Starbucks Evenings will soon be in five O.C. locations, including a cafe at Downtown Disney in Anaheim.
Starbucks Evenings features a small plates menu.
A cafe in Rancho Santa Margarita was the first in O.C. to sell alcohol.
Starbucks Evenings has been in test for five years. Starbucks now plans to expand the program across the U.S. to at least 2,000 stores by 2019.
Starbucks Evenings has been in testing for five years. Starbucks now plans to expand the program across the U.S. to at least 2,000 stores by 2019.
Three years after experimenting with wine and beer in Rancho Santa Margarita, Starbucks has confirmed plans to increase its booze offerings to hundreds of cafes across the country, including parts of Orange County.
"We're in the very early stages of considering the Orange County area for the Evenings menu," Starbucks told the Register.
Starbucks Evenings, which also sells small-plate dishes, is coming to a remodeled cafe at Fashion Island in Newport Beach. The chain also has applied for alcohol permits for cafes at 16470 Pacific Coast Highway in Huntington Beach and the Downtown Disney cafe in Anaheim, according to city records and employees.
The Surf City cafe, near Peter's Landing, would be the second Huntington Beach Starbucks to serve alcohol. A cafe at 7101 Yorktown has been selling wine for a couple of years.
As of this week, the Evenings program can be found at 70 Starbucks cafes in Washington state, Oregon, California, Chicago, Florida, Atlanta, Denver, Miami and Orlando.
The new Evenings locations in Newport Beach, Anaheim and Huntington Beach are among several hundred stores that will add beer and wine over the next 12 months. By 2019, Starbucks said, the program will expand to 2,000 stores, or nearly one-fourth of the chain's domestic locations.
The chain projects revenue from Evenings to add $1 billion to its overall sales.
In fall 2010, Starbucks began experimenting with beer and wine sales at its Olive Way location in Seattle. Since then, the pilot program has gradually expanded to cafes in cities across the U.S.
In 2015, market research firm Technomic projects adult beverage sales to increase 3.1 percent at restaurants and bars. That's slightly better than retail sales, which are expected to grow 2.4 percent.
Donna Hood Crecca, a senior director at Technomic, gave Starbucks credit for taking its time to test the Evenings program.
She said Starbucks' Evenings expansion comes as more quick-service chains add alcohol to menus. Burger King, Sonic and White Castle began testing beer sales a few years ago. This year, Irvine-based Taco Bell announced plans to open urban restaurants that serve alcohol-infused libations. The first one is expected to open in Chicago.
Hood Crecca said fast-food and fast-casual restaurants like Starbucks and Chipotle Mexican Grill add alcohol to boost dinner sales, which tend to drop after lunch.
For Starbucks, Hood Crecca said selling booze is about building the evening component and further integrating with its customers' lives, especially millennials.
Gen Y consumers have "embraced spirits, wine and beer" earlier in their lives compared to other of-age generations, she said.
Wine selections cover all the standard varietals: pinot gris, sauvignon blanc, chardonnay, pinot noir, malbec and cabernet sauvignon. The company is sourcing from vineyards in Oregon, Italy, Sonoma County, New Zealand and Argentina. Craft beer selections vary by cafe.
Besides wine and craft beer, the Evenings menu features a variety of dishes such as truffle macaroni and cheese and flatbreads. Wine and beer prices range from $6 to $15 per glass. Small-plate dishes start at $5.
The Evenings cafes in Orange County sell beer and wine after 2 p.m. on most days. Some sell after noon on weekends.


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Your smile is one of the first things people notice about you! Not only does it affect your self-image, but it has a significant impact on your overall health as well.
Dr. Farnham invites you to visit with him and discuss how we can help you experience a smile that's healthy, comfortable, and beautiful! Call Dental Designs today to schedule your consultation.
The Dental Designs team works hard to provide first rate dental care in a comfortable and caring dental office. You and your loved ones can sit back and relax with our patient amenities while our friendly team tends to your teeth. We provide a wide range of services that will keep you and your family safe and healthy.
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There are many different types of stretches and exercises for bulging discs that you can to do help your body. Bulging discs can cause you to go through a lot of pain. The healing process can be accelerated with proper techniques being put into a action.
The severity of a disc bulging is going to dictate how quickly you can heal. If you have severe damage to the tissue, you are going to have a much harder time to get back to normal. If there is just a pinched nerve or something along those lines, you could get back on track must faster. Either way, exercises can go a long way to helping repair the disc bulging.
Certain exercises can be done right at your own home with very little equipment or the need for much space at all. If you are suffering from a disc injury, you should try some of these exercises to help your spine.
In order to do this exercise, you want to first lay flat on your back and grab your legs behind the knees. Do this with both of your hands. Next, pull up on both of your legs until you reach a point where you are about ninety degrees. Then, pull very tight on your legs to feel a nice stretch on your back area. You want to repeat this process for roughly 10 to 12 repetitions in total.
One of the other popular exercises for bulging discs is going to have you using an exercise ball. You want to use the weight of your very own torso to help stretch out the vertebrae in your spine and extend it.
Start by lying face down on the exercise ball. The injured disc in your back should be right above the ball itself. Using your arms, extend them straight forward in front of your head. Then, do the same with your legs so that they are fully extended backwards. This will create a nice stretch on your back. Once this is done, relax and repeat this over the same set of 10 to 12 repetitions.
This next exercise is a side and anterior flexion. What you will be doing here, for starters, is to strand straight up. You then want to stretch the arm out on the side that you have your injured disc. Holding your arm in place, bend it to the opposite side of the injured disc in a slow motion. While your arm is in this location, bend forward just a bit. Return to the starting position and repeat for 10 to 12 repetitions.
This first extension exercise will start by having you standing up. Place the plans of your hands behind your lower back with your arms extended down. Push your chest forward to create an arch of your back. Continue pushing until you reach a natural point of extension, and then return.
This second extension exercise is an alternative or supplement to the last one we just went through. You will lie down on a flat surface with your face down to the ground. With your palms on the ground, try and pull to get your hands together near your chest. Push up on your hands, keeping your legs and your hips in the same position. This creates a bending motion at the waste. Lower back down and repeat.


Wieliczka Salt Mine was a working salt mine since the 1300's and finally closed in 1996. It is a UNESCO world heritage site and we took a tour down to the depths of 500 + feet to see some of the most amazing salt mine shafts and colossal rooms sculpted into chapels where the whole thing is salt – floors, ceiling, stairs, alter you name it was all carved out of salt rock.
Poland has its charms for example when we were on the first train to Prague a man came by through the different train cars and was selling beer out of his gym bag. You would not see that in Canada. The Polish train system is a nightmare and we missed our connection in Katowice because they changed platforms on us and only announced it in Polish so many of us were stranded in Katowice. We took a later connection which has led to another connection in Poland and I am nervous that we are going to miss another train to Prague. I will let you know.
Too many people smoke here, it is like the 60's in Canada when it was cool to smoke, yuk.
The women here love their shoes and 99% wear boots or heels everywhere and I felt really underdressed but comfortable in my black, Nike runners. They have great boots here and I had a hard time leaving this place without a few pairs, but where would I put them, I am already at my maximum weight for luggage, such a shame. Food, wine, and boots are all rather inexpensive and those are 3 of my favourite things so I can't wait to return here but I am not taking the train.


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Targeted conservation genetics of the endangered chimpanzee
African wild dogs (Lycaon pictus) from the Kruger National Park, South Africa are currently not inbred but have low genomic diversity
Christina Meiring, Haiko Schurz, … Marlo Möller
Genomic analysis on pygmy hog reveals extensive interbreeding during wild boar expansion
Langqing Liu, Mirte Bosse, … Ole Madsen
Genetic assignment of illegally trafficked neotropical primates and implications for reintroduction programs
Luciana Inés Oklander, Mariela Caputo, … Daniel Corach
Two decades of non-invasive genetic monitoring of the grey wolves recolonizing the Alps support very limited dog introgression
Christophe Dufresnes, Nadège Remollino, … Luca Fumagalli
A genomic Neolithic time transect of hunter-farmer admixture in central Poland
D. M. Fernandes, D. Strapagiel, … R. Pinhasi
Ancient admixture from an extinct ape lineage into bonobos
Martin Kuhlwilm, Sojung Han, … Tomas Marques-Bonet
Genomic skimming and nanopore sequencing uncover cryptic hybridization in one of world's most threatened primates
Joanna Malukiewicz, Reed A. Cartwright, … Christian Roos
Methodological challenges in the genomic analysis of an endangered mammal population with low genetic diversity
Lídia Escoda, Oliver Hawlitschek, … Jose Castresana
Faecal DNA to the rescue: Shotgun sequencing of non-invasive samples reveals two subspecies of Southeast Asian primates to be Critically Endangered species
Andie Ang, Dewi Imelda Roesma, … Rizaldi
Peter Frandsen ORCID: orcid.org/0000-0001-7131-09441,2 na1,
Claudia Fontsere ORCID: orcid.org/0000-0003-2233-60263 na1,
Svend Vendelbo Nielsen4,
Kristian Hanghøj2,
Natalia Castejon-Fernandez4,
Esther Lizano ORCID: orcid.org/0000-0003-3304-98073,5,
David Hughes6,7,
Jessica Hernandez-Rodriguez3,
Thorfinn Sand Korneliussen8,9,
Frands Carlsen1,
Hans Redlef Siegismund2,
Thomas Mailund4,
Tomas Marques-Bonet3,5,10,11 na2 &
Christina Hvilsom1 na2
Heredity volume 125, pages 15–27 (2020)Cite this article
Populations of the common chimpanzee (Pan troglodytes) are in an impending risk of going extinct in the wild as a consequence of damaging anthropogenic impact on their natural habitat and illegal pet and bushmeat trade. Conservation management programmes for the chimpanzee have been established outside their natural range (ex situ), and chimpanzees from these programmes could potentially be used to supplement future conservation initiatives in the wild (in situ). However, these programmes have often suffered from inadequate information about the geographical origin and subspecies ancestry of the founders. Here, we present a newly designed capture array with ~60,000 ancestry informative markers used to infer ancestry of individual chimpanzees in ex situ populations and determine geographical origin of confiscated sanctuary individuals. From a test panel of 167 chimpanzees with unknown origins or subspecies labels, we identify 90 suitable non-admixed individuals in the European Association of Zoos and Aquaria (EAZA) Ex situ Programme (EEP). Equally important, another 46 individuals have been identified with admixed subspecies ancestries, which therefore over time, should be naturally phased out of the breeding populations. With potential for future re-introduction to the wild, we determine the geographical origin of 31 individuals that were confiscated from the illegal trade and demonstrate the promises of using non-invasive sampling in future conservation action plans. Collectively, our genomic approach provides an exemplar for ex situ management of endangered species and offers an efficient tool in future in situ efforts to combat the illegal wildlife trade.
In an era of human-induced acceleration of species loss, often referred to as the sixth mass extinction era (Ceballos et al. 2015), conservation efforts to save endangered species are calling for novel approaches to mitigate the ongoing extinction crisis.
Since the discovery of the common chimpanzee (Pan troglodytes), humans have been drawn to this charismatic species. Despite our fascination, human activities have led to a drastic decline in the population size of the chimpanzee. In the last two decades, chimpanzees have been listed as 'Endangered' at the species level on the IUCN Red List, with one of the four recognised subspecies, the western chimpanzee (P. t. verus) being listed as 'Critically Endangered' in the latest assessment (Humle et al. 2016). Human encroachment on the natural range of the chimpanzee has further caused an intensified conflict between humans and chimpanzees (Hockings et al. 2015). One by-product of the human wildlife conflicts has been a rise in opportunistic trafficking of chimpanzees, which, in recent years has become more organised and systematic (Stiles et al. 2013). Besides wildlife trade, other continuous threats including habitat destruction, poaching for local consumption, and human linked disease outbreaks has led to a drastic decline in the wild chimpanzee populations (Humle et al. 2016). Together, these threats emphasise the importance of a 'One Plan Approach' conservation programme linking in situ and ex situ efforts (Traylor-Holzer et al. 2019) to prevent the predicted extinction of chimpanzees within the current century (Estrada et al. 2017).
Outside Africa, several regional chimpanzee conservation programmes exist, with the largest being the European Association of Zoos and Aquaria (EAZA) Ex situ Programme (henceforth EEP). The EEP targets the subspecies level and today, breeding programmes for two of the four recognised subspecies, the western chimpanzee (P. t. verus) and the central chimpanzee (P. t. troglodytes) have been established (Carlsen and de Jongh 2019). The primary aim of the EEP is to safeguard the survival of healthy self-sustaining populations targeting the taxonomical level of subspecies (Carlsen and de Jongh 2019). The extant EEP populations consist of wild founders and descendants thereof. However, in times before high resolution genetic technologies were available and even in its early development, knowledge of subspecies labels and relatedness between founders were inaccurate and has led to admixture of subspecies in the captive population (Hvilsom et al. 2013). Early attempts to add a genetic layer to the EEP management has confirmed that knowledge of subspecies ancestries, inbreeding and relatedness estimates are instrumental to preserve genetic diversity in captive populations (Hvilsom et al. 2013). Yet, most recent attempts based on microsatellite markers (Hvilsom et al. 2013), did not have the necessary resolution or predictive power to disentangle several generations of hybridisations in the EEP breeding population. Although we still do not know its full extent, hybridisation between neighbouring subspecies of chimpanzees has been shown to occur in the wild (Hvilsom et al. 2013; Prado-Martinez et al. 2013; de Manuel et al. 2016) and therefore, it is not unlikely that some founders in the EEP harbour shared ancestries from more than one subspecies. The current strategy in the EEP targets un-admixed breeding individuals and with the current methods, it is impossible to tell if small admixture proportions arose from an early ex situ hybridisation event followed by several generations of backcrossing or from a naturally admixed founder. Therefore, founders are potentially being wrongfully excluded from the breeding programme due to their admixed ancestry.
The scenario outlined above, is by no means exclusive to captive management of chimpanzees but extends to practically any ex situ management programme of populations based on wild born founders with a taxonomical subdivision. When morphology alone is insufficient in taxonomical delimitation between subspecies or the targeted conservation units, genetic resources becomes increasingly important. Yet, the choice of genetic resource is not always trivial. In response to a growing availability of different types of genetic resources with widely different applications, several studies have tried to develop guidelines based on the management requirements (see e.g. Grueber et al. 2019; Norman et al. 2019).
As described, the complexities in EEP management of chimpanzees requires a new rigorous solution as previous attempts using either mitochondrial DNA, or microsatellites have proven insufficient. With a genome-wide set of ancestry informative markers, we predict that it will be possible to obtain the desired depth of predictive power to infer ancestries in the present and previous generations and classify individuals with shared ancestries as either descendants of admixed founders or ex situ hybrids. This could provide the foundation of a possible reassessment of the current management strategies under the EEP and in turn, allow for inclusion of wild born hybrids in the breeding programme if these are found to resemble the diversity of the species in the wild.
In their natural range, chimpanzees have become a commodity and organised illegal trade poses a serious threat to the species. Over the period from 2005 to 2011 a reported minimum of 643 chimpanzees were harvested from the wild for illegal trade activities (Stiles et al. 2013). However, extrapolations suggest that 20 times as many individuals have become victims of the illegal wildlife trade in that relatively short time span (Stiles et al. 2013). While most of the captured individuals are sold as bushmeat, a considerable number of mostly juvenile chimpanzees end up in the illegal pet trade. When conservation authorities confiscate illegally kept chimpanzees, they are placed at wildlife sanctuaries, often arbitrarily based on availability of space and proximity to the confiscation site. Whilst some of the rescued chimpanzees require specialised lifetime care, others may be successfully reintroduced into their natural habitats after extensive preparation (Beck et al. 2007). For chimpanzees destined to lifetime care, proper management planning requires knowledge about relatedness among sanctuary chimpanzees in order to set up family groups. In cases, were chimpanzees are suitable for reintroduction, knowledge of geographical origin is essential as several studies have shown lineage-specific adaptations in all four subspecies in their respective geographical ranges (e.g. Nye et al. 2018). In the first complete geo-referenced genomic map of the chimpanzee, de Manuel et al. (2016) portrayed a strong correlation between geographical origin and genetic diversity, where the former can be inferred solely based on the latter. Employing genetic testing at the site of confiscation (e.g. airports and transport hubs) would enable conservation authorities to infer geographical origin of confiscated individuals and with time, strive to facilitate a return of these individuals to a protected area in the region where they were captured. Alternatively, confiscated chimpanzees can be sent to a neighbouring sanctuary with housing capacity, where specialised care and rehabilitation can be provided, and if possible, future reintroduction can be planned. Genetic testing at an early stage of confiscation also has the potential to understand and help break trafficking routes and enable CITES authorities to track and enforce law control in situations where chimpanzees are housed in disreputable zoos and entertainment facilities. However, to be a practical tool in conservation, the genetic test needs to maximise the inference accuracy, require very little investment, and pose as little risk to animal health as possible. These requirements limit our choice of applicable data types. With a novel SNP array design where the level of genetic information is only surpassed by costly whole genome sequencing, we argue that our approach constitutes the most cost-efficient option for conservation management in situations where funding is often scarce and demands for rigorous solutions are high.
Using a selected panel of 59,800 targeted ancestry informative markers, we demonstrate the ability to infer robust estimates of ancestry in several generations of the EEP chimpanzee breeding population. We further show how this set of ancestry informative markers can be used to determine geographical origin of confiscated individuals and demonstrate how these methodologies can readily be applied to using non-invasive sampling. In combination, these methods harbour great potential for future global management plans for the chimpanzee and provides an important exemplar for management of endangered species in general.
A total of 179 chimpanzee samples were collected and analysed in the present study (Supplementary File S1 SequencingStatistics.xlsx). For the purpose of cross-validation between sequencing batches and to test our methodology on non-invasive hair sampling, a number of individuals were sequenced in duplicates and triplicates, which lead to 167 unique individuals. 136 from the EEP population housed in 47 different European zoos and primate rescue and rehabilitation centres (Table S2), and 31 from eight sanctuaries across Africa (Table S3). To form a reference panel, we complemented the genotypes of EEP and sanctuary chimpanzees with whole genome data from 58 geo-referenced wild-born chimpanzees, representing the four chimpanzee subspecies, and additionally, one known admixed individual (Ptv-Donald) and one known descendant of wild born individuals (Ptv-Clint) (Prado-Martinez et al. 2013; de Manuel et al. 2016).
DNA extraction and library preparation
DNA was extracted using a standard phenol-chloroform protocol. Samples were quantified with a Qubit 2.0 fluorometer, Qubit® dsDNA BR Assay Kit (Thermo Fisher Scientific). DNA library preparation was carried out in three batches. For the first batch (24 samples) and the second batch (63 samples), extracted DNA was sheared with a Covaris S2 ultrasonicator using the recommended fragmentation settings to obtain a 350 bp insert size. For the third batch (92 samples) DNA was sheared using the recommended settings of Covaris S2 to obtain 200 bp insert size. The first batch of 24 libraries (with 6 more samples not used in this study) were prepared using 1.5 μg of DNA and the TruSeq DNA HT Sample Prep Kit (Illumina), following manufacturer's instructions and 14 cycles of polymerase chain reaction (PCR) amplification. The second batch of 63 samples (with 17 more samples not used in this study) were processed using 500 ng of starting DNA and following the custom dual-indexed protocol described by Kircher et al. (2012) and 12 cycles of PCR were done for indexing and amplification. The remaining 92 samples (with two more samples not used in this study) were processed using 200 ng of starting DNA following the BEST protocol (Carøe et al. 2018) with minor modifications (initial reaction volume was incremented up to 50 μl to accommodate a larger amount of starting DNA and 10 cycles of PCR amplification). For this third batch, we used inline barcoded short adaptors with the same seven nucleotide barcodes at the P5 and P7 adaptors. Clean-ups were done using homemade SPRI beads (Rohland and Reich 2012). Libraries were eluted in 25 μl of ddH2O and quantified with an Agilent 2100 Bioanalyzer using a DNA 1000 assay kit.
Target capture design
We performed a target capture enrichment experiment using baits synthesised by Agilent Technologies. We targeted 59 800 autosomal sites that were ancestry informative markers and designed using the panTro4 genome. Marker selection was done using published chimpanzee genomes (Prado-Martinez et al. 2013) and by applying a sparse PCA method on 10 Mbp bins of the genomes (Lee et al. 2012). Variant sites were then weighted to identify the most informative markers for the first two principal components (PCs) and 200 AIMs were extracted per segment. The genome was binned to have an unbiased and evenly distributed sampling of the genome and to have enough resolution to provide estimates of ancestry in highly admixed individuals.
For target enrichment hybridisation, libraries were pooled equimolarly based on a library prep method to obtain a total of 19 pools (see Supporting Information for a detailed description of the targeted enrichment hybridisation). PCR amplification product was cleaned up using our homemade SPRI beads (Rohland and Reich 2012). Each enriched sample was then quantified on a NanoDrop, BioAnalyzer and then sequenced.
Fastq filtering and mapping
Libraries were sequenced on five lanes of a HiSeq 2500 ultra-high-throughput sequencing system, one lane for 24 chimpanzee samples, 2 lanes for 63 chimpanzee samples and 2 lanes for the remaining 92 samples. Inline barcoded libraries captured in the same pool (92 from Batch 3) were de-multiplexed using Sabre software v. 1.0 (https://github.com/najoshi/sabre).
Prior to mapping, paired-end reads were filtered to remove PCR duplicates using FASTUNIQ v. 1.1 (Xu et al. 2012) and adaptors (Illuminaclip) and low quality first five bases in a read (Slidingwindow:5:20) were trimmed using TRIMMOMATIC v. 0.36 (Bolger et al. 2014). Overlapping reads were merged with a minimum overlap of 10 bp and minimum length of final read to 50 bp, using PEAR v. 0.9.6 (Zhang et al. 2014). Then, reads were mapped using BWA v. 0.7.12 (Li and Durbin 2009) to the Hg19 reference genome (GRCh37, Feb.2009 (GCA_000001405.1)). PCR duplicates were removed using PICARDTOOLS v. 1.95 (http://broadinstitute.github.io/picard/) with the MarkDuplicates option. Further filtering of the reads was done to discard secondary alignments and reads with mapping quality lower than 30 using SAMTOOLS v. 1.5 (Li et al. 2009). We then filtered for the targeted space (4 bp around the selected SNP) using BEDTOOLS intersect v. 2.16.2 (Quinlan and Hall 2010).
The total aligned reads were calculated by dividing the number of uniquely mapped reads (the remaining reads after removing duplicates) by the number of production reads. The on-target aligned reads were calculated by dividing the target filtered reads by the production reads. Then, the total coverage was calculated by dividing aligned bases by the length of the assembly (Hg19) and the target effective coverage dividing the on-target bases by the targeted genomic space. Finally, the enrichment factor of the capture performance was calculated by taking the ratio between the on-target reads by total mapped reads over the target size by genome size.
Variant discovery was performed using GATK 'Unified Genotyper' (DePristo et al. 2011) for each sample independently with the following parameters -out_mode EMIT_ALL_SITES -stand_call_conf 5.0 -stand_emit_conf 5.0 -A BaseCounts -A GCContent -A RMSMappingQuality -A BaseQualityRankSumTest. Genotypes from each sample were combined in a single VCF using GATK 'CombineVariants' (DePristo et al. 2011) with -genotypeMergeOptions UNIQUIFY –excludeNonVariant parameters. We also included the genotype information of available whole genome data of aforementioned 58 wild-born geo-referenced chimpanzees and Ptv-Donald and Ptv-Clint (Prado-Martinez et al. 2013; de Manuel et al. 2016). Unless differently stated in separate analysis, the variants with a depth of coverage less than 3, a quality score less than 30 (QUAL < 30), minor allele frequency (MAF) of 0.005 and a missingness rate of >60 % were removed using VCFTOOLS v. 0.1.12 (Danecek et al. 2011). We only kept the genotypes that were inside the target space by using the -bed option in VCFTOOLS v. 0.1.12 (Danecek et al. 2011).
Ancestry inference and inbreeding
We inferred proportions of shared ancestries in two approaches. First, to detect underlying genetic structure with a reduction of the dimensionality in the data, we performed a principle component analysis (PCA) using EIGENSOFT v. 6.1.3. (Price et al. 2006). All samples were included without pruning of sites in linkage disequilibrium or MAF, in order to avoid exclusion of fixed sites between populations. Analyses on shared ancestry in ex situ and sanctuary populations were done with reference to the genetic structure in the wild born individuals with ADMIXTURE v. 1.2 (Alexander et al. 2009). To avoid any bias introduced from a joint analysis with related individuals, each of the 167 unique individuals from the EEP and sanctuary populations were analysed separately one by one against a reference panel of all wild born individuals. After applying a MAF filter (--maf 0.05) in PLINK v. 1.07 (Purcell et al. 2007) to exclude sites polymorphic in only one individual, a set of 45,542 sites where kept for analysis. Each analysis of ADMIXTURE v. 1.2 (Alexander et al. 2009) was iterated 100 times under an EM optimisation algorithm and termination criteria of a log-likelihood increase of 10−5 between iterations. A value of K = 4 was chosen to obtain clusters in line with the four recognised subspecies of chimpanzees. To assess convergence, the 100 iterations were evaluated to ensure that iterations did not differ by more than 1 log-likelihood value.
For each of the individuals with admixture coefficients >0.99, we applied NGSRELATEv2 (Hanghøj et al. 2019) to estimate pairwise relatedness and individual inbreeding coefficients based on population allele frequencies from each of the inferred admixture clusters, after excluding MAF < 0.05 (see Supplementary Information for details along with per population and global estimates of FIS).
Hybrid classification
To further explore the ancestry sharing in the EEP and sanctuary individuals and to be able to differentiate shared ancestry originating from the founding individuals and EEP hybrids, we developed a hidden Markov model (available on GitHub http://gihub.com/svendvn/ImmediateAncestry) to allow for an inference of the posterior proportion of ancestries in the three immediate previous generations. In addition, we estimate where these immediate ancestors belong in the pedigree. For full documentation of the model, see Supplementary Information.
Re-assignment of geographical origin
We applied the methodology of ORIGEN (CRAN R package https://cran.r-project.org/web/packages/OriGen/index.html) as described by Rañola et al. (2014), to re-assign the geographical origin of confiscated sanctuary individuals. We applied the FitOriGenModelFindUnknowns parameter to the 1690 highest ranked informative markers to assign individual geographical origin onto the allele frequency surface, inferred from the wild born reference panel.
Non-invasive sampling
To test our targeted capture approach on non-invasively collected hair samples, we sequenced three individuals where we had both blood samples, whole genome reference data and hair samples. Hair samples were capture sequenced using the same methodology as described above for blood samples, except we added a pre-treatment step in the DNA extraction of hair samples to enhance lysis of keratin. Shared ancestry and geo-graphical origin was analysed as described above.
Capture sequencing and variant calling
First we quantified and assessed the performance of our capture methodology in the selected targeted space. We wanted to ensure sufficient representation of the targeted genomic regions to reliably call the selected variants. In a total of five lanes of HiSeq2500 we obtained ~1000 million production reads, and on average, each sample received five million reads. After removing PCR duplicates and considering only primary alignments with a mapping quality higher than 30, we obtained an average of 3.6 million mapped reads (74.31%) per sample (Supplementary File S1). The average effective target coverage on the 59,800 autosomal SNPs was 21.69X with 12.91% of on-target reads (four base pairs around the targeted SNP, Supplementary File S1) which fulfilled our theoretical prediction of 20X. In terms of capture performance, this last statistic is an underestimate since the full length of the capture bait is 120 base pairs and in this analysis, we only considered the four base pairs around the targeted SNP. Still, we considered it to be more accurate since it is the true space where the informative SNP falls. Lastly, to summarise the performance of the capture methodology, we computed the enrichment factor that relates the number of aligned reads on the target space divided by the production reads, with the size of the target space to the size of the whole genome. The resulting enrichment factor of 89.31X reasserts the advantages of capture to ensure enough coverage for genotyping purposes (Supplementary File S1).
Considering all samples without overlap, we obtained a total of ~150,000 genotypes. However the average number of SNPs called per sample was 30,337 sites passing the filtering steps (MAF 0.05 and max-missing 0.6, after we excluded samples '12103' and '12349' due to low coverage). The maximum number of SNPs called in one individual was 51,952 and the minimum was 10,783 (Fig. S1). Among the variation found in western chimpanzees, only a third of these were polymorphic in the western chimpanzee (Table S1), yet, of the 46,260 polymorphic sites, 15,738 were private in the western chimpanzee (Fig. S2). For fixed sites, the western chimpanzee also had the highest number of private sites (Fig. S2). Among the four subspecies, the eastern chimpanzee had the highest total number of polymorphic sites, followed by the central chimpanzee, Nigerian-Cameroon chimpanzee, and western chimpanzee, respectively (Table S1).
Population structure, ancestry, and inbreeding
The major axes of variance in EEP and sanctuary individuals were explored with a PC analysis with reference to the panel of geo-referenced individuals with known subspecies label from Prado-Martinez et al. (2013) and de Manuel et al. (2016). The first PC (PC1) explained 70.49% of the variance in our data, separating the western chimpanzees from the three other subspecies in the reference panel (Fig. 1b). With 16.53 % of explained variance, PC2 separated the Nigerian-Cameroon chimpanzee, central chimpanzee, and eastern chimpanzee.
Fig. 1: Subspecies ancestry and inbreeding in wild and captive populations of chimpanzees.
a Geographical distribution ranges of the four chimpanzee subspecies (IUCN 2015; QGIS 2018). b Population structure by principal component decomposition of sanctuary and the EAZA Ex situ Programme (EEP) populations with reference to wild born individuals. c Shared ancestry inferences of sanctuary and EEP individuals summarised from individual ADMIXTURE analysis against the reference panel of wild born individuals. Individuals from the reference panel are labelled with a subspecies ancestry prefix and known sample name in previous literature (Prado-Martinez et al. 2013; de Manuel et al. 2016), sanctuary individuals are labelled with common sample name identifiers, and individuals from the EEP are labelled by studbook number (Tables S2 and S3). d Individual inbreeding coefficients for all individuals with admixture proportions >0.99 in either of the four inferred clusters. Inbreeding estimates were estimated within each cluster independently. Clusters are colour labelled in accordance to (a–c).
The majority of the 167 tested individuals from the EEP and sanctuary populations, clustered with either of the four reference populations, while a minor part of the individuals scattered in between the defined populations (Fig. 1b). The inferred ancestries from the ADMIXTURE analysis conveyed the same patterns of genetic population structure separating the geo-referenced individuals into four distinct clusters with varying degree of ancestry sharing between geographically neighbouring subspecies (Fig. 1c). With this as a reference, we assigned the EEP and sanctuary individuals into groupings in terms of their ancestry patterns of either non-admixed or hybrids with multiple components of ancestry. Of the 167 tested individuals, 121 could be confidently assigned as non-admixed (admixture proportion from one subspecies ≥ 0.99). All 31 sanctuary individuals were assigned to subspecies level without evidence of admixture, where five clustered with the western chimpanzee, one with the Nigerian-Cameroon chimpanzee, one with the central chimpanzee, and 24 with the eastern chimpanzee. In the EEP population, we inferred the majority of the 90 non-admixed individuals to belong to the western chimpanzee (41), three with the Nigerian-Cameroon chimpanzee, 25 with the central chimpanzee, and 21 with the eastern chimpanzee. Of the remaining 46 EEP individuals, 38 were inferred to be hybrids with two ancestry components while the last eight had three ancestry components.
Of all the individuals from the EEP, sanctuary, and the reference panel with admixture coefficients >0.99, relatedness estimates were low (Figs. S3–S6) while we identified eight individuals with inbreeding coefficients above 0.2 (Fig. 1d). Within these eight individuals, all four subspecies were represented, as were wild and captive born chimpanzees.
To explore ancestry patterns in the previous three generations, we ran our ancestry classification model going back k = 3 generations and visualised the number of loci each ancestor in generation k contributed to the ancestral informative part of the genome (see Supplementary Information). In general, our method correctly estimated the expected ancestries of our reference panel individuals (Fig. 2a). Several eastern and Nigerian-Cameroonian chimpanzee individuals were estimated to contain substantial ancestry components from the mutually neighbouring central subspecies. The known hybrid Ptv-Donald (Prado-Martinez et al. 2013) was estimated by the method to be at least one-eighth central chimpanzee, yet the large proportion of loci that were assigned to the central chimpanzee in the posterior distribution might suggest that Ptv-Donald could be as much as one-fourth central chimpanzee.
Fig. 2: Hybrid classification.
Hybrid ancestry in a the reference panel, b the EEP population, and c the sanctuary population. The estimated posterior ancestries, θ is shown for the eight ancestors k = 3 generations back in time, for each individual in the three populations. The ancestors are orderedaccording to the "unphased" pedigree in the bottom of the plot. The width of each rectangle indicate the expected proportion of loci that are assigned to thatancestor (conditioned on the estimate of θ). Small widths suggest deviations from the model and features that could be improved by posterior correction.
Similar to the ancestries inferred with ADMIXTURE, our method classified a large fraction of the EEP and sanctuary individuals to have ancestors from only one subspecies in the last three generations (Figs. 1c, 2b, c). In general, individuals inferred to belong to the eastern chimpanzee had third generation ancestors of central chimpanzee ancestry (Fig. 2b, c). Similarly, four inferred central chimpanzees in the EEP population, showed small proportions of ancestry from the Nigeria-Cameroon chimpanzee. Comparably, one sanctuary individual, Edward, was inferred here as a Nigeria-Cameroon chimpanzee with a small proportion of central chimpanzee ancestry. However, performing posterior correction by replacing the low central chimpanzee ancestor with another high posterior Nigeria-Cameroon ancestor, would likely make a more accurate estimate. Among the admixed EEP individuals, our model showed similar results to those obtained with ADMIXTURE but as ancestry patterns became increasingly complex (more than two ancestral subspecies) our inferred posterior proportions became increasingly uncertain (Figs. 2b, S14). We further observed that in some cases, small deviating (possibly deep coalescing) segments could have let the model to prefer configurations in the ancestry patterns to switch halves (Fig. 2c), while the correct configuration would probably be a simple case of hybridisation in the parent generation.
Geo-localisation
Based on an allele frequency surface map, built from our reference panel of wild born individuals, we determined the geographical origin of all 31 sanctuary individuals. Generally, the inferred probabilities of geographical origin gave accurate estimates (i.e. high probabilities assigned to just one or a few adjacent grid cells) for all sanctuary individuals (Fig. 3). Also, all individuals assigned to the natural range of their inferred subspecies label. The majority of our tested sanctuary individuals belonged to the eastern chimpanzee where the geographical origins were inferred to six provinces along the eastern part of the natural range of the subspecies. Seven of the eastern individuals had low probability estimates divided over a cluster of adjacent grid cells, with the highest ranking cell assigned probability of less than 0.1. All five western chimpanzee individuals were assigned to the same grid cell in the eastern limits of their range. The single individual from the Nigeria-Cameroon chimpanzee was assigned to a locality in Cameroon while the one central chimpanzee was assigned to the coastal region of Gabon.
Fig. 3: Geographical origin estimates for sanctuary individuals.
Based on the allele frequency surface map of the reference panel, sanctuary individuals are assigned probabilities of geographical origin, here summarised from individual estimates.
Expanding our targeted capture approach to non-invasively collected hair samples, corroborated the results obtained with blood samples. ADMIXTURE estimates converged to the same result in the two sample types for all tested individuals and geographical origin was assigned to the same locality between samples (Figs. 4, S15–S19). Compared to the reference, ancestry estimates in our capture array approach did not always reveal the minor components of shared ancestries found when including all variant sites in the genome (Fig. 4).
Fig. 4: Ancestry and geographical origin estimates from non-invasive samples.
a Geographical origin estimates from hair samples based on the allele frequency surface map of the reference panel, tested individuals are assigned probabilities of geographical origin, here summarised from individual estimates with comparison to blood samples (Figs. S15–S19). b Shared ancestry estimates for hair samples compared to whole genome reference data and capture sequenced data from blood.
As an exemplar for conservation genetics of endangered species, we have designed a novel capture array that targets identified ancestry informative markers across the genomes of 24 wild born chimpanzees (Prado et al. 2013) and the PanTro4 reference genome. Acknowledging that the selected ancestry markers were derived from a relatively limited set of genomes, which could potentially introduce an ascertainment bias towards specific subspecies, we confirmed that our design has the power to correctly identify the subspecies of an extended panel of newly sequenced chimpanzee genomes (de Manuel et al. 2016) (Fig. 1). Based on this proof of concept, we sequenced 167 chimpanzees from the EEP and sanctuary populations and analysed subspecies ancestries and geographical origin. We further show how this approach can be extended to non-invasive samples with robust results.
Ancestry of the ex situ population
In our test panel of 167 chimpanzees, 136 were from the EEP population housed at 47 European zoos and rehabilitation centres. Based on information on disembarkation or place of capture, we know that the majority of chimpanzees who founded the current EEP population came from West Africa. In accordance to this, a majority of the 90 non-admixed individuals could be assigned to the western chimpanzee (Fig. 1c). Our findings confirm that for the western chimpanzee, early efforts of the EEP that sought to identify a core group of non-admixed western chimpanzees using mitochondrial DNA (Jepsen and Carlsen unpublished) and microsatellites (Hvilsom et al. 2013), have been momentarily successful. Yet, using similar methodologies, previous attempts have only managed to identify a small group of central chimpanzees since the breeding effort for this subspecies was established (Carlsen and de Jongh 2019). Here, we identify 25 central chimpanzee individuals in the EEP population that show no evidence of shared ancestry with other subspecies (Fig. 1c), and hence from a genetic viewpoint, would qualify as a suitable bolster to the current breeding population. Similarly, the 21 inferred non-admixed eastern chimpanzee individuals could form the crucial starting point from where a separate breeding effort could be established under the EEP. In contrast to this, of our tested 136 EEP individuals, only three could be assigned to the Nigerian-Cameroonian subspecies (Fig. 1c) and in general, of the four subspecies, the Nigeria-Cameroon chimpanzee is by far the least represented in the EEP population (Carlsen and de Jongh 2019). Yet, with our targeted capture approach, it will now be feasible to scan the remaining EEP population (~1000 housed individuals) for additional non-admixed chimpanzee individuals in order to explore the possibilities of creating separate breeding populations for the two remaining subspecies.
Still, with a presumed small EEP population of eastern and Nigerian-Cameroonian chimpanzees, it might prove difficult to avoid inbreeding, although our estimates suggests, that high inbreeding coefficients are not exclusive to these particular subspecies. In fact, individuals with inbreeding coefficients in the range of 0.2–0.4 were found in each of the four subspecies and includes both wild and captive born individuals (Fig. 1d). It is therefore difficult to establish whether the amount of inbreeding in EEP individuals are a consequence of breeding among closely related individuals or whether it stems from inbred founders. In a few cases, like individual '14073', we know from reliable pedigree information, that this individual is the offspring of two full-siblings (Carlsen and de Jongh 2019). For the large majority of the EEP population, this knowledge is not available or is associated with high levels of uncertainties. Together with accurate ancestry inferences, genetically-based inbreeding estimates will be of high importance in management of the breeding population as will other factors such as age, fecundity, behaviour and housing capacities.
Of our 136 tested EEP individuals, 46 were inferred to be of hybrid origin (Fig. 1c). In terms of distinguishing founder individuals with shared ancestry components (wild born hybrids) from ex situ hybrids, our ancestry analyses show that the majority of our inferred hybrids are between non-neighbouring populations in the wild (e.g. between the western chimpanzee and either of the three other subspecies) and are therefore most likely the result of hybridisation in the EEP breeding population. From a management standpoint, these should eventually be phased out of the breeding programme. Yet, some known hybrids have been allowed to breed under the current management. This has been done with the purpose to maintain population numbers in an interim period while the populations reach their target size and also to allow experienced females to pass on up-bringing behaviour to young individuals in the housed groups. To explore the extent of wild born hybrids in the EEP and the possibility of including these in the breeding efforts, we developed a new method for hybrid classification that can trace ancestry patterns three generations back. This could possibly allow us to distinguish between hybrids bred in captivity and wild born hybrids, where the latter could be included in breeding programmes, as they represent natural processes in the wild. However, two key requirements to such an inclusion are a better understanding of the extent of hybridisation in the wild and an EEP management decision on what a suitable admixture threshold would be.
As validation for the hybrid classification model (see also Supplementary Information), our method infers the known hybrid background of Ptv-Donald to have received at least 12.5% of its ancestry from the central chimpanzee, which is in the range of what was previously estimated using whole-genome sequencing data (Prado-Martinez et al. 2013). Yet, in the EEP population, only a few of the inferred hybrids fit with the expectations of ancestry patterns in wild born hybrids. The majority of the inferred hybrids include a western chimpanzee ancestry component (Fig. 2b), which is highly unlikely to occur in the wild due to the vast geographical distance to any neighbouring subspecies (Fig. 1a). Of the eight inferred hybrids with adjacent distribution ranges, one central/Nigerian-Cameroonian and seven central/eastern hybrids (Fig. 2b), we know from studbook information that all eight individuals were captive born (Carlsen and de Jongh 2019) (Table S2). The only cases where our model might have picked up remnants from natural hybridisations are the ancestry components of central chimpanzee in what we inferred to be non-admixed eastern chimpanzees using ADMIXTURE (Fig. 1c, Fig. 2b). However, this could likely be due to a general limitation of our model to separate these two subspecies due to their evolutionary close relationship and history of allele sharing (Prado-Martinez et al. 2013; de Manuel et al. 2016). Although we did not identify any wild born hybrids in the tested set of individuals, our model predictions will be highly useful in terms of pinpointing the timing of admixture and help to illuminate blanks in the studbook regarding possible sires.
Sanctuary ancestry and geographical origin
In contrast to the predominance of western chimpanzee individuals in the EEP population, the majority of the tested sanctuary individuals are inferred to belong to the eastern chimpanzee. Of the 31 tested individuals, we only find four that can be assigned to the western chimpanzee and a single individual from each of the Nigeria-Cameroon chimpanzee and the central chimpanzee (Fig. 1c). When exploring ancestry patterns in the last three generations, we obtained similar results as in the EEP population, where small posterior proportions of central chimpanzee were found in individuals of the eastern chimpanzee (Fig. 2c). This is most likely due to the limitations of our model when it comes to distinguishing shared alleles between these two subspecies, and we do not infer any geographical origin close to possible contact zones between the two subspecies (Fig. 3).
For western and Nigerian-Cameroonian chimpanzees, we obtained high probabilities in the assigned origins but with little spatial resolution. Essentially, all five western chimpanzee individuals assign to the same grid cell. As de Manuel et al. (2016) have previously shown, population structure inferred in the western and Nigerian-Cameroonian populations, may not offer enough resolution to provide fine scale determination of geographical origin. To improve origin estimates in these populations, it is crucial to obtain a better representation of georeferenced samples across their distribution ranges. This has been achieved for most of the central and eastern chimpanzee ranges, but with only one central chimpanzee individual (Doris), we cannot fully evaluate the prediction power and resolution for this subspecies. Nevertheless, the estimated geographical origin of Doris is very close to the reported confiscation site (Table S3), which gives us some assurance that future efforts to determine origins in the central chimpanzee will be possible. With a larger set of individuals from the eastern chimpanzee, we can start to appreciate the full potential of the method. The 24 analysed individuals can be assigned to geographical origins in six localities along the eastern edge of the distribution range of the eastern chimpanzee, where the majority originates from two locations in the northern and southern regions of the Democratic Republic of Congo (DRC) (Fig. 3). First of all, this might tell us that these regions are heavily affected by poaching and illegal trafficking, although the abundance of confiscation sites might also be biased by the locality of contributing sanctuaries. Only further testing of individuals from sanctuaries across the species range will allow us to assess regional threat levels. However, with the inferred origins of the eastern chimpanzee individuals all along the eastern edge of the range, we can conclude that the threats are not confined to a few regions for this subspecies but are distributed across the eastern boarders of the DRC.
When comparing the inferred geographical origins with the reported confiscation sites for all our tested sanctuary individuals (Table S3), it becomes apparent that the trafficking routes generally operate within a relatively local scale. Overall, we see that most of the tested individuals originate from locations that are within close proximity to where they have been confiscated, though with two notable exceptions, Louise and Edward. Louise was confiscated in Moscow, Russia and inferred to have originated from West Africa, while Edward was confiscated in Nairobi Airport, Kenya with inferred origin in Cameroon. This confirms that the illegal trade of wild chimpanzees spans beyond country borders and the African continent as reported in Stiles et al. (2013). Both individuals are now housed in sanctuaries where specialised care can be provided, yet, in these cases, both individuals have been placed in sanctuaries far from their geographical origin and possibly within mixed subspecies groups (other individuals from these sanctuaries have been assigned to different subspecies). Without proper knowledge of their ancestry, sanctuaries might face the same challenges as we have seen in the EEP population, with admixture of subspecies as a result of (unintended) breeding. Genetic testing at an early stage could help to ameliorate these challenges and as we have shown, our genomic approach extents to non-invasive sampling (Fig. 4), making these methods both an accurate and practical tool in conservation efforts to help combat the illegal trade of chimpanzees.
We further predict that this approach will be self-empowering as sampling gaps in the distribution range of the chimpanzee are continuously covered and DNA extraction methods for non-invasive samples improve. This will significantly advance our predictive power of geographical origin and provide valuable insight to shared ancestries in natural populations with positive knock-on effects to hybrid assessment in the ex situ populations.
Our capture array approach of targeting ancestry informative markers offers a standardised and cost-effective method that accurately guides ex situ and in situ conservation management programmes. At the current rate of decline, chimpanzees are predicted to go extinct within the current century (Estrada et al. 2017). Conservation efforts might therefore, in a foreseeable future, be obligated to supplement wild populations with individuals from the ex situ populations as a last resort to prevent them from going extinct. Should it come to this, our approach facilitates the safeguarding of genetically self-sustainable populations that will have preserved a genetic profile that resembles their wild counterparts.
The current extinction crisis however, extends well beyond chimpanzees and the demand for molecular genetics to help guide future population management programmes is immense, ranging across the taxonomical scale of birds, reptiles, amphibians, and mammals. For the latter alone, more than ten EEP genetic projects are underway and globally, regional zoo associations are undertaking molecular genetic studies for which the present study serves as an important blueprint for linking in situ and ex situ conservation efforts.
The genetic data used in the present study is a publicly accessible through the Dryad Digital Repository, https://doi.org/10.5061/dryad.31zcrjdh7.
Alexander DH, Novembre J, Lange K (2009) Fast model-based estimation of ancestry in unrelated individuals. Genome Res 19:1655–1664
Beck B, Walkup K, Rodriques M, Unwin S, Travis D, Stoinski T (2007) Best practice guidelines for the re-introduction of Great Apes. Gland, Switzerland
Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120
Carlsen F, de Jongh T (2019) European studbook for the chimpanzee Pan troglodytes. Copenhagen
Carøe C, Gopalakrishnan S, Vinner L, Mak SST, Sinding MHS, Samaniego JA et al. (2018) Single-tube library preparation for degraded DNA. Methods Ecol Evol 9:410–419
Ceballos G, Ehrlich PR, Barnosky AD, García A, Pringle RM, Palmer TM (2015) Accelerated modern human–induced species losses: entering the sixth mass extinction. Sci Adv 1:e1400253
Danecek P, Auton A, Abecasis G, Albers CA, Banks E, DePristo MA et al. (2011) The variant call format and VCFtools. Bioinformatics 27:2156–2158
de Manuel M, Kuhlwilm M, Frandsen P, Sousa VC, Desai T, Prado-Martinez J et al. (2016) Chimpanzee genomic diversity reveals ancient admixture with bonobos. Science 354:477–481
DePristo MA, Banks E, Poplin R, Garimella KV, Maguire JR, Hartl C et al. (2011) A framework for variation discovery and genotyping using next-generation DNA sequencing data. Nat Genet 43:491–498
Estrada A, Garber PA, Rylands AB, Roos C, Fernandez-duque E, Fiore A Di et al. (2017) Impending extinction crisis of the world's primates: why primates matter. Sci Adv 3:e1600946
Grueber CE, Fox S, McLennan EA, Gooley RM, Pemberton D, Hogg CJ et al. (2019) Complex problems need detailed solutions: harnessing multiple data types to inform genetic management in the wild. Evol Appl 12:280–291
Hanghøj K, Moltke I, Andersen PA, Manica A, Korneliussen TS (2019) Fast and accurate relatedness estimation from high-throughput sequencing data in the presence of inbreeding. Gigascience 8:1–9
Hockings KJ, McLennan MR, Carvalho S, Ancrenaz M, Bobe R, Byrne RW et al. (2015) Apes in the anthropocene: flexibility and survival. Trends Ecol Evol 30:215–222
Humle T, Maisels F, Oates JF, Plumtre A, Williamson EA (2016) Pan troglodytes. IUCN Red List Threat Species
Hvilsom C, Frandsen P, Børsting C, Carlsen F, Sallé B, Simonsen BT et al. (2013) Understanding geographic origins and history of admixture among chimpanzees in European zoos, with implications for future breeding programmes. Heredity 110:586–593
IUCN (2015) IUCN Red List of Threatened Species. Version 20153: www.iucnredlist.org
Jepsen BI, Carlsen F Genetic identification of West African Chimpanzee, Pan troglodytes verus, based on mitochondrial DNA analysis. Unpublished
Kircher M, Sawyer S, Meyer M (2012) Double indexing overcomes inaccuracies in multiplex sequencing on the Illumina platform. Nucleic Acids Res 40:e3. https://doi.org/10.1093/nar/gkr771
Lee S, Epstein MP, Duncan R, Lin X (2012) Sparse principal component analysis for identifying ancestry-informative markers in genome-wide association studies. Genet Epidemiol 36:293–302
Li H, Durbin R (2009) Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 25:1754–1760
Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N et al. (2009) The Sequence Alignment/Map format and SAMtools. Bioinformatics 25:2078–2079
Norman AJ, Putnam AS, Ivy JA (2019) Use of molecular data in zoo and aquarium collection management: benefits, challenges, and best practices. Zoo Biol 38:106–118
Nye J, Laayouni H, Kuhlwilm M, Mondal M, Marques-Bonet T, Bertranpetit J (2018) Selection in the introgressed regions of the chimpanzee genome. Genome Biol Evol 10:1132–1138
Prado-Martinez J, Sudmant PH, Kidd JM, Li H, Kelley JL, Lorente-Galdos B et al. (2013) Great ape genetic diversity and population history. Nature 499:471–475
Price AL, Patterson NJ, Plenge RM, Weinblatt ME, Shadick NA, Reich D (2006) Principal components analysis corrects for stratification in genome-wide association studies. Nat Genet 38:904–909
Purcell S, Neale B, Todd-Brown K, Thomas L, Ferreira MAR, Bender D et al. (2007) PLINK: a tool set for whole-genome association and population-based linkage analyses. Am J Hum Genet 81:559–575
QGIS (2018) QGIS Geographic Information System. Open Source Geospatial Foundation Project. http://qgis.osgeo.org
Quinlan AR, Hall IM (2010) BEDTools: a flexible suite of utilities for comparing genomic features. Bioinformatics 26:841–842
Rañola JM, Novembre J, Lange K (2014) Fast spatial ancestry via flexible allele frequency surfaces. Bioinformatics 30:2915–2922
Rohland N, Reich D (2012) Cost-effective, high-throughput DNA sequencing libraries for multiplexed target capture. Genome Res 22:939–946
Stiles D, Redmond I, Cress D, Nellemann C, Formo RK (2013) Stolen apes—the illicit trade in chimpanzees, Gorillas, Bonobos and Orangutans. Birkeland Trykkeri AS, Norway
Traylor-Holzer K, Leus K, Bauman K (2019) Integrated Collection Assessment and Planning (ICAP) workshop: helping zoos move toward the One Plan Approach. Zoo Biol 38:95–105
Xu H, Luo X, Qian J, Pang X, Song J, Qian G et al. (2012) FastUniq: a fast de novo duplicates removal tool for paired short reads. PLoS ONE 7:e52249. https://doi.org/10.1371/journal.pone.0052249
Zhang J, Kobert K, Flouri T, Stamatakis A (2014) PEAR: a fast and accurate Illumina Paired-End reAd mergeR. Bioinformatics 30:614–620
The authors would like to thank all institutions who provided samples for this study: The EAZA Biobank, Stichting AAP Amersfoort Zoo, Antwerp Zoo, Zoo Delle Star ad Aprilia, Royal Burgers' Zoo Arnhem, Monde Sauvage Safari, Badoca Safari Park, Barcelona Zoo, Zoo Parc de Beauval, Bioparc Valencia, Borås Zoo, Parco Natura Viva, Edinburgh Zoo, Ölands Zoo, Plättli Zoo, Givskud Zoo/Zootopia, Safaripark Beekse Bergen, Hodonin Zoo, Xanthus Zoo, Kolmården Zoo, Krakow Zoo, Kristiansand Zoo, Lagos Zoo, Le Pal Zoo, Leipzig Zoo, Liberec Zoo, Lisbon Zoo, Madrid Zoo, Magdeburg Zoo, Olmense Zoo, Zoo Osnabrück, African Safari, Plaisance du Touch, Plzen Zoo, Centro de Rescate de Primates Rainfer, Zoological Center Tel Aviv—Ramat Gan, Safari Ravenna, Zoo di Roma, Leintal Zoo, Serengeti-park Hodenhagen, Reserve Africaine de Sigean, Wilhelma Zoo, Loro Parque Zoo, Touroparc, Twycross Zoo, AAP Primadomus, Warsaw Zoo, Schwabenpark, Zagreb Zoo, Cente International de Recherches Médicales de Franceville, Chimfunshi Wildlife Orphanage, Jeunes Animaux Confisques au Katanga, Ngamaba Island Chimpanzee Sanctuary, Tacugama Chimpanzee Sanctuary, The Chimpanzee Conservation Center. We further wish to thank Tom de Jongh and Lisbeth Borbye for valuable language editing and comments to the manuscript and Abigail Ramsøe for early developmental stages of the hybrid classification model. PF is supported by the Innovation Fund Denmark doctoral fellowship programme and the Candys Foundation. CF is supported by "la Caixa" doctoral fellowship programme. TSK is funded by Carlsberg grant CF19-0712 prepared within the framework of the HSE University Basic Research Program. TMB is supported by BFU2017-86471-P (MINECO/FEDER, UE), U01 MH106874 grant, Howard Hughes International Early Career, Obra Social "La Caixa" and Secretaria d'Universitats i Recerca and CERCA Programme del Departament d'Economia i Coneixement de la Generalitat de Catalunya (GRC 2017 SGR 880). EL is supported by CGL2017-82654-P (MINECO/FEDER, UE).
These authors contributed equally: Peter Frandsen, Claudia Fontsere
These authors jointly supervised this work: Tomas Marques-Bonet, Christina Hvilsom
Research and Conservation, Copenhagen Zoo, Roskildevej 38, 2000, Frederiksberg, Denmark
Peter Frandsen, Frands Carlsen & Christina Hvilsom
Section for Computational and RNA Biology, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200, Copenhagen, Denmark
Peter Frandsen, Kristian Hanghøj & Hans Redlef Siegismund
Institute of Evolutionary Biology, (UPF-CSIC), PRBB, Dr. Aiguader 88, 08003, Barcelona, Spain
Claudia Fontsere, Esther Lizano, Jessica Hernandez-Rodriguez & Tomas Marques-Bonet
Bioinformatics Research Center, Department of Mathematics, Aarhus University, C. F. Møllers Allé 8, 8000, Aarhus C, Denmark
Svend Vendelbo Nielsen, Natalia Castejon-Fernandez & Thomas Mailund
Institut Català de Paleontologia Miquel Crusafant, Universitat Autònoma de Barcelona, Edifici ICTA-ICP, c/ Columnes s/n, Cerdanyala del Vallès, 08193, Barcelona, Spain
Esther Lizano & Tomas Marques-Bonet
MRC Integrative Epidemiology Unit at University of Bristol, Bristol, BS8 2BN, UK
Population Health Sciences, Bristol Medical School, University of Bristol, Bristol, BS8 2BN, UK
GLOBE, Section for Geogenetics, Øster Voldgade 5-7, 1350, Copenhagen, Denmark
Thorfinn Sand Korneliussen
National Research University, Higher School of Economics, 20 Myasnitskaya Ulitsa, 101000, Moscow, Russia
Catalan Institution of Research and Advanced Studies (ICREA), Passeig de Lluís Companys 23, 08010, Barcelona, Spain
Tomas Marques-Bonet
CNAG-CRG, Centre for Genomic Regulation (CRG), Barcelona Institute of Science and Technology (BIST), Baldiri I Reixac, 408028, Barcelona, Spain
Peter Frandsen
Claudia Fontsere
Svend Vendelbo Nielsen
Kristian Hanghøj
Natalia Castejon-Fernandez
Esther Lizano
Jessica Hernandez-Rodriguez
Frands Carlsen
Hans Redlef Siegismund
Thomas Mailund
Christina Hvilsom
Correspondence to Peter Frandsen or Claudia Fontsere.
The authors declare that they have no conflict of interest.
Publisher's note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Associate Editor: Xiangjiang Zhan
Supplementary File S1
Frandsen, P., Fontsere, C., Nielsen, S.V. et al. Targeted conservation genetics of the endangered chimpanzee. Heredity 125, 15–27 (2020). https://doi.org/10.1038/s41437-020-0313-0
Revised: 31 March 2020
Issue Date: August 2020
New developments in the field of genomic technologies and their relevance to conservation management
Gernot Segelbacher
Mirte Bosse
Elena Buzan
Conservation Genetics (2022)
Readers Choice: The best of Heredity 2020
For Authors & Referees
Heredity (Heredity) ISSN 1365-2540 (online) ISSN 0018-067X (print)

redpajama_set_name: RedPajamaCommonCrawl


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Home Forums > GENERAL YACHTING DISCUSSION > Technical Discussion > Engines >
MAN LE403 engines
Discussion in 'Engines' started by Sin miedo, Jun 19, 2022.
Sin miedo, Jun 19, 2022
Sin miedo New Member
does anyone know the rate of fuel burn for a pair of MAN 403LE engines running a 72 feet yacht?
bernd1972, Jun 20, 2022
bernd1972 Senior Member
Flensburg, Germany
You´re talking about the V12 or V8? 2842 or 2848?
mapism, Jun 20, 2022
mapism Senior Member
They used to build a V10 as well (2840).
In fact, if I should hazard a guess based on the vessel size, I think that's likely to be the one the OP is referring to.
A V12 would be even more appropriate for a 72 footer, but the LE403 version of the V12 is unheard of on pleasure boats, because it's a heavy duty rating engine, with a 720hp output @1800 rpm.
As opposed to 800 and 1050 hp @2300rpm respectively for the V8 and V10, whose LE403 version were both light duty.
And if that sounds like their models designation was a bit of a mess, it's because it was!
Anyway, happy to post MAN curves, if the OP specifies which engine he's interested in.
V12 are LYE and LZE, my fault. So it´s obviously a V10 or V8. If V8 (2848 LE403) itßs the medium duty version @540hp. Got plenty of info on that one, have them myself. PM?
I believe the 800hp from the V8 came later with the bigger bore and electronic injection versions. The mechanical V8 were 540hp and 680hp (LE403 and LE401)
Bernd, happy to enter in more details by PM if you wish, but I think you're mixing up the V8 numbers/ratings (or possibly kW vs. Hp, not sure).
I happen to know the 90s MAN range pretty well because I came across many of them during my last boat search, eventually chosing one powered by the D2848 LE403.
And I'm pretty sure that this version was light duty rated, capable of 800hp.
But still 100% mechanical - in fact, the V8 was the very last MAN block to be converted to CRM, skipping the semi-electronic EDC versions that they fitted on the V10 and V12 for a few years.
The predecessor D2848 LE401 was capable of 680hp instead.
BTW, MAN never increased the bore of their engines - not even in the current lineup, which is still the same old 128mm that has been around forever.
The slightly higher displacement of V8 and V12 in the second half of the noughties (by which time the V10 had already been phased out) came from a redesigned basement+crankshaft, with a longer stroke.
If you're sure to have the medium duty V8 in your boat, the only one I'm aware of was the LE405, also mechanical and rated for 650hp @2100 rpm.
But I never studied their heavy and medium duty range in detail!
Hi Mapism,
Maybe I am wrong on this and the medium duty 2848 is the LE405, however, the medium duty cersion of the 680hp defintelly comes @540hp. Also copied by Doosan (Daewoo Heavy industries), same two power ratings and all. It´s well possible that the LE 401 and LE403 are both two generations of the same 2848, however, I´m not perfectly sure with different power ratings of these two versions. From my memory both mechanical engines were 680hp in the light duty rating. But don´t make me swear it. Perhaps you could check the volume? The 2848 series was allways 14,6 liters, in all versions. But I think there was an increased volume version on the V8-block @ about 16,5 liters as well. No sure about the type code.
MAN definitetlly increased the volume, not sure if they did by increasing bore or stroke. This change was, for example, done with the step from the 2866 to the 2876 with the six-bangers.
bernd1972 said: ↑
MAN definitetlly increased the volume, not sure if they did by increasing bore or stroke.
This change was, for example, done with the step from the 2866 to the 2876 with the six-bangers.
Always stroke. As I said, bore has been 128mm for decades, and still is in current days.
With the inline 6, stroke was upped from 155mm of the 2866 to 166mm of the 2876 - hence displacement from 12 to 12.8 liters.
With the V engines, stroke went from 142 to 157mm - hence displacement from 14.6 to 16.2 liters (V8) and from 22 to 24 liters (V12).
A longer stroke version of the V10 doesn't exist, because by the time they redesigned the V engines basement+crankshaft, they also dismissed the V10.
Fiammetta42 and bernd1972 like this.
The four-stroke marine diesel D2840 LE 403 model has a bore and stroke of 128 mm x 142 mm and two banks of five cylinders arranged in a 90-degree V, giving a total swept volume for the engine of 18.27 liters.
So, does anyone know the fuel consumption at 1900 RPM?
Capt Ralph, Jun 20, 2022
Capt Ralph Senior Member
Satsuma, FL
Sin miedo said: ↑
I was wondering if you were still interested.
Guys, Lets find this kid some answers.
Fuel burn does NOT depend just on the engine model and its RPM, but also on its load at that RPM - which in turn depends on the propeller demand of the specific boat you are considering.
And if you think this is just a useless analysis paralysis, well, think again: the same engine, at the same RPM, can show huge differences in fuel burn, between two different boats.
That said, according to MAN, one D2840 LE403 at 1900 RPM should burn 25 GPH.
You can see this and other relevant numbers in the bottom table of the second page, in the attached specs sheet.
But don't take this as gospel truth, because even the manufacturers have absolutely no way to know how much their engines will actually burn on each installation, and their "prop demand" numbers are just a theoretical average.
What the manufacturer know with a good level of accuracy is the output at full load that an engine is capable to produce, and in turn its full load fuel burn. Which, just to put things into context, is a whopping 47 instead of 25 GPH, for this engine.
But you should do a bit of math to calculate that, starting from the specific consumption in g/kWh.
D2840LE40x.pdf
Sin miedo likes this.
As an aside, it's nice to see that my guess in post #3 was correct.
And FWIW, this is not a powerplant I'd be happy to have on any 72 footer, for various reasons.
But this is for another thread.
Sarnico, Jul 11, 2022
Sarnico New Member
new to the forums so hello!! I might be of help with my first post as I run two D2840 LE403 (V10, 1050 bhp) on a 60 foot open:
@30 knots cruise I see about 110 liters per hour indicated, this is at about 80/85 % load
I am not 100% sure but this should be at 1.800/1.900 rpm
I can check at the weekend if you want me to. Have a great week and kind regards, Felix
mapism, Jul 11, 2022
Hi and welcome.
Your post and your username make me guess that you're talking of a Sarnico 60 HT, right?
Very nice boat indeed, just perfect for Med cruising!
Hi Mapism, thank you!! I have the older 58 unfortunately
As you seem to know your stuff, and without wanting to hijack the thread, how much black smoke do you think is acceptable just befor the turbos spool up on these engines? Mine are putting out a fair bit...
Mmm... Really BLACK smoke, I'd say very little, if any at all.
What is pretty normal is some grey smoke upon cold start, which can last anywhere from a minute to about 4 or 5, depending on the environment temperature.
Another normal occurrence is some black soot deposit on the hull around the exhausts, after running for some time at or just above idle.
This is due to the combination of two factors: first, the engines not reaching their optimal operating temperature, and second, the fact that in most boats (though I can't remember yours specifically), at very low speed the exhaust gas can't escape from the underwater outlet, and is forced through the lateral (or transom) bypass, hence sticking a bit on the hull around it.
But after say 5 minutes or so from startup, you should only have a minimal amount of grey-ish smoke.
Otherwise, it would be a good idea to pull all the injectors, have them calibrated, and their nozzles replaced.
How many hours did your engines clock (or how many since the last injectors check, if already done in the past)?
Most MAN engineers recommend to check the injectors every thousand hours, though many agree that they can last much longer (I made that at around 1500 hours on my boat, fwiw).
Sarnico likes this.
Sin miedo, Jul 11, 2022
Sarnico said: ↑
Thank you so much for the information. The load is about is 102,000 lbs. Max speed suppose to be only 22 knots.
thank for your reply - can confirm the grey smoke on start up, usually disappears after about a minute or so. Also a little soot and yes there is a low speed bypass for the exhaust.
No smoke at all at slow speeds - just when I want to get it up on the plane it smokes just prior to the turbos spooling up. At cruise there is just a faint haze if that makes sense...
Engines have close to 700hrs and yes I think an injector check is sensible (along with A1 service). Just need to find a good MAN- competent service agent in the Adriatic which seems to be some task
My pleasure - you should be "ok" then I guess!!


text:
http://www.readwritethink.org/classroom-resources/calendar-activities/19th-amendment-constitution-gave-20280.html
The 19th Amendment to the U.S. Constitution gave women the right to vote in 1920.
E-mail / Share / Print This Page / Print All Materials (Note: Handouts must be printed separately)
Calendar Activity Type Historical Figure & Event
ABOUT THIS CALENDAR EVENT
Proposed on June 4, 1919, it took more than a year for the 48 states to ratify the 19th Amendment, which became law when the Secretary of State announced the completion of the ratification process on August 26, 1920, officially giving women in the U.S. the right to vote.
CLASSROOM ACTIVITY
Six months before the 19th Amendment established women's right to vote in the United States, the League of Women Voters was founded to help women become responsible voters. Today, the League of Women Voters works toward helping American citizens be active, involved participants in the political system—from voting to campaigning to taking a position on current issues.
Invite a representative from the local chapter of the League to talk to your students about voting rights and what they can do to be active in politics, even if they are not old enough to vote yet. Use the ReadWriteThink lessons Vote For Me! Developing, Writing, and Evaluating Persuasive Speeches and Voting! What's It All About? to explore voting with younger students.
The World Factbook: Suffrage
Compare voting rights in the United States to voting rights around the world. Students may be shocked to find that in some countries women are still not permitted to vote!
Votes for Women: Selections from the National American Woman Suffrage Association Collection, 1848-1921
In addition to historical documents related to the passage and ratification of the 19th Amendment, this Library of Congress site includes texts, photos, political cartoons, and lesson plans.
Wayback: Stand Up For Your Rights
This PBS website on civil rights includes a special feature for kids on "Women and the Vote."
This website offers information about Women's Rights National Historical Park, which is located in Seneca Falls, the home of many important sites in the history of women's rights in the U.S.
Grades 9 – 12 | Lesson Plan | Standard Lesson
Critical Literacy: Women in 19th-Century Literature
Reading historical selections will give students the perspective they need to compare the author's purpose and voice of two separate writers.
Analyzing Character Development in Three Short Stories About Women
Students read three short stories about women; discuss the development of female characters, gender differences, and society's expectations; and write scripts in which the characters discuss their similarities and differences.
From Friedan Forward—Considering a Feminist Perspective
Students write letters expressing personal views on issues like equal pay, equal education/employment opportunity, and gender roles—and receive these letters six years later.
Grades 3 – 5 | Lesson Plan | Unit
Voting! What's It All About?
Students explore a variety of sources for information about voting. They evaluate the information to determine if it is fact or opinion, and then create a graffiti wall about voting.
Grades 4 – 5 | Lesson Plan | Standard Lesson
Vote for Me! Developing, Writing, and Evaluating Persuasive Speeches
This lesson encourages students in grades 4 and 5 to think critically and write persuasively by focusing on preparing, presenting, and evaluating mock campaign speeches.
Grades 6 – 12 | Calendar Activity | September 17
Constitution Day is observed today in the U.S.
Students explore an annotated online version of the Constitution before sharing what they learned about one aspect of the document through the Trading Card interactive.
Grades 3 – 12 | Calendar Activity | March 1
March is National Women's History Month.
A scrapbook highlighting the accomplishments of famous American women is created as a class using the Alphabet Organizer and Bio-Cube.
Grades K – 5 | Podcast Episode
Celebrating Women Brave and Strong
Check out these books about two exceptional but nearly-forgotten women-Mighty Jackie: The Strike-Out Queen and Brave Harriet: The First Woman to Fly the English Channel.
Grades 6 – 12 | Podcast Episode
Feminist Books for Teens
In this episode, you'll hear about a variety of feminist books for teens—including works of realistic fiction as well as fantasy, biography, historical fiction, and satire.
© 2020 IRA/NCTE. All rights reserved.
Legal | International Reading Association | National Council of Teachers of English


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text:
The Case of Louis Victor Leborgne ("Tan")
Louis Victor Leborgne, born on July 21, 1809, was one of six children in a middle-class family. Leborgne suffered from epileptic attacks beginning about age 24, but he was able to work. He was employed as a formier, an individual who creates wooden molds for hat or shoe manufacture. He began having terrible headaches in his thirties and eventually became unable to work.
By this time, he had begun to lose his expressive language ability, although whether the loss was sudden or gradual, is not known, His family declined to support him so the Prefect of Police arranged for Leborgne's residence at Bicêtre Hospital in Paris in 1834. Ten years after his admission to Bicêtre, he gradually developed complete paralysis of his right arm followed by paralysis of his right leg.
On April 11, 1861, Louis Victor was transferred to the surgical ward of the hospital because he had developed gangrene of his right leg; the physician Pierre Paul Broca was called to see him.
Broca made note of the 50 years old patient's history, including that he had been a long-term patient at Bicêtre and that he had been admitted because he had "lost the ability to speak." On admission, it had been noted that Leborgne "differed from a sane man only in the loss of articulated speech." According to Broca, Leborgne "understood almost everything that was said to him," "but regardless of the question addressed to him, he always responded: 'tan, tan.'" This verbal stereotype was "accompanied by a gesture of his left hand." At the hospital, apparently the patient was commonly called Tan.
Leborgne was bedridden during the last 7 years of his life, finally succumbing to gangrenous bed sores. Broca reasoned from the evolution of Leborgne's symptoms that "[the] probable diagnosis was therefore: original lesion in the left anterior lobe, propagated to the striate body of the same side." Broca deduced that Leborgne's expanding hemiparesis had been an extension of the original lesion that had produced his expressive language deficit. From what was known at the time about motor control, Broca reasoned that the seat of expressive language had to be in the left hemisphere.
Leborgne died 6 days after Broca first saw him in 1861. When Broca had the opportunity to examine Leborgne's brain at autopsy, he observed "a loss of substance of the cerebral mass" in the left anterior frontal lobe to which he attributed the loss of expressive language. A "softening extended well beyond the limits of the cavity" and these, presumably newer lesions, extended to the parietal lobe, the "temporal-sphenoidal" lobe, the insula and extra ventricular nucleus of the striate body, the last of which caused paralysis of Leborgne's right limbs.
One hundred and forty-six years after Broca's report, an MRI of Leborgne's preserved brain revealed that the left hemisphere lesion extended more medially than had been appreciated by Broca. The cause of Leborgne's malady remains uncertain, although an inflammatory vascular etiology, possibly meningovascular syphilis, could explain the late progression of deficits in the same vascular territory as the initial deficits.
Importance of this case:
Broca's presentation of Leborgne and, later that same year, another patient named Lelong, supported the concept of cerebral lateralization of language.
On April 4, 1861, just one week before Broca first examined Leborgne, he had attended a meeting of the Anthropological Society of Paris and had heard Bouillaud's son-in-law, Ernest Auburtin, argue for the frontal localization of language. Auburtin described his examination of a Mr. Coulerier whose failed suicide-by-gunshot had exposed the left frontal region of the patient's brain. When Auburtin compressed Mr. Coulerier's left frontal lobe with a spatula, he observed a speech arrest; release of the spatula allowed the return of language.
When Broca presented Tan, he did not initially argue for the lateralization of language but rather for consideration of the cerebral convolutions in groups or functional regions. Broca's main achievement was that he published a total of seven additional cases of aphasia after that of Leborgne, with careful autopsy correlation that ultimately provided evidence for the left inferior frontal gyrus as the seat of language.
Broca's work opened the door to exploration of the complex neurological basis of language. However, while Broca's aphasia is named for the expressive aphasia and associated left hemisphere lesion that Broca identified in Leborgne's brain at autopsy, modern imaging has demonstrated that Leborgne's left hemisphere had more extensive damage than is now known to be sufficient to produce "Broca's" aphasia.
Excerpted from: "Six Landmark Case Reports Essential for Neuropsychiatric Literacy"
Authors: Sheldon Benjamin, M.D., Lindsey MacGillivray, M.D., Ph.D., Barbara Schildkrout, M.D., Alexis Cohen-Oram, M.D., Margo D. Lauterbach, M.D., Leonard L. Levin, M.S. L.I.S., M.A.
speech center in brain
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The Case of Solomon Shereshevsky


text:
Group Insurance Board Delays Action on Self-Insurance
December 14, Wisconsin Health News
The Group Insurance Board delayed a decision Tuesday on changes to the health plan for state employees, including whether to shift to a regional or self-insurance model.
Chair Michael Farrell said the board has directed Department of Employee Trust Funds staff and Segal Consulting to gather more information and data about the proposals. The board will reconvene in January.
"There is much complexity and volumes of information that relate to our consideration," he said. "We're not taking these decisions lightly."
During closed session Tuesday, members weighed seven scenarios offered by ETF staff, according to a memo.
One proposal could continue the current fully insured model with minimal changes. That model would have up to 16 different vendors participating.
Other scenarios under consideration would have between six and 11 participating vendors. Those proposals could be fully insured, self-insured or a hybrid. Some of them would divide the state into regions.
A seventh proposal, which is not recommended by ETF or Segal, would have one or two vendors under a self-insured model.
The board has been considering self-insurance off and on for five years according to the memo. It's also considering other changes like three-year contracts with health plans.
Phil Dougherty, spokesman for the Wisconsin Association of Health Plans, said his members "recognize that more can be done in the current competitive model to achieve the state's financial and quality goals."
"Moving to Segal's regionalization strategy would create the risk of reducing competition and choice, eliminating high quality healthcare networks and separating plan participants from their doctors and medical homes," Dougherty said in a statement. "The right answer for the state is to use the current competitive structure and Wisconsin health plans to build on what works."
Read a summary memo on the proposals and discussion.


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I have a cold and cynical side, and I have a depressed and defeated side. Often, as is true of late, I experience both in tandem. Gratitude and thanksgiving are very important things to which one must give oneself over, and I suppose you could say the season for such things comes at a very opportune time for me. There are few things that disperse egocentric maladies than gratitude.
There are few things that disperse egocentric maladies than gratitude.
Even if your spirit is robust, however, and you lack my frailties, it's just as important for you as it is for me to get it right. This isn't some Thanksgiving Day let's-go-round-the-table routine. When it comes to our giving thanks, let's not be rote and glib in our citations of generic "health" and "most people we like are not dead yet." What we want is to experience the acknowledgement, humility, and growth inherent to a sincere and vibrant spirit of gratitude. What are those specific components of life for which we are so grateful we feel as though we owe something in return? Some will inevitably be matters of fate, matters for which we have no definitive entity to whom we might owe, and for those elements we are simply to lift our hands and thoughts to the sky or to the earth, or to both at once.
That's our responsibility, here…more importantly, it's the only way we can truly connect with one another at all.
We're going to find that there are things for which specific and real other people are responsible, and it's on us to say directly to those people the thanks that are due to them. If it's appropriate to show one's gratitude with some small but tangible token, let's take the opportunity to do so. That's our responsibility, here. This is not just good form, it's the only way we ever will truly grasp the totality of our own stories – and, even more importantly, it's the only way we can truly connect with one another at all.


text:
No truth in rumours that LFC are in 'advanced talks' over Ryan Fraser
Well while that came right out of nowhere, the reports that Liverpool are in talks with Ryan Fraser of Bournemouth, it was quickly quashed with a couple of hours.
As most of these reports are, the denial came from the Liverpool Echo and simply stated that 'the Echo understands.. '. Quite who told them this is open to debate, so we'll just assume they had a line from the club.
However, such a speedy denial could also indicate that the club were hoping to get things done on the quiet, announcing the signing as soon as the window was open, and bringing in a top young winger on the cheap.
The only downside is that we're apparently looking to do a swap deal for Harry Wilson. The young Welshman is on loan at the South coast side this season and has scored four goals already, showing that he certainly has the quality to thrive at this level.
To lose Wilson would be a huge disappointment though. However, to then gain a player of Fraser's undoubted ability, with a track record of being able to hack it in the Premier League.
The 25 year old was one of the stars of the top division (outside of the top 6) last season, scoring 8 goals and providing 15 assists in the Premier League alone.
According to those same reports, with his contract being up next summer, he'll go for a cut price, so Bournemouth are keen to get what they can for a player that's destined for a bigger club.
Whether the reports are true or not, Fraser is certainly a player that Liverpool would be likely to be looking at. No doubt to the disappointment of those Reds that still think there's a chance that we're going for Mbappe.