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--------- I j I ~iP INTELLECTUAL PROPERTY PHILIPPINES i OFFICE OF THE DIRECTOR GENERAL SPECIFIQUE GARMENTS Appeal No. 14-08-44 MANUFACTIJRING, INC., Opposer-Appellee, Inter Partes Case No. 14-2006-00107 Opposition to: -versus- Application No. 4-2004-005782 Date Filed: 30 June 2004 LT2, LLC, Respondent-Appellant, Trademark: Le TIGRE & Design x-------------------------------------x DECISION LT2, LLC ("Appellant") appeals Decision No. 2007-87, dated 29 June 2007, and Resolution No. 2008-17 (D), dated 31 October 2008, of the Director of the Bureau of Legal Affairs ("Director") sustaining the opposition filed by SPECIFIQUE GARMENTS MANUFACTURING, INC. ("Appellee") to the Appellant's application for the registration of the mark "Le TIGRE & Design)". Records show that the Appellant filed on 30 June 2004 its trademark application covering goods falling under Class 25 of the Nice Classification I namely: clothing, which includes men's, women's, and children's apparel, sweaters, sweater shirts, sport shirts, athletic shirts, knit shirts, t shirts, jackets, sport coats, all-weather coats, raincoats, pants, slack jeans, trousers, jump suits, track suits, shorts, boxer shorts, swim trunks, robes; headwear, namely: knitted ski hats, hats, caps, visors; shoes, boots, sneakers and sandals. The application was published in the "IPO e-Gazette" for Trademarks on 12 April 2006. On 10 August 2006, the Appellee filed its "NOTICE OF OPPOSITION" alleging the following: 1. It manufactures and sells garments bearing the mark Le TIGRE & Design to Shoe mart, Ever Plaza, Inc. and other business entities and it assigns employees to Shoemart to oversee the sales of "Le Tigre" shirts; 2. It has adopted and used Le TIGRE & Design since 1982 or for about 24 years now; 3. The mark sought to be registered by the Appellant is identical to the mark earlier adopted and used by the Appellee, and, thus, cannot be registered; I The Nice Classification is a classification of goods and services for the purpose of registering trademarks and service marks, based on a multilateral treaty administered by the World Intellectual Property Organization. This treaty is called the Nice Agreement Concerning the International Classification of Goods and Services for the Purposes of the Registration of Marks concluded in 1957. Republic of the Philippines T1\.TT..... T I7'£"'T"TT'" T v n D r l . D v n ....... I"'l.DDT£'lD r i 4. It worked to promote and create goodwill for Le TIGRE & Design by placing advertisements on different media, namely, radio and print ads; and 5. To allow a junior user, such as the Appellant, to be the registered owner of Le TIGRE & Design would cause damage to it and the goodwill and reputation acquired by Le TIGRE & Design because of its efforts and resources will go for naught, and instead, a third person will reap the benefits thereof. The Appellee submitted the following evidence to support its opposition: 1. Letter of the Sales Manager of the Appellee, dated 13 October 1987;2 2. Affidavit of Evangeline Natad Bernal, executed on 09 August 2006;3 3. Affidavit of Estelita Ramirez, executed on 09 August 2006;4 4. Sales Invoices:" 5. Affidavit of Valentino Madriaga, executed on 10 August 2006;6 6. Affidavit of James Ong, executed on 09 August 2006;7 I 1 7. 8. Affidavit of Henry Go, executed on 10 August 2006;8 Advertising contracts and related official receipts." and 9. Patches and labels used by the Appellee." f I i The Appellant filed its "ANSWER" on 04 January 2007 alleging the following: i I l 1. Rep. Act No. 8293 ("IP Code") adopts the "First-to-File" system in the protection of intellectual property rights and vacated the reliance on the first to invent system in patents and the prior use in the case of trademarks; 2. There was no attempt on the Appellee's part to register Le TIGRE & Design which is a glaring and clear indication that it did not own, much less, create, concoct or make it; 3. It made, concocted and owned Le TIGRE & Design and it filed applications and obtained registrations thereof not only in the Philippines but in other countries; 4. Its applications/registrations in various countries are reflected in the websites of the trademark offices of some of these countries, and which evidently shows that the Appellee has no basis under the IP Code to file the opposition and that the Appellee 2 Annex "A". 3 Annex "B". ~ Annex "C", 5 Annexes "0" to "I". 6 Annex "J". 7 Annex "K". 8 Annex "L". 9 Annexes "M" to "Q". 10 Annex "R". J./ brazenly, unlawfully, illegally, fraudulently and deceitfully appropriated, adopted and used the mark; and S. It would be in the height of injustice to invalidate the registration of its mark based on the opposition of the Appellee who unlawfully, illegally, fraudulently and deceitfully appropriated, adopted and used the subject mark without its knowledge and consent causing incalculable damage, prejudice and injury to its intellectual property rights on Le TIGRE & Design. The Appellant's evidence consists of the Trademark Status Report for the Appellant's marks in various countries" and printouts from the websites of the trademark offices in various countries. 12 In sustaining the opposition, the Director ruled that the Appellee's actual and continuous use of the subject mark has resulted to its ownership thereof under Rep. Act No. 166, as amended" ("RA 166") and that the Appellee has gained vested rights as provided in Sec. 236 of the IP Code. She held that the repeal of RA 166 did not terminate all trademark rights acquired under and pursuant thereto. According to her, Sec. 236 of the IP Code draws a conclusion that the rights referred therein include the rights of owners and prior users of unregistered marks that were acquired in good faith before the IP Code took effect and includes the right to appropriate exclusive use of the mark and the right to oppose or prevent the registration of the mark in favor of others. She also held that the Appellee satisfactorily proved its superior right through actual sales in local commerce way back in 1982 by presenting sales invoices and affidavits of its employees and other disinterested persons. The Director also denied the Appellant's "MOTION FOR RECONSIDERATION" treating it as "mere scrap of paper" for lack of notice and hearing. Dissatisfied, the Appellant filed the instant appeal through registered mail on 19 November 2008. c In its "NOTICE OF APPEAL", the Appellant maintains its arguments before the Director and asserts that the Appellee cannot acquire by prior use a mark already owned by another. The Appellant points out that it owns and used Le TIGRE & Design since 20 September 1979 while the Appellee has used it only on 13 December 1991, at the earliest. According to the Appellant, mere use of a mark without registration under RA 166 will not be converted into ownership for if it so, then there will be no need for registration under RA 166 based on local use. The Appellant also alleges that the Appellee has not applied for registration based on local use under RA 166 although it allegedly used the mark in 1982 up to 1998, or for a period of 16 years, because it knows it does not own it and that if it applies for registration there will be a chance that its illegal, fraudulent, unlawful, deceitful and even its criminal act may be found out. Thus, says the Appellant, the Appellee not having secured any registration under RA 166, it has not gained for itself any vested rights under Sec. 236 of the IP Code and therefore is bereft of any protection from the new law. In 11 Exhibit "1". 12 Exhibits "2" to "9". 13 AN ACT TO PROVIDE FOR THE REGISTRATION AND PROTECTION OF TRADEMARKS, TRADE-NAMES AND SERVICE MARKS, DEFINING UNFAIR COMPETITION AND FALSE MARKING AND PROVIDING REMEDIES AGAINST THE SAME, AND FOR OTHER PURPOSES. .V f contrast, the Appellant contends, its trademark applications or registrations in 38 countries should be considered as a very strong if not irrefutable proof of ownership of the mark. On 12 December 1998, this Office issued an Order calling the attention of the Appellant to the requisites for the filing and perfecting an appeal to the Director General and required the said party to submit within 3 days from receipt thereof a legible copy of the NOTICE OF APPEAL, proof that it served copies of the appeal upon the Director and the Appellee, 3 legible copies of the appealed decision, and a valid certification of non-forum shopping. The Appellant was warned in the Order that failure to comply therewith would cause the dismissal of the appeal. The Appellant filed a "COMPLIANCE" on 18 December 2008. Upon evaluation of the submission, this Office issued the Order of 15 January 2009, stating that the Appellant failed to attach to the COMPLIANCE copies of the appealed decision and that the certificate of non-forum shopping was not signed by the Appellant but by its counsel, and that in the interest of justice, this Office was giving the Appellant a final period of 3 days from receipt thereof to submit a copy of the assailed decision and proof that it has authorized its counsel to execute the certificate of non-forum shopping. The Order cautioned the Appellant that failure to comply shall cause the dismissal of the appeal. On 23 January 2009, the Appellant filed, through its counsel, a "COMPLIANCE AND MANIFESTATION'. Consequently, on 05 February 2009, this Office issued another Order that with the filing of the COMPLIANCE AND MANIFESTATION on 23 January 2009, the Appellee and the i J Director are required to file within 30 days from receipt thereof comment to the appeal and to ! ] 1 forward records ofthe case, respectively. On 03 March 2009, the Appellee filed its "COMMENT (To the Appeal)" alleging that the the decision of the Director is final and executory as it was filed out of time and not signed by a duly authorized representative. According to the Appellee, the Appellant's COMPLIANCE AND MANIFESTATION must be stricken off the records because it does not bear the required information of the Appellant's counsel's compliance to the Mandatory Continuing Legal Education ("MCLE"). Also, the Appellant asserts that it has duly acquired ownership of Le TIGRE & Design by prior use even without a registration thereof and has a vested right therein. It argues that the law on trademarks adheres to the principle of nationality and territoriality and the registration in foreign countries is not registration in the Philippines. The repeal of RA 166, the Appellee contends, did not terminate all trademark rights acquired under and pursuant to the said law and that it has proven its superior right through actual sales in local commerce way back in 1982 while the Appellant has not presented a single evidence on its actual use of the mark either in the Philippines or abroad. Meanwhile, on 05 March 2009, the Director forwarded the records of the case. Thus, this Office issued its Order of 09 March 2009 giving the parties 15 days from receipt thereof to file their respective memorandum. On 30 March 2009, the Appellant filed a motion for extension to file memorandum, while the Appellee filed its memorandum the following day. Finally, on 03 April 2009, this Office issued an Order granting the Appellant's motion for extension or until 09 April 2009. The Appellant did not file its memorandum. r i \ .... This Office noted that the Appellee has raised two pivotal technical issues: first, that the appeal was filed out of time, and second, that the Appellant's counsel did not comply with the Supreme Court's requirement regarding the disclosure of compliance with MCLE in the pleadings filed in this appeal proceeding. On the first issue, records show that the Appellant received a copy of Decision No. 2007-87 on 06 July 2007. Pursuant to Sec. 2 of the Uniform Rules on Appeal, the deadline for the Appellant to file a motion for reconsideration or an appeal was on 05 August 2007. Considering however, that 05 August 2007 was a Sunday, the Appellant filed its motion for reconsideration on the next working day on 06 August 2007. In addition, the records show that the Appellant received on 18 November 2008 a copy of Resolution No. 2008-17 (D) which denied its motion for reconsideration. Contrary to the Appellee's allegations, the Appellant filed the instant appeal not on 18 December 2008 but on 19 November 2008. It is settled in our jurisprudence that when a party files a motion for reconsideration on the last day to perfect an appeal, the appellant still has one day from )' receipt of the denial of the motion within which to perfect an appeal." Going now to the second issue, it is the Appellee's argument that the Appellant's COMPLIANCE AND MANIFESTATION filed on 23 January 2009 must be stricken off the records i as the same does not bear the information regarding the MCLE Compliance of the Appellant's counsel, hence, the appeal should be dismissed. In this regard, the Supreme Court Resolution on Bar Matter No. 1922, dated 03 June 2008, explicitly provides, among other things: The Court further Resolved, upon the recommendation of the Committee on Legal Education and Bar Matters, to REQUIRE practicing members of the bar to INDICATE in all pleadings filed before the courts or quasi-judicial bodies, the number and date of issue of their MCLE Certificate of Compliance or Certificate of Exemption, as may be applicable, for the immediately preceding compliance period. Failure to disclose the required information would cause the dismissal of the case and the expunction ofthe pleadings from the records. (Underscoring supplied) Following its publication for 60 days in a newspaper of general circulation, Bar Matter No. 1922 was supposed to take effect on 25 August 2008. However, in its Resolution En Bane, dated 02 September 2008, the Court resolved to amend the effectivity date of the implementation of Bar Matter No. 1922 to 01 January 2009. The requirement under Bar Matter No. 1922 covers all pleadings filed in courts and quasi judicial bodies. Based on the clear and unequivocal wordings of the Supreme Court, the dismissal of the case and expunction of the pleadings from the records for non-compliance are mandatory. Hence, the Appellant's counsel in all the pleadings filed on behalf of the Appellant from 01 January 2009 onwards was required to indicate the information regarding said counsel's MCLE Certificate of Compliance or Certificate of Exemption. 14 See Lloren, et al us. De Veyra, et al, G. R. No. L-13929. 28 Mar. 1962. • I J I A scrutiny of the records shows that none of the pleadings submitted or filed by the iI Appellant starting January 2009 bear the required information on its counsel's compliance to MCLE. ! This Office also noticed that while the issue was raised by the Appellee in its comment to the 1 appeal, a copy of which was duly served upon the Appellant, the Appellant did not exert effort to I address such deficiency and submit the required information nor indicate or disclose them in its I subsequent pleadings. I Thus, pursuant to the Supreme Court Resolution on Bar Matter No. 1922, this Office is constrained to strike out the Appellant's COMPLIANCE AND MANIFESTATION, filed on 23 January 2009 and other subsequent pleadings. I 1 Consequently, it is deemed that there has been no compliance by the Appellant to this Office's Order of 15 January 2009. This notwithstanding the amendments to the Uniform Rules on Appeal which took effect on 25 February 2009. 15 Although the amendments apply retroactively to all pending appeals in the Director General as of 25 February 2009, and dispensed with the requirement of certification of non-forum shopping and reduced the number of copies of the I appealed decision to be attached to the appeal memorandum from three to two, among other things, the deadline for the Appellant to submit compliance to the Order of 15 January 2009 had lapsed before the amendments took effect. Moreover, the expunction of the Appellant's COMPLIANCE AND MANIFESTATION means that the Appellant is still to be considered to have failed to submit the required number of copies of the appealed decision. t J 1 WHEREFORE, premises considered, the appeal is hereby DISMISSED for the reasons 1 discussed above. Let a copy of this Decision as well as the trademark applications and records be furnished and returned to the Director of the Bureau of Legal Affairs for appropriate action. 1 1 Further, let also the Director of the Bureau of Trademarks and the library of the Documentation, I 1 Information and Technology Transfer Bureau be furnished a copy of this Decision for information, 1 guidance, and records purposes. I \ ! SO ORDERED. DEC 0 82881 Makati City ! i I I I I 15 Promulgated though Office Order No. 12, S. 2009.
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Residence Inn Cranbury South Brunswick
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NYC-the day Bill went crazy with credit card!
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A foreigner in Our Own Country!
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Things are going a little more smoothly this AM
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Anyway, I better try and sum this post up. It's getting long. Two more panels with Daniel Gillies and Torrey DeVitto. Daniel is of course super funny and Torrey is just lovely! Check out my pics!
I hit the airport again the next day bound for San Francisco! ...

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Would the Lib Dems manage to keep Nick Clegg as leader during coalition negotiations with Labour?
Labour and the SNP have competing visions for Scotland and the UK – could they really find common ground in Westminster?
Hung parliament preparations.
Will the decision whether or not to go into coalition in 2015 depend more on money than on principle?
Will the Lib Dems eschew a Lib-Lab pact and allow Labour to struggle on as a minority? And why?
Some Conservatives are nervous about what their party will look like beyond coalition with the Lib Dems. Both their and voters' views jar with the PM's insistence that he doesn't want another coalition.
Harriet Harman may hand over the DCMS brief if she takes a leading role in negotiations with the Lib Dems.
Inflation alone will ensure that the allowance rises to over £11.3k and minimum wage workers will still be paying tax.
As Andrew Adonis argues, successful reforms are incremental and build on existing best-practice, rather than trying to reinvent the wheel.
The planned change to the definition in the Energy Bill would cut the number of households classed as fuel poor from 3.2 million to 2.4 million.


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Donna DodsonChief, Computer Security DivisionNational Institute of Standards and TechnologyCommerce Department
In recent years, NIST has had a big say in how government uses technology and secures its IT, so it's a testament to Donna Dodson's quiet leadership that its influence is now even greater.
She has displayed a clear understanding of how to engage with the White House, intelligence agencies and industry on cybersecurity. Dodson became the glue that held all the sides together. In the process, she made NIST's security policies, best practices and tools more effective.
Many organizations have been vying for leadership of the cybersecurity debate and could have submerged NIST's more practical sensibility. By speaking authoritatively and carrying a padded stick, Dodson has become one of the most trusted government advisers on cybersecurity.
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About the Author
Brian Robinson is a freelance writer based in Portland, Ore.
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Unisys Federal Systems' Venkatapathi Puvvada, this year's Eagle award winner for industry, stresses the importance of community.
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The spy agency wants to better integrate cybersecurity into its traditional human intelligence operations.
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NH Nacional
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Clean rooms, gorgeous lobby, wonderful breakfast, beautiful view from terrace in the room. Not much else I could ask for.
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Vehicles equipped with a standard sunroof glass panel are being recalled because the glass can break without warning. Dealers will replace the sunroof glass on affected vehicles to correct this concern.
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text: Quick View Eat. Nourish. Glow.: 10 easy steps for losing… by Amelia Freer January 1, 2015 | Kobo Edition (eBook) Available for download from $15.99 Nutritional therapist and healthy eating expert Amelia Freer dispatches fad diets to the distant past while guiding you to a rejuvenated future. Amelia Freer brings a fresh and unique voice to the field of holistic health. In this, her first book, she explains her 10 steps and provides over 25… See Details $15.99 was $15.99


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This is a thick, comfortable rug. However this is not a lilac and cream colored rug. Nor is it yellowish cream. The lilac shade is good but the other is really beige or even a greenish beige. So the rug is cool in tone and not as interesting as the photo suggests. I'm keeping it because it's so hard to return, and I had factored in keeping a quieter version of what I'd hoped for, knowing that at least it would have some charm and not compete with the art or decorations in the room. I'm not giving it a higher rating due to the unprofessionalism of the inaccurate photographic work and what a huge hassle that is in the buying of rugs.
We bought this rug to complement our neutral oatmeal-colored couches in the living room. It's a lovely, thick wool rug that has added a nice splash of subtle color to the room. I think the colors seen online are accurate- very light lilac and light creamy yellow diamond pattern. 'Feels nice on foot. Minimal shedding in the first week, as expected.
Adds character and warmth to the living room. Love the colors.
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European Highlights » Review by dylan
This tour was amazing, saw some amazing sites and met some great people. I would recommend this to anyone!
Review written Oct 2012
My Contiki was: Amazing, Fun, Superbevent 0.3
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The making of Harry Potter
Enjoy the most memorable sets from all of the films.
Save £5 compared to buying tickets on the day, and get priority entrance by buying from the VisitBritain Shop.
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Not "a" judge, there have been quite a few now including the appellate courts that agreed with that first judge, and now before 9 more, the highest in the land.<quoted text>
So a judge bought it......andddddddd....that means.....?
<quoted text>
Far easy to dismiss my point, than offer an opposing view.
But it doesnt matter, even if they chose the wrong way, we put overturning prop8 on the next ballot...and poof... gone in a walk, dont even have to spend much money on it.
This is happening there is no stopping it now, even your media prophet has stated this now.


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|hotels > United States > Texas > Dallas > hotel search
Comparison of hotel rates in Dallas
|Get the best rates in Dallas. Please enter your hotel search details. We will then search for hotels on the websites listed opposite so you can compare the cheapest rates available. Comparison results are in price oder.
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The speech and hearing impaired women who gathered at Church Park Presentation Convent School to celebrate International Women's Day on Sunday communicated their need to be understood and interpreted.
The women were participating in an event organised by Deaf EnAbled Foundation, which began reaching out to women in Chennai three years ago. The 100 women participants spent the morning playing indoor games. Post-lunch, M. Ramya, Director, Communication, made a presentation on women achievers. She spoke about Myanmar's leader Daw Aung San Suu Kyi and hearing and speech impaired women such as Vaishnavi Jayaraman, a single mother who runs a petrol bunk in Coimbatore. "They symbolise confidence and independence which women from districts sometimes lack. We want the women to come forward, become achievers," she adds.
Srimathi Jayakumar, 32, works in the back office of an international bank. Though she has been in the company for five years she does not benefit from the company's in-house training programmes. "I want to grow in my profession, but as there are no interpreters I lose out on training sessions," Ms. Jayakumar mimes. "If others get three weeks of training to update their knowledge, I get only one week and it is not enough," she adds.
S. Rajeswari, 23, of Tindivanam could not study beyond standard VII as her parents would not send her to a residential school. Rajeswari works in a shoe-making unit for a daily wage of Rs.70.
S. Anbuvizhi, 25, who works as a maid in an expatriate's house in Puducherry wants an interpreter who would explain to her parents about her wish to marry. "The groom's parents have agreed to the proposal but my parents do not understand my needs. I need an interpreter to explain to them my feelings," she says. She proposes to start an empowerment programme in Puducherry.
Sunday's event, which was organised by women, aims to encourage women to learn from the achievers to take up vocational courses, learn practical life skills, English and to operate computers.
The Foundation on its part is evolving a uniform sign language incorporating the various 'dialects' used by the speech and hearing impaired persons across the country. "We are lobbying for interpreters in hospitals, training programmes and for inclusion of sign language as a scheduled language," says Ms. Ramya, who hails from a family of persons with speech and hearing impairment.
Keywords: Women's Day


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Tirupati Hotels
123 Tirupati Hotels List ranked by travellers
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Rank 1
Upadyaya Nagar, Karakambadi Road,
"Ours was a spiritual cum holiday tour to Tirupathy with my father in law, mother in law, my wife..." More ›Murali krishna, Delhi, Today
6 Photos
Rank 2
Shilparamam, Tiruchanoor Road
"We were a mixed batch. My friend who is 82 years of age and my friend's son who is 14 years old...." More ›Kabita, Bangalore, 5 days ago
India Says Yes Award
Winner for categories:
6 Photos
Rank 3
Near Ramanuja circle, Renigunta road, Tirupati
"Bliss is rightly located and did not have any issues in booking and check-in. Their service was..." More ›Shivva kumar, Pune, Today
8 Photos
Rank 4
"Fantastic ambiance, lovely hospitality, features are very new and just 2 year old hotel. Food..." More ›Madhu babu, Delhi, 4 weeks ago
India Says Yes Award
Winner for categories:
7 Photos
Rank 5
Opp. APS RTC Bus Stand, Renigunta Road, Tirupati
"I visited Fortune Kences with my family for visiting the temple. The hotel is very near to the..." More ›Prathibha, Chennai, 2 days ago
India Says Yes Award
Winner for categories:
7 Photos
Rank 6
209, T.P.Area, Tirupati
"If you are visiting Tirupati and have an early morning Darshan, then this hotel should be good..." More ›Hasbriale, Delhi, 2 weeks ago
India Says Yes Award
Winner for categories:
5 Photos
Rank 7
G Car Street, Tirupati
"This was quite interesting to stay in this hotel with good environment. Good servicing with..." More ›V shehi patap purshotum, Ahmedgarh, 2 months ago
4 Photos
Rank 8
33-37, Renigunta Road, Tirupati
"Tirupathi is a holy place and pilgrimage center. Most of the time you are required to stay..." More ›Vs, Secunderabad, Today
India Says Yes Award
Winner for categories:
10 Photos
Rank 9
Opp APSRTC Bus Stand, Tirupati
"It is probably one of the most decent hotels in the city and is pretty affordable and reasonable...." More ›Sunil, Mumbai, 6 days ago
4 Photos
Rank 10
14/37, Karakambadi Road, PO Akkarampall, Tirupati
"Recently I stayed in this hotel for one day. Rooms are spacious, but bathroom is not clean and..." More ›Venkoba rao, 1 week ago
India Says Yes Award
Winner for categories:
16 Photos
Rank 11
No: 13-6-771/20, opp. APSRTC Bus Stand, Tirupati
"We celebrated our engagement function in Udayee international. Function went very nice and grand...." More ›V bhaskar reddy, Delhi, Today
India Says Yes Award
Winner for categories:
8 Photos
Rank 12
177. T.P. Area, Tirupati
"I visited Tirupati many times and never had an easy darshan like this time, thank God for that. I..." More ›Yeshwanth, Nov 8, 2014
India Says Yes Award
Winner for categories:
5 Photos
Rank 13
Near RailwayOver Bridge, Renigunta Road, Tirupati
"The hotel is located very close to the railway station and the bus station. The rooms are..." More ›Sirivasan, Adra, 3 months ago
12 Photos
Rank 14
Beside Railway Overbridge, Renigunta Road, Tirupati, Chittoor Dist.
"This was booked on one of our Thirumala temple visit. Was conveniently located decent place to..." More ›B, Mumbai, 3 weeks ago
3 Photos
Rank 15
514 , Chalapathi Towers, Near Ramanuja Circle
" It was well-located. Food served was good. Service was fantastic. Everything is perfect." More ›Aryabhanu, Jul 10, 2014
5 Photos
Rank 16
14-3-303 , A ,D.R.Mahal Road, Opp Second Choultry
"The hotel though three star, extended excellent facilities which included regular power supply..." More ›Thiyagu, Vellore, Sep 8, 2014
6 Photos
Rank 17
"The place is very good or I can say excellent. The food service is also good. Moreover the food..." More ›�r s meena, Delhi, 2 weeks ago
5 Photos
Rank 18
14-2-118/119, T.P Area, Opp Govindaswami Temple, Tirupati
"I stayed in the hotel on religious purpose with my family. Overall, experience in the hotel was..." More ›Haranand, 3 months ago
5 Photos
Rank 19
S.No.270/5, Beside SSB Kalyana Mandapam, Tiruchanoor Road
"The overall stay was excellent and it was a family trip for relaxation. The hotel was..." More ›Sandip, 3 months ago
5 Photos
Rank 20
Byepass Road, Tirupati
"It was one the most awesome hotels in my life, that I have experienced ever. The employees of..." More ›Satpreet, Chennai, Aug 18, 2014
India Says Yes Award
Winner for categories:
15 Photos
Rank 21
No: 13-3-37, Behind Venkateswara Theatre, Old Tiruchanoor Road
"There is no good hotels near by and you need to cross the railway cross to find good hotels...." More ›Bharat rajan, Delhi, 5 days ago
5 Photos
Rank 22
19-9-29/A, Tiruchanur Road, Sankarambadi Circle, Tirupati
"Hotel was conveniently located. I reached there comfortably. It was around one km away from bus..." More ›Jaychandra, 3 months ago
4 Photos
Rank 23
"I visited Tirupati for religious purpose. The hotel was nice and there were all facilities in..." More ›Shakshi, Indore, 1 week ago
5 Photos
Rank 24
4-70/1, Padmavathi Puram,
"Parking space not available. The staff service was good. No issues in check-in and check-out...." More ›Jayamanickam, Chennai, Sep 6, 2014
India Says Yes Award
Winner for categories:
5 Photos
Rank 25
Tirumala Byepass Road, Opp: T T D Srinivasam Complex, Tirupati
"It is located just a little far from Tirupati bus stand. It is a budget fitting hotel. You can..." More ›Surendra .b, Hyderabad, 2 months ago
India Says Yes Award
Winner for categories:
4 Photos
Rank 26
217, T.P.Area, Opp: APSRTC Bus Stand, Tirupati, Chittoor Dist.
"I accidentally visited this hotel, as my friend already checked in there. The rooms are quite..." More ›Himanth, Madanapalle, Sep 10, 2014
3 Photos
Rank 27
349, G car Street, Opp Rly Station, Tirupati
"Hotel Annapurna located opposite to Railway Station, T. P Area. The hotel is 18 kms away from..." More ›Nagalakshm, Bangalore, 1 week ago
6 Photos
Rank 28
20-3-133/2� Siva Jyothi Nagar, Near�Postal Colony, Tirupati
"I have booked room online for 4 members. Office is average. No food facility but we can get food..." More ›Chandrahas, Hyderabad, 1 week ago
3 Photos
Rank 29
No.650, Siva Jothi Nagar, Temple Valley road Bypass
"I stayed at Elite hotels for a single night. My flight got delayed but they extended my check out..." More ›Reenu, Delhi, Aug 21, 2014
5 Photos
Rank 30
6-8-995, Opp: Nandi Circle, Near Kapila Theertam, Srinivasa Nagar, N.G.O.'s Colony,
"We had good time and enjoyed. Staff also gave right guidelines as how to reach for temple and..." More ›Paresh, Mar 14, 2013
Hotels in Tirupati with Tariff Rates
- Tirupati is a vibrant pilgrim town, home to the famous Venkateswara Swamy Temple. It attracts pilgrims in large numbers and they can choose to stay in either hotels or guesthouses. Pilgrims visiting Tirupati spend an average of 2 nights in the destination and have a hotel budget of INR 1300 per person/night. People travelling in groups prefer hotels like Haritha Srinivasam Complex, Fortune Kences and Bhimas Deluxe Hotel, as they are located close to the temple. Since a large number people visit the destination in groups, budget hotels are popular. Ram Bagicha Guest House - I, Vishnu Nivasam Guest House, Reddy Bhavan and Sushma Hotel among other options are top budget places to stay as ranked by HolidayIQ.com Travellers. Hotels located near the bus stand are also recommended by HolidayIQ.com Travellers. Places to stay near the APSRTC bus stand include Fortune Kences, Grand Hotel and Udayee International.
- There are 106 hotels in Tirupati reviewed by actual travellers. The top rated Tirupati hotels are Hotel SLV Grand, Fortune Select Grand , Hotel Minerva Grand. The most sought after hotels, based on these reviews are Group stay hotels, Family stay hotels, Couple stay hotels. The lowest available tariff for Tirupati hotels stands at Rs.100 and highest hotels rate is approximately Rs.4000. The cheapest Tirupati hotel deals are available at Rs.113. You can search for Tirupati Hotel
(74), Tirupati B&B/Guest house/Hostel/Lodge(28), Tirupati Resorts(2), Tirupati Apartment/Serviced apartment(2).
There are different types of hotels in Tirupati available for families, lovers, kids, friends and specially for newly married couples who are looking for perfect honeymoon destination.
Star Hotels in Tirupati
There are 2 Three Star Hotels in Tirupati and most popular 3 star hotels in Tirupati are Fortune Kences, Sindhuri Park Hotel.
Hotels in Tirupati by PriceThere are 99 Budget hotels in Tirupati which are priced below Rs. 2500. Tariff for hotels in Tirupati starts from 100. Cheap hotels in Tirupati may provide basic amenities like a private bedroom, colour TV, Telephone, AC, 24-hour front desk and housekeeping staff on call. These hotels are affordable and value for money.
Popular Locality Hotels in TirupatiThe top 5 localities for hotels in Tirupati to stay are Tiruchanoor Road hotels, Railway Station hotels, APSRTC Bus stand hotels, Bye Pass Road hotels, Renigunta Road hotels.
Hotels near Main Attractions in TirupatiThe most popular attractions in Tirupati are Deer Park, Sri Venkateswara Temple, ISKCON Tirupati, Sri Govindarajaswami Temple, Sri Kapileswaraswami Temple and you can find 16 hotels near Deer Park, 16 hotels near Sri Govindarajaswami Temple, 16 hotels near Sri Kapileswaraswami Temple.
Hotels near Main Transportation PlacesThere are 2 railway stations in Tirupati which are Tirupati Main railway station, Tirupati West Halt railway station. You can find 22 hotels near Tirupati Main railway station, 22 hotels near Tirupati West Halt railway station.
The Airport in Tirupati is located in/near Tirupati, Tirupati and there are 22 hotels in Tirupati near airport travellers can choose from.


text:
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We recently announced the successful deployment of our Convergent Charging Solution at Telstra. Telstra is not only the largest CSP down under, but also has a growing international presence with operations spanning 15 countries. So it's not surprising that they…
sector news Monetization of the MomentEMI describes the hot topic of MWC for operators and vendors: Real-time. Operators see the subscriber/consumer/customer is making personalized, real-time decisions that form their digital lifestyle experience, and the operator has a micro-opportunity to discern personas and context in order to monetize the moment.
event Mobile World Congress Shanghai
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white paper Analysys Mason White Paper: Beyond the Digital Façade
Planning to become a Digital Service Provider? Analysys Mason reveals in this whitepaper why BSS plays a central role.
blog Who Tied a Bucket To My Speedboat? (part two)
In part one, I looked at why several of the traditional approaches to billing are stalling development at CSPs. Think of these approaches as, well, 'buckets' tied behind an imaginary speedboat that longs to accelerate towards profitability from Digital Services…if…


text:
Michael E Braun
Street Address:
816 31st Ave SW
Suite A
Minot, ND 58701-7031
Mailing Address:
816 31st Ave SW Ste A
Minot, ND 58701-7256
Main road located behind the Dakota Square Mall and next to Gate City Bank.
Phone: (701) 838-0713
Toll Free: (800) 452-8968
Fax: (701) 852-9051
Street Address:
16 2nd St NW
Tioga, ND 58852
Located on the corner; West of Tioga Motor
Phone: (701) 664-2008
My background and achievements
- Minot State University, BS - Business Management
- State Farm Agent since 1999
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- Joel Haman - Office Manager
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Our mission is to help people manage the risks of everyday life, recover from the unexpected and realize their dreams.
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Email Me
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text:
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Continue Shopping.
iHerb Customer Reviews
Can't believe what happened at DAY 1
1 person found the following review helpful
Posted by MichaelLabib on Dec 10, 2011
Hi, I have been using Glucosamine, fish oil, the french miracle piascledine and some other supplements to help relief my constant knees pain. Last week I decided on trying collagen, I made the order through iherb.com and at the same day I stopped the piascledine. The pain started to come back once again until yesterday the pain was reduced again to 0% when I take the first 2 capsules of BioCell Collagen dr's best. I can say that the pain was gone after 10 hours of my first dose, in fact I can't believe what happened. I just wake up few minutes ago, in the middle of the night, to write this review and WOW my dry eye condition has dramatically been improved. This is one of the best supplements, if not just the best, that you can take if you have cartilage or joints problems. I'm still going to take Glucosamine and Fish Oil, and maybe I will continue taking piascledine but after 1 month until I'm really sure that BioCell collagen is working that great. One more thing, don't believe that researches that say that Hyaluronic Acid is not absorbable by our body. It's not the case with the low molecular BioCell Hyaluronic Acid, but that holds true with non-BioCell hyaluronic acid which I have tried for 10 days without any results. WARNING: THIS PRODUCT INCREASED MY BLOOD PRESSURE BY 40-50 POINTS, IT MIGHT NOT BE THE CASE FOR YOU. Anyway, the best thing that you can do is try it and take your blood pressure daily for the first 2 weeks. I'm a hypertension patient, so I do think it only might raise my blood pressure and not people with no blood pressure problems. I regret that I had to stop this great product :(
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Even though I still surf the internet for prices and products I invariably find myself returning to iherb for the very highest quality products and the best prices.
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Has really helped with some worn out joints


text:
Is this a bowl lift mixer or a tilt-head stand kind, sorry I'm a little confused.
i'm so happy i got this stand mixer!...
With features like the soft start electronic control to help prevent ingredients from splashing out of the bowl, this refurbished KitchenAid professional HD series 5-quart stand mixer is a must for any kitchen.
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ITEM#: 12994755With features like the soft start electronic control to help prevent ingredients from splashing out of the bowl, this refurbished KitchenAid professional HD series 5-quart stand mixer is a must for any kitchen. This stand mixer features a handsome black onyx finish and constructed from metal for superior durability and ease of cleaning.
- 5-Quart stainless steel bowl with handle
- 475 Watts
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- Stainless steel wire whip
- Five (5) rubber feet keeps unit stable and protects countertops
This high-quality item has been factory refurbished. Please click on the icon above for more information on quality factory-reconditioned merchandise.
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nice !
i'm so happy i got this stand mixer!Read More
KitchenAid 5-quart Stand Mixer (Refurbished)
Love this mixer! It is attractive and durable. Have already recommended to friends.Read More
Excellent KitchenAid
I was scare of buying Refurbished but now I realize I was scared for no reason. I am more than happy with my purchase.Read More
Like new. Wish it came with a pouring shield.
Love this mixer, use it all the time. Head doesn't tilt. So having to buy pouring shield separately. Otherwise, the best purchase/investment I've ever made.Read More
Best purchase i ever made
I previously owned a Kitchenaid Artisan. I received it 12 years ago as a wedding gift. 4 years ago I lost it in a divorce and I hear its still going strong. So after a long time of mashing potatoes, mixing batter and dough by hand, I decided to get a professional model. And might I add, what a great addition to my kitchen. Looks great, way more powerful than my previous tilt head. As far as factory refurbished you ask? You'd never know the difference. Besides the box and manual clearly stating its a refurb, there wasn't a nick, scratch, mark or scuff on it and it performs flawlessly. For the same price as new KA entry level model, this is a steal and well worth a second look if your on the fence.Read More
Buy Refurbished!
I have purchased many refurbished items in the past and I've never been disappointed with quality. Well, the same is true of this Professional HD stand mixer. Prior to this purchase, I'd had the Kitchen Aid mixer on my wishlist for months! I purchased this model for it's power kneading function for breads, doughs, etc and it has not disappoint. Don't hesitate to purchase - the price and features make this unit a great deal!Read More
Questions & Answers
Hi sham, the bowl elevates with a handle. I hope that this information was helpful. Thank you for shopping with us.
Hello miked251, it depends on the KitchenAid attachment, some KitchenAid attachments are compatible with any KitchenAid Stand Mixers, some are not. We hope this helps and thank you for shopping with us.
does tjos model have a two prong or three prong plug? what is the plastic material in the deails
This has a three-prong plug. There is plastic in the housing and base.
Does this model actually have 475 watts?
Yes bluesky8, this has 475 watts. I hope this helps, thanks for shopping with us.
Does this Item #: 12994755, KitchenAid RKG25H0XOB Onyx Black 5-quart Pro HD Series Stand Mixer (Refurbished), have metal or plastic gears? Thank you.
Hello rainbowbreeze, The gears are all steel. I hope this helps, thanks for shopping with us.
Shopping Guides
Kitchen Mixer Buying Guide
Many cooks' favorite kitchen appliance, the mixer, is used for many kitchen tasks. Mixing up dough for freshly baked bread or whipping up a quick batch of cookies for the kids is simple when you have a kitchen mixer. Stand mixers and hand mixers are very popular items with bakers. Mixers are designed with the cook in mind, which means they can come with many different options. This kitchen mixer buying guide will guide you through the available options.
How to Clean Electric Mixers
Electric mixers are perhaps the most used of all small kitchen appliances. From mixing cake batters to kneading homemade bread to mashing potatoes, electric mixers make the cook's job much faster and easier. Knowing how to clean electric mixers is particularly important, whether you own a stand mixer or a hand mixer. When dried food collects in crevices or mixer attachments, it can interfere with the proper functioning of your appliance and cause your mixer to stop working.
Types of Kitchen Mixers
Whether you're baking or whipping up pancake batter, you need mixers you can rely on. Fortunately, you can choose between a hand mixer for light tasks, a professional mixer if you bake daily and other mixers when you shop online. This guide will help you find the best mixer for your lifestyle.
Food Processor Buying Guide
What exactly can you use a food processor for? It can be confusing to know what you're supposed to do with all the different attachments. It seems simpler to just keep using your old blender to do whatever it is a food processor does. A food processor is so much more than just a mixer. When you use your food processor, you'll save time, kitchen space and dishes. Another perk to these food processors is the wide variety of colors and finishes available. Not only will you create amazing food; you'll also get the option of matching your food processor to your picture perfect kitchen. It may seem a bit intimidating at first to have so many options, but once you get to know your food processor, you'll find that it is more than just a vegetable chopper. Don't let your uncertainty keep you from investing in a quality food processor for your kitchen.
Tips on Buying Mixers
What's your kitchen style? Are you a baker who whips up cakes and pies from scratch? Do you make homemade bread and pasta? Or have you never met a boxed cake mix you didn't like? The way you cook and the food you like to eat will determine the best mixer for your kitchen. Follow these tips to make sure the one you buy meets your needs and your tastes.
How to Use a Pedometer
Many fitness experts encourage people to take 10,000 steps a day. Doing this can aid in weight management, lower your blood pressure and cholesterol and increase your overall health. Wearing a pedometer is an excellent means of tracking the steps you take through the day, whether your goal involves sports or simply increased fitness and exercise. Knowing how to use your pedometer properly is key to helping you reach your 10,000 steps a day.
5 Great Kitchen and Bath Projects
A lot of home improvements revolve around kitchen and bath remodels. These are the most used rooms in the house, so it makes sense to make them as comfortable and attractive as possible. Here are a few project ideas to help you re-imagine your kitchen or bath.
5 Things to Look for in Kitchen Faucets
When it comes to plumbing fixtures, few see more use than bathroom and kitchen faucets. Because they're so heavily used, durability is a primary feature to consider when selecting a sink faucet. There are plenty of functional faucet features to choose from. You'll also want to consider the style of your kitchen, including the sink and surrounding area. Here are a few features you may want to keep in mind as you look for a kitchen faucet that fits.
Top 5 Home Appliances for the Kitchen
Make preparing and reheating meals even easier when you buy home appliances for your kitchen. While your refrigerator and stove do most of the hard work, there are a few appliances that can simplify your life and save you valuable time -- and you can often buy home appliances at incredibly affordable prices online. Read on for the top five home appliances you can use in the kitchen.
How to Use a Laminator
Once reserved solely for schools and offices, a laminator has become small and compact enough for use in your own home for a variety of different projects. Whether you want a hot laminator for sealing documents or a cold laminator to protect your photos, the following tips will help you use a laminator to create durable projects and preserve the items you already have.
Best Uses for Night Vision
Human eyes are not adapted to see in the dark, making natural night vision difficult. Night vision devices, however, aid your eyesight in low lighting by amplifying the surrounding light. Night vision devices can also magnify objects so they will appear closer or detect body heat through infrared light, but the main use of night vision is to see things that your eye wouldn't normally be able to see in the dark or in low light. Many hobbies, and even some jobs, require night vision.
Top 5 Uses of a Binder
Binders are an essential office supply item. They are used by virtually everyone: busy professionals, students and teachers. Binders are even used at home for personal art and scrapbooking projects. They come in a variety of colors and sizes. When working with office supplies, like a binder, it may help to purchase additional binder accessories, such as sheet protectors, sticker pages, photo pages and dividers, to help you stay organized. With the right office supplies and a binder, you'll find solutions for projects in and out of the office. Here are the top five uses for a binder.
5 Uses for Stacking Chairs
When you want seating that's both compact and functional, stacking chairs are a good solution. Stackable chairs don't have moving joints, making them extra sturdy. You can unstack them quickly for an outdoor party or for entertaining guests and then stack them up and tuck them away in a garage or corner of the home just as quickly. Whether you're looking for a set of durable patio chairs for the summer or comely chairs to use in the living room year-round, stackable chairs may be what you're looking for. Keep reading to find out about five different uses for stacking chairs that you could implement around your home.
How to Use Table Runners
Table runners complement your table settings by adding color, texture and a unifying element to your tablescape. Table runners add a touch of elegance, bringing a sense of tradition to an occasion. The rules regarding the use of table runners and other kitchen linens are fluid and are better described as guidelines. When it comes to kitchen linens and the design of your dining room table, you can be as formal or as casual as you like.
How to Use a Chopping Board
A chopping board is one of the most basic pieces of equipment in the kitchen. Chopping boards, also known as cutting boards and butcher blocks, protect your countertops from scratches and your knives from excessive dulling. Cutting boards are also much more sanitary than chopping your ingredients on the countertop or a paper plate or napkin. Used properly, a chopping board can also be a tremendous help in preparing your meals.
How to Use a Portable Air Conditioner
Nothing will bring more immediate relief to a hot summer day than a portable air conditioner. If you can't take the heat, get out of the kitchen. Then, go back into the kitchen with a portable air conditioner. A room air conditioner is perfect for that one room that struggles to cool down, and they can give your home an extra boost of cool air on blistering days. Portable air conditioners are reasonably easy to use, but there are still a few things you should know. Using portable air conditioners correctly will ensure they are able to reliably lower the temperature in any given room. Here are a few tips on using your portable air conditioner.
How to Use Measuring Cups
Measuring cups are one of the workhorses of cooking essentials in the kitchen stable, and for good reason. Pinching, dashing and tossing can work well for seasoning, but when you are crafting a cake, baking bread or creating a rich sauce to drizzle over an entree, you need the correct cooking essential tools to measure the ingredients or disaster can quickly unfold. Using measuring cups properly will allow you to get the most out of your favorite recipes.
How to Use a Blender as a Juicer
Making smoothies and milkshakes is easy with the right blender to mix everything together into a smooth consistency. A juicer, on the other hand, separates juice from fruit and vegetables by squeezing it. If you want the pure vegetable or fruit juice of a vegetable or fruit, you will probably need to buy a juicer, such as an orange juicer, rather than a kitchen blender. For thicker smoothies and drinks that still contain fruit pulp, however, blenders will work well. Read on for tips on using your food blender as a juicer for a variety of fruits.
Top 5 Wedding Gifts for the Kitchen
Newly married couples are often starting fresh, so kitchen items are a necessity, and they're among the most preferred gifts for the wedding. These are the best kitchen gifts to give the bride and groom to help them get started on their new life together.
Top 5 Timesaving Kitchen Appliances
Kitchen appliances help you take care of your family and impress your friends, but you have more things to do than slave away in the kitchen. At Overstock.com, we know how busy you are; that's why we offer a wide selection of the high-quality, timesaving kitchen appliances you need at prices you'll love. Read on to discover the top five kitchen appliances renowned for saving you time.
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text:
Page 1
Resequencing data provide no evidence for a human
bottleneck in Africa during the penultimate glacial
Per Sj¨ odin1, Agn` es E Sj¨ ostrand1,2, Mattias Jakobsson1,3, and Michael G B Blum4∗
1Department of Evolutionary Biology, Evolutionary Biology Centre, Uppsala University,
Norbyv¨ agen 18D, 75236, Uppsala, Sweden
2MNHN-Universit´ e Paris 7, Centre National de la Recherche Scientifique, UMR 7206
Eco-anthropologie et Ethnobiologie, Paris 75005, France
3Science for Life Laboratory, Uppsala University, Uppsala, Sweden
4Universit´ e Joseph Fourier, Centre National de la Recherche Scientifique, Laboratoire
TIMC-IMAG UMR 5525, Grenoble, 38041, France
∗Author for correspondence (firstname.lastname@example.com)
Keywords: human origins; African genetic diversity; penultimate glacial age; approximate
Bayesian computation; African bottleneck; oxygen isotope 6
Running head: No bottleneck at the origin of humans
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Based on the accumulation of genetic, climatic and fossil evidence, a central theory in
paleoanthropology stipulates that a demographic bottleneck coincides with the origin of our
species Homo Sapiens. This theory proposes that anatomically modern humans – which
were only present in Africa at the time – experienced a drastic bottleneck during the penul-
timate glacial age (130-190 kya) when a cold and dry climate prevailed. Two scenarios have
been proposed to describe the bottleneck, which involve either a fragmentation of the range
occupied by humans or the survival of one small group of humans. Here, we analyze DNA
sequence data from 61 nuclear loci sequenced in three African populations using Approximate
Bayesian Computation and numerical simulations. In contrast to the bottleneck theory, we
show that a simple model without any bottleneck during the penultimate ice age has the
greatest statistical support compared to bottleneck models. Although the proposed bottle-
neck is ancient, occurring at least 130 kya, we can discard the possibility that it did not
leave detectable footprints in the DNA sequence data except if the bottleneck involves a less
than a 3 fold reduction in population size. Finally, we confirm that a simple model with-
out a bottleneck is able to reproduce the main features of the observed patterns of genetic
variation. We conclude that models of Pleistocene refugium for modern human origins now
require substantial revision.
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1 Introduction
Quaternary ice ages have deeply affected the genetic diversity of many living organisms
(Hewitt 2000) and humans may be no exception. There is a variety of facts to suggest that
humans drastically decreased in number during the MIS6 time period (Marine Isotope Stage
6), which corresponds to the penultimate major ice age 190-130 kya (Lahr and Foley 1998).
The marine isotope stages correspond to the different stages of mass-ratio between18O and
16O stable isotopes found along marine stratigraphic sequences. The mass-ratio between18O
and16O being proportional to the temperature at which the sediments were formed, these
stages correspond either to glacial periods or interglacial periods (Shackleton et al. 1977).
MIS6 had very similar conditions to the last ice age (MIS2), though it was a bit colder
(Petit et al. 1999) and the ice sheets extended over a slightly greater area than during MIS2
(Hetherington and Reid 2010). During MIS6, deserts, savanna and open dry forest widened
whereas rain forest receded (Van Andel and Tzedakis 1996; Dupont et al. 2000) resulting
in open grassland that would not protect early humans against predators (Lahr and Foley
There are several lines of genetic evidence for a bottleneck: all present-day mitochondrial
sequences coalesced into a single individual—the 'mitochondrial Eve'—at the onset of MIS6
(Gonder et al. 2007; Behar et al. 2008); numerical simulations tuned to reproduce autosomal
genetic diversity also point to a bottleneck during MIS6 with a human effective population
size smaller than 2,000 individuals at the bottleneck's inception (Fagundes et al. 2007); a
bottleneck during MIS6 can account for the 8 fold difference between the times to the most
recent common ancestor of mitochondrial and autosomal genes (Blum and Jakobsson 2011);
and the 1.5-4 times lower level of genetic diversity in humans compared to great apes can
been attributed to a bottleneck (Kaessmann et al. 2001). Finally, the timing of the bottleneck
is consistent with the available evidence on the morphological evolution of humans, which
indicates that the first known specimen of Homo sapiens emerged during or just before MIS6
(White et al. 2003; McDougall et al. 2005). Because there is a temporal match between
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the evidence for the first specimen of Homo sapiens and the MIS6 bottleneck, which would
have promoted population differentiation, some authors refer to the MIS6 bottleneck as a
speciation bottleneck (Garrigan and Hammer 2006; Fagundes et al. 2007).
Two scenarios have been proposed for how the climate change during MIS6 affected the
human population (Lahr and Foley 1998). In the 'fragmentation' hypothesis (see figure 1),
the human population in Africa would have been divided into several geographically restricted
groups or refugia with no gene flow between them. Extensive aridity in Africa during MIS6,
far more severe than during the Last Glacial Maximum (Casta˜ neda et al. 2009), would have
caused the contraction and fragmentation of available areas that could sustain humans. The
alternative 'founder' hypothesis postulates that of all the hominid populations living in Sub-
Saharan Africa, only one is ancestral to modern humans. Candidates for a refuge during
the long cold MIS6 are the south coast of South Africa with its unusual confluence of plant
diversity, coastline richness, and moderate climate (Marean et al. 2007; Marean 2010) as
well as East Africa where the earliest well-dated anatomically modern humans have been
described (McDougall et al. 2005).
Compared to non-African populations, African populations are good candidates for de-
tecting a potential bottleneck 130 kya because non-African populations have been through
a more recent out-of-Africa bottleneck, which masks patterns of earlier bottlenecks (Voight
et al. 2005; Amos and Hoffman 2009). Furthermore, the San and the Biaka Pygmy carry pa-
ternal and maternal lineages belonging to some of the deepest clades known among modern
humans and may therefore carry important information for reconstructing ancient human
demography (Knight et al. 2003; Behar et al. 2008; Henn et al. 2011).
In this study, we search for evidence of a bottleneck during MIS6 using sequence data
from 61 autosomal 20kb DNA fragments typed in the African Biaka, Mandenka, and San
populations (Wall et al. 2008; Hammer et al. 2010). We investigate three different demo-
graphic models: a model without a bottleneck, a model with a bottleneck at MIS6, and a
model where the human population was fragmented during MIS6.
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2 Material and methods
We summarized the information contained in the sequence data using summary statistics that
are informative with respect to ancient demography (table 1) (Depaulis et al. 2003; Voight
et al. 2005). The summary statistics were computed for each of the 61 autosomal sequences
(supplementary tables 1-3, Supplementary Material online) and the means and variances were
retained (table 1). We evaluated the statistical support of a bottleneck during MIS6 using
Approximate Bayesian Computation, (ABC; Beaumont et al. 2002; Csill´ ery et al. 2010). The
ABC approach relies on comparing the observed values of the summary statistics to summary
statistics simulated under the two versions of the bottleneck model ('fragmentation' and
'founder') as well as under a 'no bottleneck' hypothesis that does not include any bottleneck
during MIS6 (figure 1). The choice of mutation rate can impact the results and because
there is a 2-3 fold difference between the mutation rate estimated from mother-father-child
trios (or quartet) (∼ 10−8/bp/generation; The 1000 Genomes Project Consortium 2010;
Roach et al. 2010) and the mutation rate derived from human-chimp comparisons (∼ 2.5 ×
10−8/bp/generation; Nachman and Crowell 2000; Fagundes et al. 2007; Gutenkunst et al.
2009; Laval et al. 2010), we considered both estimates of the mutation rate in our analyses.
(a) Sequence data
The data comprise 61 re-sequenced independent intergenic regions from the autosomes, which
are in areas of medium or high recombination at least 100 kb from the nearest gene, and the
sequence data was generated by Wall et al. (2008) and Hammer et al. (2010). The sequenced
regions were chosen to minimize any potential confounding effects of natural selection (Wall
et al. 2008). Each region encompasses approximately 20kb and generally consists of three
2kb sequence fragments, separated by 7kb of unsequenced DNA. We considered three African
populations: Mandenka (16 sampled individuals), Biaka (15 sampled individuals), and San
(9 sampled individuals) from Namibia. The sequences of two common chimpanzees, available
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in the database, were used as outgroups.
(b) Statistical Analysis
For each population, we used ABC to find the range of demographic parameters that yield
summary statistics similar to the empirical summary statistics computed for the three pop-
ulations. We chose to use the following summary statistics: mean and variance over the 61
loci of the number of SNPs, mean and variance of Tajimas's D, mean and variance of Fu and
Li's D∗, mean and variance of Fay and Wu's H, and mean of Hudson's estimate of the re-
combination rate (Hudson 1987). Although phased haplotypes are provided in the database,
we do not consider haplotypic summary statistics since these are known to be sensitive to
recent admixture (Lohmueller et al. 2010). For each scenario of human demographic history,
we simulated 100,000 multilocus summary statistics as follows.
• Generate the demographic parameters according to the prior distributions given in
table 2.
• For each of the 61 loci, generate a sample using the software ms (Hudson 2002). The
ms software is based on the coalescent framework; it first generates a random geneal-
ogy of the sample under a particular demographic model and mutations are thereafter
randomly placed on the genealogy. For each set of demographic parameters, we gener-
ated 61 autosomal sequence-regions with the same sample sizes and the same sequence
lengths as in the empirical data (see tables S1-S3).
• Compute the summary statistics, using the libsequence C++ library (Thornton 2003),
for the simulated sequences in order to obtain the (Euclidean) distance between ob-
served and simulated summary statistics. Before computing the distances, standardize
the summary statistic so that they have a variance equal to 1.
When performing parameter inference, we retained the half percent of the simulations
(500 out of 100,000 simulations) with the shortest distances between simulated and observed
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statistics. After this rejection step, we used heteroscedastic regression adjustment to account
for the difference between simulated and observed summary statistics (Blum and Fran¸ cois
2010). The adjustment step reduces the variance of the accepted parameter values, which is
typically inflated compared to an ideal sample based on exact matches (i.e. null distances)
(Csill´ ery et al. 2010).
To perform model selection, we simulated 100,000 datasets for each of the three models
and computed the three posterior probabilities with two different methods. In the first
method, we retained the half percent of the 300,000 simulations with the shortest distances
and assumed that the proportions of accepted simulations in each model are proportional
to the posterior probabilities (Pritchard et al. 1999). The second—more sophisticated—
method fits a multinomial logistic regression Pr(Y |s) between the summary statistics s and
a trichotomous variable Y ∈ {1,2,3} that corresponds to the model index. Inserting the
values of the observed summary statistics sobsinto the regression equations gives the posterior
probabilities Pr(Y = i|sobs),i = 1,2,3, for the three models (Beaumont 2008).
We checked for population structure (Pritchard et al. 2000) within each population be-
cause the three investigated demographic models assume homogenous populations, but we
found no indication of stratification.
To assess if there is enough information in the genetic data to distinguish between the three
models, we considered a leave-one-out method. For each model ('no bottleneck', 'founder'
or 'fragmentation'), we repeated 5,000 times the following procedure: i) pick the simulated
genetic data from a particular simulation among the 100,000 simulations of the correspond-
ing model, ii) use the summary statistics of the chosen simulation as target instead of the
empirical data, iii) estimate the model probabilities using the remaining 299,999 simulations.
Using the method based on multinomial logistic regression, we assign each of the 3 × 5,000
simulated cases to the model with the greatest posterior probability. All ABC analyses were
performed using the R abc package (Csill´ ery et al. 2011).
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We set the prior distribution of the demographic parameters (table 2) so that the bot-
tleneck ended 130 kya and started at a time uniformly distributed between 150 and 190
kya. In all three demographic models, we assumed that Sub-Saharan African populations
experienced demographic expansion (Voight et al. 2005; Cox et al. 2009). For the muta-
tion rate, we chose two different priors, each accounting for the variability of mutation rates
across loci. For the first prior, we estimated a mutation rate for each of the 61 markers by
computing the mean number of nucleotide differences between two chimp sequences and the
human sequences. We assumed a genetic divergence of 6 millions years between human and
chimpanzee, and a generation time of 25 years. We then fit a Gamma distribution to the
61 estimated mutation rates. In units of generations, we obtained a Gamma distribution
with a shape parameter of 17.86 and a scale parameter of 1.62 × 10−9(mean = 2.9 × 10−8
mutations/bp/generation). For the second prior, we chose a Gamma distribution parameter-
ized so that the mean mutation rate corresponded to 1.10 × 10−8/bp/generation (The 1000
Genomes Project Consortium 2010) and the variance is the same as in the first prior. We
further assumed a Gaussian distribution, with mean fmeanand variance fvar, for the ratio
f of effective recombination rate to effective mutation rate (Plagnol and Wall 2006). The
prior for fmeanand fvarwere uniform between 0.2 and 2.0 and between 0 and 1.0 respectively.
Recombination rates were assumed to be constant within loci but to vary across loci.
3 Results
We first evaluated the relative statistical support of the 'founder', the 'fragmentation', and
the 'no bottleneck' models. For each of the three African populations and both mutation
rates, we find that the 'no bottleneck' model has the highest posterior probability (table
3 and supplementary table 4, Supplementary Material online). When using multinomial
logistic regression to evaluate posterior probabilities, the probability of the model without a
bottleneck is at least 79% for each African population.
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The potential bottleneck that we investigate is ancient, occurring more than 130 kya, and
it is therefore uncertain if there is enough information in the sequence data to distinguish
between the demographic models. The out-of-Africa and the out-of-Beringia bottlenecks left
detectable signals in human genetic diversity, but they occurred much more recently than
MIS6 (Voight et al. 2005; Amos and Hoffman 2009). We consider three potential lines of
evidence to assert that there is sufficient information in the data to discard a bottleneck
during MIS6. We define the bottleneck intensity, b, by
NB= NA× 10b, (1)
where NB is the total population size during the bottleneck and NAdenotes the ancestral
population size before the bottleneck. A value of b = −1.5 corresponds to an extreme
bottleneck with a 30 fold reduction of population size whereas there is no bottleneck when
b = 0.
First, if MIS6 is too ancient to affect the distribution of genetic variation in modern
populations, the expected posterior distribution of model parameters that are related to this
period of time should be the same as the prior distribution of these parameters. This is not
what we observe. Although the prior distribution for b is uniform between −1.5 and 0, the
peak of the posterior distribution is typically very close to b = 0 and values of b smaller than
−0.5 are unlikely (P < 8% for each population when averaging over models and mutation
rates) (figure 2 and supplementary figure 1, Supplementary Material online). Second, we
investigated whether the correct model was inferred when the genetic data was simulated
from that particular model. For a severe bottleneck, a detectable signal is still present in
the sequence data: when b = −1.5, we correctly assign 97% of the cases when simulations
from the founder model are used as target data, and this number decreases to 79% for the
fragmentation model (figure 3). For a mild bottleneck, a 3 fold reduction of population size
(b = −0.5), these proportions drop to 32% and 50% for the founder and the fragmentation
models. Figure 3 shows that for a bottleneck with b = −0.5, 20% of the founder and 38% of
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the fragmentation simulations are incorrectly assigned to the 'no bottleneck' model. Thus,
there is a non-negligible chance that the model without a bottleneck receives the largest
support if the data were generated under a bottleneck model with b ≥ −0.5. Third, for
the empirical data, recall that the support for the 'no bottleneck' model was at least 79%
for each population and both mutation rates (table 3). For the particular 20% (founder
model) and 38% (fragmentation) of the mild-bottleneck simulations (b = −0.5) that were
incorrectly assigned to the 'no bottleneck' model, we investigated the relative support for the
three different models. We found that a strong support ≥ 79% for the 'no bottleneck' model
occurs in less than 1% of these simulations giving evidence that the observed data are also
inconsistent with a mild-bottleneck (b = −0.5).
To understand where the signal against a bottleneck comes from in the genetic data, we
searched for the summary statistics that penalize the bottleneck models. The mean Fay and
Wu's H = 0.58 of the San sample is large compared to the values simulated by all three
demographic models suggesting that there is a deficit of high-frequency mutations compared
to intermediate frequency mutations in the San sample (figure 4). There is a gradient among
models, with the 'founder' model generating mean Fay and Wu's H least similar to empirical
values, followed by the 'fragmentation' model, and the 'no bottleneck' model generates values
that are most similar. For the Biaka and the Mandenka samples, the variance of the Tajima's
D over the 61 loci is small (table 1) even when compared to the simulations performed without
a bottleneck. A typical footprint of bottleneck is, in fact, an unexpectedly large variance
of Tajima's D (Haddrill et al. 2005; Voight et al. 2005). Once again, we find a gradient
among models: the 'founder' model generates the most different and largest values, the 'no
bottleneck' model generates the most similar values, and the 'fragmentation' model is in-
between. These are the summary statistics that most clearly distinguish between bottleneck
models and a model without a bottleneck, but other summary statistics also add weight to
the inference (supplementary figures 2-3, Supplementary Material online).
We finally tested if the different demographic models, after having been fitted to the three
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populations, were able to reproduce the observed summary statistics. We simulated 2,000
replicates of the summary statistics using demographic parameters that were drawn from
their posterior distribution. Since there are too many summary statistics to easily visualize,
we applied principal component analysis to graphically project the posterior distributions of
the summary statistics in two dimensions (Cornuet et al. 2010; Berlin et al. 2011). These
posterior predictive checks show that each demographic model provides a good fit to the
summary statistics computed from the empirical data (figure 5). To explore the match
between observed and simulated summary statistics individually, we used two-sided posterior
predictive P-values (Gelman et al. 2003) as an exploratory tool (supplementary table 5,
Supplementary Material online). For the model without a bottleneck, most posterior P
values are found to be larger than 5% for both mutation rates confirming that this simple
demographic model provides a good fit to the summary statistics. The exceptions are the
variance of the number of SNPs for the San, the mean of the Tajima's D for the Biaka and
the variance of the Tajima's D for the Mandenka. The models that include a bottleneck do
not provide a better fit to these summary statistics for which the posterior P values are also
smaller than 5% for one of the two mutation rates (see supplementary table 5, Supplementary
Material online).
4 Discussion
Our study shows that autosomal sequence variation from two hunter-gatherer populations
(San and Biaka) and one agriculturalist population (Mandenka) do not provide evidence
for a bottleneck during the cold period of MIS6. Using an ABC approach, we find that
the simple demographic scenario without a bottleneck during MIS6 (the 'no bottleneck'
model in figure 1) is preferred to the alternative models (the 'founder' and 'fragmentation'
versions of a bottleneck) for all three populations. Although the three considered models
can be almost perfectly fitted to reproduce the observed summary statistics (figure 5), the
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fitted bottlenecks involve a less than a 3-fold reduction during MIS6 (figure 2) making them
indistinguishable from the model without a bottleneck. With a less than a 3-fold reduction
of population size, there is insufficient signal in the summary statistics to clearly discriminate
between the demographic models (figure 3). A reduction in population size by a factor of 3
would still be a noteworthy event, but in comparison with the bottlenecks associated out-of-
Africa migration or the peopling of the Americas, it would not stand out as an extraordinary
event, and, as we have demonstrated, it would not leave substantial traces in population-
genomic patterns of variation. More severe bottlenecks are, however, detectable. Assuming
an effective population size of the order of N = 10,000 individuals corresponds to autosomal
gene genealogies going back, on average, 2 × 2N = 40,000 generations. Since the end of
MIS6 occurs about 5-6,000 generations ago, the bottleneck would have taken place in the
most recent quarter of the genealogical tree. Intuitively, a moderate to strong bottleneck
occurring during MIS6 would leave a detectable signal in the data because bottlenecks distort
the shape of the gene trees (e.g. Galtier et al. 2000).
(a) Pleistocene refugia hypothesis
The lack of evidence for a severe human bottleneck during MIS6 suggests a revision of the
current Pleistocene refugium model of anatomically modern human origins (Lahr and Foley
1998) and, more generally, has implications for evaluating the importance of ice ages on speci-
ation (Klicka and Zink 1997). Of particular relevance is the 'Pleistocene refugia hypothesis'
(Haffer 1969). It assumes that during several dry climatic periods of the Pleistocene, the
tropical forest was divided into a number of smaller forests which were isolated from each
other by tracts of nonforest vegetation, and that the remaining forests served as refugia for
numerous populations (Haffer 1969). Fragmentation of the African forest is debated, and
simulations of paleovegetation during the glacial ages suggest that African tropical forests
were not severely displaced by expanding grasslands during glacial ages (Cowling et al. 2008).
Nonetheless, the fact that the mitochondrial TMRCAs of many subspecies of orangutans and
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common chimpanzees overlap with the cold MIS6 has been interpreted as that Pleistocene
refugia may have promoted speciation and diversification in the Hominidae family (Arora
et al. 2010; Bjork et al. 2011). As shown in this study, however, the occurrence of a mito-
chondrial TMRCA during MIS6 is not sufficient evidence for a severe reduction in population
size associated with glacial refugia.
(b) Population size and density during the Pleistocene
A small human effective population size, on the order of 10,000 individuals, which is smaller
than the effective population size of most great apes, has been interpreted as a result of a
very long history, starting ∼ 2 mya, of a small population size, coined as the long-necked-
bottle model (Harpending et al. 1998; Hawks et al. 2000). Our findings are consistent with
this hypothesis, but, depending on the mutation rate, we find either an effective population
size of NA= 12,000 (95%C.I. = 9,000 − 15,500 when averaging over all three demographic
models) using the mutation rate calibrated with the human-chimp divergence or an effective
population size of NA= 32,500 individuals (95%C.I. = 27,500−34,500) using the mutation
rate given by whole-genome trio analysis (The 1000 Genomes Project Consortium 2010)
(supplementary figure 4 and table 6, Supplementary Material online). Not surprisingly, the
estimated effective mutation rates θ = 4NAµ are comparable for the two mutation rates we
considered, and are equal to 1.4 × 10−3/bp/generation (95%C.I. = (1.1 − 1.7) × 10−3).
Relating the estimated effective population size to the census population size during the
Pleistocene is a difficult task because there are many factors affecting the effective population
size (Charlesworth 2009). Nevertheless, based on published estimates of the ratio between
effective and census population size, a comprehensive value on the order of 10% has been
found by Frankham (1995). This 10% rule roughly predicts that 120,000 − 325,00 individ-
uals (depending on the assumed mutation rate) lived in Sub-Saharan Africa some 130 kya.
Assuming that the range of humans extends over all the 24 millions km2of Sub-Saharan
Africa, the density of humans at that time would have been extremely low between 0.5 and
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1.4 individual per 100 km2, which is even lower than the lowest recorded hunter gatherer
density of 2 individuals per 100 km2reported for the !Kung (Kelly 1995) and the density of
3 individuals per 100 km2estimated for Middle Paleolithic people (Hassan 1981). However,
this discrepancy disappears if humans were restricted to an area some 3-6 times smaller than
the entire Sub-Saharan Africa.
(c) Joint population analysis and demographic assumptions
Our approach to investigate the potential MIS6 bottleneck considered the three African
populations separately and do not account for their shared ancestry. Although additional
information regarding divergence times, in particular, can be obtained from a joint analysis
of the populations, subpopulation specific analyses are common in modeling studies of pop-
ulation size change and provide robust results regarding the out of Africa bottleneck (Marth
et al. 2004; Voight et al. 2005; Li and Durbin 2011). Additionally, the time period of MIS6
is likely more ancient than the divergence of the San population (110-130kya, Gronau et al.
2011; Veeramah et al. 2011) so that all three populations should be equally affected by demo-
graphic changes during MIS6. Using worldwide whole-genome sequence data, Li and Durbin
(2011) showed that all African and non-African populations are very similar in their esti-
mated effective population size history before 150kya. However, because it has been argued
that local samples cannot be regarded as being drawn from a panmictic population (St¨ adler
et al. 2009), we also considered simulations of a divergence model without a bottleneck where
the first divergence involves the San population and the second one involves the Mandenka
and the Biaka populations (Veeramah et al. 2011). We find that there is little variation of
the population-wise summary statistics as function of the divergence times (supplementary
figure 5, Supplementary Material online), indicating that using a divergence model has lit-
tle practical impact, at least for the summary statistics we considered. Furthermore, when
pooling all three populations together, the mean Tajima's D and the mean Fu and Li's D∗
decrease (Table 1), which can be accounted for by a divergence model (Figure 6). The three
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population-wise means of Tajima's D are between −0.23 and −0.32 and it decreases to −0.77
when pooling the three populations together. Similarly the three population-wise means of
Fu and Li's D∗are between −0.14 and −0.23 and it decreases to −0.99 for the pooled sample.
That these summary statistics decrease with the number of pooled populations is explained
by the increase of low frequency polymorphisms as we pool more and more populations (Ptak
and Przeworski 2002; Hammer et al. 2003). Similar to the population-wise analysis, the sum-
mary statistics (Tajima's D and Fu and Li's D∗) in the pooled sample can be explained by
a demographic model without an ancestral bottleneck.
Another potential limitation of our analysis is the rather crude modeling of population
change and there are more general models of piecewise constant trajectories that have been
investigated (Minin et al. 2008; Li and Durbin 2011). This being said, accounting for such
detail requires more complex models and the fine details of the demographic history will
be difficult to detect with genetic data. Providing a fine-scale resolution of demographic
history may give the false impression that we can reconstruct extremely detailed fluctuations
of population history but estimates of these fluctuations should not be over-interpreted as
some of them may be non-significant (see Figure 2 of Li and Durbin 2011).
(d) African bottlenecks and population genetic data
The fact that African resequencing data are compatible with a demographic model that
does not include an ancestral bottleneck confirms previous findings obtained with autosomal
sequence data (Pluzhnikov et al. 2002; Adams and Hudson 2004; Voight et al. 2005; Cox
et al. 2009; Laval et al. 2010), autosomal SNPs (Marth et al. 2004) and mtDNA (Atkinson
et al. 2009). Similarly, a recent analysis found that complete individual genomes from the
San, Bantu and Yoruba populations were consistent with a population expansion in Africa
(Gronau et al. 2011). Furthermore, based on two diploid Yoruban genomes, Li and Durbin
(2011) found a population expansion during MIS6 confirming the absence of a bottleneck
during this time period. However, a peculiarity of their analysis is that the Yoruba pop-