 11:00am-12:00am, Tue-Wed 11:00am-2:30am, Thu 11:00am-3:00am, Fri 11:00am-4:00am, Sat 11:00am-3:00am.-- Rating: 4.5 HappyCow(s)
Saran Indian Cuisine
5157 N Lee Hwy ~ PH: 703-533-3600
Moderate-- Lacto, Indian, Buffet, Catering, Take-out--- Additional Info: Vegetarian Indian restaurant featuring a lunch buffet. Some dishes contain dairy - vegans to ask. Tue-Sun 11:30am-9:30pm.Closed Mon.-- Rating: 4.5 HappyCow(s)
1028 N Garfield St ~ PH: 571-257-5231
Inexpensive-- American, Fusion, Fast food, Take-out, Non-veg--- Additional Info: Serves meat, veg options available. Fast causal eatery serving bowl-based entrees with choice of toppings, including lightly cooked veggies and house-made sauces. The Mediterranean and sweet chili pineapple signature bowls are both vegan, and the other signature bowls can be made vegan. Or build your own bowl.. Drinks include whole fruit smoothies. Bright space, on a busy corner. Opened 2015. Accepts credit cards. Mon-Sat 11:00am-9:00pm.-- Rating: 1 HappyCow(s)
Busboys and Poets - Shirlington
4251 S Campbell Ave ~ PH: 703-379-9756
Moderate-- Vegan-friendly, Lacto, Ovo, American, Beer/Wine, Non-veg--- Additional Info: Serves meat, veg options available. Its second location, opened in Aug 2007. Offers many vegan choices, marked on the menu, including vegan brunch eggs and egg wrap, soy yogurt and granola (for brunch), vegan pizza and calzone, quesadilla, falafel, hummus, portabello sandwich, veggie burger, PB & banana sandwich, baked tofu cake, vegan dessert. Has Wi-Fi. Accepts credit cards. Mon-Thu 8:00am-12:00am, Fri-Sat 8:00am-1:00am, Sun 8:00am-11:00am.-- Rating: 4 HappyCow(s)
Cibo Express
Ronald Reagan National Airport, Terminal A ~ PH: 718-656-6210
Inexpensive-- Vegan-friendly, Fast food, American, Take-out, Non-veg--- Additional Info: Serves meat, veg options available. Food counter at the airport. Offers prepared & packaged foods with items from local food companies. Has vegan sandwich, soy milk for coffee, Asian-style noodles. Accepts credit cards. .-- Rating: Not rated yet
Clarendon Silver Diner
3200 Wilson Blvd ~ PH: 703-812-8600
Moderate-- Lacto, American, Take-out, Non-veg--- Additional Info: Serves meat, veg options available. Reported has veggie food. Accepts credit cards. .c-- Rating: Not rated yet
Dama Pastry and Restaurant
1503 Columbia Pike ~ PH: 703-920-3559
Inexpensive-- Vegan-friendly, Take-out, Ethiopian, Non-veg--- Additional Info: Serves meat, veg options available. Ethiopian restaurant in the DC area. Offers vegan versions of some traditional meat dishes. You can get a veggie steak sandwich, kitfo (chopped seitan with greens and tofu soft "cheese"), dulet (chopped seitan, with peppers, onions, and spices from the kitfo), tibs (larger pieces of seitan sauteed in either mild spices or berbere sauce), doro wat (seitan on a chopstick "bone" with a tofu-based "hard boiled egg" in berbere sauce). The veggie combo varies day to day and is enough food for 2 or 3 people. Casual place, more about the food than the ambiance. Service sometimes be slow, but the food usually comes quickly. Note: The bakery next door usually has 1-3 kinds of vegan cake, and always has vegan cookies plus baklava (made with honey). Soy milk is available to make vegan cappuccino and other coffee drinks. Accepts credit cards. Mon-Sun 11:00am-10:00pm.-- Rating: 4 HappyCow(s)
Delhi Dhaba
2424 Wilson Blvd (at Barton) ~ PH: 703-524-0008
Moderate-- Lacto, Indian, Beer/Wine, Non-veg--- Additional Info: Serves meat, veg options available. Three item combo with rice or naan, salad, and raita for about $8. Accepts credit cards. .Call for hours - tell us.-- Rating: 2 HappyCow(s)
549 S 23rd St ~ PH: 703-271-6040
Moderate-- Vegan-friendly, Beer/Wine, Ethiopian, Eritrean, African, Non-veg--- Additional Info: Serves meat, veg options available. Offers a limited number of dishes, but does have vegan choices which are clearly marked. Staff is knowledgeable about vegan diet. Has outdoor seating. Accepts credit cards. Mon-Sat 11:00am-11:00pm.-- Rating: 4 HappyCow(s)
Galaxy Hut
2711 Wilson Blvd ~ PH: 703-525-8646
Moderate-- Vegan-friendly, Ovo, Lacto, American, Fast food, Beer/Wine, Non-veg--- Additional Info: Serves meat, veg options available. Must be 21+ years old to enter. All veg but will serve meat upon request. Little retro diner, bar, and concert venue tucked away in the side streets of the Clarendon area. Began offering an all-veg menu in fall 2012; offers vegan choices for all dishes (BBQ sandwich, cheesteak, chili, fries, etc). The atmosphere is dark and slightly indie-dive-y. Art adorns the walls and you have the option of eating at several retro gaming tables. Live music nights. Opened another location in June 2012 called Spacebar in Falls Church. Has outdoor seating. Accepts credit cards. Mon-Sun 5:00pm-2:00am.-- Rating: 4 HappyCow(s)
House of Steep
3800 Lee Hwy Ste D ~ PH: 703-567-1589
Moderate-- Thai, Japanese, Ovo, Non-veg--- Additional Info: Serves meat, veg options available. Tea house offering a large variety of teas and a menu of food which includes oatmeal, veggie wasabi dumplings, seaweed salad, edamame, hummus, pita chips, miso soup, veggie wrap, rice bowls plus baked goods. Has outdoor seating. Wheelchair accessible. Accepts credit cards. Mon-Fri 7:00am-8:00pm, Sat 8:00am-8:00pm, Sun 10:00am-5:00pm.-- Rating: 3 HappyCow(s)
Le Pain Quotidien
2900 Clarendon Blvd ~ PH: 703-465-0970
Moderate-- Vegan-friendly, Ovo, Lacto, Organic, American, Western, Bakery, Non-veg--- Additional Info: Serves meat, veg options available. International restaurant chain featuring artisan bakery and communal table. Menu includes several ovo-lacto vegetarian dishes as well as vegan ones (denote by carrot sign). Accepts credit cards. .-- Rating: 4 HappyCow(s)
Matsutake Sushi
Ronald Reagan National Airport, Terminal B/C ~ PH: 703-417-0521
Moderate-- Japanese, Beer/Wine, Take-out, Non-veg--- Additional Info: Serves meat, veg options available. Pre-security sit-down sushi restaurant. Vegans could get a 18-piece veggie roll combo and inari. A couple noodle and rice dishes are or can be veg, and there's a vegetarian dinner box with tofu teriyaki. Accepts credit cards. .-- Rating: 4 HappyCow(s)
South Block Co.
3011 N 11th St ~ PH: 703-741-0266
Inexpensive-- Lacto, Ovo, American, Fast food, Juice bar, Take-out, Non-veg--- Additional Info: Serves meat, veg options available. Juice and smoothie bar offering coffee drinks, cold-pressed juices, smoothies, wraps, breakfast and acai bowls. Uses organic produce. Keeps on hand almond, soy, and coconut milk for drinks. Accepts credit cards. Mon-Fri 7:00am-8:00pm, Sat-Sun 9:00am-5:00pm.-- Rating: 4 HappyCow(s)
4075 Wilson Blvd ~ PH: 703-522-2016
Moderate-- Vegan-friendly, Ovo, Lacto, Salad bar, American, Take-out, Non-veg--- Additional Info: Serves meat, veg options available. Fast casual salad bar chain restaurant. Offers create-your-own salads, juices, wraps. Has outdoor seating. Accepts credit cards. Mon-Sun 11:00am-10:00pm.-- Rating: 3 HappyCow(s)
2710-B Washington Blvd ~ PH: 703-528-2828
Moderate-- Vegan-friendly, Ovo, Lacto, Pizza, American, Take-out, Delivery, Non-veg--- Additional Info: Serves meat, veg options available. Franchise pizzeria. Offers a Berkeley vegan cheese pizza with veggie burger crumbles. The veggie sandwich and veggie calzone could be made vegan. Has gluten-free crust. Wheelchair accessible. Accepts credit cards. Mon-Thu 10:00am-9:00pm, Fri-Sat 11:00am-10:00pm.-- Rating: 3 HappyCow(s)
Health Food Stores, Vegan Shops, Vegan Bakeries, Vegetarian B&Bs, Veg Catering, Veg*n Organizations: (2 Listings)
Whole Foods Market
2700 Wilson Blvd ~ PH: 703-527-6596
Part of large natural foods supermarket chain. Has fresh fruits and vegetables including organic produce, deli and hot food bars, vitamins, supplements, personal care products, grocery, and much more. Accepts credit cards. .Open daily.-- Rating: 4 HappyCow(s)
1025 N Fillmore St Ste G ~ PH: 571-970-6460
Bakery also making vegan cupcakes, chipwiches (chocolate chip cookie ice cream sandwich), cookies, and cream pies. Accepts credit cards. Mon-Thu 10:00am-8:00pm, Fri 10:00am-10:00pm, Sat 10:00am-8:00pm, Sun 10:00am-6:00pm.-- Rating: 4.5 HappyCow(s)
It is advised to call ahead before you visit any listing in the restaurant directory to confirm the open hours, etc.
If you appreciate this free service and would like to offer your support, please contribute online at: www.happycow.net/supporter.html
All contents copyright © 1999-2015 HappyCow's Healthy Eating Guide. All rights reserved.


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GeForce 5900 XT Problem
I've recently purchased a PNY Technologies GeForce 5900 XT (64MB) graphics card (AGP2/Pro) and everything works mine with a single 512MB ram chip (Twin Mos 512MB PC3200 400 MHz).
The problem occurs when i try to install a second 512MB ram chip (also Twin Mos 512MB Pc3200 - the next serial No. in the batch) and find that the graphics start corrupting on games (Star Wars Battlefront, Vice City, etc).
I'd be very grateful for any help in solving this problem.
My system is a Athlon XP2800 on a Asus A7n8X deluxe mobo.
Many thanks.
Re: GeForce 5900 XT Problem
Re: GeForce 5900 XT Problem
Thanks for the reply and re-direction.
Have swapped out my graphics card - same problem. I think I've tied the roblem down to the RAM (wrong forum again?). From reading the other forum posts it looks like the Twin Mos RAM I've installed doesn't like my A7N8X Mobo or vice versa.
Spoke to the shop i bought the ram from and they reckon it won't work in dual mode unless i purchase specific 1040 dual mode packs (2 x 512MB). The shop suggested i try it single line (slots 1 and 2 rather than 1 and 3 or 2 and 3). Will give it a go.
Re: GeForce 5900 XT Problem
a 5900xt with 64mB? On agp pro? That's very weird!
Looks like your ram is the problem.
Re: GeForce 5900 XT Problem
Its 128MB AGP - should have written in original post.
Re: GeForce 5900 XT Problem
sounds like a RAM problem
Re: GeForce 5900 XT Problem
Definitely a RAM problem.
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The North Branch wrestling banquet on April 5 recognized the team earning the Academic Gold Award. Named All-Conference were seniors Eric Bergstrom, Jake Pilz and Zack Sundly, along with juniors Reid Johnson and Aaron Senne. Senior Sam Peltier earned All-Conference Honorable Mention. Pilz was awarde...Read More
2013 season
The North Branch-Chisago Lakes FIRST Robotics Team 3038 has been busy the past month getting ready for the 2013 season. The team was selected to be a Beta Testing Team for new software and hardware to be used during the upcoming season. "This is pretty exciting for us and will give us a chance to...Read More


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Miss Hudson, in your own native city of San Francisco, there's an art gallery in the Legion of Honor in which there's an oil painting of Casanova. It's quite obvious that you have never seen this painting. For your information, Miss Hudson, this is what Casanova looked like. He had big ears, a scar over one eye, a broken nose, and a wart on his chin, right here. I suggest, Miss Hudson, that when you return to San Francisco, you visit this gallery and see this painting!


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Original recipe yields 12 servings
Amount per serving ({{servings}} total)
Based on a 2,000 calorie diet
Ready In
This made absolutely the best chicken wings I have ever put in my mouth! It was the biggest hit at our New Year's Eve party! Everyone wanted the recipe!!!
This is a good sauce, a tasty marinade, but not hot what so ever. The chicken came out very tender. I will spice it up next time, thanks Nancerini!
This sauce was incredible!! Thanks for the recipe!


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Narrow down the 415 results:Map View
x The average starting price for a place to stay in Istanbul is £54 per night.
Muvezzi Street Besiktas IstanbulOutstanding 100%
Yerebatan cad. Salkimsogut sok. No:6 Sultanahmet- Address 2 :Outstanding 100%
Küçük Ayasofya Mah., Demirci Reit Sok 28Outstanding 100%
Inebey Mahallesi Gazi Mustafa Kemal Pasa Caddesi No:11 YenikapiOutstanding 100%
Cankurtaran Mahallesi Akbıyık Caddesi No 24 Fatih /SultanahmetOutstanding 100%
Yavuzsultanselim Mah.Vakıf Mektebi Sok.No:1 FatihOutstanding 98%
Alemdar Mahallesi, Zeynep Sultan Camii Sokak No 21Outstanding 96%
ku.ishu.kh Very helpful and friendly staff, great view from the terrace. Easy access to most major historical sites! Really enjoyed my stay there.
Divanyolu Caddesi No.10 Sultanahmet SultanahmetOutstanding 96%
sangeeta.y If you are looking for a cosy little place right in the heart of the main part of the city then this is the place to book. All the main places in Sulatnhamet are at a stones throw from this hotel. The surrounding area is simply beautiful with Aya Sophia & Blue Mosque & other buildings & monuments. The terrace resturant offers great view outside. I did not use the resturant so cannot comment on food. There are a lot many cosy and good resturants around the hotel. They offer airport pick & drop at resonable rates as well. Overall, a nice & cosy place for budget travels.
alemdar mahallesi divanyolu caddesi numara 70/4 suOutstanding 96%
Akbiyik Caddesi Bayramfirin Sok no:12 SultanahmetOutstanding 95%
nicknkelly Lovely hostel, newly renovated bathroom, very clean - but in comparison to other hostels in the area you pay substantially more for it.
Bereketzade Camii sk. No:3, Galata, BeyogluOutstanding 95%
dreisman This is a great place to stay. Perfect location, homey atmosphere, clean, great view of the city from a roof patio. The staff was super helpful and friendly. It would be the first place I would look into when I return.
Binbirdirek Mah. Dizdariye Yokusu Sok. No:3 SultanahmetOutstanding 95%
Akbiyik Cadessi Bayramfirin Sokak No 7, SultanahmetExcellent 94%
baluba I stayed here in January 2009 and absolutely loved it! At 49 1/2 I was a little concerned about the age restriction thinking it would be full of teenagers, but this was not the case and I certainly was not made to feel 'old'! I had booked a mixed dorm as I was on a relatively tight budget but, being low season there were not many guests so I was automatically upgraded to a female dorm with en suite bathroom which I had to myself for the first 2 nights. The dorms are basic, beds are supplied with one sheet, a pillow and a blanket. The central heating was a bonus as in January it is damned cold. The shower had oodles of hot, powerful water and all of the toilets have their seats covered with some weird contraption which revolves around leaving you with a perfectly clean piece of plastic on which to park your bum. No towels but lots of toilet paper...and a hairdryer!!! The upstairs bar cum breakfast room cum lounge area is absolutely gorgeous with a large sumptiously cushioned seating area where you can while away hours watching tv, playing cards, chess or backgammon or chat with fellow travellers over a cold beer. Beer by the way is 7L for a 500 ml bottle. There is limited food available but it is cheap and filling. The staff couldn't have been more friendly or helpful, they successfully managed to arrange for me a visit to the Florence Nightingale Museum which involved photocopies of my passport, a fax to the Army headquarters and several phonecalls, none of which they charged me for. It really felt like a home from home and even after just 4 nights, I did not want to leave. One point though, if you arrange an airport transfer, make sure they fully understand which airport you are arriving at otherwise, like me, you could face a bus, a metro, the fernicular, a tram and finally a walk to get there. If you choose to go this way, it is actually very easy and will cost you no more than £9.00
Akbiyik cad (street) Amiral Tafdil sk. no.6 Sultanahmet / IstanbulExcellent 94%
IGITIHI I was pleasantly surprised by this hostel! Very clean, great breakfast, nice beds and friendly stuff. The showers were very comfortable and the lounge one of the best I have seen. It's also situated at a very convenient place. I really enjoyed my stay and would definitely stay again and recommend it to anybody.
Create your own travel plan including Istanbul as one of your stops. You can use it to store all your research (even hotels found on other websites!)
Do you have questions about Istanbul? You might try asking in our active travel forums and see if anyone else has been there.
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Revision as of 15:43, 6 March 2009 by 184.108.40.206 (Talk)
Estë (Q: "Rest") was a lady of the Valar, the spouse of Irmo, who dwelt with him in the gardens of Lórien in Valinor. "Grey is her rainment, and rest is her gift." She was known in Sindarin as Îdh and in Nandorin as Eord. She was often called "The Gentle". She was the "healer of hurts and weariness."
See Also
|Manwë · Ulmo · Aulë · Oromë · Mandos · Irmo · Tulkas
|Varda · Yavanna · Nienna · Estë · Vairë · Vána · Nessa


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The Olympics are set to open July 27 in London. Special preparations are being made throughout the urban center, and the city will soon be swamped with thousands of competitors and spectators eager to watch the games. Among those excited for the beginning of the athletic tradition is Noor al-Malki, an athlete from Qatar.
Noor runs the 100-meter dash and will be the first female athlete to represent Qatar in the Olympics. The Guardian reports that the athlete runs the sprint distance in just under 13 seconds, which is not a time that will place Noor anywhere near winning a medal, compared with record times under 11 seconds. Instead, the goal of the Qatar government and Noor herself is to get the country on the map and to stand for Muslim women playing sports on the international stage. She will be joined by two other athletes, Nada Arkaji, a swimmer, and Bahia al-Hamad, a rifle shooter.
Qatar's vision and intiative took some wrangling. They had to convince the Olympic officials to include Noor in order to open up the stage for Qatari representation. The stance taken by Qatar also differs sharply from Saudi Arabia, which has denied women the space to play Olympic sports. Many commentators had expected Saudi Arabia to follow Qatar's example in sending female athletes to the Olympics this year, but the head of the Saudi Olympic games committee stated, "At present, we are not embracing any female Saudi participation in the Olympics or other international championships."
In the 2008 summer Olympics held in Beijing, Qatar, Brunei and Saudi Arabia were the only three countries that failed to send female athletes to compete. Both Qatar and Brunei are sending contestants this year. Human Rights Watch has noted that Saudi Arabia does not allow women to play most sports, even within the confines of schools.
Women have faced an uphill battle throughout the history of the Olympic games in every country. Female athletes started to compete in the Paris Olympics in 1900 on a limited basis. Women's weightlifting was not even on the Olypmic docket until 2000 when it made its debut. Sprint running, like the sport Noor competes in, was banned for women between 1928 and 1960 because women were considered too feeble to complete the 800-meter sprint.
Even countries which have been sending women to the Olympic games for a number of years, such as India, are only now sending new female athletes to compete for the first time in certain sports. For example, Geeta Phogat is the first Indian female athlete in wrestling to qualify for the Olympics.
Her athletic prowess and abilities are often considered unfeminine in her local community. Phogat has stated that many community members "said nobody will marry us because we would have disfigured ears," the Times of India reports.
Phogat and Noor al-Malki are breaking traditional molds of femininity which include marriage and children at a relatively young age. They are also breaking Olympic traditions which have been weighted against female athletes throughout the decades. Avery Brundage, a well-known president of the IOC in the 1950s and 60s allowed that women could do sports "of all kinds" but only under "the proper supervision." This type of thinking has clearly affected women's participation and chances in the Olympic Games, even in recent years.
Admittedly, many women athletes have become celebrated icons in Olympic events. Just consider the love and support gymnast Shawn Johnson enjoyed during the 2008 Olympics. However, women continue to be sidelined in many sports outside of the gymnastics arena, a sport that is often stereotypically associated with feminine charm and ability. It appears that pressure and expectations of the inclusion of women in Olympic events helped to bring about changes in Qatar and Brunei. Human Rights Watch hopes to pressure Saudi Arabia to reconsider its position on female sports participation for future events, but this year it looks as though the Saudi women will have no representatives.
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Read more: athletes, islam, london, olympics, Qatar, sexism, women
Photo Credit: Smithsonian
Good for them but Saudi is still in the dark ages or worse.
Good for Qatar. As for Saudi Arabia, why should it be too late to kick them out? Just refuse to let their athletes and support staff into the country. Problem solved. Maybe by next Olympics they'll have noticed what century this is.
Gabriela M. wrote:
The Olympics Committee should have a rule that says if a country doesn't want to send female athletics to compete in the Olympics it can't send men either.
They do have such a rule, Gabriela, but they have never properly enforced it until now.
Saudi Arabia stalled until it was too late for the Olympics committee to do anything, then refused to admit women. I'm not sure if they'll be banned in the future.
Not only is Qatar sending female athletes to the Olympics for the first time, but so is Brunei. I'm glad to see it!
Before the Olympics was about money and politics anyone could compete. I would like to see a return to those true values where we applaud those who take part; without restrictions put on anyone, by anyone.
Congrats to her. I hope she does well!
The Olympics Committee should have a rule that says if a country doesn't want to send female athletics to compete in the Olympics it can't send men either.
It's just like Linda said, it many parts of the world it's women who have to do the heavy lifting and long walks to get water and firewood while the "strong" men stay home in the shade, drinking tea and chatting. Yes, we're so fragile and they're so strong and brave!
Of course the "weaker" sex, you know the one that carries and delivers babies, does all the heavy work like fetching water and heavy household and farm chores, is incapable of participating in sports becaust they're so fragile.
thanks for sharing
Disclaimer: The views expressed above are solely those of the author and may
not reflect those of
Care2, Inc., its employees or advertisers.
What cage Laara? Dancing around in circles? Clearly you must have seen a different video!
The dream is that there is a utopian vision of the small farmer supplying his neighbors. Reality is…


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3DMark03 - damage control for the GeForce FX?
View Single Post
Team Rainbow
Join Date: Jul 2002
Location: FRANCE
Posts: 2,273
Originally posted by Nutty
Rumours are the retail version of the card will be much quiter fan wise, and I bet the drivers improve considerably.
What the chances that nvidia unveil "free" AA with newer drivers on the hardware at some point?! Now that would be impressive. I still think GF-fx's AA quality is fine. The ppl slating it seem to have no understanding of equal comparisons, and seem to be completley blind to me.
Good to see someone with the same thinking as me. No one has said anything about Nvidias 4xs FSSA, and from what we have see so far, it looks better than 6x2 and 8xs, so thats obvious there are driver problems there.
I think the GF-FX will do well, but lets not forget, 3dmark will push todays cards, remember 3dmark2001 and the GF3/8500 and now look at it with the 9700/GF-FX. Still I cant wait to see what Im missing out on by not having a 9700 or GF-FX
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About Aptina™ A-Pix™ Technology
Aptina A-Pix is a series of advanced pixel technologies, featuring lightguide and deep photodiode, and 65 nanometer pixel design rules that cost-effectively advance pixel performance. The new, third generation Aptina A-Pix enhances quantum efficiency and minimizes crosstalk to capture sharp images with vibrant colors even in the low-light conditions that challenge traditional sensors. Aptina has already shipped millions of imaging products containing earlier generations of Aptina's reliable and established FSI technology. Aptina A-Pix makes it possible for mobile phone cameras to capture quality images to compete with digital still cameras, and enables a new class of hybrid camera that combines digital still image capture with advanced high-performance HD video.
Gennadiy Agranov, Aptina's VP of Imaging Technology discusses Aptina™ A-Pix™ technology


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Is it a dial around the lens?
Just posted: Hands-on preview of the Nikon Coolpix P330
We've just posted a hands-on preview of Nikon's latest small form-factor compact, the Coolpix P330. Externally almost identical to its predecessor the P310, the P330 features a larger 1/1.7" BSI-CMOS sensor and raw capture mode - two additions that should make it much more popular with enthusiasts, as well as putting it in direct competition with Canon's popular PowerShot S110. We've spent a little time with the P330, and for more details, images and specifications, click the links below to read our hands-on preview.
Click here to read our hands-on preview of the Nikon Coolpix P330
Jun 10, 2015
Apr 13, 2015
Dec 16, 2014
Nov 27, 2014
Oops, my bad thought this was the cool pix a.
I wonder if Fuji is going to fight back with a law suite for copying that style of sensor. This camera is clearly trying to copy the x series. Maybe Fuji is going to comeback and make a dslr.
P3300 "Test" Photos - Full res images
Has anyone found full-sized sample images from the P330?
I found these, two of them are full res and they look fantastic, similar to the P7700 and nicer to my eyes than the S110.
If it has manual exposure control in video mode like the P7700 it should be an excellent all-around shooter. And if IQ is as good as the class leading P7700, I may just have to pick one up. I only wish they had made the lens faster like the one on the excellent MX-1, but then it could never be this small.
They need to make camera compete with rx100.
Unfortunately they can't because a 1" sensor Nikon compact would cannibalize their Nikon 1 line-up. It's tough when you have competitive products in a variety of sensor format.
Fine looking camera, but with the RX100 at just roughly $80.00 more, it's becoming very hard to be a pocket camera!
RX100 appears to be selling for ~$650, while the P330 is expected to initially list for $379.
"RX100 appears to be selling for ~$650, while the P330 is expected to initially list for $379."
It'll drop in price VERY quickly, as was the case with earlier P3x0 models.
And we know that the RX100 will NEVER drop in price. Sony doesn't play that.
Personally, I've owned an RX100 and it's an OK camera, but unless you need such small camera, spending $650 which is MILC, a used X100 or a mid-level DLSR type money on a P&S, is a tough sell.
And after spending hours editing 36 mp D800 files, the last thing I want to do is edit 20 mp RX100 RAW files. If the shooting experience were better, maybe. But when it shoots like a regular P&S, is slippery, and has no VF or tilt LCD, all the IQ in the world can't make up for it.
Will be my next camera if it supports remote release for interval shooting. Sadly I don't think it will support that feature.
Any chance we can get the P7700 review finished before this one is reviewed? Its only been 6+ months now...
Looks like a great improvement over the previous model. Just got an S100 on deep discount though. A very nice cam also except for operational speed. Maybe the Nikon will be faster.
Aftermarket finger grips are available for these pocket cams and they work wonderfully.
"Looks like a great improvement over the previous model. Just got an S100 on deep discount though. A very nice cam also except for operational speed. Maybe the Nikon will be faster."
Nikon's P3x0 series has always been pretty responsive and, in this regard, pleasure to shoot with (unlike with, say, the Canon G1X). Surely the P330 won't be an exception.
Thanks for the comment. I'll probably check one out.
The picture you have posted above looks like the most uncomfortable grip I have ever seen...
Of course nobody holds light compact cameras like that...
...unless you are like me and need to operate the camera with just one hand from pulling it out, shooting, and putting it away. In this regard it works great and was a primary requirement. The movie mode button, however, is awkwardly placed on the back for one hand operation.
They need to make camera compete with rx100.
About time they brought RAW to this camera. Now it competes directly with the Canon S95, S100, S110 etc.
f/1.8-f/5.6 ? I'm no Olympus fan, but XZ-10 will destroy it in low-light, despite the smaller sensor.
Perhaps, but you'll have better DOF control with the Nikon, at least at the wide end. And if it performs like the P7700, you should have better high ISO ability, but of course the faster max aperture at full tele on the Olympus will offset the high ISO advantage for the Nikon.
The XZ-10 does look nice, but it remains to be seen if IQ will be as good as cameras like the P7700 or MX-1. The samples from Olympus's website did not impress but I still have my eye on this camera mainly for it's size, and fast lens.
That said, I'm not the type who needs the smallest of the small. A camera like the MX-1 or X20 feels nicer in my hands than slippery little cameras like the RX100 or S110.
But for me, having f/2.7 @ 140mm eq is much more attractive than 50% larger sensor. The number f/5.6 is so unpleasant to read and think of.
Ad "smallest of the small": well, pocketability is the main point of both P330 & XZ-10. If the strict pocketability is not a hard constraint, then there are multiple options (P7700, MX-1, XZ-2, X10, X20, EX2F ...)
And then there is RX100
you didn't get it. it's a fade-in effect.
I waited for this type of camera from Nikon (APS-C). But I'm really disappointed to see the final product. Well, maybe Nikon wants it to look like ordinary point and shoot cameras out there. I love the sensor size but I hate the look of this camera.
I think you're thinking of the Coolpix A.
Finally Larger sensor .
Yes, finally larger sensor in a toy housing. It will be better if the housing of this camera is something similar to Fujifilm X100. But that's Nikon.
Connect with dpreview


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I am kind of caffeine addict and especially when I am in US: The coffee there is good. LOL. From HB I didn't find very many coffee shops so I got my satisfaction daily from Starbucks. Which was totally ok for me. The staff there was fun, very nice types, absolutely fantastic ones. Too bad that I didn't asked that one cute guy out...just kiddin' !
But a tip for you. Starbuck is there and a lot better than some other coffee shops along the Main Street.
Address: 221 Main Street
Get Food, Shop and Buy Art at Farmer's Market
HB has their twist on the common farmer's market. Farmer's Market and Art A-Faire occurs each Friday at Pier Plaza, 315 Pacific Coast Highway.
Having a farmer's market near the beach is always convenient. It starts at PCH/Main st. This is a great way to have a picnic at the beach. Get the picnic basket for collecting exotic fruits, veggies and gourmet snacks.
Along with the food, there are vendor booths with fresh sunflowers, glassware, craft items, purses and much more.
Art-A-Faire features vibrant original frames of California skyline, hand-made sculpture and various trinkets of artistic quality.
Parking is at 1st street where there is a lot.
- Arts and Culture
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Dress Casually
At HB you can enter almost anywhere in your bikinis. Casual, comfortable clothing is fine all year round. In fact, Hawaiian shirts always look good—and right at home—in Huntington Beach. Jackets and ties are optional even in the most elegant of restaurants. During evenings and winter days when the ocean breezes turn cool, you'll want a jacket, sweater or sweatshirt. Dress in layers so you can remove clothing as the day warms up and replace it as the day cools off, particularly if you plan to stay on the beach.
Skatin' HB!
After the actual surfing, skating is a way to get around in HB. It is more like a life style transportation in this nice beach town, where everything and everybody are very casual and relaxed.
Take a skateboard and enjoy this town, boards are allowed along the 8.5 mile paved beach trail. Then there are two skate parks for enthusiastics.
One skate park is located in front of Huntington Beach High School near the corner of Main Street and Yorktown Avenue. The second skate park is visible from Golden West Street near Warner Avenue at Murdy Park. Knee pads and helmets are required in both locations.
Salute Duke
This is a tribute to an acclaimed surfer, Duke Kahanamoku by the main strip and surfer hall of fame. Walk up to duke and give your salute. Bow down!
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The present invention relates to a method for adjusting the sensitivity of a microbial culture to an antibiotic. The method employs a substance derived from a bacterial species with sensitivity to the antibiotic. Such bacterial species include the same species used in the culture or a different species.
1. A method for detecting the presence of at least one antibiotic in a test sample, whereby the at least one antibiotic includes a first antibiotic from a beta-lactam structural class and an antibiotic from another structural class or a second antibiotic from a beta-lactam structural class, the method comprising:
a) combining the test sample with both a microbial culture and a microbial receptor, the culture comprising a microbial species susceptible to at least one member of the beta-lactam structural class and susceptible to at least one member of the additional antibiotic structural class or the second antibiotic from a beta-lactam structural class, the microbial receptor characterized by its sensitivity to the beta-lactam structural class, the microbial receptor having been extracted from a bacteria with sensitivity to the beta-lactam structural class, whereby contacting the susceptible microbial species with an antibiotic to which the culture is susceptible causes growth inhibition in the culture;
b) growing the culture; and
c) determining an amount of culture growth,
wherein a lack of culture growth reflects a presence of an antibiotic and
wherein the microbial receptor selectively reduces test sensitivity to the first antibiotic beta-lactam structural class without substantially reducing test sensitivity to the antibiotic from another structural class or the second antibiotic from a beta-lactam structural class.
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This application is based on and claims priority from U.S. Provisional Patent Application No. 60/523,065, filed Nov. 18, 2003, and U.S. Provisional Patent Application No. 60/574,252, filed May 25, 2004, both of which are hereby incorporated by reference.
Antibiotics used in animal feed or to treat farm animals, such as dairy and beef cattle and swine, occasionally contaminate the food supply. The hazards associated with these undesirable residues include allergic reactions, propagation of resistant organisms and other long-term health risks. As a result, government agencies such as the United States Department of Agriculture and the United States Food and Drug Administration require monitoring of such residues in foods. Foods such as milk, meat, poultry, seafood and animal feed are routinely tested for the presence of such residues. We disclose herein a user-friendly method for detecting a variety of residues in a single, broad-spectrum test.
Various animal parts and excretions can be tested for the presence of antibiotics. Tests for antibiotics in kidney include tests for hog kidney and veal bob kidney. Problems with some of the available tests, particularly in the pork industry, include sensitivities that do not match well with governmental or industry limits. Some tests are over-sensitive to certain antibiotics and under-sensitive to others.
Sensitivities of currently available microbial growth inhibition tests, are based primarily on parameters that affect test sensitivity to all drugs. Such parameters include growth organism used, amount or concentration of growth organism used, vessel dimensions, media volume, media type, mix of nutrients, incubation time and temperature. As a result, if a test is adequately sensitive to some drugs, but overly sensitive to other drugs, other methods for sensitivity adjustment may be needed.
Another problem with some tests is that they may require cumbersome extraction procedures or use of organic solvents that must be removed from the sample prior to test operation. This adds a cumbersome procedural step. Some tests are also procedurally cumbersome, requiring incubation of a sample swab on a lawn of bacterial culture and inoculation of an agar surface with spores just prior to running the test.
In addition to problems with slow testing time, over-sensitivity, under-sensitivity and extraction, none of the currently available tests are provided in an all-in-one format in which all test reagents and sampling devices are provided in one test instrument.
We disclose herein various embodiments of a test apparatus and method to detect antibiotics in a test sample using growth inhibition of a microbial culture. Some embodiments include all of the reagents for a microbial culture in a ready-to-use format. The culture can include, for example, nutrients, agar, one or more buffers, spores and color indicator. Alternatively, any one or more of such components can be provided separately, for example in a separate reagent compartment hereinafter referred to as a "niblet". In an embodiment of the ready-to-use format, the user only has to add the sample, or sample extract, and incubate for a prescribed time period, for example about 1.5 to 4 hours, at a prescribed temperature, for example about 55-70 degrees C., and observe the results.
If extraction is required, an extractant can be provided separately from the other reagents. In an example the extractant is in a niblet. The niblet containing the extractant can be included in a test unit including other test components. The test unit can include the niblet contained within a vial attached to the bottom of the test unit. The seals of the niblet can be puncturable membrane seals such as metallic foil seals or foil-like seals or plastic or plastic-like seals. Puncturing the seals allows the extractant to contact the culture. The user only has to add the sample to the extractant, contact the extracted sample with the culture, and incubate for the prescribed time period at the prescribed temperature. The extractant can be a buffer solution capable of extracting antimicrobial drugs from a kidney sample, such as a bovine or porcine kidney sample. The extractant can also be a reagent for merely removing the sample from a swab and transferring the sample into the culture.
Some embodiments include two or more buffers (hereinafter referred to as a polybuffer) having multiple pKa values. A polybuffer can improve pre-test and/or post-test stability of the culture and other aspects of test performance. Post-test stability allows results to be retained at room temperature for an extended period, for example 6-8 hours or more, without a change in test result. The polybuffer can be premixed within the culture or provided separately such as in a niblet for later addition.
An embodiment includes reducing sensitivity of the microbial culture to certain inhibitors of culture growth. In some growth inhibition assays, the inhibition of microbial culture by antibiotics from a sample, or reversal of such inhibition, can be used as an antibiotic screening mechanism. The sensitivity of such tests can be adjusted by contacting the culture with at least one adjustment reagent. Examples of adjustment reagents include binding protein, enzyme, chemical analogue and/or antibodies that bind to, or otherwise inhibit the effectiveness of, antibiotics. The adjustment reagent can be used to adjust the sensitivity of the test for at least one antibiotic or other culture growth inhibitor. An example of a microbial enzyme useful as an adjustment binder is an antibiotic receptor/binder from the cell wall of microbes such as Bacillus (Geobacillus) Stearothermophilus (B. st.). The adjustment reagent can be applied in a variety of ways, for example combined within a culture or broth, applied on top of a solid culture, such as an agar matrix, added on a sample swab or contained in a dilution buffer.
Possible adjustment reagents include antibiotic receptors/binders isolated from bacteria, monoclonal and polyclonal antibodies. When the adjustment reagent is an antibiotic binder, such as a binding protein, for example a bacterial antibiotic receptor or antibody, the adjustment reagent can reduce the availability of antibiotics capable of inhibiting the growth of the particular culture used, such as by binding to the antibiotic. Similarly, adjustment reagents can include substances that, rather than bind with the antibiotic, reduce the influence of the antibiotic on the growth organism.
Some embodiments include a test apparatus with a solid or semi-solid culture within a vial. The culture can include all or some of the test biochemicals including agar, nutrients, color indicators, one or more buffers and spores. In an embodiment, the culture, prior to the addition of test sample, is adjusted to a pH of greater than about 8. Such a pH can help avoid culture contamination, for example with mold, and lengthen pre-testing shelf-life. The vial can include a sealed bottom end and a membrane seal over the top end. In test operation the user can puncture the seal with, for example, a pipette tip or swab and then dispense the sample, for example a 200 microliter sample of milk, into the vial.
Some embodiments include the complete test unit containing all of the reagents, premixed together and ready for use in a culture and, in addition, a sampling instrument, such as a swab or probe and optional extractant. The probe can be used to puncture the one or more seals separating the optional extractant from the culture. The probe can be used to absorb and apply the sample to the reagents within the test apparatus and can comprise an absorbent material such as an absorbent, fibrous, cotton-like or cotton material. An example of such a sampling instrument is a swab or probe in the format of a POCKETSWAB (POCKETSWAB is a registered trademark of Charm Sciences, Inc. Lawrence, Mass.). The format of the POCKETSWAB provides the advantage of controlled movement of the swab in a test device that provides physical support for the swab. For example, the swab can be controllably moved into the reagent compartment containing an extraction buffer. Using the POCKETSWAB device the swab can remain in the extractant for the time desired. In addition to the convenience of delivering the swab to the customer in an all-in-one device, the POCKETSWAB provides the physical support so that the swab will remain within the extractant or buffer solution without the user providing external support to the swab while soaking. After sample extraction, the swab or swab tip is moved longitudinally through a seal thereby allowing the sample to flow onto or into the culture.
In a particular embodiment, the test method is used to reduce sensitivity of a microbial culture to the beta-lactam family of antibiotics. A beta-lactam binding protein is used to selectively reduce test sensitivity to the beta-lactams. Test sensitivity will be mostly unaffected relative to antibiotics or inhibitors other than beta-lactams. In an embodiment, the beta-lactam binder is a receptor from B.st. and the culture is made of spores of B.st.
An embodiment described herein involves a user-friendly method, device and kit for the detection of a broad range of residues of antibacterial compounds in a sample such as an agricultural product. Antibiotics that may be detected include beta-lactams, sulfonamides, tetracyclines, macrolides, aminoglycosides, quinolones and amphenicols. In an embodiment, the user of the test adds the sample, incubates and observes the results.
A useful mechanism of antibiotic detection is microbial growth inhibition. Examples of microbes useful in such an application include: B.st.; B. subtilis; B. megaterium; S. aureus; Ps. Aeuginosa; E. coli; and B. licheniformis. Generally microbes which exhibit detectable growth inhibition in the presence of antibiotics may be useful. Other examples of microbial growth inhibition tests include those described in U.S. Pat. Nos. 5,354,663 and 5,489,532, the teachings of which are incorporated herein by this reference. Microbes, such as B.st., that sporulate are particularly useful.
One of the benefits to microbial inhibition tests is that they can be broad spectrum compared to family or antibiotic specific antibiotic binding based tests such as those utilizing antibodies or other bacterial binders/receptors. For example, both beta-lactam and sulfonamide antibiotics will cause some inhibition of growth of B.st. The sensitivity of the currently available microbial inhibition tests are based on parameters including particular growth organism used, concentration of bacteria or spores, the nutrients provided and the incubation time and temperature used. Thus, in the example of B.st., using one of those parameters to adjust test sensitivity may result in reduction of sensitivity to all or multiple of the antibiotics to be detected. It may be desirable, however, to reduce test sensitivity of a microbial growth inhibition assay to only certain antibiotics, or other inhibitors such as only those for which the test is overly sensitive. For example, using B.st., it may be desirable to reduce test sensitivity to only beta-lactams and not sulfonamides or other drugs or inhibitors.
In an embodiment, a binding protein isolated from bacteria, sometimes referred to as a receptor, antibiotic receptor, antibiotic binder or bacterial receptor, is utilized as an adjustment binder alone, or in combination with specific antibodies. Bacterial proteins that are sensitive to multiple antibiotics, such as a family of antibiotics, for example the beta-lactams, can allow sensitivity adjustment of multiple antibiotics to which the particular binding protein is sensitive. Antibiotic binders, such as antibodies, that are more specific, allow more specific sensitivity adjustment.
An antibiotic bound by such an adjustment reagent may be rendered unavailable, or less available, to inhibit bacterial growth. Although not wishing to be constrained by theory antibiotic binders being employed on top of agar, or other solid culture, may create large molecular weight substances that do not easily diffuse into a solid culture. Adjustment reagents, such as binders, combined into a solid culture or liquid medium, may inactivate, weaken or otherwise interfere with the antimicrobial properties or affect of the antibiotic. As such, substances other than antimicrobial binders may be useful to adjust test sensitivity including enzymes, such as beta-lactamase, that destroy antibiotic activity or otherwise inactivate antibiotics or substances that compete with antibiotics in the bacterial cell, such as analogues. One such substance is para-aminobenzoic acid (PABA), an analogue of sulfonamides with which sulfonamides compete in certain bacterial metabolic pathways. Similarly useful adjustment reagents may include substances that change the structure of an antibiotic or otherwise reduce the activity or make inactive the target antibiotic or antibiotic family. Antibiotic binders may also include non-viable bacteria or bacterial extract such as cell wall extract.
Adjustment reagents, including antibiotic binders, can be either specific to a particular drug for which the unadjusted test is overly sensitive, or have affinity to multiple drugs. The adjustment reagent may be added with the sample or pre-mixed into a culture. One example of an adjustment reagent for multiple drugs is a microbial receptor, such as the beta-lactam receptor or receptors isolated from B.st. such as described in U.S. Pat. Nos. 4,239,745 and 4,239,852 the teachings of which are hereby incorporated by reference. Other possible sources of beta-lactam receptors include organisms from the genus Bacillus including B. subtilis, B. megaterium, and B. licheniformis. Other possible sources include S. aureus, Ps. Aeuginosa, B. licheniformis, and E. coli. Examples of useful specific adjustment reagents for particular drugs include monoclonal or polyclonal antibodies. Other examples of using antibodies for sensitivity adjustment are provided in U.S. Pat. No. 6,319,466, the teachings of which are incorporated herein by this reference.
Antibiotic binders can be isolated from bacteria and purified using known techniques. In an embodiment, beta-lactam binder isolated from B.st. is used to adjust the sensitivity of a microbial inhibition assay using B.st. as the growth organism. In such an embodiment, test sensitivity to all of the beta-lactams is reduced while test sensitivity to other antibiotics to which B.st. is sensitive remains relatively unaffected. This is useful, for example, when target sensitivity is European multi-residue levels (MRLs) and/or U.S. safe levels, both of which regulatory levels allow beta-lactam in tissue that may be above the levels of detection of an unadjusted B.st. microbial growth inhibition assay.
Other antibiotic binders that may be useful include receptors isolated from various part of bacterial or other cells including the ribosome or part of ribosome to which certain antibiotics can bind. Examples of antibiotics that bind to or otherwise inhibit ribosome function include tetracyclines, sulfonamides or fluoroquinolones. Bacterial growth inhibition by antibiotics such as tetracyclines, sulfonamides and fluoroquinolones may be adjusted by the addition of such binders. In addition, in some embodiments the adjustment reagent/binder can be an isolate from the same organism used to produce the culture.
Other useful methods of adjusting sensitivity include adding dead cells and/or crude extract that contains the desired binders/receptors.
In one embodiment, depicted in the figures, a solid culture is provided in a reagent chamber such as a vial 4 or vial-like device. Located in the same device can be a niblet 5 containing an appropriate extractant or liquid diluent 11. In an embodiment, an antibiotic binder is added to the top of the solid culture 15. In another embodiment, an antibiotic binder is mixed into the solid culture 15. In a further embodiment, the antibiotic binder is stored separately and added to the solid culture 15, along with the sample addition, prior to sample addition or soon thereafter. In another embodiment, the antibiotic binder is included pre-applied to the sampling swab 1, for example by covalently binding to the swab 1.
In an embodiment for detection of antibiotics in a sample requiring extraction, an extractant is used prior to adding the sample to the culture. An extractant that will separate antimicrobial drugs from a sample, or from a swab or other sampling device, avoids complicated extraction methods or use of organic solvents. An example of useful extractants include a combination of Trizma Base and Potassium Phosphate, such as Potassium Phosphate Monobasic in water at a pH of about 7 to about 8, for example approximately 7.5. Such an extractant buffer can be provided separately from the culture or, for ease of use, in an all in one test device such as the POCKETSWAB format 8. Using the POCKETSWAB format 8, after sample extraction the extractant (containing sample) is simply combined with the test reagents for incubation. Such extractants may extract antibiotics from the sample or merely serve to transfer the sample, including antibiotic, from the swab or other sampling device into the culture.
The POCKETSWAB format 8 is described in U.S. Pat. No. 5,965,453, which is incorporated herein by this reference. In the POCKETSWAB format 8, the extractant can be within a niblet 5. The niblet 5 can be situated within the device, above the culture 15. The niblet 5 can be sealed on the top 13 and bottom 14 ends such as with a membrane seal, for example, puncturable foil seals, on both ends to retain the buffer or extractant therein. When using the POCKETSWAB format 8, the swab 1 is removed and contacted with the sample. After sample contact, the swab 1 is used to puncture the first seal 12 (the vial 4 seal) and then the niblet top seal 12 thereby placing it within the niblet 5 and in contact with the extractant. After sufficient extraction time, the second sealed end 14 (the bottom end) of the niblet 5 is punctured allowing the contents to flow onto the culture 15. The sample is incubated at above room temperature, for example between about 60 degrees C. to about 70 degrees C., such as about 64 degrees C. Spore germination and/or bacterial growth, or lack thereof, is determined by observation of changes in growth indicators within the culture 15.
Possible growth indicators include indicators that undergo a detectable change as a function of the growth or inhibition of the culture, such as pH indicators and oxidation/reduction (redox) indicators. For example, indicators that change color in the presence of an acid or base such as to a yellowish color if the environment is acidic or purple/blue color if basic or neutral. In the case of bacterial growth, or bacterial growth following germination of spores, acid is produced. Thus, indicators will reflect creation of an increasingly acid environment as bacterial growth proceeds. If bacterial growth or spore germination is inhibited, for example by antibiotics or other inhibitors in the sample, the environment will be less acidic. The color of the acid/base indicator will reflect that relatively low acidity.
Some embodiments utilize a polybuffer. An example of such a polybuffer is a combination of buffers, one with a pKa of above 7, for example about 8 to about 11, and another with a pKa of below 7, for example about 4.5 to about 6.3. The polybuffer can be included in the culture, in a separate niblet or provided separately for later addition. When premixed in the culture, the polybuffer can be used to both stabilize the reagent system prior to test operation and stabilize results after the testing is complete.
In a particular example of a polybuffer combined with the culture, borate and succinate are used. Borate helps provide a high pH environment to stabilize the culture during pre-test storage. Succinate helps provide a low pH environment to stabilize the pH of the system after test operation. For example, in a negative sample the pH of the culture will be reduced as spores germinate and bacteria multiply. After test completion the color of the culture will reflect the test results. The buffer with pKa of below 7, for example succinate, will help stabilize the pH of the now acidic environment, thereby minimizing or preventing further color change. External pressure for pH change, for example decreased temperature after the test is removed from an incubator and the test returns to room temperature, may cause the culture to become more basic and, therefore, the test will appear more positive. A buffer with pKa below 7 will help stabilize the result even upon return to room temperature. In another embodiment Trizma Base is the high pKa buffer and can be combined with a low pKa buffer such as succinate.
Choice of buffers will be governed by a variety of factors. One factor is avoiding buffers that are particularly sensitive to the temperature changes within the test. Another factor is the starting pH of the particular buffer. For example, Trizma Base may provide less temperature stability as compared to borate. Trizma Base, however, can provide a more basic starting pH and, therefore, possibly better test sensitivity and stability as compared to borate. In an embodiment Trizma Base is used to adjust the pH of the pre-use culture to a pH of greater than about 8, for example about 10.5.
Adjusting the pH of the starting culture to a relatively high pH is also a method to improve test sensitivity and/or shelf life. A high starting pH, for example above about pH 7.5, for example about pH 8 or above, such as in the range of about pH 7.5 to about pH 11, may help prevent premature spore germination. The high pH environment may also help avoid mold contamination. Generally, the high pH environment may help extend the shelf-life of the culture prior to use.
In another embodiment multiple test samples can be tested using a test plate for example a 96 well test plate. In such an embodiment media culture and adjustment binders can be provided together in the well. In another embodiment multiple vials are supplied, for example attached to each other by a perforation or breakable plastic, so that one or multiple tests can easily supplied to the user.
The following culture can be used for detection of antibiotics and other inhibitors in a variety of matrices including, for example, urine, milk, water, poultry, seafood, feed and feed extracts and meat, such as kidney samples.
A Bromocresol purple (BCP)/Tris Solution was prepared by combining 25 mL of a Trizma Base solution (TBS), the solution including 2.5 grams Trizma Base in 100 mL of reverse osmosis/deionized water (RO/DI Water), with 100 milligrams of BCP and mixing well. The media was prepared by dissolving 5 grams glucose and 1 gram Mueller Hinton Broth (Mueller Hinton broth includes, in purified filtered water, 2 grams per liter (g/L) beef extract, 17.5 g/L casein hydrolysate acid and 1.5 g/L starch, pH 7.3 at 25 degrees C) into 100 mL RO/DI Water. Then a 0.01 mg/mL solution of trimethoprim in RO/DI Water was prepared and 1.2 mL was added to the media. Next 20 mL of BCP/Tris (sterile filtered through 0.45 micron filter) was added to the media.
Agar was prepared by combining 0.3 grams Difco Bacto-Agar (Item #0140-01), 0.225 NaCl and 17.485 RO/DI Water. The mixture was heated to 95� C. and then removed from the heat and allowed to cool to 75� C. Next 6 mL of media (prepared as described above with BCP/Tris) was added to the agar and mixed for 5 minutes. The mixture was cooled to 57� C. and 1 mL spore solution (concentration of 1 billion cfu/mL) was added and mixed together for 5 minutes. 0.200 mL was dispensed into the bottom of a vial 4.
The dispensed vial 4 was prepared as described in example 1. An extractant was prepared by adding 4.8 grams of 47.2% Trizma Base and 52.5% Potassium Phosphate Monobasic to 1000 mL RO/DI Water (pH should be 7.5+/−0.10). The extractant was sealed within the niblet 5 and the niblet was added to the vial 4. The vial 4 was heat sealed with foil.
(Note: this procedure is described below using a format in which the swab is provided packaged within the test unit such as in the PocketSwab.)
The single service kidney swab procedure was run by first making a 3 inch incision into kidney. The swab 1 was then withdrawn from the test unit, the test unit in the form of a POCKETSWAB, by gently pulling and twisting the handle 2 out of the test unit body 7. The swab 1 was then inserted into the incision of the kidney and allowed to sit for 15 minutes to allow full absorption of liquid into the swab 1. The swab 1 was reinserted by gently pushing down and twisting to engage the threads 6 and screwing the handle down slowly about halfway. The swab tip 3 punctured the seals 12, 13 & 14 immersing the swab tip 3 into the extractant within the niblet 5 where it sat for 2 minutes. The swab 1 was then screwed down all the way. After shaking and tapping the vial 4 to get residual liquid into the bottom of the vial 4 and in contact with the culture 15, we placed the vial 4 into a heat block set at 67� C. and incubated for 2.5 hours (if urine is tested instead of kidney, incubate for 4 hours). After 2.5 hours the vial 4 was removed from the incubator and vial color was observed in comparison to reference colors. If the negative control did not match the color specified on the protocol incubation was continued for an additional 10 minutes.
Using the above procedure with the further addition of various amounts of beta-lactam receptor units to the top of the culture, from 0.3 units to 5.0 units, test sensitivity to penicillin G, was reduced by varying amounts dependent on the amount of receptor added. See example 5 below for receptor unit determination.
The culture is prepared as described in example 1. The extractant is prepared as described in example 2. The test is run the same as in example 3 except that the swab tip 3 was allowed to sit in urine sample for 10 seconds to allow full absorption into the swab tip 3 followed by incubation for 4 hours.
Culture is prepared as in Example 1. Prior to dispensing 200 microliters into the vial 4, antibiotic binder is prepared, in this example an antibiotic binder from the cell wall of B.st., also known as receptor. Varying amounts of inhibitory receptor units are mixed into the culture. Receptor is defined as protein removed from cellular membrane of B.st. A receptor unit (1 U) is the amount of receptor that will, in the test, reverse the culture growth inhibition of 12.5 ppb of penicillin G.
Receptor is useful in a variety of forms including purified form, in the form of cell paste in which receptor is a component of the whole cell or in the form of broken cell fragments.
Results from experiments are shown in Tables 1, 2 and 3.
Results in Table 1 are from tests using the culture described in Example 1 and with the addition of receptor added into rinse buffer (rinse buffer composition is 4.8 grams of 47.2% Trizma Base and 52.5% Potassium Phosphate Monobasic to 1000 mL RO/DI Water) at 0, 0.3 units (U), 1.0 U, 2.0 U, 5.0 Upper test. Sample contained the following amounts of penicillin G: 0, 5 parts per billion (ppb), 10 ppb, 12.5 ppb, 25 ppb and 50 ppb. As shown, 1.0 U was able to reverse the result of a 12.5 ppb penicillin G sample. Results are either positive (+) or negative (−).
Table 2 data shows that when adding 1 U or receptor sensitivity to drugs other than beta-lactams results did not change. Results are either positive (+), negative (−) or borderline between positive and negative (+/−). Abbreviations are penicillin G (PenG); sulfamethazine (SMZ); sulfadimethoxine (SDM); tylosin (TY); gentamicin (G); oxytetracyclines (OT).
The results in Table 3 show that the receptor can be added to various locations or components within the test to achieve the same or similar result. Results shown are from negative sample and samples containing 12.5 ppb penicillin G and 50 ppb penicillin G. Results are either positive (+) or negative (−).
The following culture can be used for detection of antibiotics and other inhibitors in a variety of matrices including, for example, urine, milk or kidney samples and can be used alone in a test container or in the single service test unit or in sensitivity adjustment examples.
BCP/Borate/Succinate Solution was prepared by adding 3.8 grams of Borate and 6 grams Succinate to 100 mL of reverse osmosis/deionized water (RO/DI Water) in 125 mL flask. 50 milligrams of BCP was added to a 50 mL conical tube and 5 mL of Borate/Succinate solution was added to the 50 milligrams BCP and mixed.
To prepare media 5 grams glucose and 1 gram Mueller Hinton Broth was dissolved in 100 mL RO/DI Water. 1.2 mL of a 0.01 mg/mL solution of trimethoprim in RO/DI Water was prepared and added to the media. A 20 mL of BCP/Borate/Succinate was added to the media and the media was sterile filtered through a 0.45 micron filter and then cooled to 4 degrees C.
Agar was prepared by combining 0.3 grams Difco Bacto-Agar (Item #0140-01), 0.225 NaCl and 17.485 RO/DI Water. The mixture was heated to 95� C. and then removed from the heat and allowed to cool to 75� C. Next 6 mL of media (prepared as described above with BCP/Tris) was added to the agar and mixed for 5 minutes. The mixture was cooled to 57� C. and 1 mL spore solution (concentration of 1 billion cfu/mL) was added and mixed together for 5 minutes. 0.200 mL was dispensed into the bottom of each of the vials.
After dispensing, tests a run test to determine if sensitivity is adequate. If sensitivity is not adequate the procedure is repeated using either more spores (under sensitivity) or less spores (if oversensitive).
The following Table 4 shows results from antibiotic detection tests using culture described in Example 6. In test operation 200 microliters of various milk sample, spiked with known concentrations of antibiotics, were pipetted into test vials and incubated for the prescribed time (in this test 2 hours 10 minutes) and prescribed temperature (64 degrees C+/−2 degrees C.). Incubation was in the Charm I Inctronic incubator. Results were recorded immediately after test completion (Color 1) and after being left at room temperature for 16 hours (Color 2). Results listed are in parts per billion (ppb). 4 samples were run at each concentrations except for raw negative milk (16 samples) and 3 ppb pen G (2 samples) and amoxicillin (2 samples). Abbreviations are pencillin G (PenG); sulfamethazine (SMZ); sulfadimethoxine (SDM); tylosin (TY); gentamicin (G); oxytetracycline (OT); pirlimycin (Pirl); amoxicillin (Amox); ampicillin (Amp); cloxacillin (Clox); ceftiofur sodium (Ceft); cephaprin (Ceph); tilmicosin (Til); sulfadoxine (SDN); neomycin (Neo). Color result 1 and 2 is a negative result, 3 is a borderline result and 4 and 5 is a positive result.
The following culture can be used for detection of antibiotics and other inhibitors in a variety of matrices including, for example, urine, meat, poultry, seafood, milk or kidney samples and can be used in a variety of formats including single service or sensitivity adjustment examples.
A Trizma/Succinate solution was prepared by adding 2.5 grams Trizma Base and 6 grams Succinate to 100 mL of reverse osmosis/deionized water (RO/DI Water) in a 125 mL flask. 40 milligrams of BCP (see example 1 for BCP preparation) was then added to a 50 mL conical tube and 25 mL of the Trizma/Succinate solution was added to the 40 milligrams BCP and mixed.
The media was prepared by dissolving 5 grams glucose and 1 gram Mueller Hinton Broth in 100 mL RO/DI Water. 1.2 mL of a 0.01 mg/mL solution of trimethoprim in RO/DI water was added to the media. 20 mL of the BCP/Trizma/Succinate preparation was added to the media and the media was sterile filtered through a 0.45 micron filter and cooled to 4 degrees C.
0.3 grams Difco Bacto-Agar (Item #0140-01), 0.225 NaCl and 17 mL of RO/DI water was combined and mixed and heated the mixture to 95� C. The mixture was removed from the heat and allowed it to cool to 75� C. and then 6 mL media (with BCP/Tris) was added and mixed for 5 minutes. The mixture was cooled to 57� C. and 2 mL spore (concentration of 1 billion cfu/mL) was added and mixed for 5 minutes. 0.200 mL was dispensed into the bottom of each vial.
After dispensing, tests were run to determine if sensitivity was adequate. If sensitivity is not adequate the procedure is repeated using either more spores (if under sensitive) or less spores (if oversensitive).
The following Table 5 shows results from antibiotic detection tests using the culture described in Example 6. In test operation 200 microliters of various milk samples, spiked with known concentrations of antibiotics, were pipetted into vials and incubated for the prescribed time (in this test 2 hours 30 minutes) and prescribed temperature (64 degrees C. +/−2 degrees C.). Results were recorded immediately after test completion and 16 hours later. Incubation was in various Charm Inctronic I incubators. Concentrations listed are in parts per billion (ppb). 4 samples were run at each concentration except 16 negative samples were run and 2 samples each for 3 ppb pen G and amoxicillin. Abbreviations are pencillin G (PenG); sulfamethazine (SMZ); sulfadimethoxine (SDM); tylosin (TY); gentamicin (G); oxytetracycline (OT); pirlimycin (Pirl); amoxicillin (Amox); ampicillin (Amp); cloxacillin (Clox); ceftiofur sodium (Ceft); cephaprin (Ceph); tilmicosin (Til); sulfadoxine (SDN); neomycin (Neo).
The following culture can be used for detection of antibiotics and other inhibitors in a variety of formats and matrices including, for example, urine, milk or kidney samples.
BCP/Borate/Succinate solution was prepared by adding 3.8 grams of Borate and 2.7 grams Succinate to 100 mL of reverse osmosis/deionized water (RO/DI Water) in a 125 mL flask. 100 milligrams of BCP was added to a 50 mL conical tube and 25 mL of the Borate/Succinate solution was added to the 100 milligrams BCP and mixed.
To prepare the media 5 grams glucose was dissolved into 100 mL RO/DI water. 1.2 mL of a 0.01 mg/mL solution of trimethoprim in RO/DI water was added to the media along with 20 mL of the BCP/Borate/Succinate preparation. The media was sterile filtered through a 0.45 micron filter and cooled to 4 degrees C.
0.3 grams Difco Bacto-Agar (Item #0140-01), 0.225 NaCl and 18 mL of RO/DI water was combined and mixed and the mixture heated to 95� C. The mixture was then cooled to 75� C. afterm which 6 mL media was added and mixed for 5 minutes. The mixture was then cooled to 57� C. and 1 mL spore (concentration of 1 billion cfu/mL) was added and mixed in for 5 minutes. 0.200 mL was dispended into the bottom of each vial.
After dispensing, run tests to determine if sensitivity is adequate. If sensitivity is not adequate repeat procedure using either more spores (if undersensitive) or less spores (if oversensitive).
To efficiently dispense culture (such as is required in the previous examples), containing spores of Bst, into the bottom of the vial 4, the agar must be heated to approximately 57 degrees C. Dispensing is done rapidly, for example in less than one hour, preferably in 45 minutes or less, so that the culture can be quickly cooled. If the culture is not quickly cooled, spores, for example B.st. spores, may germinate prematurely. That is, spores will germinate prior to application of the sample. Excess premature germination will reduce test sensitivity.
Another method for preventing premature spore germination, which can be used alone or in conjunction with rapid dispensing, is to increase the pH of the culture, for example in the range of about pH 7.5 to about pH 11. The increase in pH provides non-optimal conditions for spore germination. The increase in pH also allows more stability over long storage times.
In some embodiments and examples, such as example 4, in which the sample matrix being tested is variable, a dilution buffer can be used to standardize the sample matrix. In example 4, a test for urine was described. In this example, to standardize the urine samples prior to testing the sample is diluted approximately 1 to 20 with a mixture of extractant (4.8 grams of 47.2% Trizma Base and 52.5% Potassium Phosphate Monobasic to 1000 mL RO/DI Water-pH should be 7.5+/−0.10) combined with about 8-10 grams per liter beef extract. Other samples, including feed and water can be similarly diluted (for example water was diluted 1 to 5 feed 1 to 30). By diluting the sample, test sensitivity is similarly reduced.
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Home / Store / Journals / Electronic Articles / Fusion Science and Technology / Volume 64 / Number 2
L. M. Garrison, G. L. Kulcinski
Fusion Science and Technology
Volume 64 / Number 2 / August 2013 / Pages 216-220
Format:electronic copy (download)
Single crystal tungsten samples with orientation (110) were irradiated in the Materials Irradiation Experiment with normal incidence 30 keV He+ at 900 aC. Samples were mechanically polished and then electropolished with a KOH solution before irradiation to 3×1017 to 6×1018 He+/cm2. With increasing fluence sample surface pore size increased from ~20 nm to more than 100 nm. Mass loss also increased with fluence to a maximum of 15 g/m2 lost for the highest fluence sample.
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